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 COMU
UNIC
CAÇÃO ORA
O AL
OR
RAL C
COM
MMU
UNIC
CATIO
ON
PR
RESE
ENT
TACION ORA
O AL
CO.001
AL TIME PCR
P
FOR ANTEMOR
A
RTEM DIA
AGNOSIS IN
I HUMANS AND HIGH
H
REA
THR
ROUGHPU
UT RABIES
S VIRUS SCREENIN
S
NG
Zaratte-Segura P1,1, Bastida-Gonzalez F1, Ellison J2, Gallardoo-Romero N3, Loparev V4,
Velaasco-Villa A2 - 1Institutto Politecnicco Nacionall, MEX - ES
SM-UPIBI, 2Centers foor
Diseaase Controll and Prevenntion, Atlannta USA - Rabies
R
Sectioon, 3Centerss for Diseasse
4
Conttrol and Prevvention, Attlanta USA - POX Secttion, Centers for Diseaase Control and
Preveention, Atlaanta USA - Biotechnoloogy Core Faacility
a most deevastating diseases,
d
typpically a 1000% case
Rabies is one off the oldest and
A
rabies diaggnosis in hum
mans is impportant to in
nstitute infection
morttality rate. Antemortem
contrrol procedurres and avoid further exxposures off health perssonnel and other
o
people
who may have been
b
in conttact with salliva of the patient,
p
to determine
d
eppidemiologiically
uals were exxposed to thhe same sourrce, to mon
nitor diseasee progression
if othher individu
durinng experimeental therapeutic interv
ventions, to prevent
p
cross infection
n during orggan
transsplantation, and if negaative, to examine other differentiall diagnoses. Four samples
are reecommendeed for antem
mortem testiing of rabiees in humanss (saliva, seerum, nuchaal
skin biopsy and cerebrospinnal fluid) inn which antiibodies, viraal antigens and
a nucleic
mmunofluoreescence andd
acidss are detecteed using neuutralization, indirect annd direct im
reverrse transcripption PCR methods.
m
Thhe turnarounnd time to provide
p
conclusive resuults
on alll four sampples analyzeed by all tecchniques is approximate
a
ely 24 to 488 hrs. Real time
t
PCR
R appears to be one prom
mising techhnique to exppedite resullts on nucleic acid deteection
and aaccurately quantitate
q
viiral loads inn patient sam
mples. How
wever, there is no univerrsal
prim
mer or probe that may deetect the bro
oad lyssavirrus diversity
y described to date, whhich
decreeases the seensitivity off the test draamatically.
The oobjective off this study was the dessign of a sennsitive and specific reaal-time PCR
R
assayy to detect and
a quantitaate a broad variety
v
of raabies virusees in antemoortem and
postm
mortem sam
mples from human/anim
h
mal tissues aand bodily fluids.
f
Meth
hods. Moree than 2500 complete nu
ucleoproteinn sequencess of RABV representattive
of a gglobal specttrum of variiants from the
t GenBannk and CDC
C databases were
w analyzzed
to deesign 12 setss of broadlyy reactive prrimers and probes.
p
Twoo primer setts and probees
from
m the literatu
ure were conncomitantly
y consideredd. A total off 14 primer sets and proobes
weree tested both
h in silicon and
a with a panel
p
of 20 viral isolatees (represen
nting a broaad
varieety of RABV
V circulatinng in bat and
d carnivoress worldwidee) using endd point reveerse
transscription PC
CR and real time PCR assays
a
run inn both a Ligght Cycler 480
4 (Roche,,
Germ
many) and CFX96
C
Touch™ Real-T
Time PCR (Bio-Rad,
(
U
USA)
Detection System
m for
compparative purrposes. A drroplet digitaal PCR systtem QX100 (Bio-Rad Laboratories
L
s,
Inc) w
was used too assess the number of copies of am
mplicons peer sample. To
T confirm
detecction, PCR products
p
weere sequencced and idenntified in thee BLAST NCBI
N
databaase.
Resu
ults. The priior primer sets
s and probbes from thhe literature had limitedd sensitivityy
circuumscribed too RABV cirrculating in respective regions of Asia
A and Affrica. None of
the pprimer sets and
a probes we
w designedd was able to
t detect alll viruses tessted alone.
How
wever, a min
nimal set of five primerrs and probees used conccomitantly in
i separate
reacttions were able
a to detecct all RABV
V included in
i this studyy. The systeem was senssitive
enouugh to detectt up to 5 coppies of ampplicon.
Concclusions. Real time PC
CR is a highly sensitivee and specifiic techniquee yet there is
i no
singlle set of brooadly reactivve primers and
a probes capable
c
of detecting
d
all the existinng
lyssaavirus diversity. We described fivee broadly reeactive prim
mer sets that were used
concomitantly inn a 96 well platform too detect all RABV
R
repoorted globallly. This form
mat
mans and for high throuughput screeening
is suiitable for raabies antemoortem diagnnosis in hum
of fieeld samples. Additionaal research on
o a quantitaative assay will not onlly be able too
assesss the numbber of copiess of amplicoons, but alsoo correlate with
w an estiimated totall
numbber of viablle viral partiicles.
Refeerence
Wachharapluesaddee S, et al. Expert Revv Mol Diagnn. 10(2):207
7, 2010.
Coerrtse J, et al. J Clin Micrrobiol. 48(11):3949, 20010
CO.003
nitoring of a French bat colony shown natu
urally infectted by EBL
LV-1 from
Mon
20099-2012. Disccovery of a new infecttion case in
n the colony
y three years after thee
first positive ra
abies diagnostic.
Picarrd-Meyer E1,2 - 1C. Borrel, D. Jouann - CPEPES
SC Lorrainee, France, 2A.
A Servat,
M.W
Wasniewski, M. Moinet, F. Boué, F.
F Cliquet - ANSES - Nancy
N
Laborratory for
Rabies and Wild
dlife
The ppassive surv
veillance off bat rabies was
w improvved in Francce since 200
00, thanks too a
Natioonal bat rabbies networkk constitutedd by Veterinnary Services and bat handlers
h
froom
the C
Chiroptera group
g
(Sociéété Françaisse pour l’Etuude et la Prrotection des Mammifèères).
To date,59 Eptesicus serotiinus were sh
hown infectted by the European
E
Baat Lyssaviruus
type 1 (EBLV-11) in France out of 950 cases reporrted throughhout Europee. In the conntext
urveillance, we reported for the firrst time in Juune 2009 inn France, in the
of the passive su
c
sixx positive cases
c
villagge of Ancy sur Mosellee (located inn the north eeast of the country),
in a ccolony of Eptesicus
Ep
serrotinus natuurally shownn infected by
b EBLV-1.. The field
studiies, carried out
o from 20009-2012 onn 186 testedd bats with vaccinated
v
b specialissts
bat
show
wed a fall off the seropreevalence byy 5. In 2009,, 45% bloodd samples were
w shown
posittive with a modified
m
FA
AVNt adaptted for bat micro-samp
m
les for the presence
p
of
neutrralizing EBL
LV-1 antiboodies againsst 8% in 2011. Three years
y
after thhe first repoort of
the E
EBLV-1 infe
fection in thee colony off Ancy Sur M
Moselle, a new
n positivee case was
reporrted into thee reproductiion colony at
a the end of July 20122. The colonny was
consttituted by 80 bats in M
May and by 46
4 animals aat the end of July. Simiilarly to 20009,
the ccase was repported on a juvenile
j
fem
male and thee isolated viirus was EB
BLV-1b.
Thannks to the ed
ducation of bat workerss to the riskks of bat rabbies, a new positive
p
case
was ddetected on
n July 23, 20012 in Easteern France on
o a Myotis naterreri. This
T speciess was
recenntly shown infected by BBLV (Bookeloh Bat L
Lyssavirus) in Germanyy. To date, this
is thee first case reported
r
in France on the
t species M
Myotis nateerreri. This case was foound
in a vvillage distaant of 40 km
m from Ancy sur Moselle. The moonitoring of the colony from
Ancyy sur Mosellle with the results of fiield studies investigated from 20099-2012 willl be
preseented and diiscussed as well as the new case of
o infection diagnosed on
o Myotis
naterrreri.
CO.004
VEL LYSSA
AVIRUS FROM
F
A MINIOPTE
M
RUS SCH
HREIBERSII BAT IN
N
NOV
SPA
AIN.
Arécchiga N1,2, Vázquez-Mo
V
orón S1,3, Berciano J1,3, Nicolás O4, Aznar C1,3, Juste J5,
6
2
Rodrríguez C , Aguilar
A
A , Echevarría
E
J1 - 1Instituuto de Salud
d Carlos III - Centro
Nacional de Miccrobiología,, Majadahon
nda, Madridd, España, 2Centro Méddico Nacionnal
xico - Unidaad de Investtigación Méédica en Inm
munología, 3Centro de
Sigloo XXI, Méx
Invesstigación Biiomédica enn Red de Eppidemiologíía y Salud Pública
P
(CIB
BERESP),
Espaaña, 4Centroo de Recupeeración de Fauna
F
de Vaallcalent, Lleeida, Cataluuña, Españaa,
5
6
Estaación Biológ
gica de Doññana (CSIC)), Sevilla, Andalucía,
A
E
España,
Unniversidad de
d
Alcaalá de Henarres, Madrid,, España
t Spanishh Rabies Surrveillance P
Program, a bat
b carcass was
w receiveed on
In the frame of the
th
National Ceenter of
Marcch 12 , 2012, in the Ceentro Nacionnal de Micrrobiología (N
Micrrobiology) (CNM).
(
Thee bat was foound in the city
c of Lleid
da, in July 2011,
2
and itt was
takenn to the Willdlife Care Center
C
of Vallcalent
V
(L
Lleida, Catallonia). The bat died sooon
after admission. Two differrent RT-PCR
R generic methods
m
for the Lyssaviirus genus (1,2)
(
d
B
Brain
and ttwo commeercial rabies antisera forr antigen deetection werre used for diagnosis.
smeaars were possitive for booth, FAT annd RT-PCR,, as well as the oropharringeal swabb for
RT-P
PCR. The baat was morpphologically
y and moleccularly iden
ntified as Miiniopterus
schreeibersii (3). To determiine the idenntity of the virus,
v
a frag
gment of thee nucleoprottein
gene (N) was seequenced. Dataset
D
repreesentative of
o all Lyssavviruses, inclluding the
w used forr the phyloggenetic reco
onstruction. The topoloogy
recenntly described IKOV, was
obtaiined by Bay
yesian Inference (BI) sh
howed a new
w virus relaated more too IKOV andd
WCB
BV than any
y other lyssaavirus incluuded in phyllogroups I and
a II. Thesse results suuggest
a new
w virus, nam
med Lleida bat lyssavirrus (LLEBV
V), taking inn considerattion the locaality
wherre the bat was found. Inn Europe, frrom 1977 too 2011, a tottal of 988 cases of bat
rabiees were repoorted; Eptessicus serotinnus and E. issabellinus which
w
accouunt for moree
than 95% of the cases are considered
c
thhe major naatural reserv
voirs of EBL
LV-1. Seveeral
Myottis spp. are reservoirs
r
f EBLV-22, BBLV, ARAV, and KHUV(4).
for
K
In
I Spain,
EBLV-1 has onlly been desccribed in E. isabellinuss (5). Interestingly, the lowest
b LLEBV was with EB
BLV-1. Thee LLEBV has
h been dettected
nucleeotide identtity shown by
on M
M. schreibers
rsii such as WCBV.
W
M
Miniopterus
g
genus
preseently belonggs to the
Vesppertilionidaee family as the
t other baat genera linnked to lyssaviruses in Eurasia
(Epteesicus, Myootis and Murrina). Howeever, recent molecular analyses haave postulated
the ggroup as an independennt monospeccific Minioppteridae fam
mily (6). M. schreibersiii is a
migrratory bat, widely
w
distriibuted throu
ughout Soutthern Europpe and Eurassia. This batt
speciie gathers inn caves in laarge numbeers (thousannds) for winttering, mov
ving in sprinng to
different and som
metimes disstant summeer roosts forr reproductiion. Due to its migratorry
habitts and typically large siize of popullations of thhis bat, it is quite probaable that oncce an
infecctious agentt is introduced, it may spread
s
quickkly within populations.
p
. The
evoluutionary relationships between
b
thee new LLEB
BV with WC
CBV and IK
KOV need to
t be
clariffied to deterrmine whethher they will form one or more ph
hylogroups. Further
analyyses are in process
p
to assess
a
this quuestion andd to establishh a probablee potential role
r
as a hhuman path
hogen. Moree studies muust be done in order to evaluate eccological asppects
of LL
LEBV circu
ulation.
Nidiaa Aréchiga Ceballos is post-doctorral fellow granted
g
by thhe Consejo Nacional de
d
Cienncia y Tecno
ología (CON
NACyT) Mexico.
M
Thiss research was
w financiaally supporteed by
the pproject SAF 2009-091772 of the Geeneral Reseaarch Program
m of the Sppanish Minisstry
of Sccience and Education.
E
References:
11.
22.
33.
44.
55.
66.
Echevarrría JE, et all. J Clin Miccrobiol.; 39: 3678-83. 2001.
2
Vázquezz-Morón S, et al., J Virrol Methodss. 135: 281--87. 2006.
Ibáñez C,
C et al.,Actaa Chiropterol.; 8: 277––297. 2006.
Schatz J.et al., Zoonnoses Publicc Health. Inn press.
Vázquezz-Morón S, et al., Emerrg Infect Diis.; 17(3): 52
20-23. 2011
1.
Hoofer, S. R y Van Der Bussch
he. Acta Chhiropterol.. 5,
5 supplemeent:1–63. 20003.
CO.005
MAN RABIIES IN TH
HE UNITED
D STATES: EVALUA
ATION OF
F CLINICA
AL
HUM
FIND
DINGS FR
ROM 1960––2010
Bass JM1, Petersen BW1, Mehal
M
JM2, Blanton
B
JD1, Rupprechht C1 - 1CDC
C - Poxviruus and
2
ogens and Pathology
P
Rabies Branch, CDC - Divvision of Hiigh-Consequuence Patho
kground: Clinical
C
diaggnosis of huuman rabies is challeng
ging in the United
U
Statees
Back
(U.S.) due to thee rarity of human
h
casess, non-speciific symptom
ms, and infrrequent
p
exxposure history. The traaditional preesentations of rabies,
attainnment of a potential
furioous and paraalytic, are reeported in reegions of enndemic caniine rabies, but
b have nott
beenn systematically charactterized amoong rabies caases in the U.S.
U This sttudy aims too
exam
mine the clinnical characcteristics of patients in the
t U.S. thaat are associiated with raabies
to aidd in diagnossis and survveillance.
hods: Patiennt data weree extracted from an exiisting databbase associatted with
Meth
sampples submittted for rabiees diagnosiss to the Poxvirus and Rabies
R
Brancch at the CD
DC.
A dee-identified dataset consisting of reecords for all
a patients diagnosed
d
w rabies ffrom
with
19600-2010 and patients
p
ruleed-out for rabies from 2007-2010 was provid
ded. Patientss
with at least onee symptom reported
r
weere includedd in analysiss. Chi-squarre was usedd for
tests of associatiion.
ults: Of the 252 patientts in the dataset, clinicaal symptoms were repo
orted for 2466;
Resu
103 ((41.9%) of 246 were diiagnosed with rabies. S
Symptoms significantly
s
y associatedd with
rabiees (p<0.01) were localizzed pain or paresthesiaa (OR 10.4, 95% CI 5.6
6-19.1),
hydrophobia (OR 9.9, 95% CI 4.3-22.2
2), dysphaggia (OR 3.1, 95% CI 1.8
8-5.2), localized
2 95% CII 1.7-5.1), annd aerophobbia (OR 15..3, 95% CI 1.9-121.3). The
weakkness (OR 2.9,
preseence of agitation or a foocal neuroloogic sign (ddysphagia orr localized pain,
p
paressthesia, or weakness)
w
h a combined specificcity of 95%
had
% and a likelihood ratio of
1.8 fo
for rabies. Symptoms siignificantlyy associated with patiennts for whom
m rabies waas
ruledd-out by laboratory diaggnosis incluuded: confuusion (OR 2..9, 95% CI 1.5-5.4),
malaaise (OR 3.33, 95% CI 1.9-5.6), seizzure (OR 3.1, 95% CI 1.8-5.4) andd headache (OR
4.1, 995% CI 2.4-8.8).
h
a, and aerop
phobia should be
Concclusion: Foocal neuroloogic signs, hydrophobia
emphhasized in thhe evaluatioon of patiennts for rabies and the suubmission of
o samples for
fo
laborratory diagn
nosis. Thesee clinical associations may
m also bee applied to a formal caase
definnition to impprove reporrting of hum
man rabies where
w
laboraatory diagno
osis is limitted.
Ongooing analysiis of clinicaal data from
m patients tessted for rabiies may be utilized to
develop algorithhms of signss and sympttoms predicctive of rabies diagnosis.
R
or Epidemiolog
E
gy and Surveillance of Rabies
R
Topiic: Human Rabies
nancial Asssistance: Jeennifer Basss performedd this work as
a a fellow in
i the
Honoors and Fin
CDC
C Experience Applied Epidemiolog
E
gy Fellowshhip. Brett Petersen
P
inittiated this
projeect as an offficer in CDC
C’s Epidem
mic Intelligennce Servicee.
CO.006
mans
Cliniical Featurres of Dog- and Bat-accquired Raabies in Hum
Udow
w SJ1, Marrrie RA1, Jacckson AC1 - 1Universityy of Manitooba - Internaal Med.
(Neuurology)
nces in rabiees due to caanine and baat rabies viruus variants have been
Cliniical differen
notedd, but no deetailed studiies have beeen reported to support these
t
observ
vations. Usiing
PubM
Med and thee MMWR we
w identified
d 120 case rreports of raabies from the
t USA,
Canaada, Europee, and Asia. We system
matically absstracted seleected clinicaal features,
resullts of investigations, inccubation tim
mes and durrations of illlness. Detaiils about clinical
featuures were reecorded. Casses were claassified as ddog- or bat-aacquired baased on repoorted
anim
mal exposuree or viral vaariant typingg by molecuular or monooclonal antiibody
charaacterizationn. Categoricaal variabless were summ
marized as frequency
f
(%
%), and
contiinuous variaables were summarized
s
d as mediann (interquarttile range [IQ
QR]). We
comppared bat- and
a dog-acqquired cases using chi-ssquare or Fiisher’s exacct tests for
categgorical variaables, and Mann
M
Whitn
ney U tests for
f continuo
ous variablees. Of 120 ccases,
38 (332%) were dog-acquire
d
ed and 54 (445%) were bat-acquired
b
d. Survivorss and cases
acquuired from aerosolized viral
v
exposu
ure or tissuee/organ trannsplantation were excluuded.
The median
m
incuubation timees for dog- and bat-acqquired rabiees were 63 (IQR 42.75, 108)
and 552.5 (IQR 27.25,
2
92.5) days, respeectively (p=
=0.074). Thee median du
urations of
illnesss for dog- and
a bat-acqquired rabiess were 17 (IIQR 11.75, 23) and 14 (9.25,18.5))
days, respectiveely (p=0.2011). There waas no differeence in patiients with bat- and dogp
m
malaise,
enccephalopathhy,
acquuired rabies in terms of the presencce of fever, prodromal
sore throat, cran
nial nerve abbnormalitiess, hemipareesis or seizu
ures. Clinicaal manifestaations
w more common
c
in bat- than dog-acquiredd rabies inclluded a locaal prodromee of
that were
sensoory or motoor symptomss (p=0.026)), hemisensoory abnormaalities (p=0.042), tremoor
(p=0.003), and myoclonus
m
(p=0.009). Neither
N
trem
mor nor myyoclonus waas observed in
og-acquired rabies. Aerophobia andd facial or pharygneal
p
patieents with do
spasms werre
moree common in
n dog- thann bat-acquireed rabies (pp=0.007 andd p=0.029, respectively
r
y).
Hydrrophobia waas more com
mmon in doog-acquired rabies (p=0
0.054). Therre was no
difference betweeen dog- annd bat-acquiired rabies in
i terms of results
r
of diiagnostic
a
inn
invesstigations suuch as skin biopsy, saliivary analyssis or the deetection of antibodies
serum
m and cerebbrospinal fluuid (CSF). The
T CSF protein was higher
h
for baat rabies (799;
IQR 52, 109 mg
g/dL) than dog
d rabies (331; IQR 26,, 48, mg/dL
L,; p=0.012). In summarry,
m
local symptoms,
s
bat-aacquired rabbies is assocciated with more
tremor, andd myoclonuss,
wherreas dog-acq
quired rabiees has more hydrophobbia, aerophoobia, and pharyngeal orr
faciaal spasms. We
W speculatee that these clinical diffferences maay reflect diifferences inn the
routee of viral en
ntry of the raabies virus variants
v
intoo the nervouus system because
b
fundaamental diffferences in the neuropaathology orr viral distribbution havee not been
identtified. Bat raabies virus variants maay also havee greater efffects on the blood-CSF
F
barrier by affectting endotheelial cell perrmeability tthrough unkknown mechhanisms.
CO.007
mune respon
nses in hum
man CNS during
d
rabiees virus ind
duced encephalitis
Imm
Frankka R1, Batteen B2, Shiehh WJ2, Niezzgoda M1, Zaki
Z S2, Ruppprecht C1 - 1Centers foor
Diseaase Controll and Prevenntion - Poxvvirus and Raabies Brancch (PRB), 2Centers
C
for
Diseaase Controll and Prevenntion - Infecctious Diseaases Patholoogy Branch (IDPB)
Undeerstanding the
t mechaniisms of rabiies virus cleearance from
m the CNS will
w be a
signiificant step towards
t
thee treatment of
o clinical rabies.
r
Althoough a few animal studdies
have provided innsight abouut immune reesponses inn the CNS duuring rabiess encephalittis,
ffindata about the saame are verry scarce forr humans. Inn our study, formalin-ffixed, paraff
NS) tissues from patiennts who succcumbed to
embeedded, centrral nervous system (CN
rabiees followingg infection with
w RABV
V variants coommon to thhe tricoloredd bat
(Periimyotis subfflavus) in thhe United States, caninne RABV prresent in Haaiti, and cannine
RAB
BV from Afg
fghanistan, as
a well as tissues from a patient whho recovereed from clinnical
laborratory confiirmed rabiess following cat bite in Colombia,
C
but
b succumb
bed as a ressult
of seecondary meedical intervvention, werre subjectedd to comparrative immuunohistologiical
evaluuations idenntifying partticular imm
mune cell poppulations asssociated with rabies
encepphalitis. A non-enceph
n
halitic brain from an inffluenza patiient was useed as a negaative
contrrol. Populattions of B-cells (CD20)), T-cells (C
CD3), and macrophages
m
s (CD68), as
a
well as the preseence of rabiies virus anttigens were compared using
u
a sem
mi-quantitatiive
n Antiviral Response RT²
R
scalee. In addition, gene exppression anaalyses, usingg the Human
Profiiler™ PCR Array, focuusing on thee expressionn of 84 key genes
g
invollved in the
innatte antiviral immune
i
ressponse, were performedd. No rabiess virus antiggens were
deteccted in the brain
b
tissue of the patieent who survvived cliniccal rabies orr in the conttrol
brainn. T-cells annd macrophaages were abundant
a
in the parench
hyma in all rabies patieents,
but B
B-cells weree detected only
o
in the perivascular
p
r tissue of thhe putative rabies
r
survivor,
and rrabies patiennts infectedd with canin
ne RABVs. Few
F T- andd B- cells, annd only locaal
microoglia cells, were detectted in the innfluenza pattient. Differrences in thee expressionn of
multiiple genes associated
a
w innate immunity,
with
i
aas well as innflammatoryy responsess,
weree identified, suggesting the importaance of theiir role in rabbies encephalitis and viiral
clearrance from CNS
C
tissue..
CO.008
Firstt Myotis lucifugus Raabies Virus Variant Deetected in a Human
Orciaari L1, Brow
wn C2, Lijew
wski V2, Fraanka R1, Jacckson FR1, Niezgoda M1, Tack D1,
1
1
Yageer PA , High
htower D , Rupprecht C1 - 1Centers for Diseaase Control and Prevenntion
- Rabbies Program
m, 2Massachusetts Dep
partment of Public Heaalth
m from Baarnstable County, MA was evaluaated at Masssachusetts
A 633 year old male
tertiaary care faciility for posssible strokee and encephalitis. Althhough the patient’s firsst
sympptoms were joint stiffneess, within 2 days the ppatient was exhibiting signs
s
of
hydrophobia, an
nd acute proogressive enncephalitis. Serum, CSF
F, nuchal (skin) biopsyy, and
f
the pattient were sent to CDC
C for rabies diagnostic
d
t
testing.
No
salivva samples from
rabiees virus IgG
G or IgM anttibodies werre detected in serum an
nd CSF by the
t indirect
fluorrescent antibbody (IFA) test, and noo viral neutrralizing antiibodies werre detected in
i the
serum
m or CSF saamples by thhe rapid fluuorescent foocus inhibitiion test (RFFIT). Rabiees
viruss antigen waas detected in nuchal biiopsy samples using diirect fluoresscent antiboody
(DFA
A) test. Nested (and hem
minested) RT-PCR
R
am
mplicons were producedd from skin and
salivva using mulltiple rabiess virus nucleeoprotein geene primerss sets. Sequeence analyssis of
the eentire nucleo
oprotein genne and compparisons wiith samples in the CDC
C database and
a
Genbbank indicatted that the rabies viruss variant waas associateed with Myo
otis sp bats.
Furthher analysiss of phylogeenetic trees (1000) by Neighbor
N
Jo
oining, Maxximum
Parsiimony and Maximum
M
L
Likelihood
indicated
i
thhe variant was
w most parrsimonious with
the ccommon “litttle brown bat”
b Myotis lucifugus. Postmortem
P
m brain tissuues were possitive
for raabies virus antigen
a
by the
t direct flluorescent antibody
a
test. Antigenicc typing witth
monooclonal antiibodies to thhe rabies virrus nucleopprotein was consistent with
w the
previious results of a bat rabbies variant,, but lackedd the resoluttion of geneetic typing
methhods. Sequen
nce analysis of the RT-PCR ampliicons from the complette nucleoprootein
gene were consiistent with the
t previouss findings of the variannt seen in M.
M lucifugus.
h had kno
own encountters
Althoough M. luccifugus is coommon in thhe US and ffrequently has
with humans and animals, this
t is the fiirst documented case of this rabiess virus variaant in
man. In conntrast to thiss unique findding, the rabbies virus variant
v
assocciated with a
a hum
solitaary bat with
h rare knownn human orr animal enccounters Lassionycteris noctivaganss
(silveer-haired baat) has beenn responsiblle for most hhuman rabies cases in the
t USA ovver
the laast 2 decadees.
CO.009
CEPHALIT
TIS OF UK
KNOWN ET
TIOLOGY
Y AND HUM
MAN RABIES IN TH
HE
ENC
UNITED STAT
TES, 1999-2008
H
R1, Mehal J1, Petersen
P
BW
W1, Recuencco S1, Tack D1,
Voraa NM1,2, Blaanton JD1, Holman
1 1
2
Ruppprecht C - CDC, Epiidemic Intellligence Serrvice
Enceephalitis is a severe neuurologic synndrome caussed by a varriety of infeectious and noninfecctious patho
ologies. In many
m
instancces a definitive etiologgy of encephhalitis is nott
identtified, but a study of enncephalitis in
i Californiaa found thatt 0.3% of caases referredd for
furthher evaluatioon to a speccialized diag
gnostic facillity (Glaser,, Gilliam et al. 2003) w
was
due tto rabies. Unnder recognnition of hum
man rabies in the Uniteed States by
y healthcaree
proviiders may account
a
for missed
m
diag
gnoses of raabies. The purpose of th
his investigaation
was tto estimate the numberr of encephaalitis-relatedd deaths in the
t United States
S
and the
t
propoortion due to
t rabies speecifically.
uman mortaality data in the United States betw
ween 1999-2
2008 and
We rreviewed hu
identtified encep
phalitis-assoociated deathhs using ICD-10 codess. Codes were categorizzed
basedd on if an ettiology of encephalitis
e
o not. We also
a reviewed all
was listed or
laborratory-confiirmed humaan rabies cases within tthe United States
S
and itts territoriess that
weree reported to
o the Centerrs for Disease Control aand Preventtion (CDC) during this
samee time period.
e
-associated deaths werre identifiedd, of which 8726
8
A tottal of 12,457 cases of encephalitis
(70%
%) were of unknown
u
etiiology. Tweenty (0.2%) cases weree coded as raabies. Durinng
this ssame time period,
p
27 coonfirmed caases of rabiees were reported to CD
DC. Of thesee 27
casess, 6 of the exposures occcurred in countries
c
othher than thee USA. Doggs were the most
m
comm
monly reporrted exposuure among im
mported casses. Of the 21
2 indigenoous cases, 144
weree attributablee to bats, 4 to tissue/organ transplaantation, 1 to
t a dog expposure in Puuerto
Rico and 2 from
m unknown sources.
s
f
sug
ggest that thhe majority of encephaalitis-associaated deaths in the Uniteed
Our findings
Statees were of unknown
u
etiiology. Amoong the connfirmed casees of rabies,, the majoritty
who acquired raabies withinn the United States acquuired the inffection from
m wildlife.
t indigenoous cases occcurred throough an unkknown expoosure. Healthhcare
Nearrly 10% of the
proviiders shouldd consider evaluating
e
f rabies whhen a patiennt develops acute
for
progrressive enceephalitis of unknown etiology
e
despite routinee evaluation
n or if clinical
histoory is suggesstive.
Glaseer, C. A., S. Gilliam, ett al. (2003).. "In search of encephaalitis etiologgies: diagnostic
challlenges in thee Californiaa Encephalittis Project, 1998-2000.." Clin Infecct Dis 36(6)):
731-7742.
CO.110
MAN RABIIES IN RIO
O CASCA MUNICIPALITY, 20
012: CASE REPORT,, SIX
HUM
YEA
ARS AFTER
R THE LA
AST RECO
ORD IN MINAS GERA
AIS, BRAZ
ZIL
Santaana LF1, Toourinho BD2, Dias GMF2, Pinto JE
EG2, Amânccio FF2, Brito MG2, Cuunha
MMS
SF2, Bento LJ2, Rodriggues DR2, Trindade
T
ML
L2, Ferraz ML
M 2 - 1Secreetaria de Esttado
2
da Saaúde de Min
nas Gerais, Secretaria de Estado da
d Saúde dee Minas Gerrais
Rabies is a virall disease thaat affects thee central neervous systeem (CNS) of mammals and
ost 100% off cases. In 2012,
2
six yeaars after thee last record
d of a humann
it’s ffatal in almo
case in Minas Gerais,
G
the trransmissionn has been confirmed thhrough aggrression by
pidemiologgical
vamppire bat. Thhe objective of this repoort is to desccribe the cliinical and ep
charaacteristics of
o this case. On 06/19/22012, the Heealth Deparrtment of Minas
M
Geraiss
Statee has been notified
n
by the
t Regionaal Health Deepartment of Ponte Nov
va of a
suspeected case of
o human raabies from Rio
R Casca municipality
m
y. He was a 32-year-oldd
malee patient, farrmer, residinng in the ruural area. Thhe patient suuffered bitteen by a bat in
i his
rightt hand after trying to reemove it from the back of a calf in the month of May. At that
time the patient did not seekk medical care
c and the event was not notifiedd to health
griculture deepartment. The
T calf dieed about 10 days after the
t incidentt and
serviice or the ag
no m
material was sent for labboratory exaamination. S
Since 06/111/2012, the patient
p
succeessively sou
ught care beecause of a barbed
b
wiree cutting on
n the same hand
h
attackeed by
the bbat, which began to shoow dormanccy. Then he developed lip deviatio
on, fever andd
vomiiting, and cllinical progression tow
ward paresthhesia, insom
mnia, agitatio
on, confusioon,
sore throat and drooling.
d
Teetanus, musscle twistingg and tonsilllitis were diiagnostic
hypootheses raiseed by the muunicipal heaalth servicees. The aggrression was reported onnly
after assistance at Ponte Noova municip
pality, and ssuspected hu
uman rabiess was notified.
B Horizonnte
On thhe same dayy, the patiennt was transsferred to thhe referral hoospital in Belo
muniicipality forr specializedd care. Patieent sedationn was maintained, with antiviral
admiinistration and
a control of symptom
matic complications. Thhe patient deeveloped
hypeernatremia, ventilator-a
v
associated pnneumonia and
a cardiac arrests. Theere was
antem
mortem labo
oratory confirmation of rabies infeection by Po
olymerase Chain
C
Reacttion
(PCR
R) in cerebrospinal fluiid and salivaa sample, annd genetic sequencing
s
indicated thhat it
was ccompatible with Desmodus rotunddus genetic variant. Deespite speciffic treatmennt, the
patieent died on 06/28/2012
0
due to refraactory circuulatory shocck. Postmortem examinnation
was pperformed by
b direct im
mmunofluorescence reaaction, with infection co
onfirmationn at
CNS
S fragments.. During thee conduct off the case, epidemiolog
e
gical investiigation was
proceeeded by medical
m
reporrts and recoords reviewiing, family interviews and
a active
searcch of exposeed. In assocciation, heallth educationn, social mo
obilization and
a intersecctoral
workk actions weere developeed. Throughh the describbed informaation it is evvident the
impoortance of health system
ms being abble to properrly suspect, notify, andd investigatee,
added to need to
o maintain surveillance
s
e and controol activities of rabies in
n Minas Gerrais
state.
CO.111
unity in Reelation to Su
usceptibilitty, Diagnossis and
Meaasures of Raabies Immu
Prevvention
Hanllon CA1 - 1Director,
D
Kaansas State University
U
Rabies Laboratory, Maanhattan, Kaansas
USA
A
n
of animals ino
oculated inteentionally with
w
A nuumber of repports describbe limited numbers
rabiees virus thatt do not become ill and may have llittle to no detectable
d
a
antibody
whhile
otherr conspecifiic individuaals succumb
b to the samee inoculum.. This review
w will detaiil
methhods used in
n measuringg immunity and optimal method vaalidation, inn relation to
potenntial identiffication of baseline
b
suscceptibility, enhanced disease
d
diagnnosis, and
optim
mal preventiion practicees for human
ns and anim
mals. Potenttial factors for successfful
rabiees virus infeection includde: 1) protection of infe
fected neuro
ons from host immunemediiated destruuction by thee inhibition of virus-meediated apopptosis and overexpress
o
sion
of im
mmune-subv
versive moleecules resullting in killiing or inactiivation of 'p
protective' T
cells migrating into
i
the infeected nervouus system; 2)
2 an hypothhesis that leethality resuults
m neuronal dysfunction
d
inhibiting proteins
p
reqquired for neeuronal maintenance; 33) a
from
positt that virus phosphopro
p
otein and hoost cell dyneein light chaain 8 interacction may
regullate viral rib
bonucleoprootein linkingg to cell trannsport; 4) a thesis that innate
immuunity, paraddoxically, faavors rabiess virus neuroo-invasivenness throughh enhanced
infilttration and promotion
p
o CD8(+) T cell eliminnation; and then alternaatively that ; 5)
of
type I IFN produuced in the rabies viruss-infected nnervous systtem reducess neurod partially protects
p
from
m fatal infection. It is clear
c
that innnate immunne
invassiveness and
cells detect pathhogens, mosst likely inclluding rabiees virus, viaa pattern reccognition
m
recepptions, suchh as Toll-likke receptors and others. Pathogen-aassociated molecular
patteerns activatee receptors which
w
inducce productioon of pro-innflammatoryy cytokines and
signaals to activaate inflammaation. In adddition, these receptors are requiredd for an adaaptive
immuune responsse. Innate annd adaptivee immune reesponses actt as two inteerlocking
defennse lines. Foollowing a rabies
r
exposure, virus m
may be initiially suppreessed by innnate
immuunity accom
mpanied by effector T cell
c recruitm
ment for activation of adaptive
a
immuunity. Alterrnatively, thhe uniquely high mortallity rate of successful
s
r
rabies
virus
infecction may bee due to viruus-host inteeractions thaat remain laargely a mysstery. The
accurrate and preecise measuurement of an
a adaptive immune ressponse mayy be definedd by
experimental meethods or well-describe
w
ed methods,, such as thee rapid fluorescent focuus
v
in some laborratories. Thee interpretattion
antibbody test, ass performedd and fully validated
of finndings baseed on these various
v
metthods shouldd be in relattion to clinical observaations
and ccollaborativ
ve investigattion of uniqque, isolatedd, novel finddings. The fine-tuning,
f
interaaction, and timing of innnate and addaptive host responses and the meethods used for
detecction and measurement
m
t, will requirre dedicatedd investigattion towardss optimal diisease
preveention.
CO.112
FERENCE
E IN INTRA
ACELLUL
LAR LOCA
ALIZATION AND EX
XPRESSIO
ON
DIFF
LEV
VEL OF RE
ECOMBIN
NANT RAB
BIES G-PRO
OTEINS OF
O STREET
T VIRUS
(KYOTO STRA
AIN) AND FIXED VIIRUS (CVS
S-26 STRA
AIN) EXPR
RESSED IN
N
K293T CEL
LLS
HEK
Ham
mamoto N1, Kaku
K
Y1, Noguchi
N
A1, Morikawa S1, INOUE
E S1 - 1Natioonal Institutee of
Infecctious Diseaases - Veteriianry Sciennce
bies virus (R
RABV) has been mainlly studied with
w fixed viiruses
Virullence of rab
(labooratory straiins) having different deegrees of paathogenicity
y, however, virulence iss
muchh different between
b
fixxed viruses and
a street viiruses (wildd strains). Highly
H
attenuuated
fixedd strains of RABV
R
doess not cause lethal infecction with prrofound infl
flammation
accom
mpanied wiith apoptosiis and neuraal degenerattion in the central
c
nervoous system
(CNS
S). Inductioon of innate immune responses in CNS
C
is a haallmark of in
nfection witth
highlly attenuateed strains, whereas
w
neurral damage is absent orr minimal annd innate
immuune responsses are not induced
i
in animals
a
infeected with street
s
virus. In the streeet
viruss infected ceells, intracytoplasmic virion
v
maturration takenn place in thhe ER/Golgii
appaaratus is com
mmonly obsserved and budding
b
of virus
v
from cellular
c
plassma membrrane
is lesss frequent than
t
in the fixed
f
virus infected
i
cellls. Since RA
ABV G-pro
otein is critical
for innduction off virus neutrralizing antibbody, profoound expresssion and buudding of viirions
on ceellular surfaace in fixed virus infectted cells miight be a maajor target of
o the host
immuune system. Thus diffeerent pathoggenicity betw
ween street virus and fixed
fi
virus m
might
be asssociated wiith differentt localizatio
on of virion maturation. However, little is knoown
abouut molecularr mechanism
m of virion maturation
m
T
both of fixeed and streeet viruses. To
eluciidate this, we
w have exprressed G-prroteins of CVS-26 straiin (fixed virrus) and Kyyoto
strainn (street viru
us) in HEL2293T cells upon
u
transfe
fection of th
he pCAGGS
S (CXN2)
plasm
mid bearingg G-genes off CVS-26 annd Kyoto sttrains, respeectively. Inttracellular
expreession and localization
l
n of G-protein of each strain
s
was thhen examineed by
fluorrescence anttibody technnique and Western
W
blott analysis using anti-G mAb (No.##7-19, kinndly providded from Drr.Kawai). Coonfocal laseer scanning microscopyy showed CVSC
26 G
G-protein waas mainly loocalized on plasma mem
mbrane, whhile Kyoto G-protein
G
was
predoominantly localized at perinuclearr membranee region. CV
VS-26 G-prootein was shhown
to bee expressed in abundance than Kyooto G-protein in HEK2293T cells by
b Western blot
b
analyysis. These results indiccate maturaation site of infectious RABV
R
solely determinned
by loocalization of
o G-proteinn. Four amin
no acids in a signal pep
ptide (SP) of
o G-proteinn of
CVS
S-26 were diistinct from
m those of Ky
yoto. The number
n
of puutative N-liinked
glycoosylation sittes was 3 annd 2 on G-pproteins of CVS-26
C
andd Kyoto, resspectively.
Furthher analysiss is requiredd to elucidatte whether these
t
differeences affectt intracellulaar
locallization andd expressionn level of RA
ABV G-prootein.
CO.113
R
viru
us-induced Oxidative Stress
S
Rolee of Mitochondria in Rabies
Alanndijany T1, Kammouni
K
W2, Roy Chhowdhury SK
S 3, Fernyhhough P4, Jaackson AC2 1
Univversity of Manitoba
M
- Medical
M
Miccrobiology, 2University
y of Manitooba - Internaal
3
Med. (Neurologgy), St. Bonniface Hosppital Researcch Centre - Neurodegennerative
versity of Manitoba
M
- Pharmacolog
P
gy and Therrapeutics
Disorders, 4Univ
ouse model of rabies with
Neurronal processs degeneration occurs in an experrimental mo
hindllimb footpaad inoculatioon of the chhallenge viruus standard--11 (CVS) strain.
s
CVS
S
infecction of prim
mary mousee and rat dorrsal root gannglion (DRG
G) neurons has associaated
axonnal degeneraation with axxonal swelllings and neeurite outgro
owth reducttion. The CV
VSinducced axonal swellings feeature proteein adducts of 4-hydroxxy-2-nonenaal (4-HNE),, a
markker for lipid peroxidatioon, indicatinng a criticall role of oxidative stresss. Western
immuunoblottingg analysis inndicated thatt adducts off 4-HNE ex
xpression is also increassed
in thee CVS-infected rat adrrenal medullla (PC12) cell line. Mittochondrial dysfunction is
one oof the most important causes
c
for overproducti
o
ion and accu
umulation of
o reactive
oxyggen species (ROS). We investigateed the effectts of CVS innfection on several
mitocchondrial parameters inn different cell
c types (D
DRG primaary neurons, PC12, mouuse
neurooblastoma (MNA),
(
andd baby hamsster kidney (BHK-S13)) cells) at 72
2 hrs post
infecction. The biochemical activity of electron traansport systeem (ETS) complexes (II, IIIII, aand IV) and Krebs cycle enzymes (citrate syntthase and malate
m
dehyddrogenase) were
evaluuated using a spectrophhotometric approach.
a
K
Krebs cycle enzyme acttivities weree not
affeccted in CVS
S- versus moock-infected
d cells. Com
mplex I activ
vity was siggnificantly
increeased in all CVS-infect
C
ted cells verrsus mock-innfected conntrols. Comp
plex I was
increeased by 30--35% in CV
VS-infected DRG and PC12
P
cells, whereas it was
w increassed
by 655-75% in MNA
M
and BH
HK-S13 cellls. These vaalues were proportiona
p
al to the
susceeptibility off the cells too CVS infecction suggessting a direcct effect of the
t CVS
infecction on Com
mplex I. Coomplex II-IIII activity was
w normal in
i the infectted cells.
Com
mplex IV acttivity was uppregulated in
i all types of CVS-inffected cells. However, the
t
increease did not relate to thhe susceptibiility of the cells
c
to the infection, su
uggesting aan
indirrect effect. We
W postulatte that enhannced Complex IV activvity in CVS-infected ceells
may play a role in avoidingg apoptosis. NADH, whhich is a Coomplex I-subbstrate, leveel
m
ed PC12 ceells. NAD+ level
was ssignificantlyy higher in CVS-infectted versus mock-infect
in CV
VS-infectedd PC12 cellss was similaar to that in mock-infeccted controlls. Despite tthe
increeased activitty of ETS complexes, CVS
C
infectiion reducedd the intracellular level of
ATP in PC12 ceells. The redduced ATP level in CV
VS-infected DRG neuroons may expplain,
t reductioon in the neuurite outgrow
wth that waas previouslly observed.. We
at leaast in part, the
prediict that a hig
gh mitochonndrial innerr membranee potential iss generated in CVS
infecction becausse of increassed proton pumping
p
accross the mitochondriall inner
mem
mbrane due to
t higher acctivity of Coomplex I and IV, and decreased
d
prroton
consuumption as indicated by
b reduced intracellular
i
r ATP level. Induction of a high
mitocchondrial membrane
m
potential pro
omotes electtron leakagee, primarily
y at the Com
mplex
I sitee, leading too ROS overggeneration and
a oxidativve stress.
CO.114
n RABV infection
Paraadoxical rolle of IFN in
Lafon M1 - 1Insttitut Pasteurr Paris Frannce
BV infectionn is sensitivee to type 1 IIFN
A serries of expeeriments cleearly indicatte that RAB
signaaling and th
hat P and N protein-meddiated IFN evasion is efficient
e
to promote
p
virrus
repliccation. Nevvertheless, in
i the coursse of infectioon, the IFN induction in
i the wholee
RAB
BV-infected nervous syystem, NS, is far from being
b
abrogated. Indeedd, after injection
of RA
ABV (Challenge Viruss Standard, CVS strain)) into the hiindlimbs of mice, a
progrressive infeection withinn the spinal cord and thhe brain is accompanie
a
d by a robuust
innatte immune response
r
chharacterized by a type 1 IFN responnse. It may not be
surprrising that IF
FN can be produced
p
in
n the NS durring infectioon because the mechannisms
evolvved by RAB
BV to escappe the IFN response aree restricted to
t infected neurons,
n
thee
only cell type ex
xpressing thhe P and N proteins.
p
Thhese mechan
nisms cannoot operate inn
n express any viral proteins, gliaal cells beinng rarely inffected
glial cells becauuse they do not
heless, glial cells are innnate-immunno-competent cells andd they do noot
in vivvo. Neverth
needd to be infected to mounnt an innate immune reesponse sugg
gesting thatt non infecteed
glial cells may be
b the sourcce of hetero
ocellular IFN in the RA
ABV -nfecteed NS. Onee can
o the heteroocellular IFN
N in RABV
V infection is.
i Beside
wondder what thee function of
intrinnsic antiviraal propertiess, IFN also controls thee expression
n of a large number of IFN
stimuulated genes (ISG). The ligand of the Program
mmed deathh protein-1, (PDL-1) (aalso
nameed B7-H1), is an ISG which
w
expreession is uprregulated inn RABV-inffected NS annd
whicch has been demonstratted to be a critical
c
factoor for RABV
V neuroinvaasiveness.
Thus, it can be proposed thhat RABV evades
e
the antiviral
a
effe
fect of IFN in
i the infectted
m the heterocellular IFN
N to facilitatte its
neuroons, whereaas RABV beenefits from
progrression in thhe NS .
CO.115
ALYSIS OF
F RNA EXP
PRESSION
N BY BLOO
OD MONO
ONUCLEA
AR CELLS
ANA
STIM
MULATED
D BY HUM
MAN RABIE
ES CSF
Willooughby RE1, Jia S1, Heessner M1 - 1Medical College
C
of Wisconsin
W
- Pediatrics
o effective antivirals, survival froom rabies haas been corrrelated withh the
In thhe absence of
appearance of neutralizing antibody within
w
7 dayss of hospitallization. Thhis adaptive
t earlier innnate immuune responsee to rabies, which can be
b
humooral responsse follows the
subvverted by thee rabies viruus phosphopprotein. Unnderstandingg the cerebrrospinal fluiid
(CSF
F) environm
ment affectinng innate an
nd adaptive immunity to rabies viruus is key too
imprroving rabiees survival. To
T sensitiveely detect thhe presence of cytokinees, chemokiines
and oother imporrtant immunne modulato
ors (small nuucleotides and
a lipids) inn CSF, we
emplloyed a nov
vel bioassay whereby a well-controolled periphheral blood mononuclea
m
ar
cell ((PBMC) po
opulation off a healthy blood
b
donor is used asa sensitive biosensor
b
thhat
transscriptionally
y responds to
t these diluute disease-aassociated factors.
f
Thee readout is a
compprehensive genome-sca
g
ale array. We
W examineed 7 control CSF and 133 CSF sampples
from
m 6 patients with
w laboratory-confirm
med rabies, dating from
m hospital days
d
4 to 26.
Dog and bat rabbies were eqqually repressented. CSF
F was incubbated with reeporter (PBMC)
m PBMC waas then extrracted, labelled and anallyzed using
for 9 hours, totaal RNA from
Affyymetrix Hum
man Genom
me U133Pluss2.0 array. Unsupervise
U
ed analysis separated raabies
CSF from controols but did not
n clearly group
g
rabiees samples by
b patient, suggesting
s
thhat
ments are greeater determ
minants of thhe
rabiees disease itself and asssociated medical treatm
innatte immune response
r
to rabies measured in CS
SF than are intrinsic
i
host variables. In
general, interferron-inducedd genes weree up-regulatted while cy
ytokine gennes were dow
wnman PBMC respondingg to human rabies.
r
regullated in hum
CO.116
CROARRA
AY ANALY
YSIS OF CE
ENTRAL NERVOUS
N
S SYSTEM
MIC
ASSOCIATED
D WITH TH
HE INFILT
TRATION OF MICR
ROGLIA IN
N MICE
OWING SIG
GNS OF PA
ARALYSIS
S AFTER THE
T
INTR
RAMUSCU
ULAR
SHO
INOCULATIO
ON OF RV (CVS-11 STRAIN)
1
UE S , Udaa A1, Sugiurra N1, Hamaamoto N1, Park
P
CH2 - 1National In
nstitute of
INOU
Infecctious Diseaases - Veteriianry Sciennce, 2Kitasatto Universitty - Veterinaary Patholoogy
e
year. As
A
Rabies occurs inn worldwidee and more than 70 0000 people diee of rabies every
n
l symptomss were preseented
the ddisease proggresses of paatients, morre specific neurological
incluuding such as
a insomniaa, anxiety, coonfusion, sllight or parttial paralysiis, excitationn,
halluucinations, agitation,
a
diifficulty swaallowing, annd hydropho
obia (fear of
o water).
Patieents show seevere paralyysis graduallly and evenntually deadd after a com
ma. Mice
inocuulated intram
muscularly (i.m.) with CVS-11 (fiix strain) shhowed the seevere hind limb
l
parallysis on 7 daays and thenn dead evenntually, how
wever mice inoculated
i
w CVS-111
with
intraccerebrally (i.c.)
(
were dead
d
withouut limb parallysis (Kojim
ma et al. 20009 and Sugiiura
et al.. 2011). Forr understandding of hindd limb paralyysis, mice inoculated with
w CVS-11,
h
i.m. aand i.c. wass comparativvely analysed by microoarray and hisotopatho
logy. Brainns and
spinaal cords of mice
m were separatively
s
y collected after
a
7 days of the postiinoculation of
i.m. aand i.c.. Virral antigens was similaarly observeed in both off brains andd spinal cordds in
micee inoculatedd i.m. and i.cc.. Pathologgically, micrroglia was infiltrated in
n spinal cordds in
micee inoculatedd i.m. not buut i.c.. In microarray, exxpression leevel of genees was
norm
malized withh each mockk. After com
mparative annalysis of geene expresssion in micee
inocuulated i.m. and
a i.c., signnificantly (ffold changee >2, /p/<0.005) changedd genes werre
exam
mined by Ing
genuity Pathhway Analy
ysis (IPA). As
A the resullts, calcium
m ion relatedd
gene and immun
ne responsee genes incluuding inflam
mmations, chemotaxis,
c
inflammatiion
w obvioussly up-regullated in i.m. in both of brains and spinal cordss.
and aapoptosis were
Addiitionally, the /p/ valuess of these in spinal cordds were obv
viously loweer than thosee of
brainns. Moreoveer, there is significant
s
changes
c
of S
Stat4, Ifng, Irf7 and Il12 which is the
t
centrral regulatioon factors off those resp
ponses. The evoked stroong immunee responses
assocciated with the infiltrattion of microglia in CN
NS of mice infected
i
i.m
m. suggest a
reasoon of damagge developeed severe paaralysis in mice
m inoculaated with CV
VS-11, i.m..
This work was supported
s
by a grant-in
n-aid for thee Health andd Labour Sccience Reseaarch
M
of Health, Lab
bour and Welfare
W
of Jaapan.
Grannt from the Ministry
CO.117
OTEOMICS
S ANALYS
SIS OF HU
UMAN RAB
BIES CSF
PRO
Willooughby RE1, Halligan B2, Green A2 - 1Medical College of
o Wisconsiin - Pediatriics,
2
Meddical Collegge of Wisconsin - Bioteechnology and
a Bioengiineering Cen
nter
ncompletelyy understood. Wildtypee rabies infeection
The ppathogenesiis of humann rabies is in
is miinimally cyttolytic or innflammatoryy, and does not includee major disru
uption to the
bloodd brain barrrier. Cerebroospinal fluidd (CSF) froom human raabies patiennts is therefoore
ideallly suited foor direct anaalysis by pro
oteomics appproaches thhat may subbstantively
eluciidate the im
mmune respoonses, alteraations in meetabolism an
nd fundameental cellularr
mechhanism that contribute to rabies paathogenesis and recovery.
We eexamined 200 control CSF and 13 CSF
C samplees from 6 paatients with laboratory-confi
firmed rabies, dating froom hospitall days 4 to 59.
5 Dog and
d bat rabies were
w equallly
repreesented. Try
ypsin digestts of CSF prroteins weree analyzed by
b liquid revversed-phasse
chrom
matography
y followed by
b mass speectrometry uusing a Therrmo LTQ-O
Orbitrap. Peeptide
identtifications were
w filteredd at a False Discovery Rate
R (FDR)) of < 5% an
nd proteins were
requiired to havee two or more peptide forms
f
obserrved to be coonsidered. No
N rabies viirus
derivved peptidess were deteccted in any sample. In aggregate,
a
the
t abundannce of 180
proteeins were statistically significant between
b
patiients and coontrols (p < 0.05) whenn
correected for mu
ultiple testinng and 36 proteins werre more thann 2 fold incrreased in
patieents and 64 proteins
p
weere detected only in pattients only. Some of thee groups thaat
thesee proteins were
w involveed in innate and acquireed immunity
y, complem
ment, proteasses,
strucctural proteins, synapticc granules, energy
e
metaabolism, inn
nate immun
nity and
natriuuresis.
CO.118
T
BE THE
T
LAST
T ATTEMP
PT OF THE
E HOST TO
TH17 CELLS: COULD THEY
EAR THE RABIES
R
VIIRUS?
CLE
Fernaandes ER1, Guedes F1, Martins JF
F2, Pagliari C3, Duarte MIS
M 3 - 1Insttituto Pasteuur de
2
São P
Paulo, Insttituto de Inffectologia Emílio Ribass - Departam
mento de Paatologia,
3
Facuuldade de Medicina
M
daa Universidaade de São Paulo
P
- Deppartamento de
d Patologiaa
Introoduction: following
f
ann antigenic stimulus naaive CD4+ T lymphocyytes becomee
activvated, expan
nd and differentiate intoo T helper ssubtypes Thh1 or Th2 lyymphocytes.
Receently, a new
w subtype naamed Th17 has
h been prroposed. Sim
milar to the other subtyypes
of im
mmune respoonse, Th17 cells requirre specific ccytokines annd transcription factorss for
their differentiattion. TGF-β
β along with
h IL-6 are crrucial cytok
kines in thiss process, while
w
L-21 has a role
r in the amplificatio
a
on of the Thh17 responsee and IL-23 is responsiible
the IL
for thhe maintenaance of diffeerentiated Th17
T
cells. Although
A
thhe role of Thh17 cells is not
yet fully
fu understtood, data from
fr
the liteerature sugggest that thesse cells havve importantt role
in hoost defense against
a
micrroorganism
ms, in particuular when thhe Th1 and Th2 type
immuunity is not efficient too clear the pathogen.
p
Aim: to evaluate and quanttify the cellss expressingg IL-6, IL-117 and TGF-β in specim
mens
us system inn human rab
bies cases trransmitted by
b dogs.
of ceentral nervou
Mateerial and methods:
m
sixx fragmentss of central nervous
n
sysstem (cortexx, hippocam
mpus,
basall ganglia, ceerebellum, medulla
m
obllongata andd spinal cord
d) were seleected from each
e
speciimen of the four humann rabies casses transmittted by dogss. By immunnohistochem
mical
reacttion with thee use of Streptavidin-bbiotin-peroxxidase methood it was exxamined thee
expreession of cyytokines IL--6, IL-17 an
nd TGF-β. All
A immunostained cellls were
quanntified using
g a grid-scalle in an areaa of 0.0625 mm2 considdering 40 fields in eachh
fragm
ment of the CNS (10 fields in menninge and 300 fields in parenchyma)
p
). Results were
w
expreessed in num
mber of cellls per mm2.
2
Resu
ults: it was observed
o
hiigh expressiion of TGF--β (586.68 cells/mm
c
), followed by IL6 (2228.79 cells/m
mm2) mainlly in the parrenquimal region and th
he presencee of cells
expreessing IL-17
7 primarily in meningeeal (187.21 cells/mm2)..
Discu
ussion and conclusion
n: consideriing that the cytokine microenviron
m
nment will direct
d
the tyype of imm
mune responsse against innfection, if there is a prredominancce of cytokinnes
such as IL-1 andd IL-6, theree is a proinfflammatoryy profile, if there
t
is an in
ncreased
GF-β and IL
L-10, we caan suggest an
a immunoreegulatory profile; howeever,
expreession of TG
the ccombination
n of cytokines can geneerate other profiles
p
of thhe immune response inn an
attem
mpt to comb
bat the infecctious agent. The concoomitant pressence of cellls expressinng
TGF-β, IL-6 andd IL-17 sugggest a Th177 pattern of immune ressponse, whiich would be
b an
mpt by the host
h to clearr the rabies virus
v
after the
t profiles of Th1 and Th2 immunne
attem
respoonse have faailed viral elimination.
e
CO.119
ANIM
MAL MOD
DELS AND
D BIOLOG
GICS EVAL
LUATION:: EXPERIM
MENTAL
RAB
BIES VIRU
US INFECT
TION AND
D DOSE TIT
TRATION
N OF CL184
4
MON
NOCLONA
AL ANTIB
BODY COM
MBINATIO
ON IN THE
E SYRIAN HAMSTER
Tayloor ST1, Ellison JA1, Frranka R1, Marissen
M
WE
E2, Rupprechht C1 - 1Cen
nters for Dissease
Conttrol and Prevvention, Attlanta, GA, USA
U
- Diviision of Higgh-Consequeence Pathoggens
& Paathology, 2Crucell
C
Hollland BV, Leeiden, The Netherlands
N
s
ute progressive encephaalitis responnsible for ov
ver 55,000 human
h
fatallities
Rabies is an acu
i
woound
each year. This zoonosis is preventablee, if promptt medical inntervention includes
A
ely 10 millioon people
care and both acctive and paassive immuunization. Approximate
xis (PEP) annnually. Th
he World Heealth
receiive rabies poost-exposurre prophylax
Orgaanization reccommends the adminisstration of hhuman and/oor equine deerived antirabiees immune globulin
g
(HR
RIG and ER
RIG) as welll as cell cullture vaccinne for moderrn
PEP in humans. However, in
i many devveloping reggions wheree canine rabbies is enzoootic,
nization aree necessary due to the cost
c prohibiitive,
alternnative soluttions for passsive immun
limitted supply of
o HRIG andd ERIG. Suuch disparitiies have pro
ompted the developmen
d
nt of
anti-R
RABV mon
noclonal anttibody (mA
Ab) cocktailss that can bee produced on an indusstrial
scalee with consiistent potenccy and decrreased produuction costss in compariison to HRIIG
and E
ERIG. To assess the effficacy of a mAb combination in raabies PEP, we
w evaluateed the
use oof CL184, a 1:1 proteinn mixture raatio of two human
h
anti--RABV mA
Abs
(CR557/CR4098)) produced on the PER
R.C6® humann cell line, in
i the Syriaan hamster
modeel. In separaate experim
ments, femalee hamsters were
w divideed into grouups and
inocuulated on Day -1 into thhe gastrocnnemius musccle with a leethal dose of
o a geneticaally
distinnct carnivorre or bat RA
ABV isolatee (Asian dogg or Parastrrellus hespeerus,
respeectively). On Day 0, HR
RIG at 20 IU
U/kg (n=211) or CL184
4 at 6 μg/kg,, 12 μg/kg or
o 16
μg/kgg (n=21/gro
oup) was addministered to groups at
a the site off inoculation
n. In each
experiment, a co
ontrol groupp (n=12) and a vaccine only group
p (n=21) recceived a placcebo
7 14, and 28,
2 hamsterss in experim
mental groupps received a
inocuulation. On Days 0, 3, 7,
50μl dose of com
mmercially available RABV
R
vacciine. High mortality
m
waas observed in
both placebo and vaccine only
o
groups by Day 40. Preliminarry data from
m the Syrian
ments demonnstrate these animals are
a a suitablee model and
d suggest thhat
hamsster experim
CL1884 may be a non-inferiior alternativve for HRIG
G in rabies PEP
P scenarios.
CO.220
ALYSIS OF
F RABIES VIRUS GL
LYCOPRO
OTEIN SEQ
QUENCES
S IN
ANA
REL
LATION TO
O THE PR
ROPOSED USE OF MONOCLO
M
ONAL ANT
TIBODIES
FOR
R POST-EX
XPOSURE PROPHYL
LAXIS
Kuzm
mina N1, Ellison J1, Orrciari L1, Maarissen W2, Kuzmin I1, Rupprechtt C1 - 1CDC
C,
2
Cruccell Holland
d
mune globullin (RIG) foor post-expo
osure prophy
ylaxis (PEP
P) is
The ddemand forr rabies imm
signiificant. Unfo
fortunately, the cost of RIG
R is prohhibitive for many
m
patiennts in
developing coun
ntries. Severral monoclo
onal antiboddies (MAbs) which neuutralize rabiies
h
been prroposed as a replacemeent for convventional RIIG due to thhe
viruss (RABV) have
abilitty of their laarge-scale production
p
a a reducedd cost. In thee present stuudy, we
at
generated 487 RABV
R
glycooprotein (G)) sequencess from a varriety of virall lineages, and
a
he dataset with
w 154 com
mplete and 115 partial G sequencees available in
supplemented th
Bank. The objective
o
waas to evaluaate variabilitty of knownn MAb-bind
ding epitopees on
GenB
the G
G, which maay preclude virus neutrralization. The
T analysiss demonstrated that binnding
site oof MAb CR
R57 (amino acids
a
226-2231 of the G ectodomaiin) is very conservative
c
e.
The ssubstitution
ns detected (such
(
as K2226R in seveeral raccoonn and Africaan dog RAB
BV
isolaates; L231P//S in severaal skunk, racccoon, and various
v
bat RABV lineeages; etc.) did
d
not ppreclude viru
us neutralizzation from previously published studies.
s
No substitutionns
that aabolished binding of MAb
M CR57 in
i escape mutant
m
studiees were deteected in
naturrally occurrring field RA
ABV isolatees. In contraast, numerous substituttions were
deteccted in the binding
b
site of MAb CR
R4098 (AA
A 330-338 off the G ectoodomain).
Exam
mples includde a K330N
N substitutio
on in a bat issolate from Brazil; V33
32I/F
substtitutions in several RAB
BV lineages, associateed with big brown
b
bats;; N336D in
severral viruses associated
a
w big broown bats in North
with
N
America, in Sou
uth-African
monggoose RAB
BV, in one African
A
and one Koreann dog RABV
V isolate; N336G/S
N
in
severral raccoon isolates; E3337D in sevveral caninee RABV from Serbia an
nd in the souuthcentrral skunk RA
ABV isolates; I338T inn the canyonn bat and Arctic
A
RABV
V isolates.
Subsstitutions in position 3336, particulaarly the N3336D, were detected
d
earlier in escappe
viruss studies and
d precludedd neutralizattion of suchh viruses by MAb CR40098.
Neveertheless, noo isolates haad substitutiions in bindding sites foor MAbs CR
R57 and CR
R4098
simuultaneously. There is noo reason to expect
e
that any of the viruses
v
from
m our study
woulld escape neeutralizationn by a combbination of these
t
MAbss in vivo. Thhe situationn is
different for HuMAb RAB1 (also referrred to as 177C7). We confirmed nu
umerous
3 which may
m abolish
h binding off MAb RAB
B1 as
substtitutions, paarticularly inn position 336,
was sshown prevviously by escape virus generation. The RAB1
1 was propoosed as a sinngle
MAbb for use in human rabiies PEP, claaiming that there
t
are noo natural RA
ABV isolatees
whicch harbor critical substiitutions in itts binding site (the com
mbination 3336D-346K in
i the
G ecttodomain). We encounntered this combination
c
n in the majoority of viruuses from onne of
the liineages asso
ociated withh big brownn bats distribbuted broad
dly in Northh America. Our
O
findings clearly demonstratte that the proposed
p
usee of a singlee MAb for rabies
r
PEP is
i
i
al recommenndations.
inapppropriate, inn line with internationa
CO.221
a effective as liveNeuttralization antibodies in combination of MCP-1 are as
atten
nuated rabiies virus in
n preventing mice from
m developin
ng rabies
Fu Z1, Huang J2, Li G2, Zhaang G2, Zhoou M2 - 1Unniversity of Georgia - Huazhong
H
Agriccultural Unniversity, 2University
U
off Georgia
virus that causes
ABV) is a neurotropic
n
c
fatal disease in humans
h
andd
Rabies virus (RA
mals. Curren
ntly there is no cure for rabies oncee clinical sig
gns appear. It has beenn
anim
hypoothesized thaat once the virus enterss the centrall nervous syystem (CNS
S), neutralizzing
antibbodies in thee periphery cannot crosss the bloodd–brain barrrier (BBB) into
i
the CNS.
Previious studiess have demoonstrated thhat treatment with live-aattenuated RABV
R
via the
t
intraccerebral rou
ute 5 days after
a
infectioon with wildd-type virusses can leadd to the clearrance
not oonly the atteenuated, butt also the wiild-type viruus. Direct administratioon of liveattennuated RAB
BV stimulateed high leveels of neutraalization anttibodies and
d enhanced the
BBB
B permeabiliity. Howeveer, direct inttracerebral administrattion of live-attenuated
RAB
BV possessees safety conncerns. In thhe present sstudy, neutraalization anntibodies weere
admiinistered in conjunctionn with a cheemokine, MCP-1
M
(know
wn to enhannce the BBB
B
perm
meability), in
nto mice aftter infection
n with wild--type virus. Significanttly more
proteection was found
f
in micce treated with
w this com
mbination when
w
compaared to treattment
with neutralizatiion antibodiies alone wiithout MCP
P-1. Furtherm
more, the co
ombined
ment with neutralizatio
n
on antibodiees and MCP
P-1 is as effeective as thee live-attenuuated
treatm
RAB
BV in prevennting mice from
f
develooping rabiess. These stuudies furtherr demonstraate
that eenhancemen
nt of the BB
BB is criticaal for immunne effectorss in the periphery to ennter
into tthe CNS to clear RABV
V.
CO.222
uman monooclonal anttibody com
mbination foor rabies poostDeveelopment of CL184 hu
expoosure proph
hylaxis, froom preclinical design to
t clinical evaluation
e
Mariissen WE1, Niezgoda
N
M2, Ellison J2, Franka R2, Kuzminaa N2, Kuzm
min I2, Tayloor T2,
2 1
2
Ruppprecht C - Crucell Hoolland bv - Project
P
Mannagement, Centers
C
for Disease Coontrol
and P
Prevention
t
The ccurrently reecommended prophylaxxis for indivviduals expoosed to rabies virus is the
combbined adminnistration off rabies vaccine and rabbies immunne globulin (RIG).
(
How
wever, limiteed supply haampers the availability of RIG, paarticularly in
n enzootic aareas.
To ciircumvent the
t global RIG
R limitatio
on we aimed to developp a human monoclonal
m
l
antibbody combin
nation, CL1184, for rabiies post-expposure prophylaxis (PE
EP) that wouuld
replaace the plasm
ma origin RIG.
R
CL184 consists off two human
n IgG1 mAbbs, CR57 annd
CR40098, which are directedd against noon-overlappping rabies virus
v
(RV) glycoprotein
g
n
epitoopes. Previoously, we haave shown thhat the in viitro breadthh of neutraliization of CL184
againnst a large panel
p
of streeet RV of vaarious anim
mal origins as
a well as in vivo protecction
by C
CL184 in a Syrian
S
hamsster rabies challenge
c
model was coomparable to results
obtaiined with hu
uman RIG. A detailed preclinical
p
selection prrocedure waas applied too
estabblish the CL
L184 antiboddy combinaation. Effortts on RV su
urveillance to
t ensure
adeqquate coveraage by CL1884 continuee. In additionn, encouragging data froom the Phasse I
(US and India) and
a Phase III (US and Philippines)
P
clinical evaaluation of CL184 havee
beenn obtained. In
I preparatioon of the pivvotal Phasee III evaluations for CL
L184, a finall
Phase IIb evaluaation has beeen executed
d for which data analyssis is ongoin
ng. The futuure
C
may help to ensuure consisteent supply of
o pivotal liffe-saving
availlability of CL184
bioloogics to rabiies endemicc areas and could
c
substaantially conntribute to thhe reductionn of
humaan rabies deeaths, when combined with
w educattional measuures and effforts to elim
minate
caninne rabies.
CO.223
r
nt rabies viiruses for both
b
GM--CSF or flaagellin imprroves the efficacy of recombinan
pareental and orral immuniizations
Fu Z1, Zhou M2, Zhang G2, Ren G2 - 1Huazhong
H
A
Agricultural
l Universityy - Universiity of
Georrgia, 2Univeersity of Geoorgia
p
stuudies indicaated that reccombinant rabies virusees expressinng chemokinnes
Our previous
and ccytokines (iincluding GM-CSF)
G
coould enhancee the immunnogenicity by
b inducingg
innatte immunity
y and recruiiting/activatting dendritiic cells and B cells. In this study,
bacteerial flagelliin was cloned into the rabies
r
viruss genome annd recombin
nant virus
rLBN
NSE-Flic was rescued. To comparre the immuunogenicity of rLBNSE
E-Flic with
recom
mbinant virrus expressinng GM-CSF
F (rLBNSE
E-GMCSF), mice were immunizedd with
each of these reccombinant rabies
r
virusses by i.m. oor the oral rooute. The paarental viruus
BNSE) withoout expressiion of any foreign
f
moleecules was included fo
or comparisoon.
(rLB
The ii.m.-immunnized mice were
w bled att three weekks after the immunizatiion for the
meassurement off virus neutrralizing antiibodies (VN
NA) and theen challengeed with 50 LD50
L
CVS
S-24. The orrally immunnized mice were
w boosteered after th
hree weeks and
a then bleed
and cchallenged one
o week affter the boooster immunnization. It was
w found thhat both thee
recom
mbinant virruses LBNS
SE-GMCSF and LBNSE
E-Flic induced higher levels
l
of VN
NA
and pprotected more
m
mice aggainst rabiess challenge than the paarental rLBN
NSE in bothh the
i.m.- and the oraally immuniized groupss. Together, these studiees suggest that
t recombbinant
xpressing GM-CSF
G
or flagellin aree better vaccines than the
t parent virus
v
rabiees viruses ex
for bboth parentaal and oral im
mmunizatioons, most likkely by recrruiting/activvating dendrritic
cells.
CO.224
PER
RSISTENCE
E OF ANT
TI-RABIES
S NEUTRA
ALIZING ANTIBODI
A
IES IN A
RUR
RAL AMAZ
ZONIAN COMMUNI
C
ITY VACC
CINATED WITH PURIFIED VERO
CEL
LL RABIES
S VACCIN
NE (PVRV) FOLLOW
WING VAM
MPIRE BAT
T RABIES
OUT
TBREAK
Medeeiros R1, Maestri
M
A2, Le
L Guern A--S3, Martoreelli L4, Alm
meida M4, Kataoka
K
APA
AG4,
5
5
5
5
5
3
Houiillon G , Jussot V , Zoccchetti C , Bosch
B
Castellls V , Rasu
uli A , Chteo
oui M , Jalllet
C3, B
Behillil S3, Assef
A
R2, Rodrigues
R
L2, Lima R6, Rotivel Y7, Tordo N8 - 1Universiddade
Fedeeral do Para,, Brazil, 2Unniversidadee Federal doo Para, 3Instiitut Pasteurr, Paris, 4Cenntro
de Controle de Zoonoses,
Z
S Paulo, 5Sanofi Pastteur, Lyon, 6Secretaria de Saude do
São
7
8
P
Innstitut Pasteuur - Virologgy
Para,, ) Institut Pasteur,
man rabies trransmitted by
b vampire bats is a connstant threaat in the Am
mazonian reggion,
Hum
regullarly exceedding in fatallities the claassical dog transmitted
t
rabies in reecent years. In
20044-5, several outbreaks have
h
occurreed in Para, B
Brazil. In May-June
M
20005, followiing
15 deeaths, mostlly children, in Augustoo Correa, a rural
r
commuunity of 53,,000 peoplee
dispeersed along the Amazon estuary, 3,500
3
subjeccts aged from 2 to 60 years
y
receiveed
puriffied Vero ceell rabies vaaccine (PVR
RV Verorabb®, Sanofi Pasteur,
P
Fran
nce) for posst- or
pre-eexposure pro
ophylaxis. The
T presenttation will summarize
s
t follow-uup of this
the
comm
munity duriing 4 years after
a
vaccinnation. The persistence
p
of anti-RABV neutraliizing
antibbodies (VNA
Abs) levels was evaluaated by RFFIT, FAVN, and Plateliia ELISA (B
BioRAD
D®) against the PV andd CVS referrence strainss as well as a vampire bat
b isolate.
Subjeects with VNAb
V
levelss <0.5 IU(EU
U)/mL weree boosted. From
F
a totall of 507 subbjects
incluuded in the study,
s
428 (84.4%)
(
werre availablee all along thhe follow-u
up, an excelllent
adherence consiidering the isolation,
i
diispersion, annd nomadicc live of the communityy.
markably, 5%
% to 7% of the
t surveyedd populationn were poteentially re-eexposed onee to
Rem
severral times eaach year throough animaal bites (maiinly dogs also bats, catss, monkeys)).
The ppersistence of the WHO
O minimum
m “seroproteective” level of VNAbss (≥0.5 IU/m
mL)
four years after vaccinationn was observ
ved in 85.8%
% of the non boosted population
p
((346
s
diifference inn VNAb perrsistence pro
ofiles were
patieents). Interesstingly, no statistical
obserrved in pre--exposure (445 subjects)) and post-eexposure (30
01 subjects)) treated
popuulations. Gloobally, the VNAb
V
levell and persisttence were better in yo
oung populaation
than in elderly, and
a in femaales than in males, partiicularly the 16–40 yearrs old maless
otection ratees. No interrference was observed
whicch showed loower GMT and seropro
betw
ween anti-maalaria treatm
ment and PV
VRV immunnogenicity. At the methhodologicall
levell, RFFIT annd FAVN reesults appearred highly concordant.
c
The concorrdance was
loweer with the ELISA
E
resullts that show
wed a globaal increase in GMT valu
ue over the
yearss paralleled by a decreaase in statisttical correlaation with RFFIT
R
(Pearrson’s
correelation coeff
fficient = 0.882 in 2007 to
t 0.42 in 20009). A hun
ndred serum
m samples were
w
seleccted random
mly each yeaar to evaluatte the concoordance of RFFIT
R
resullts using thee PV
strainn versus a lo
ocal vampirre bat isolatte.
In suummary, this study dem
monstrated persistence
p
o anti-rabiees VNAbs in
of
i the vast
majoority of vacccinees (PVR
RV Verorabb®) from thiis communiity at repeatted risk of
vamppire bat bitees.
CO.225
uation of raabies virus variants
v
Rabiies Surveilllance in thee United Sttates- evalu
Dyerr J1, Blanton
n JD1, Ruppprecht C1 - 11Centers foor Disease Control
C
and Preventionn Divission of Highh-Consequeence Pathoggens and Patthology, Po
oxvirus and Rabies Brannch,
Rabies program
m
6
rabid animals reppresenting a
Durinng 2011, 499 states and Puerto Ricoo reported 6,031
1.9%
% decrease from
f
the 6,153 rabid an
nimals reporrted in 2010
0. Relative contribution
c
ns by
the major
m
animaal groups weere as follow
ws: 1,981 raaccoons (322.8%), 1,6277 skunks
(27.00%), 1,380 bats
b (22.9%
%), 427 foxees (7.1%), 303 cats (5.00%), 65 catttle (1.1%), and
a
70 doogs (1.2%). Comparedd to 2010, a significant increase was reported among rabiid
skunnks. Canine rabies viruss transmission has beenn eliminatedd in the Unitted States since
20044 and monitooring the raabies virus variant
v
assocciated with rabid domeestic animalls is
criticcal. We evalluated rabiees diagnostic submissioon data for the
t US from
m 2008-20111 for
reporrted rabid dogs,
d
cats annd coyotes. A total of 11,546 rabid cats,
c
dogs and
a coyotes were
reporrted, with raabies virus variants
v
chaaracterized iin 35%. Catts compriseed the majorrity
of rabbid animalss not characcterized. No canine rabiies virus variants were reported. Most
M
rabidd domestic animals
a
werre infected with
w the rabbies virus vaariant circullating in thee
predoominant meesocarnivoree reservoir from
f
the geeographic arrea of subm
mission.
How
wever, isolated cases asssociated witth bat rabies virus variaants were reeported. Theese
findings highlight the need for enhanceed surveillaance to monnitor the circculation of
mine emergennce of new
rabiees virus variiants in locaal carnivore populationns to determ
rabiees virus variiants. State health
h
depaartments maay not test suuspect rabidd animals unnless
a hum
man exposuure occurs. Moreover,
M
v
variant
typinng is not peerformed onn all sampless
thouggh CDC proovides rabiees virus charracterization, if requested. The pu
ublic health
impliications of host
h shifts and
a potentiaal spillover of rabies viirus variantss from wildllife
to doomestic anim
mals reinforrces the neeed for additional laboratory diagnoostics.
CO.226
BIES AND RABIES PROBLEM
P
MS IN NIGE
ERIA
RAB
Ogunnkoya AB1,22,3,4,5,6 - 1Ahhmadu Belloo Universityy, Zaria, Niggeria - Veteerinary
Mediicine, 2Osinnubi, MOV - Rabies Reesearch Uniit, Centers for
f Disease Control
C
andd
Preveention, Atlaanta, G.A-U
U.S.A, 3Garbba, A - Diaggnostic and Extension Dept,
D
Natioonal
4
Veterinary Reseearch Instituute, Vom- Nigeria,
N
Auudu, SW - Dept.
D
of Vet.. Medicine, Fac.
Y - Diagnosstic
of Veet. Medicinne, Ahmadu Bello Univversity, Zaria-Nigeria, 5Atuman, YJ
and E
Extension Dept,
D
Nationnal Veterinaary Researchh Institute, Vom- Nigeeria,
6
Ehim
miyein,AM - Dept. of Vet.
V Mediciine, Fac. off Vet. Mediccine, Ahmad
du Bello
Univversity, Zariia-Nigeria
Officcial reportinng of rabies in Nigeria started in 19912. The National Veteerinary Research
Instittute (NVRI)), Vom, witthin the last 77 years, cconfirmed 4809 cases of
o animal rabbies
in Niigeria. Rabiies control through
t
imm
munization programs
p
has crashed woefully
w
annd
conseequently, th
he rabies situuation has become
b
chaaotic and confounding. Locally, onnly
2,1377,615 dosess of dog antii-rabies vacccines were produced between
b
195
56 and 20055
(averrage 43,6255 per year) by
b NVRI, Vom.
V
Dog poopulation in
n Nigeria is currently
estim
mated at 8 million.
m
In thhe last 20 yeears of reseaarch and folllow ups onn rabies and
assocciated probllems, only 10%
1
of the dogs’
d
popullation receivved anti-rabbies
immuunization. Within
W
the immunized dogs, rabiess outbreaks occurred frrequently.
Eviddences on th
he trend of rabies cases recorded (11983-1991) confirmed 40%-60%
increease in rabiees positive cases
c
for evvery decade in Nigeria. Molecular epidemiology
and pphylogenetiic analysis study
s
of som
me dog rabiees isolates in
i Plateau State
S
confirm
med
the fl
flow of rabiees virus from
m neighboriing and far North Africcan countriees into Nigeeria.
Somee studies off prevalencee of rabies antigens
a
in the
t brain and saliva of apparently
a
healtthy dogs slaaughtered foor human co
onsumption in Nigeria; revealed a 28% prevallence
of rabbies antigen
n in the connsumed dogss in North-W
West, 31% from
f
North-East and 244%
from
m North-Cen
ntral regionss of Nigeriaa. Similarly, 6%-8% of the dogs haad rabies anntigen
in theeir saliva att the point of
o slaughter..
A stuudy of the epidemiolog
e
gy of rabies in wildlife in Bauchi State,
S
revealled the preseence
of rabbies antigen
n in mongooose (11%), jackals
j
(9%
%), squirrels (8.3%), hyydrax (17%)) and
wild cats (16%). This suggeests an ongo
oing spreadd of rabies within
w
the wild
w animals in
Nigeeria.
Concclusively, raabies is a prroblem in Nigeria
N
such that even th
he apparenttly healthy dogs
d
slaugghtered for human
h
conssumption haarbor the virral antigen in the brainns and salivaa.
This is an obvioous public health risk an
nd may havve serious im
mplication. The
T low nuumber
of Niigerian dogs immunizeed (10% insttead of 70-880%) leavess the countrry with
abunndance of ep
pizootic sibllings for rabbies outbreaaks. The wid
de spread off rabies in thhe
wildllife is an em
merging proof of the role of wildliffe in the epiidemiology of rabies inn
Nigeeria. It is ourr recommenndation thatt a RITA moodel to rabiees control as
a employedd in
the A
Americas bee applied in West Africca especiallyy Nigeria.
Keyw
words: Rab
bies, Rabies problem, Rabies virrus, Appareently health
hy dogs,
Vacccination, Nigeria
CO.227
ANSLOCAT
TION OF DOG
D
RAB
BIES IN ISR
RAEL BY TOURISM
T
M
TRA
Daviid D1, Dverees N1, Yakoobson BA2, Dvorkin Z3, Davidson I4, Yagil J5, Oved Z6 1
Kim
mron Veterinnary Institutte - Rabies Laboratory, 2Kimron Veterinary
V
Institute
I
Direcctor, 3Direcctor of Veterrinary Serviices, Jerusallem district, Israel, 4Kiimron
5
Veterinary Instittute, Bet Daagan 50250, POB 12, Israel,
I
Bet Zait,
Z
Israel, 6Veterinaryy
o Israel, Beet Dagan, 50
0250, Israell
Serviices, State of
mic and strayy
Rabies is enzoottic throughoout the Middle East. Inn Israel rabiees is endem
miliaris) form
m the main reservoir annd transmittter. Since 2004
2
and to the
dogs (Canis fam
w to the
preseent the statee of Israel haas been forcced to cope with a rabiees strain new
counntry, designaated V7. Beecause dogs are in closee contact wiith people thhe new V7
strainn that circullated at the Northern reegion of Israael possessees a serious health threaat to
humaans.
In the present coommunicatiion we repo
ort a tourism
m type of rab
bies transloccation, in which
w
mily from Jeerusalem toook their unv
vaccinated dog
d to the northern
n
Israael.
a fam
On 19 Decembeer, 2011 a doog was diaggnosed positive for rabiies in the Issraeli Nationnal
ory at the Kimron
K
Veteerinary Instittute. A casee investigatiion revealedd that
Rabies Laborato
d belongiing to a fam
mily living inn
on 133 Decemberr, a 3 years old Goldenn Retriever dog
Jerussalem showed clinical symptoms
s
of
o inappetennce, salivatiion and incooordination.. The
dog w
was vaccinaated twice against
a
disteemper and parvo
p
virus Duramune
D
Max®
M
(Forrt
Dodgge, Iowa, USA)
U
but nott against rabbies. On 15 December the dog was admitted to
t a
privaate veterinarry clinic in Jerusalem and
a under cllinical exam
mination it showed
s
unusually alert behaviour
b
a reactionn to externall stimuli. Th
and
he dog show
wed no clinical
ggression duuring the perriod of illneess. On 16 December
D
t dog show
the
wed
sympptoms of ag
ulteriior deterioraation of clinnical sympto
oms with coonvulsions and unconsciousness aand
was ttreated with
h diazepam (Assival, Teva). As noo health imp
provement was
w seen, thhe
dog w
was euthaniized on 18 December
D
a was trannsferred to the
and
t Kimron Veterinary
Instittute.
Rabies was diag
gnosed by direct
d
fluoresscence assaay, and was confirmed by
b isolationn of
mily of suckkling
the raabies virus in tissue cuulture and itss inoculatioon into a fam
micee. Reverse transcriptase
t
e - PCR andd direct sequuencing weere applied to
t a 469 basse
pair ((bp) G-L in
ntergenic reggion fragmeent and to thhe entire 1350 bp of thee nucleoprootein
gene . A phylog
genetic tree showed
s
thatt the Jerusaalem dog's sequence belonged to thhe V7
t circulatted in northeern Israel. D
During the period
p
of 20004 to 2011, 181
genetic variant that
rabiees viruses beelonging to V7 geneticc variants weere isolatedd from rabidd animals in
northhern Israel. The Jerusallem dog's raabies virus isolate
i
was the only example of thhe V7
genetic variant that
t was eveer isolated from
f
an anim
mal in Centtral Israel. The
T Jerusaleem
was probably bitten byy the rabid stray dog thaat circulated
d on the easstern coast of
o the
dog w
Lakee Kinneret loocated in Northern
N
Israael. After ann incubationn period of about
a
2 monnths
the cclinical symp
ptoms appeeared in Jeruusalem.
As a consequencce of this raabies diagnoosis, post-exxposure vacccination waas given to the
t
o veterinariaans and an additional
a
1 people, who
18
w had com
me into conttact
two oowners, two
with the dog in Jerusalem.
J
V
Various
san
nitary measuures were im
mposed in Jerusalem,
J
ssuch
nt of measurres against stray
s
animalls, boostingg of domestiic dog
as reinforcemen
mals can seerve
vacciination and quarantine of unvaccinnated dogs. Because doomestic anim
as a bbridge betw
ween wildliffe rabies reservoirs andd humans, thheir vaccinaation greatlyy
effecctive as public health toools that aree available to
t safeguard
d the humann health.
CO.228
ANSICIÓN
N EPIDEMIIOLÓGICA
A DE LA RABIA
R
CA
ANINA EN MÉXICO
TRA
Varggas FP1, Guttiérrez VC1, Chávez IA
AF1, Lezanaa MAF1 - 1Secretaría
S
dee Salud/Méxxico CEN
NAPRECE
México com
mo resultado de las camp
pañas masivvas de vacuunación antirrábica caniina a
En M
partirr de 1990 y que se han mantenido a la fecha, el registro de
d casos con
nfirmados por
p
laborratorio por IFD
I
en estaa especie obbservó un deecremento sostenido
s
añño con año;
compparando estee comportam
miento en dos
d períodoss, refiere quue en el prim
mero de los 90’s
se accumularon 13,811
1
casos en contrasste se reducce para el seegundo del 2000
2
a 921 casos
con ddecremento del 93.3%,, se describeen a continuuación algunnas variablees
epideemiológicass asociadas a ellos.
oblación, enn la década de los 90’s los casos predominaro
on en municcipios
Denssidad de po
con ccaracterísticcas urbanas por arriba del
d 80%, afe
fectando colonias o locaalidades
popuulosas de claase media baja,
b
el restoo se ubicó enn localidadees de municcipios con
caraccterísticas ru
urales; en ell período deel 2000 se innvierte estaa proporciónn y predomiinan
los casos en áreaas marginaddas de las grrandes ciuddades y de tiipo rural. Deestaca que los
l
p
a basureros
b
descontroladdos,
casoss de los últimos 3 años comprendeen lugares próximos
vivieendas en desspoblado dee alta marginnación careentes de servvicios públiicos básicoss
(callees, electricidad, agua potable
p
y dreenaje) lo quue favorece exceso de perros
p
en
situaación de callle.
l década dee los 90’s lo
os casos ocu
urrieron en 31
Entidades federrativas afecctadas, en la
s divide el país, contraastando tress entidades que
q acumullan
entiddades de las 32 en que se
56% del total dee los casos; en el períod
do del 20000 se reduce a 24 las entiidades que
d acumulaan el 65% del
d total.
regisstraron casos, de éstas dos
s
como
Dosiis aplicadass de vacunaa antirrábicca canina, a través de centros de salud
puntoo de referenncia para ubbicar en la loocalidad loss puestos o brigadas
b
de vacunaciónn ya
identtificadas por otras jornaadas de saluud por la comunidad; en la décadaa de los 90’ss se
apliccaron 101.3 millones dee dosis, paraa el períodoo del 2000 se incrementtó a 196.2
milloones de dosiis.
demiología molecular,, de la transferencia de
d tecnologíía del CDC
C, a partir dee
Epid
19966 con la caraacterizaciónn antigénicaa con panel de
d 8 anticueerpos monoclonales, see
corrooboró en alggunas muesttras (160) de
d perro o dee personas fallecidas
f
por rabia
transsmitida por este animall según interrrogatorio, la
l presenciaa de la variaante V1 (perrro)
en ell 95% de lass muestras y el resto V33 (zorrillo) y V11 (murrciélago). A partir del año
a
20066 se realizó el
e secuenciaamiento nuccleotídico en
e algunas de
d estas mueestras, se utiilizó
comoo referenciaa la clasificaación que hiizo el CDC (doce linajees de virus de
d rabia en
perroo), corroborrando que enn la región centro
c
del país
p continúúa presente el
e linaje D5 por
persiistir el foco enzoótico en
e el Valle de
d México, de manera aislada con
n casos únicoos
los liinajes D8 enn el foco ennzoótico de Puebla,
P
D9 en el foco enzoótico
e
de
d Oaxaca y con
casoss permanenntes en los últimos añoss el linaje D10 en el surreste en el foco
f
rábico en
Yucaatán y Chiappas, recienteemente se activó
a
el D1 (asociado perro-coyot
p
te) en Nuevvo
Leónn en la fronttera con Esttados Unido
os (Texas).
CO.229
TECTION OF
O THE DOG/COYO
D
OTE VARIIANT OF RABIES
R
VIIRUS IN THE
T
DET
MEX
XICO-US BORDER
B
Jaram
millo E1, Viillarreal JZ1, Galindo EIG
E 1, Carrannza P2, Melééndez A3 - 1Laboratorioo
Estattal de Saludd Pública - Secretaría
S
dee Salud del Estado de Nuevo
N
Leónn. Guadaluppe,
2
Nuevvo León, México, Cenntro de Investigación Biomédica del Noreste - Instituto
Mexiicano del Seeguro Sociaal. Monterreey, NL, Méxxico, 3Laboratorio de Rabia
R
- Instiituto
de D
Diagnóstico y Referenciia Epidemioológicos. México, DF
Curreently in the border states of northeern Mexico,, samples frrom skunks and dogs thhat
have the V-1 anntigenic variiant of the raabies virus have been identified,
i
h
however,
geenetic
I 2010, 4 cases
c
of hum
man rabies were
w
analyysis indicatees that they are strains of skunks. In
regisstered in Meexico, whilee in Nuevo Leon
L
there w
was 1 case of canine raabies transm
mitted
by innsectivorouss bats. It hass been almo
ost a decadee since the apparent
a
non
n-transmission
of the (V-1) variiant dog/coyyote, in thee border bettween Mexico (Coahuilla and
Tamaaulipas) andd USA (Texxas). Surveiillance data suggest thaat this variannt of caninee
rabiees virus is no
ot longer inn circulationn in the Unitted States of America, however, thhe
last ddetection waas registeredd in March 2004 in thee US-Mexicco border. In
n addition, one
o
dog ccarrying thee rabies viruus was deteccted in Junee 2011 in the town of Sabinas
S
Hidaalgo,
NL., which is ab
bout 130 km
m from the US-Mexico
U
border. This finding iss very impoortant
becauuse the (V-1) variant (ddog/coyote)) could be ppresent on thhe border off both counttries.
The ggeography of
o this regioon is very siimilar to Teexas, which may permitt the free
moveement of caarnivorous species
s
on both
b
sides off the borderr. Thereforee, the objective
of this study waas to identifyy the (V-1) variant of rabies virus in the dog from
f
Sabinaas
Hidaalgo, NL. Byy using direect immunoofluorescencce, antigenic characteriization withh a
panel of 8 monooclonal antibbodies, RT--PCR and nuucleotide seequencing techniques, the
t
709ppb (751nt to 1460nt) annd 420pb (992nt to 1411nt) fragmeents from thhe semi-variiable
regioon of the virral N proteinn were anallyzed. Accoording to thee antigenic characteriza
c
ation,
the vvariant found was the V-1,
V while thhe moleculaar study wass also positiive for this
variaant. Our resu
ults suggestt the same liineages pubblished by Velasco-Vill
V
la et al., 20005 in
the aanalyzed sam
mple. This study
s
demon
nstrates the actual prev
valence of th
he V-1 variaant
(dog//coyote) in the US-Mexxico borderr and warns about the risk for transsmission off the
V-1 variant
v
to humans,
h
as well
w as to doomestic andd wild animaals.
Acknnowledgements: We arre grateful too Miguel Angel
A
Zunigaa, Isabel Agguilar Tavitaas
and A
Alma Lilian
na Lizarán Meneses
M
forr their suppoort in the diiagnosis of rabies
r
viruss.
Finanncial support: This worrk was supp
ported by thhe Rabies Prrogram of thhe Ministry of
Heallth of Nuevo
o Leon and Health Servvices of Nuevo Leon.
CO.330
PORTANCIIA EPIDEM
MIOLÓGIICA DE LO
OS MURCIIÉLAGOS
IMP
INSE
ECTÍVOROS EN LA
A TRANSM
MISIÓN DE
EL VIRUS RABIA A FELINOS Y
OTR
ROS MURC
CIÉLAGOS CASERO
OS EN EL VALLE DEL CAUCA,
COL
LOMBIA.
Núñeez CM1, Páeez MA2, Heernández RC
C3, Escobarr DH4, Boneelo PA1 - 1Grupo
G
VIRE
EM.
Labooratorio de Virología.
V
U
Universidad
d del Valle - Valle del Cauca.
C
Coloombia,
2
Laboratorio Virrología. INS
S-Bogotá - Bogotá. Coolombia, 3Unnidad Ejecuutora de
S
Saneeamiento del Valle - Vaalle del Cauuca. Colombbia, 4Secretaaría Departaamental de Salud
del V
Valle del Caauca - Valle del Cauca. Colombia
d años see ha destacaddo la imporrtancia epideemiológica de los
En loos últimos diez
murcciélagos inseectívoros coomo transm
misores de laa rabia a feliinos y otross murciélagoos
insecctívoros/caseros en Collombia. En junio del 2012, despuéés de 21 año
os sin registtrarse
casoss de rabia enn humanos en el Deparrtamento deel Valle del Cauca, se presentan
p
doos
muerrtes en mujeeres mordiddas por gato infectado con
c la variannte antigéniica 4 (VAg44),
cuyoo reservorio es el murciiélago insecctívoro Tadaarida brasilliensis. Objjetivos.
Determinar las asociacione
a
s entre espeecies de murrciélagos, su comportaamiento, la
e
gico con énfasis en riessgo
transsmisión del virus entre ellos y su significado epidemiológ
para la población humana y mascotas. Metodologgía. Durantee el periodo
o diciembre 1999
or el prograama de vigillancia
a junnio 2012, fueron capturrados más de 1.321 murrciélagos po
de raabia en el deepartamentoo del Valle del
d Cauca. El
E diagnóstiico de rabiaa se hizo porr
inmuunofluoresceencia directta e inoculacción en ratoones, utilizaando tejido encefálico
e
d los
de
murcciélagos cappturados. Laa tipificación viral se hiizo por inm
munofluoresccencia indirrecta
usanddo anticuerppos monocllonales. Ressultados. See detectaronn dos ejempplares de
Eptessicus brasilliensis posittivos para raabia en los años
a
2000 y 2002, y do
os casos máás en
especcímenes de E. brasiliennsis y Molossus molosssus, en el 20008. Se encontró virus rabia
VAgg3 en gato añño 2009 y VAg4
V
en huumano morddido por gatto año 2012
2. Se encontrraron
distinntas especiees de murciéélagos, com
mo E. brasiliiensis, M. molossus,
m
Myotis
My
nigriccans,
Glossophaga sooricina, Nocctiliio albiveentris y Carrollia perspiicillata, com
mpartiendo
r
de V
VAg 3 y 4, en murciélaagos M.moloossus
refuggios en casaas. Se detecttaron virus rábicos
y E.bbrasiliensis así como enn gato y doss humanos. Conclusion
nes. En Vallle la presenncia
de laas VAg 3 y 4 del virus rábico
r
en murciélagos
m
insectívoros/caseros, probablemen
p
nte,
ha sido facilitadda por la defforestación de
d los hábittats naturalees de estas especies;
e
asíí
d arquitectuura urbana que
q provee un
u hábitat artificial
a
quee posibilita el
comoo el estilo de
contaacto físico entre
e
las esppecies, aumeentando la pprobabilidad
d de transm
misión de rabbia
entree estas y en las personaas que habitaan las casass invadidas. Ante las dificultades para
p
contrrolar la rabiia en murciéélagos y la falta
f
de herrramientas ad
decuadas, laa vigilanciaa
contiinua de la ennfermedad en los murcciélagos, basada en el diagnóstico
d
y la tipificaación
de loos virus rábiicos por laboratorio, enn los asentam
mientos hum
manos y alreededor de ellos,
e
la vaacunación prreventiva enn animales domésticos
d
y de produucción, así como la
educación de la comunidadd para la conncientizacióón del riesgo
o y la recoleección pasivva de
u análisis, se conviertenn en las mejjores herram
mientas paraa prevenir laa
muesstras para su
transsmisión a huumanos. Aggradecimieentos y Finaanciación. Secretarías
S
de Salud
Depaartamental Valle
V
y Munnicipal Cali, Unidad Ejjecutora de Saneamientto Valle,
Labooratorios dell Instituto Nacional
N
de Salud y Unniversidad del
d Valle.
Núñeez MC. Infeectio. 2012; 16(1): 23-229.
CO.331
CONDARY TRANSM
MISSION OF RABIES
S IN LATIN
N AMERIC
CA
SEC
2
Kotaait I1, Carrieeri ML2, Ruppprecht C3 - 1Instituto Pasteur,
P
In
nstituto Pastteur - Seçãoo de
Diaggnóstico, 3Centers for Disease
D
Prevvention and Control
mpared to prrimary pathways amonng reservoirss, secondaryy transmissiion of rabies
Com
viruss has not recceived muchh attention from
f
researrchers or pubblic health professiona
p
als,
becauuse spilloveer of infectioon from hem
matophagouus and non-hematophaggous bats too a
potenntial vector is believed to be uncom
mmon. Thiss review seeeks to remin
nd those
workking on Latiin Americann rabies conntrol program
ms of the poossibility off a bat-cathumaan pathway. Followingg the increasse in the conntrol of canine rabies inn most Latinn
Ameerican counttries, epidem
miological surveillance
s
e also focuseed on hemaatophagous and
a
non-hhematophaggous bats annd the use of
o molecularr techniquess in the charracterizationn of
rabiees. At least eight
e
cases of secondarry transmisssion to hum
mans were id
dentified in Latin
L
Ameerica from 2001 to 20122: one in Brrazil (2001), two in Costa Rica (20001), four inn
Coloombia (two in
i 2008 andd two in 201
12) and one in Ecuadorr (2009). In each case, the
t
epideemiologicall investigatiion implicatted a cat as the
t vector. The
T antigennic and geneetic
analyyses identifiied variantss maintainedd by the hem
matophagouus bat Desm
modus rotunddus.
Fruitt-eating batss in the genuus Artibeus may also be
b affected by
b a variantt similar to that
t
of D.. rotundus. Such fruit bats
b may bee found in urrban areas. Such affectted species can
c
transsmit rabies virus
v
to felidds, which arre importannt predators of bats. Therefore, in cases
c
of huuman rabiess following aggression by cats in areas
a
that are otherwisee free of cannine
rabiees (variants 1 and 2) buut where theere are rabiees epizooticss in sentinells such as
herbiivores, the hypothesis
h
o secondarry transmisssion of bat rabies
of
r
virusees should allways
be innvestigated.
CO.332
ndards and Assays forr Rabies Serology
Stan
Moore S1, Hanlo
on C1 - 1Kaansas State University
U
- KSVDL/R
Rabies Laborratory
oassays are currently ussed to meassure humoraal immunity
y to the rabiees
Seveeral immuno
viruss. A standarrd or referennce serum iss a consistennt componeent of all asssays to
standdardize the results
r
and control the assay perfoormance. Tw
wo human innternationall
standdard rabies immune
i
gloobulin (SRIG) referencce serum preeparations are
a recognizzed
by thhe World Heealth Organnization (WH
HO): the firrst internatioonal WHO SRIG(WHO
O 1)
with a potency of
o 59 IU andd the secondd WHO SR
RIG(WHO 2)
2 with a pottency of 30 IU.
RIG is also known
k
as RIG
R Lot R-33 in the Unitted States, distributed
d
b
by
The WHO 1 SR
ogics Evaluaation and Research
R
(CB
BER). The WHO
W
2 SR
RIG is distribbuted
Centter for Biolo
by N
National Insttitute for Biological Staandards andd Control (N
NIBSC) in thhe UK. Theese
standdards are used globallyy to promotee uniform pootency meaasurement off RIG produucts
used for prophylaxis, indiviidual vaccinne responsee, and diseasse diagnosiss in a standaard
onsistent an
nd accurate
internnational uniit (IU/mL). Because it is importannt to have co
assiggnment of IU
U/mL valuees for rabiess immune gllobulin (RIG
G) productss and reliablle
vacciine response measurem
ments for thee evaluationn of vaccinees, the SRIG
G in use for a
particcular metho
od should bee routinely evaluated for
fo potency against
a
a recognized
internnational staandard. Prevvious studiees in 1997 and
a 2006 haave indicated that WHO
O1
SRIG
G has lost po
otency in coomparison with
w WHO 2 SRIG. Fuurther potenncy comparison
studiies have sup
pported this finding. Too determine the differennce in potenncy betweenn the
two S
SRIG prepaarations andd their potenncies in diffe
ferent assayss, a comparison study was
w
perfoormed at KS
SU. Three potency
p
leveels of each SRIG
S
and four
fo rabies virus
v
neutrallizing
antibbody (RVNA
A) positive serum sampples were teested in two
o rapid fluorrescent focuus
inhibbition assays (RFFIT), differing inn challenge virus
v
strain and cell typpe, and two
ELIS
SA assays, one
o indirectt and one blocking. Staatistical anallysis revealeed there is no
n
signiificant difference overaall in the meeasurementss when eitheer WHO 1 or
o WHO 2 are
a
used as the SRIG
G in the RF
FFIT assays.. However, a trend wass clearly seeen in higherr
mL values obbtained wheen WHO1 was
w used as the SRIG to
t obtain thee IU/mL vallues.
IU/m
Addiitionally, fo
or some of thhe samples a significannt differencee in IU/mL was found. The
compparison of inndirect ELIISA results, where the kkit standardd is used to calculate
c
the
EU/m
mL values, revealed
r
a significant difference between
b
WH
HO 1 and WHO
W
2
meassurements at
a potency leevel 2.0 IU//mL; with WHO
W
1 highher in EU/m
mL value thaan
WHO
O 2. The pootential for obtaining
o
inncongruent m
measuremen
nts with usee of differennt
SRIG
G preparatio
on and the performance
p
e of each SR
RIG in diffeerent method
ds should bbe
consiidered when
n selecting assays standdards/controols and in th
he interpretaation of rabbies
serollogy results.
CO.333
a ELISA to
t detect raabies antiboodies in willd (foxes an
nd raccoon
Evalluation of an
dogss) and domeestic carnivvores (dogss and cats)
Cliquuet F1, Guioot AL2, Scheereffer JL1, Tribout L1, Wasniewsski M1, Mähhar K3 - 1Naancy
laborratory for raabies and wildlife, Frannce, 2CPB - Conseils en
n Pharmaciee et Biologiie 3
Francce, Estoniaan Veterinarry and Foodd Laboratoryy, Virology
y-Serology department
d
Estonnia
W
and E
European Commission)
C
) have prom
moted
The iinternationaal organizations (OIE, WHO
the uuse of serolo
ogical testinng in additioon to other sspecific requuirements as
a an alternaative
to thee quarantinee for free movements
m
o pets in ceertain countrries. They have
of
h
also
recom
mmended thhat countriees involved in oral vacccination proogrammes ag
gainst rabiees
shouuld monitor the
t efficacyy of the cam
mpaigns by testing
t
a cerrtain numbeer of field
sampples collecteed from the target species to checkk their immuunity againsst rabies. Thhe
WHO
O/OIE referrence tests (the
( FAVN test and thee RFFIT) are time-conssuming,
expensive, requiire highly-trrained technnicians, the maintenancce of cell cuultures,
h a high conntainment leevel and vacccinated tecchnicians too handle livee
laborratories with
rabiees virus. In addition,
a
sinnce they aree based on cell
c culturess, they are sensitive to aany
cytottoxic produccts and conttaminating agents present in sampples. We havve evaluatedd the
perfoormances off a commerccial ELISA (BioPro Raabies ELISA
A Ab kit, Czzech Repubblic),
a bloocking ELIS
SA that deteects rabies virus
v
antiboddies, in refeerence to thee FAVN tesst.
The sspecificity assessed
a
onn 315 samplees from unvvaccinated dogs
d
and caats was 100%
%. A
total of 701 sam
mples from vaccinated
v
d
dogs
and catts were tested using thee FAVN tesst and
ELISA. The overall agrreement betw
ween the tw
wo tests wass found equaal to 86%.
the E
Conssidering sam
mples from wildlife,
w
a total
t
of 188 sera from foxes
f
and raaccoon dogss
weree sampled in
n a rabies-frree country. The specifficity reacheed 100% in those sera taken
t
from
m naïve anim
mals. Overalll, a high cooncordance ((95%: 648 out
o of 682 sera)
s
was
obserrved betweeen the BioP
Pro ELISA and
a the FAV
VN test, whhich was sim
milar in seraa
from
m red foxes (95.1%:
(
3888 out of 408
8 sera) and raccoon
r
doggs (94.9%: 260
2 out of 2274
sera)). The concoordance betw
ween tetraccycline and seropositiviity results was
w also
evaluuated. The overall
o
agreeement with
h tetracyclinne results waas excellentt in the fox for
f
both the BioPro ELISA (955.9%) and thhe FAVN teest (91.8%).. Concordannce was sligghtly
w a value of 82.8% for
f the BioP
Pro ELISA and
a 78.4% for
f
loweer in the racccoon dog, with
the F
FAVN test. Rabies antibbodies weree equally deetected withh the BioPro
o ELISA in
anim
mals vaccinaated with diffferent types of vaccinees (SAG2 or
o VRG vacccine baits) and
a
in higghly haemo
olysed sera.
In ouur hands, thee BioPro EL
LISA is a vaaluable alteernative to thhe FAVN teest for assesssing
rabiees antibody titres in vacccinated petts and in foxx and raccooon dog popu
ulations forr the
follow up of oraal vaccinatioon campaignns efficacy.. An inter laaboratory co
ollaborative
d in the nextt future to assess
a
the reeproducibilitty of the BioPro ELISA
A for
studyy is planned
laborratories invo
olved in thee monitoringg of oral vacccination prrogrammes..
CO.334
TECTION OF
O RABIE
ES VIRUS - SPECIFIC ANTIBO
ODIES IN WILD
W
DET
MAM
MMALS FROM A RA
AINFORE
EST AREA,, SÃO PAU
ULO, BRAZ
ZIL USING
G
RFF
FIT, SFIMT
T AND ELIISA TECH
HNIQUES
Arauujo DB1, Waasniweski M2, Rodriguues CS3, Cam
mpos ACA3, Martorellli LFA4, Kattaoka
4
5
3
APA
AG , Cunha EMS , Durrigon EL , Favoretto
F
SR
R3,6 - 1Universidade dee São Paulo Núclleo de Pesqu
uisas em Raaiva - Laborratório de Virologia
V
Cllínica e Mollecular, 2Naancy
3
Labooratory for Rabies
R
and Wildlife,
W
Universidad
U
de de São Paaulo, 4Centrro de Controole
5
6
de Zooonoses de São Paulo - COVISA, Instituto Biológico,
B
I
Instituto
Paasteur de São
Pauloo
The eemergent im
mportance of
o rabies in wild
w animalls in Brazil demonstrattes the necessity
of coontinuous ep
pidemiologiical surveilllance in these animal sppecies aimin
ng the
development of better strateegies for thee preventionn and contro
ol of the dissease. The use
u of
bloodd serum sam
mples from several wildd species caaptured in a native Rain
nforest areaa in
the N
North coast of São Paullo State, Braazil, was ann excellent opportunity
o
for the reseearch
of rabbies virus circulation
c
a
among
wildllife in the reegion, and also
a to comppare differeent
technniques for antibodies
a
detection. In this study w
we used thee “Rapid Flu
uorescent Foocus
Inhibbition Test – RFFIT, thhe Simplified Fluoresceent Inhibitioon Microtestt – SFIMT and
the E
Enzime Link
ked Immunoosorbent Asssay – ELIS
SA techniquues for the detection
d
of
rabiees virus-speccific antibodies in terreestrial wild mammals. Out
O of 139 samples, 155
(10,88%) presentted positive titers for RFFIT
R
(“goldd standard” for detectioon of rabiess
viruss neutralizinng antibodiees), 50 (35,99%) positivee titers for SFIMT
S
and 02 (1,43%))
posittive titers foor ELISA. When
W
comparing RFFIT
T and SFIM
MT, 100 (72%) sampless
preseented conco
ordant resultts when connsidering poositive and negative
n
titeers. These
resullts are an ev
vidence of raabies virus circulation between thee wild anim
mal species
(mainnly opossum
ms, capuchiin-monkey and
a coati) inn the studieed area, even
n when
consiidering the low concorrdance betw
ween RFFIT and SFIMT
T. The disco
ordant resullts
betw
ween ELISA
A and RFFIT
T or ELISA and SFIMT
T, (99,3%), can be due to the fact that
t
the E
ELISA kit used
u
was devveloped for vaccinatedd foxes, and when consiidering the
Brazzilian fauna, which pressent a great species varriety withouut the use off oral
vacciination, the efficacy off the techniqque could bee affected. This
T result indicates
i
the
impoortance of coontinuous research regarding a bettter knowledge of the role
r presented by
wild animals in rabies circuulation and transmissio
t
on in Brazil. Epidemiolo
ogic studiess in
v
info
formation reegarding thee
different regionss of the Couuntry could provide a valuable
c
of thhe disease, and
a also aim
ming the staandardizatioon and validdation
preveention and control
of the different diagnostic
d
s
serologic
tecchniques, esspecially coonsidering thhe great andd
o animals present
p
in ou
ur Country.
uniquue variety of
nts: Gaia Consultoria Ambiental,
A
Fundação de
d Amparo a Pesquisa do
d
Acknnowledgmen
Estaddo de São Paulo
P
(FAPE
ESP) and Coordenaçãoo de Aperfeiiçoamento de
d Pessoal de
d
Níveel Superior (CAPES)
(
CO.335
malies In th
he Rabies Indirect
I
Flu
uorescent Antibody
A
T Confou
Test
und Accurrate
Anom
Anteemortem Diagnosis
D
off Human Rabies
R
Ruddd RJ1, Wongg SJ2, Appleer KK1 - 1Rabies
R
Laborratory, Wad
dsworth Labboratories - New
Yorkk State Heallth Dept, 2Wadsworth
W
C
Center,
Diaggnostic Imm
munology Laboratory
L
New
w York Statee Health Deppt
m diagnosis of rabies in
n humans em
mploys techniques that require
The aantemortem
accurracy, speed and sensitivity. A com
mbination of
o histochem
mical, in vitrro virus
isolaation, immunnologic andd molecularr amplification proceduures are utilized in an effort
e
to diaagnose the disease.
d
Prresent day technology
t
offers a pottentially lifee-saving
treatm
ment for a disease
d
that was consid
dered invariaably fatal on
nce clinicall signs
develop. This new
n developpment adds to the need for a rapid diagnosis as
a early in thhe
p
Thhe techniquues offering diagnosis within
w
hourss are
coursse of clinicaal signs as possible.
the ddirect fluoreescent antiboody test on skin and thee indirect flluorescent antibody
a
proceedure on ceerebral spinaal fluid and
d serum. We
W describe examination
e
n by indirecct
fluorrescent antibbody assay of cerebrall spinal fluidd and serum
m taken from
m patients with
w
viral encephalitiis or a presuumed viral infection
i
froom an agentt other than rabies viruus. A
F samples frrom viral enncephalitis ppatients werre tested by
y the rabies
total of 135 CSF
m
indirrect fluoresccent antiboddy procedure. A majorrity of the spinal fluidss tested, from
patieents with encephalitis, presented
p
im
mmunoglobbulins that bound to anttigens preseent in
cell cculture subsstrate. Mostt notable weere the reacctions on kiddney cells provided
p
froom
sera or spinal fluuid obtainedd from patieents diagnossed with thee flavivirus infections
T majorityy of reactionn patterns were
w
Powaassan Viruss or West Niile Virus. The
recoggnizably diffferent than what is seeen with speccific anti-rab
bies antiboddies. Howeever,
resullts indicate that
t false poositive resullts could occcur when innterpreting the
t rabies
indirrect fluoresccent antiboddy procedure. A stainiing pattern appearing
a
similar to
speciific anti-rab
bies stainingg was observved in 7 of the
t 135 spinnal fluids exxamined. The
T
potenntial for falsse positive results
r
docu
umented in tthis work offers weighht to the
argum
ment that taandem posittive results from
f
two diiagnostic test platformss are essentiial
whenn diagnosing
g rabies in the
t human patient.
p
CO.336
NT AND GE
ENETIC LINEAGE
L
IN
I
UNIQUE RABIES VIRUS VARIAN
ECTIVOROUS BATS
S Histiotus velatus, BR
RAZIL.
INSE
Kataaoka APAG1, Favorettoo SR3,2, Marrtorelli LFA
A1, Campos ACA4, Oliv
veira RN2, R
Rosa
1
1
4
1
4
AR , Almeida MF
M , Araujoo DB , Sodrre MM , Rodrigues CS , Sacramennto DRV5,
Duriggon EL4 - 1Centro
C
de Controle
C
de Zoonoses-C
COVISA-PM
MSP, 2Instiituto Pasteurr de
Sao P
Paulo, 3Universidade de
d Sao Pauloo - ICB - Microbiologi
M
a, 4Universidade de Saao
5
Pauloo, Genomic Engenharria Moleculaar
a
ely one-thirdd of the Braazilian mam
mmal fauna and
a the Rabbies
Bats represent approximate
m 41 of the 167 speciess of bats preesent in the country. A
viruss has been issolated from
Rabiies virus ind
dependent sppecies-speccific variant was detecteed in 16 inssectivorous bats
b
Histiiotus velatuss in the Souutheast of Brazil from 1995
1
to 20009. The antig
genic
charaacterizationn was made by monoclo
onal antiboddies panel from
fr
Centers for Diseasse
Conttrol and Prevvention (CD
DC - Atlanta, USA) and the genetiic characterrization wass
perfoormed by seequencing of
o carboxi-teerminal porttion of nuclleoprotein followed
fo
by
Maxiimum Likellihood (ML
L) genetic annalysis withh GARLi software. Thee antigenic
charaacterizationn made in 122 of these saamples show
wed a uniqu
ue profile prreviously
descrribed only for
f the insecctivorous baats Histiotuss velatus (po
ositive reacctivity only with
w
MAbb C12 from the utilizedd panel). The entire 16 samples positive to rab
bies virus were
w
genetically charracterized annd they werre segregateed in the inddependent monophyleti
m
ic
b
values. These sequences showed a minimal
m
aveerage
clustter supported by high bootstrap
intrinnsic distance whiting group
g
(1.3%
%) but they presented
p
low similarityy when
comppared to oth
her lineages circulatingg in bats andd other wild animal lineeages from
Brazzil and world
dwide with a range of 8.8%
8
to 25.4%. The an
ntigenic site of the
nucleeoprotein att residue 377 to 379 (baased on PV strain) anallysis showeed a residue TEV
(Thr--Glu-Val) liike a some insectivorou
i
us bats and different to
o vampire baats lineage,
marm
mosets lineaage and terrestrial cyclee related sam
mples. The PV strain shhows the am
mino
acidss residues TDV
T
(Thr-A
Asp-Val), D.. rotundus isolates show
w AET (Alaa-Glu-Thr) and
Marm
moset lineag
ge show thee amino acid
ds residues TEA (Thr-G
Glu-Ala). This
T antigennic
variaant and geneetic lineage has been iddentified in a large areaa covering various
v
kilom
meters and different
d
bioomes for at least 14 yeaars betweenn the states of
o Minas Geerais
and S
Sao Paulo exclusively
e
i this bat species.
in
s
Surpprisingly thhe system do
ocumentatioon
not ddescribe thiss antigenic variant
v
and genetic lineeage found before in otther bat species
and tthe Histiotu
us velatus baat species neever ever foound beforee with a diffferent antigeenic
variaant and geneetic lineage.. The fact of this lineagge has been isolated on
nly in this sppecies
besiddes long tem
mporal space and geogrraphically distal
d
to eachh other, assoociated withh
phyloogenetic ressults and previously anntigenic dataa suggest strrongly that this rabies virus
v
lineaage is associiated to Histiotus velatuus.
CO.337
TECTION OF
O RABIE
ES VIRUS IN
I ORGAN
NS OF BAT
TS OF GEN
NUS
DET
ART
TIBEUS BY
Y MEANS OF HEMI--NESTED RT-PCR AND
A
REAL
L TIME RT
TPCR
R TECHNIQ
QUES
Scheeffer KC1, Fahl
F WO1, Iaamamoto K1, Carnieli Jr
J P1, Carrieeri ML1, Olliveira RN1, Ito
2 1
2
FH - Instituto Pasteur de São
S Paulo, Faculdade de Medicinna Veterinárria da
Univversidade dee São Paulo
Moleecular technniques have been used increasingly
i
y as tools foor the diagnnosis by
deteccting the rab
bies virus geenome. This study aim
med to detectt the presennce of rabiess
viruss in the wash
h of skull annd in different organs oof the genuss Artibeus bats
b using thhe
hemii-nested RT
T-PCR (hnR
RT-PCR) andd Real Timee RT-PCR molecular
m
techniques. From
approoximately 4,000
4
specim
mens of batss received at
a the Instituute Pasteur for
f rabies
diagnnosis, 30 baats of the geenus Artibeuus were seleected, with records
r
of positive
p
resuults
for raabies by thee traditionall techniquess of direct flluorescent antibody
a
tesst (FAT) andd
inocuulation of murine
m
neurooblastoma cell
c line (N22A). Salivarry glands, urinary
u
bladdders,
kidneeys, lungs, and
a also thee washes off the skullcaaps of the sppecimens weere collected.
The sscraping off the skull was
w performeed with the aid of sterille pipette tipps and then
washhed with 1,0000μL dilueent composeed of 0.85%
% saline soluution, suppleemented wiith
2% B
Bovine Fetaal Serum, freee of rabiess virus speciific antibodiies and conttaining 0.1%
%
Genttamicin Sulffate. The urrinary bladdders were diluted using the same diluent
menttioned abovve, to 1:20 (w
w/v) and othher organs w
were diluted
d 1:10 (w/vv). The
extraaction of tottal RNA waas carried ouut using the TRIzol® annd the reverrse transcripption
was ffollowed byy PCR and hnRT-PCR
h
using primeers specific for the genne encoding the
N prootein. From
m the producct derived byy the reversse transcripttion, the Real Time RT
TPCR
R technique was
w run by using primeers and probbes specificc for the antiigenic variaant 3
of rabbies virus. When
W
evaluuated the tottal samples analyzed, th
he overall results
r
of thee
sensiitivity for booth the hnR
RT-PCR andd Real Timee RT-PCR teechniques was
w 86%. A
compparison betw
ween the hnnRT-PCR annd Real Tim
me RT-PCR
R techniquess performedd by
Fisheer's exact teest has reveaaled that thee proportionn of positivees detected for
f the washhing
of the skull was similar to that
t of the organs
o
exam
minations (P> 0.05). In relation to tthe
f
in hnnRT-PCR annd Real Tim
me RT-PCR
R techniquess were 100%
% in
posittive results found
brainn washes, 900% and 93.333% in the salivary glaands, 83.33%
% and 90% in urinary
bladdders, 80% and
a 93.33% in kidneys,, and 76.67%
% and 50% in lungs. Thhese resultss
sugggest that bothh the hnRT--PCR and thhe Real Tim
me RT-PCR
R techniquess can be useed as
compplementary methods foor the rabiess diagnosis aand the techhniques are sensitive
enouugh for use in
i studies off pathogeneesis. The Reeal Time RT
T-PCR technnique performed
in thiis study pro
oved effectivve in detectting the rabiies virus in different orrgans and exxtra
neuraal tissues with
w the advaantage of beeing a fasterr and more sensitive
s
prrocedure.
CO.338
NTIFICAT
TION OF THE
T
SPECIES OF RE
ESERVOIR
RS AND HOSTS
H
OF THE
IDEN
RAB
BIES VIRU
US AND OT
THER PAT
THOGENS
S BY SEQU
UENCING OF THE
CYT
TOCHROM
ME-B MITO
OCHONDR
RIAL DNA
A GENE
Carnnieli Jr P1, Batista
B
HBC
CR2, Scheffeer KC2, Fahl WO2, Lim
ma JYO2, Olliveira RN2,
2
2
2
2 1
Castiilho JG , Iam
mamoto K , Carrieri ML
M , Kotait I - Institutto Pasteur, Brasil
B
Diaggnóstico, 2Innstituto Pastteur - Diagnnóstico
on of animaal species th
hat transmit pathogens such
s
as the rabies viruss is
The iidentificatio
of the utmost im
mportance foor public heealth and thee natural hisstory of infeectious and
o decompoosing
contaagious diseaases. Diagnostic laboraatories very often receivve mauled or
anim
mal carcasses, particularrly of bats, rendering
r
m
morphometr
ic identificaation unviabble.
The eexistence off different regional
r
nam
mes for the same animaal, morpholoogical
variaability and th
he lack of staff
s
trainedd in zoologiccal identificcation constiitute a serioous
probllem for epiddemiologicaal surveillannce. Molecuular techniqques are used
d routinely and
effecctively in syystematics, evolution
e
annd ecology to identify species
s
andd can even be
b
used to identify hybrids thaat originated
d from geneetically closee animals, in
i which thee
b morphom
metry. Somee mitochonddrial DNA (m
mtdifferences veryy often go unndetected by
A) genetic markers,
m
succh as contrrol region seequences annd the geness encoding
DNA
cytocchromes b and
a c, are frrequently ussed in the geenetic identtification off species. Many
of the genetic seequences for these genees are storedd in public-domain webbsites such as
Bank, allow
wing new seqquences to be
b compareed with existing ones in
n databases. The
GenB
objecctive of thiss study is to build a database with genetic sequuences from
m the
cytocchrome b geene of rabiees reservoir species for use in the identificatioon of these
speciies. mt-DNA
A fragments were amplified and sequenced as described previously by
Carnnieli et al. (2
2008), usingg the primerrs 5’CGA
ACTAATGA
ACATGAA
AAAATCAC
CCGTTG-33’ (sense) annd 5’TAT
TTCCCTTT
TGCCGGTT
TTACAAGA
ACC-3’ (anntisense) described by Martins
M
et al.
a
(20077). Sixty-sixx mt-DNA samples froom differentt species of wingless Brazilian
B
mam
mmals and fiifty-four sam
mples from different sppecies of chhiropterans were
w
sequeenced. Anaalysis of thee genetic seq
quences froom these win
ngless mam
mmals
highllighted the problem
p
of genetic idenntification oof species as
a only a few
w sequencess of
mt-D
DNA from wingless
w
maammals of Brazil
B
were found in GeenBank. Fo
or example, there
are seven speciees of marmoosets (genuss Callithrix)) but mt-DN
NA sequences for only some
osited in GeenBank. Hoowever, the cytochromee b gene seqquences obttained
of them are depo
m bats in thiss study, togeether with morphometr
m
ric identification carried out in parrallel,
from
allow
wed us to naame the species with ceertainty. Froom the fifty-four mt-DN
NA sampless
from
m chiropteran
ns, nineteenn species froom eight genera and four differentt families were
w
identtified. Thus, the method describedd here is effiicient in thee identificatiion of anim
mal
speciies and the search for samples
s
of mt-DNA
m
in Natural Hisstory Museu
ums and Zooos
may complemennt and certiffy unequivo
ocally the seequences in the databasse under
consttruction.
Finanncial Suppoort: Institutoo Pasteur, Brazil
CO.339
YLOGENETIC ANAL
LYSIS OF RABIES VIRUS
V
IN THE
T
STAT
TE OF RIO
O
PHY
GRA
ANDE DO SUL, SOUTHERN BRAZIL
Batissta HBCR1, Oliveira RN
N1, Carnielii Jr P1, Ferrreira JC2, Ro
osa JCA2, Castilho
C
JG1,
1
1
3
4
3
1
Fahl WO , de Paaula FC , Sales EF , Paacheco SM , Maletich DJ , Carrierri ML , Roeehe
P
- Virrologia, 2FE
EPAGRO Saúde Animaal, Instituto de
PM5, Kotait I1 - 1Instituto Pasteur
Pesqquisas Veterrinárias Desidério Finam
mor, Eldoraado do Sul, RS, Brazil, 3Universiddade
Fedeeral do Rio Grande
G
do sul(UFRGS
s
S), Porto Aleegre, RS, Brrazil, 4Instittuto Sauver,
5
Portoo Alegre, RS, Brazil, Universidad
U
de Federal do
d Rio Grannde do sul(U
UFRGS), Poorto
Aleggre, RS, Braazil and FEP
PAGRO Saúúde Animall, Instituto de
d Pesquisass Veterináriias
Desiddério Finam
mor, Eldoraddo do Sul, RS,
R Brazil
v
(RAB
BV), a membber of the
Rabies is a worlldwide zoonnosis causedd by rabies virus
us, family Rhabdoviridaae. In naturre, RABV iss maintained
d in cycles with
w
Lyssaavirus genu
distinnct natural reservoirs.
r
I the urbann cycle, the main reservvoir for the virus is the
In
domeestic dog, on the other hand, in thee sylvatic cyycle differen
nt species can
c be the
reserrvoir. In Lattin Americaa, the main natural
n
RAB
BV reservoiir is the haeematophagous
bat D
Desmodus rootundus. Hoowever, RA
ABV lineagees adapted to
t different bat
b species,,
incluuding insecttivorous andd frugivorouus bats, have been frequ
uently reporrted. The R
RABV
lineaages isolatedd from non haematophaagous bats aare geneticaally distinct from the RABV
R
lineaages whose natural
n
reseervoirs are haematopha
h
agous bats. In
I the State of Rio Grannde
do Suul (RS), souuthern Brazil, urban rabbies has nott been deteccted since 19988.
Neveertheless, raabies remainns endemic in haematopphagous an
nd non haem
matophagouss bat
speciies. The preesent work reports
r
the first
f
phylogenetic analyyses on RAB
BV isolatess
from
m the State of
o RS, for thhat, a total of
o 30 rabies virus (RAB
BV) isolates sent to rabiies
diagnnosis were analyzed.
a
T isolates were recoveered from different
The
d
batt species
(Taddarida brasiiliensis, Myootis nigrican
ns and Histtiotus velatuus), from herbivores
(boviines and bu
uffalo) and carnivores
c
(domestic doog and cat).. The bat species were
identtified with the
t aid of a morphologiical dichotoomous key. For the phy
ylogenetic
or
analyysis, total RNA
R
was exxtracted from
m original brains
b
(herbiivores and carnivores)
c
infeccted mice (b
bats) with Trizol and suubmitted to reverse trannscription/p
polymerase chain
c
reacttion (RT-PC
CR) with priimers targetting a initial portion off the nucleopprotein genee
(N). P
Phylogenetic analysis of
o the sequeenced fragm
ments revealled the occuurrence of four
fo
RAB
BV lineages, named afteer its naturaal hosts: Dessmodus rotuundus (haem
matophagouus
bat), Tadarida brasiliensis
b
(insectivoro
ous bat), Myyotis nigrica
ans (insectiivorous bat)) and
A RABV isolates from
m herbivores belonged to
Histiiotus velatuss (insectivoorous bat). All
the hhaematophaggous bat Deesmodus rottundus lineaage. The tw
wo RABV isolates from
m
carniivores clusteered within the Tadarid
da brasiliennsis lineage,, revealing two occasioonal
spilloovers from insectivorouus bats to domestic petts, thus not compromis
c
ing the statuus of
“urbaan rabies freee” of the area.
a
These findings
f
higghlight the importance
i
of the
identtification off RABV lineeages and itts value as an
a aid to suppport rabiess surveillancce.
Finanncial support: Instituto Pasteur.
CO.440
UACIÓN EPIDEMIO
E
OLÓGICA DE LA RA
ABIA EN CHILE.
C
20000-2011
SITU
Yungg V1, Favi M1, Fernanddez J1 - 1Insttituto de Saalud Públicaa de Chile
Chile, el añoo 1990, se deetecto el últtimo caso de rabia idenntificado com
mo variantee
En C
caninna desde enttonces esta variante noo circula en el país, la im
mportancia de los anim
males
silvestres en la transmisión
t
de la rabia fue reconoccida en 1985, cuando se
s detectó poor
mera vez rabiia en murciéélagos insecctívoros de la especie Tadarida
T
brrasiliensis. El
E
prim
reconnocimiento de los murcciélagos com
mo reservorrio de la enffermedad hiizo que se
amplliaran las accciones de vigilancia
v
eppidemiológiica hacia esas especies
caraccterizándosee el patrón epidemiológ
e
gico de la raabia por unaa endemia en
e quirópterros.
Desdde el año 20000 al 2011,, se analizo un total de 32802 muestras para diagnóstico
d
de
rabiaa, de estas 979 fueron positivas
p
(3,,0%), 976 murciélagos
m
insectívoro
os, 2 gatos y 1
perroo. Según la distribuciónn geográficaa de casos, estos
e
se registraron en las regioness
centrrales del paíís, y no se han
h encontraado muestraas positivas al virus ráb
bico en las
regioones extrem
mas.
A traavés de tipifficación antigénica y geenética se han
h identificcado 4 variaantes viraless que
son llas responsaables de la trransmisión de la rabia,, los princippales reservoorios silvesttres
circuulando en ell país son murciélagos
m
de la especiie Tadaridaa brasiliensiis, Myotis
chilooensis, Lasiu
urius cineriius y boreallis y finalmeente Histiottus macrotuss. La especiie
Tadaarida brasiliensis repreesenta el 91,,1% de los casos
c
positivvos
Los eestudios de caracterización antigénnica y genéttica nos han
n permitido tener un
conoocimiento más
m amplio de
d la epidem
miología de la rabia
El Prrograma de Control de Rabia conteempla la edducación de la poblacióón para evitaar el
contaacto con muurciélagos y el reporte de
d cualquieer mamíferoo sospechoso
o, la eliminaación
de coolonias de murciélagos
m
se realiza solamente
s
een casos de detección
d
de
d especímennes
posittivos, en razzón del importante rol que
q esta esppecie desarrrolla en la mantención
m
d
del
equillibrio ecológgico y dadoo el bajo porrcentaje de positividad
p
a rabia (alrrededor de 2%)
2
en caapturas masivas de estaa especie.
CO.441
MOL
LECULAR
R CHARAC
CTERIZAT
TION OF RABIES
R
VIIRUS AND
D OTHER
VIRA
AL AGENTS ISOLA
ATED FRO
OM BATS IN VENEZU
UELA.
BOY
YER L1, PU
UJOL F2, HID
DALGO M1, PAPO S3 - 1Nationaal Institute of
o Agricultuural
2
Reseearch (INIA) - Rabies Laboratory,
L
Venezuelaan Institute of Scientific Investigattion
(IVIC
C) - Molecuular Virologgy Laboratoory, 3National Institute of Integral Agricultura
A
al
Heallth (INSAI) - Departmeent of contro
ol and preveention of Raabies wildliife
Batss (Chiropterra) are reserrvoirs for zooonotic diseeases, includding Rabies, Hendra, Nipah,
N
SAR
RS-CoV, Ebola virus. Hence
H
their importance
i
as a potentiial reservoirr hosts of
virusses affectingg human andd animal heealth.
In ouur country, there
t
is no knowledge
k
of bats as reeservoir forr viruses exccept rabies. The
aim of
o this invesstigation waas the moleccular characcterization of
o rabies virrus and otheer
viral agents, isollated from bats
b in Veneezuela.
m
characteriza
c
ation was baased on: viruuses with im
mpact in pubblic health,
The molecular
persiistence in ho
osts and enddemic areass. A total off 54 bats were collectedd in differennt
states and years.. Those werre identifiedd and classiffied into: 12
2 vampires, 29 frugivorrous
orous belongging to diffe
ferent familiies, genera and
a species. They weree
and 113 insectivo
autoppsied to collect tissues from differrent organs iincluding brain tissue of
o livestockk
posittive to rabiees virus. Diffferent systeems were ussed for PCR
R to detect DNA
D
and RN
NA
viral genomes. Samples
S
weere amplified, moleculaarly characteerized and sequenced
s
t
to
W were ablle to detect 8 Herpesvirruses and 4
identtify the phylogeny of each virus. We
Polyomaviruses in trachea and lungs samples from
m different bat species and one
Astrooviruses in an intestinee of an insecctivorous baat. Eight Rabbies isolates were grouuped
in thee genus Lysssavirus gennotype 1. Foour of them
m characterizzed as antigeenic variantt 3
(Desmodus rotuundus).
t first andd paramounnt
The ddetection off these virall agents in thhe Venezueelan bats is the
inforrmation for the study off these unknnown agentts, which coould pose grreat risk to
humaans and liveestock healtth in our couuntry.
nowledgem
ments: MCT
TI-Misión Ciencia,
C
Vennezuelan Insstitute of Sccientific
Ackn
Invesstigation (IV
VIC): Molecular Virolo
ogy Laborattory, Nationnal Institutee of Agricultural
Reseearch (INIA): Rabies Laboratory, National
N
Insstitute of Integral Agriccultural Heaalth
(INS
SAI). Fundiing: IVIC.
her C, Rev.M
Med.Vir, 177:67,
Almeida M, Rev.Inst.Med.trop.S.Paullo; 53:31, 20011; Calish
ue, J.Gen.V
Virol, 90:8833, 2009; Dee Mattos C,, J.Clin.Miccrobiol, 34:11553,
20077; Chen Zhu
19966; Olivier D,
D Plos/ONE
E, 4:e2057, 2008;
2
Richter R, J.Geen.Virol, 90:44, 2009;
Won
ng S, Rev.M
Med.Vir, 17:667, 2007.
CO.443
d Temporaal Dynamiccs of Rabiess in China
The Spatial and
China CDC - Institute fo
or Viral Dissease
Tangg Q1, Yu J1, Hao L1, Raayner S2, Liang G1 - 1C
Conttrol and Prevvention, 2Chinese Acaddemy of Sciences - Wuuhan Instituute of Viroloogy
nd Objectivves: Recentt years havee seen a rapid increase in the numbber
Background an
g
distribution
d
of the viruss. In
of rabbies cases in China andd an expanssion in the geographic
spite of the serioousness of the
t outbreakk and increaasing numbeer of fatalitiies, little is
wn about thee phylogeoggraphy of th
he disease inn China. In this study, we
w report ann
know
analyysis of a sett of Nucleoccapsid sequeences consisting of sam
mples colleccted throughh the
trial Chinese Naational Survveillance Syystem as welll as publiclly available sequences. This
presents the most compprehensive dataset
d
from
m China to date,
d
compriising
sequeence set rep
210 ssequences (including
(
5 new samp
57
ples) from 115 provincees and coverring all epiddemic
regioons. Using this
t dataset we
w investiggated geneticc diversity, patterns off distributionn,
and eevolutionary
y history.
Resu
ults: Our an
nalysis indiccates that thee rabies viruus in China is primarily
y defined byy two
cladees that exhibbit distinct ppopulation subdivision
s
n and transloocation patterns and thaat
contrributed to thhe epidemicc in differennt ways. Thee younger clade originaated aroundd
19922 and has prroperties thaat closely match
m
the observed spreead of the reecent epidem
mic.
The oolder clade originated around
a
1960 and has a dispersion pattern thatt suggests itt
repreesents a straain associateed with a prrevious outbbreak that reemained at low
l levels
throuughout the country
c
andd reemergedd in the curreent epidemiic.
Concclusions: Our
O findings provide new
w insight innto factors associated
a
w the receent
with
epideemic and arre relevant to determiniing an effecctive policy for controllling the viruus.
CO.444
ds: Prioritizzing human rabies biologics in limited supply scenariios
Playying the odd
Recuuenco S1, Vo
ora NM1, Rupprecht
R
C1 - 1CDC - R
Rabies Proggram
Limiitations in th
he availabiliity and acceess to humaan rabies bioologics in en
nzootic regiions
resullt in most raabies deathss in the deveeloping worrld. Efforts to
t supply modern
m
rabiees
vacciines and im
mmune globuulin (RIG) have
h
improvved availabiility, but coost and the laack
of strructured proograms in many
m
countrries remain major obstaacles to provviding optim
mal
care. Proposed policies
p
to provide
p
rabiies post-expposure prophhylaxis (PE
EP) at no cosst to
ugh governm
ment prograams are challlenged by the
t limited supply of raabies
the ppatient throu
bioloogics that prroviders aree able to obttain. In manny cases, thee demand foor biologics
exceeeds the limiited supplies and nation
nal rabies programs aree therefore forced
f
to raation,
resullting in delaays or compplete failuress in provisioon of adequuate PEP. Op
ptimal PEP
invollves the usee of rabies im
mmune globbulin and vaaccine. Whiile WHO reecommendaations
for P
PEP are com
mprehensivee, those recoommendatioons offer noo guidance on
o managem
ment
of rabbies exposu
ures when thhere are lim
mited suppliees of biologgics in the coountry nor iif
theree is only vacccine availaable but no RIG.
R
Complex operatio
onalization issues, suchh as
to hoow to approaach prioritizzation whenn both nervoous tissue annd modern vaccines cooexist
in a ccountry, or how to optiimally integ
grate privatee distributioon of rabies biologics, are
a
not ppart of the WHO
W
guidannce documeents. We preesent a propposal on how
w to develoop
recom
mmendationns and guidelines to deeal with thesse scenarioss accounting
g for local rabies
r
epideemiology, patient
p
age and
a body sizze, delays after
a
exposuure, and culttural and soccial
issuees. Several Old
O and New
w World coountry casess are presented to highllight how thhese
challlenging circcumstances might be managed
m
andd overcome..
CO.445
IMM
MUNE RES
SPONSE OF
O BALB/C
C MICE IM
MMUNIZED
D WITH VERO
V
CEL
LL
RAB
BIES VACC
CINE AND
D BpMPLA
A-SE ADJU
UVANT
Frazaatti-Gallina NM1, Silvaa ABP, Rinaaldi DP, Sillveri A, Raw
w I, Menezees CRB 1
Instiituto Butanttan - Laboraatório de Raaiva
The pprophylaxiss is an impoortant strateg
gy to controol of humann and animal rabies diseease.
The vvaccine from
m Vero celllular culturee for humann use is efficcacy and safety. Howevver,
becauuse the techhnology useed to producce this vacciine is expen
nsive this prroduct costss
abouut ten dollarss. This cost makes them
m impossiblle the use of this vaccin
ne type in poor
p
counntries where the animal rabies conttrol is ineffiicient and thhere many cases
c
of hum
man
rabiees. Rabies disease
d
is ressponsible foor about 55,,000 deaths per year in the world. The
objecctive of thiss study was evaluate the humoral immune
i
respponse of miice (Balb/c))
immuunized withh three diffeerent doses of
o Vero rabies vaccine associated with the
BpM
MPLA-SE ad
djuvant. Thiis adjuvant is a productt obtained from
f
Bordettella pertusiis.
Three groups off ten mice were
w immunnized with tw
wo doses off 500µl (G1), 250µl (G2) or
µl (G3) of Vero
V
cell rabbies vaccinee (IB-lot 11003075) mixxed with BpM
MPLA-SE
125µ
(10µ
µg/dose). Th
hree groups control (Gcc) received only
o
rabies vaccine. Th
he immunizzation
a samples were taken ten days after
a
the last dose injeccted
occuurred on dayys 0 and 21 and
and oon days 60, 120 and 1880 to determ
mine the titerrs of neutraalizing antibbodies for raabies
viruss in BHK21 cells (RFFIT). The av
verages of thhe neutraliziing antibodies titers fouund
in thee samples from
fr
each grroup ten dayys after finiished the im
mmunization
n were 39.2,,
33.1,, and 20.4 IU
U/ml for grroups G1, G2
G and G3 respectively
r
y. The resultts obtained on
day 1180 were 17
7.1 IU /ml (G1),
(
10.6 IU
U/ml (G2) and
a 9.8 (G33). In the control groups the
averaages of the antibodies
a
t
titers
were: 29.7 (Gc1),, 26.9 (Gc2)) and 22.2 IU/ml
I
(Gc3))
after immunizattion and 10.7 (Gc1), 9.55 (Gc2) andd 8.5 IU/ml (GC3) on day
d 180 (Gcc3).
w that the addjuvant BpM
MPLA-SE increased thhe humoral immune
i
Thesse data show
respoonse for rabbies vaccine in Balb/c mice
m indepeendent of thee volume off vaccine
utilizzed to immuunize the annimals. The results founnd are very important to
t reduce the
numbber of dosess and the voolume of Veero cell rabiies vaccine utilized
u
in the
t
immuunization ag
gainst rabiees.
Finanncial Suppoort: Butantann Foundatioon
CO.446
FETY AND IMMUNO
OGENICIT
TY OF THE
E PURIFIE
ED VERO RABIES
R
SAF
CCINE NEX
XT GENER
RATION IN
I CHINES
SE PEDIATRIC (≥ 10
0 YEARS)
VAC
AND
D ADULT POPULAT
P
TIONS
Pichoon S1, Li R2, Li J3, Shuu JD1, Chavand P1, Minnutello M1, Guinet-Morrlot F1 - 1Saanofi
Pasteeur, 2Guanggxi Center foor Disease Prevention
P
aand Controll, 3National Institutes ffor
Foodd and Drug Control
C
kground: The
T Purifiedd Vero cell Rabies
R
Vacccine Next Generation
G
(PVRV-NG
(
G) is a
Back
highlly purified vaccine
v
devveloped withh innovativee technologgy and humaan and anim
mal
origin componennts-free medium. It waas shown to be at least as
a immunoggenic as
p
a similar safeety profile in
i a phase III clinical stuudy conductted
Veroorab™ and presented
in Frrance (pre exxposure reggimen). A phhase III clinnical study was
w perform
med in Chinnese
pediaatric (≥ 10 years)
y
and adult
a
populaations in sim
mulated postt-exposure regimen
r
to
furthher documennt PVRV-N
NG in compaarison to Veerorab™.
Meth
hods: This was
w a randoomized, blin
nd-observerr, controlledd study in heealthy subjeects
agedd 10 to 17 yeears (pediatrric cohort) or
o ≥ 18 yearrs (adult cohhort). Particcipants receeived
five ddoses by inttramuscularr route of PV
VRV-NG or
o Verorab™
™ (ratio 2:1 in each agee
groupp) at D0, D3, D7, D14 and D28 ass per recom
mmendation for post-exp
posure
prophhylaxis (Essen schedulle). No rabiees immune--globulins were
w adminiistered
concomitantly with
w the firstt vaccine doose.
munogenicityy was evaluated at D0, D14 and D42 by meassuring the leevel of rabiees
Imm
viruss neutralizinng antibodiees (RVNA) using the raapid fluoresscent focus inhibition
i
teest.
Testiings were performed att the Nationnal Institute for Food annd Drug Co
ontrol (Beijinng).
Safetty was evaluuated with a list of preddefined soliicited injecttion site and
d systemic
reacttions during
g the period between D0 and D14 and
a during the seven days
d
after the 2
last ddoses; any adverse
a
evennts until 28 days after the
t final dose and any SAE until 6
montths after thee final dose were also recorded.
Resu
ults: 816 paarticipants were
w enrolled; 408 in eaach age grouup correspo
onding to 2772 in
PVR
RV-NG grouup and 136 in
i Verorab™
™ group. Thhe predefineed criterion
n for noninferriority in terrms of propoortions of participants w
with RVNA
A titers ≥0.55 IU/mL at D14
D
(befoore the 4th in
njection) waas met in thhe per-protoocol analysis set and coonfirmed in the
full-aanalysis set populationn, in each off the 2 age groups.
g
All subject
s
had titer ≥
0.5 IU
U/mL at D442 whateverr the age group and thee vaccine received.
PVR
RV-NG was safe and weell tolerated
d after each vaccination
n and its saffety profile was
simillar to Verorrab™ in term
ms of solicited injectionn site and syystemic reaactions, as well
w
as unnsolicited ad
dverse evennts. There were
w no serioous adverse events relaated to
vacciination. Noo safety signnal emergedd during thee course of the
t study.
Concclusions: The phase IIII clinical stu
udy results showed thaat PVRV-NG
G is at leastt as
immuunogenic ass Verorab™
™ and presennted a similar safety prrofile, when
n administerred
accorrding to thee ESSEN reggimen in peediatric (≥ 10
1 years) annd in adult populations.
p
Takeen together with
w the ressults of the previous
p
Phhase II cliniccal study, thhis confirmss that
PVR
RV-NG is an
n improved and highlyy purified altternative foor rabies pree- and postexpoosure prophyylaxis.
CO.447
VELOPME
ENT OF AN
N IN VITRO ELISA ASSAY
A
FO
OR THE
DEV
QUA
ANTIFICA
ATION OF THE IMM
MUNOGEN
NIC GLYCO
OPROTEIN G PRES
SENT
IN V
VACCINE BATCHES
B
S: COMPA
ARAISON OF
O IN VIV
VO AND IN
N VITRO
VAC
CCINE POT
TENCY TE
ESTS
1
1
Jallett C , Castel G , Chteouui M1, Tordoo N2 - 1Instiitut Pasteur - Unit Antiiviral Strateegies,
2
Instiitut Pasteurr - Virology
wards limitin
ng and even
n sometimess
Sincee several yeears, there iss a global teendency tow
waivving the use of animal experiments
e
s in Researcch and Produuction. How
wever for
Hum
man and Veterinary rabiies vaccine producers and
a controlllers, the in vivo
v NIH asssay
still remains
r
thee standard rooutine potenncy test befoore batch reelease. Neveertheless,
fundaamental stuudies have been
b
accumuulated to coorrelate the structural
s
prresentation of
the main
m rabies antigen, thee glycoproteein G, and itts immunoggenicity. Thhe G proteinn
contaains two maain antigenic sites: site II requires a folding off the G ectoodomain to bring
b
in proximity pep
ptides distannt in the primary sequeence of the protein;
p
sitee III which is
i
d
allong the ecttodomain buut also requuires a folded conformaation of the
less dispersed
ectoddomain. Botth sites are recognized by specificc monoclonaal antibodiees (mAbs) thhat
are inn general neeutralizing viral
v
infectiion. Amongg them mAbb D1 (IgG I isotype),
direccted against site III is sppecific of thhe trimeric state
s
of the glycoproteiin (it recognnizes
the nnative but noot mercaptoo-ethanol annd/or SDS-trreated G) which
w
is pressumed to bee the
mostt immunogeenic form off the antigenn. mAb D1 has been exxtensively used
u
to evaluuate
the sttability of G trimers (Jallet et al., 1999, J. Virrol, 73: 225-33; Desmeezieres et al..,
20033, Virus Ress. 91: 181-7; Sissoeff ett al, 2005, J.
J Gen. Virool, 86: 2543-52). It has also
beenn proven suittable in ELIISA to monnitor the connsistency off the lot to lo
ot rabies vaaccine
produuction and to
t evaluate the glycoprrotein conteent (Fournier-Caruana et
e al, 2003,
Bioloogicals, 31 : 9-16). Since the end of
o the 90t's, the French National Regulatory
R
Authhority in chaarge of hum
man rabies vaaccine conttrol (ANSM
M) has decideed to use thhis
ELIS
SA test insteead of the siingle radial immuno-diiffusion assay (SRD) too monitor thhe
consiistency of production
p
o rabies vacccines. The ELISA hass been transferred from
of
m
Pasteeur Institutee to ANSM,, improved, optimised aand then vaalidated. Sinnce 2001, thhe
consiistency of production
p
h been esttablished onn around 1000 batches while
has
compparing the NIH
N assay and
a the ELIS
SA test. Thee results aree homogenoous betweenn
both methods.
ve of replaccing in vivo by in vitro assays, vacccine samplees have beenn
In the perspectiv
e
oof the glycopprotein conttent was asssayed
artifiicially altereed by heatinng and the evaluation
by m
mAb-D1 ELIISA. This assay
a
was sh
hown to be sensitive
s
ennough to dettect vaccinee
alteraations and to
t discriminnate betweenn low and high-potent
h
potency
p
battches. This type
t
of EL
LISA assay may have a promisingg future for w
waiving in vivo rabies potency tesst and
prom
mote in vitro
o antigenic/iimmunogennic quantificcation/qualification of the
t G proteiin for
vacciine batches.
CO.448
VING TOW
WARDS TH
HE REPLA
ACEMENT
T OF THE NIH TEST
T
MOV
Morllot FG1, CH
HABAUD-R
RIOU M1, CHAPSAL
C
J 2 - 1SAN
JM
NOFI PASTE
EUR - R&D
D,
2
SAN
NOFI PAST
TEUR - IO
Potenncy testing of inactivatted rabies vaaccines is trraditionally performed by an
intraccerebral (IC
C) challengee method onn mice. The method waas originallyy developedd by
the N
National Insstitutes of Health
H
(NIH)) in the 1950s for poten
ncy testing of
o inactivateed
rabiees vaccines for human uuse. The NIIH test is wiidely recogn
nized and iss currently
requiired by the World
W
Heallth Organizaation (WHO
O) and Pharrmacopoeiass for rabies
vacciines releasee. Neverthelless, the NIH
H challengee method prresents a num
mber of
limittation and isssues:



As a biological test the NIH meethod is higghly variablee, making thhis test
inapprop
priate for baatch-to-batch
h consistenccy analysis
A large number of mice is requuired
This testt is time connsuming and
d labor intennsive for booth the manuufacturer annd
the authoorities that release
r
the batches.
b
For all
a these reaasons, regulaators, experrts and manuufacturers are
a seeking an alternative
methhod to the exxisting NIH
H test that guuarantees thhe potency of
o the rabiess vaccine to be
admiinistered forr pre or post exposure vaccination
v
n.
A reccent internaational workkshop of thee NICEATM
M and ICCV
VAM was held
h in Ames,
Iowaa, USA on October
O
11-13th, 2011 on "Alternaative Methoods for Hum
man and
Veterinary Rabiies Vaccine Testing" with
w particular focus on Rabies Vacccine Potency
w as folloows:
Testiing. The maain conclusiions of this workshop were




For inacttivated veteerinary rabiees vaccines,, the Serum Neutralizattion Test (S
SNT)
serologiccal method described inn the Ph. Euur. Monograaph 0451, should be
immediaately considdered for prooduct speciffic validatioon by vaccinne manufactturers
for both adjuvanted and non addjuvanted vaaccines.
As humaan rabies vaaccines in soome regionss (e.g., U.S. and EU) arre simpler
productss (non-adjuvvanted, mon
novalent), manufacture
m
ers are encou
uraged to
develop and implem
ment an in vitro antigenn quantificattion methodd to replace the
a
quanntification methods
m
currrently usedd by
mouse chhallenge tesst. In vitro antigen
rabies vaaccine manuufacturers as
a in-processs tests inclu
ude ELISA and
a Single
Radial Im
mmunodiffuusion (SRID
D) Test.
Final pro
oduct in vitrro methods will requiree identificattion and usee of approprriate
reagents (e.g. monooclonal antibbody) with specificity
s
f the neutrralizing epitope
for
of the virrus-associatted trimericc form of glyycoprotein G.
Validatioon of in vitrro replacem
ment tests wiill need to in
nclude idenntification off
sub-poteent lots. For validating in vitro metthods for pootency testinng of humann
rabies vaaccines, it may
m be neceessary to com
mpare in vittro results to
o adequate
serologiccal titers in humans.
In the context off the Purifieed Vero Rabbies Vaccinne next geneeration (PVR
RV-NG)
ng the needd for
development, Saanofi Pasteuur has set upp an in-housse ELISA teest answerin
m
to thee NIH potenncy test.
an allternative method
The ddescription of the correesponding ELISA
E
methhod for rabiies glycopro
otein G
quanntification an
nd the data supporting the alternattive test, toggether with the proposeed
globaal strategy for
f implemeenting this ELISA
E
test in replacem
ment of the NIH
N test, wiill be
preseented.
CO.449
ELECTRO
OCHEMILU
UMINESC
CENT ASSA
AY FOR ANALYSIS
A
OF RABIE
ES
AN E
VIRU
US GLYCO
OPROTEIIN IN RABIES VACC
CINES
Smitth TG1, Ellisson JA, Carrson WC, Ma
M X, Rupprrecht C - 1Centers
C
for Disease
D
Conntrol
and P
Prevention
n
to
o evaluate the
t immunoogenicity of inactivatedd
Vacccine potencyy testing is necessary
rabiees virus (RA
ABV) vaccinne preparatiions before human or veterinary
v
appplication.
Curreently, the NIH
N test is reecommendeed by the WHO
W
expert committee to evaluate
intra-- and inter-llot variationn of RABV vaccines; however,
h
nuumerous disadvantages are
inherrent concernning cost, number of annimals and biosafety
b
reequirementss. As such,
numeerous in vitrro methods (e.g. antigeen-capture E
ELISA) havve been propposed for thhe
evaluuation of vaaccines baseed on RABV
V glycoprottein (G) quaality and quaantity whichh
correelates with vaccine
v
poteency. In this study an antigen-capt
a
ture
electtrochemilum
minescent (E
ECL) assay was develooped utilizin
ng three murrine anti-RA
ABV
G moonoclonal antibodies
a
(m
mAb) to quaantify RAB
BV G in twoo commerciaally available
inacttivated RAB
BV vacciness, one experrimental vacccine, and three
t
purifieed RABV G
prepaarations. Th
he first mAbb was speciffic for a connformationaal epitope soo that only
immuunogenic, natively
n
foldded G was captured
c
in the
t assay. Additionally
A
y, two mAbs that
bind non-compeeting linear epitopes weere employeed to evaluaate the overaall quantity of
V G and forr detection. Vaccine effficacy was also
a assesseed in
nativve and denattured RABV
vivo using pre-eexposure vacccination off mice folloowed by perripheral RAB
BV infectioon.
n
g antibody ((VNA) titer of 4.2 IU/m
ml and proteected
Puriffied G inducced a virus neutralizing
100%
% of immun
nized mice; while, an exxperimentall vaccine with
w low quaality and quaantity
of G induced a VNA
V
titer >0.03
>
IU/mll and only protected
p
211% of immuunized micee.
h
tthat in vivo efficacy maay be prediccted
Thesse preliminaary results suupport the hypothesis
from
m the in vitro
o measurem
ment of RAB
BV G using the ECL asssay. Based upon these
resullts, the ECL
L assay mayy have utilityy in measurring potencyy of RABV vaccines.
CO.550
RAB® EFFIICACY IN
N SKUNKS (Mephitis mephitis)
m
A
AND
RACC
COONS
ONR
(Proccyon lotor)
Know
wles MK1, Fehlner-Gar
F
rdiner C1, Beresford
B
A2, Rosatte R3 - 1Canadiian Food
Inspeection Agen
ncy - Centree of Expertise for Rabiees, Ottawa, Canada, 2Artemis
A
3
Techhnologies In
nc., Guelph, Canada, Ontario
O
Minnistry of Nattural Resouurces - Wildlife
Reseearch and Developmentt, Peterboroough, Canadda
RAB®, a reccombinant human
h
adenovirus type 5 vector ex
xpressing raabies
ONR
glycooprotein, haas been usedd under experimental permit
p
in thee Canadian provinces
p
o
of
Ontaario, New Brunswick annd Quebec for wildlife rabies oral vaccinationn programs..
Priorr to its use in
n the field, a series of trials
t
were conducted
c
in two terresstrial wildliffe
vectoors to determ
mine the rabbies virus neutralizing antibody reesponse to
ONR
RAB®. Eigh
hty-three % of skunks (10/12)
(
andd 75% of racccoons (8/122) seroconvverted
®
withiin 6 weeks after
a
consum
mption of ONRAB
O
inn an Ultralite bait (ULB
B) at a dose of
109.2 TCID50/mll in 1.8 mL. In the subsequent effi
ficacy trial, all
a skunks (n=28)
(
that
B®-ULB weere protectedd from lethaal rabies chaallenge, whhile
consuumed a singgle ONRAB
86% (12/14) of the unvacciinated contrrols succum
mbed to rabiees. In addittion, pre-exiisting
a
type 2 or hu
uman adenoovirus type 5,
neutrralizing antiibody to eithher canine adenovirus
achieeved by intrramuscular inoculation
i
with the viruuses 28 d prrior to
of skunks w
admiinistration of
o ONRAB® per os at 10
1 8.4 TCID50/ml, had noo effect on the
t antibodyy
®
®
respoonse to ONR
RAB . Theese series off experimennts demonsttrated that ONRAB
O
-U
ULB
show
ws promise over
o
previouus vaccine/b
bait combinnations as itt elicited a measurable
m
immuunological response
r
in both skunkks and raccooons, and prrovided prottection agaiinst
experimental letthal rabies virus
v
exposuure in skunkks. Further,, results of these
t
studiees
f
perform
mance is unnlikely to bee affected by
y pre-existinng immunitty to
sugggest that its field
otherr adenoviruses.
CO.551
duction and
d evaluation
n of a chroomatograph
hically puriified Vero cell
c rabies
Prod
vacccine (PVRV
V) in China using micrrocarrier teechnology
DC - Institutee for Viral
Tangg Q1, Yu P1, Huang Y1, Zhang Y2, Liang G1 - 1China CD
Diseaase Controll and Prevenntion, 2Beijiing, China - Liaoning Chengda
C
Biiology Co.
b cases evvery year; thhus,
Chinna is a high population country wiith millions of animal bite
it is nnecessary to
o explore annd develop more
m
effecttive and prooductive rab
bies vaccines for
humaan use. To establish
e
a safe,
s
effectiv
ve, inexpennsive and higgh-yield rabbies vaccinee, a
non-aadjuvant puurified Veroo cell rabies vaccine prooduced in thhe SPEEDA
A PVRV
microocarrier bio
oreactor wass developedd by Liaoninng Chengdaa Biology Co. Ltd. in China.
C
This vaccine waas producedd using Vero
o cells that were
w cultureed in a micrrocarrier
bioreeactor. A miicrocarrier bioreactor
b
containing
c
225 g/L of Cy
ytodex-1 waas used for
perfuusion culturre. The Veroo cell culturre density was
w up to 1.2
2–1.5 × 107 cells/ml,
virusses could bee constantlyy harvested for
f 18–22 d,
d and the resulting vacccine immunnizing
potenncy was ≥ 4.5
4 IU/ml. Vaccine
V
safeety and imm
munogenicitty post-imm
munization were
w
also assessed. A total of 602 volunteerrs were enroolled and divvided into tw
wo groups tthat
V
PVRV on days
d
0, 3, 7,, 14
weree vaccinatedd with eitherr SPEEDA PVRV or VERORAB
and 228. All subjects vaccinated with SPEEDA PV
VRV showed no seriouss local or
systeemic adverse effects. The positive conversionn rate of seruum neutralizzing antiboddies
againnst the rabiees virus reacched 100% in both the test and conntrol groupss (inoculateed
with VERORAB
B PVRV) att 14 d and 45
4 d after vaaccination, and
a no signnificant
f
betweeen the neuttralizing anttibody geom
metric meann titers (GM
MTs)
difference was found
ps. SPEEDA
A PVRV is appropriatee for mass production
p
a shows
and
of the two group
a immunoogenicity foor human poost-exposure prophylaxxis of
satisffactory clinnical safety and
rabiees.
CO.552
nk rabies in
n Texas; A retrospecttive look
Skun
Abboott S1, Meseenbrink B1, Mapston M1, Bodenchhuk M1, Oerrtli E2 - 1US
SDA-Wildlife
Serviices, 2Texass Departmennt of State Health
H
Servvices
A is ubiquittous, with thhe majority of the state within the
Skunnk rabies in Texas, USA
rangee of the Souuth-Central skunk rabiees distribution. Statewide public health
h
surveeillance inddicates a cycclic trend, with
w peaks inn total skunk rabies casses
approoximately 22
2 years apaart. We exaamined publlic health caase-reports from
f
1960-22006
to ideentify trendds, with the ultimate gooal of develooping a preddictive moddel for skunkk
rabiees epizootics. Cases weere plotted by
b county, by
b year and
d certain trennds were
obserrved. Somee counties regularly rep
ported skunnk rabies casses while many
m
others
reporrted no casees for severaal years. We
W also exam
mined rainfaall data from
m4
repreesentative coounties to determine
d
iff there was a correlationn between rainfall
r
and
skunnk rabies casses. This paaper presentts the resultts of these innvestigation
ns and preseents
oppoortunities for further invvestigationss.
CO.553
ulation Stru
ucture of tw
wo Rabies Hosts in Allaska
Popu
Clem
ment CJ, Him
mschoot E1, Goldsmith
h EW1, Hunndertmark KJ
K 1, Huefferr K1 1
Univversity of Alaska
A
Fairbbanks
N
andd Western Alaska,
A
but not
n
Rabies, is widesspread in arcctic and redd foxes in Northern
endemic in Interrior Alaska.. Areas withh endemic rabies overlaap with the habitat of thhe
c
f of endeemic
free
Arctiic fox whilee regions soolely inhabitted by the reed fox are considered
rabiees. It is thereefore not knnown if red foxes servee as competeent long-terrm reservoirrs for
rabiees or supporrt only spill over infectiions. Three strains of rabies
r
virus are present in
Alaskka with disttinct geograaphic distribbution. Furthhermore cliimate chang
ge is expecteed to
alter the distribu
ution of thesse two speciies in Alaskka, expandinng the rangee of the red fox
t arctic fo
ox. We assesssed the poppulation stru
ucture of the
into tthe historic habitat of the
two m
major hosts in Alaska, red and arcctic foxes inn the contextt of rabies strain
s
distriibution in Alaska.
A
In coontrast to prrevious studdies on popuulation struccture of Arcctic
foxess in North America,
A
wee found signnificant struucture in thee population
n of arctic fooxes,
whicch correlatess with the phylo-geograaphic distribbution of raabies strainss in Alaska. Red
foxess also show
wed evidencee of only lim
mited gene flow
f
betweeen regions of
o Alaska.
CO.554
E GERMAN
N RACCOON (PROC
CYON LOT
TOR) POP
PULATION
N AS
THE
POT
TENTIAL RABIES
R
R
RESERVOIR SPECIE
ES
Vos A1, Finke S2, Habla C1, Freuling C2, Teifke J2, Köllner B2, Müller T2 - 1IDT
H, 2Friedricch Loeffler Institute
I
- Federal
F
Reseearch Instituute for Anim
mal
Bioloogika GmbH
Heallth
t red fox (Vulpes vulp
lpes) was coonsidered th
he only reseervoir speciees for
Untiil recently, the
terrestrial wildliife rabies inn Europe. Hoowever, sinnce the late 1980s
1
anothher host speecies
N
and East Europe; the raccooon dog (Nyyctereutes procyonoides).
has eemerged in NorthInterrestingly, seeveral other potential raabies reservoir species occur in Eu
urope but
withoout evidencce that thesee animals play any role in the spreaad of the dissease; goldeen
jackaals (Canis aereus)
a
in Sooutheast Euurope, the sm
mall Indian mongoose (Herpestes
auroppunctatus) inhabiting parts
p
of the Adriatic cooastal regionn, and the raaccoon (Proocyon
lotorr) in most paarts of Euroope. Especiaally in (sem
mi-) urban arreas in Germ
many, extrem
me
high raccoon poopulation deensities havee been obseerved (appro
ox. 100 anim
mals/km²). To
T
p
thhat raccoonns in Germanny could beecome a reservoir speciies in
invesstigate the possibility
case of re-emerg
gence of rabbies in Germ
many the suusceptibility of the ´locaal’ raccoon
popuulation was investigatedd. Wild caught animalss were inocuulated with the most likkely
lyssaavirus varian
nts to infectt the Germaan raccoon ppopulation. It was show
wn that the
Germ
man raccoonns were fullly susceptible for a dogg and raccooon rabies virrus (RABV
V)
variaant. Five of 6 raccoons inoculated with a fox R
RABV isolaate showed subsequenttly
cliniccal signs. Inn contrast too foxes, non
ne of the inffected raccooons succum
mbed to rabiies
after infection with
w European Bat Lysssavirus Typpe 1 (EBLV--1); althouggh all these
onverted. Thhe most likeely event thaat a Germann raccoon will
w come in
raccooons seroco
contaact with a lyyssavirus is through contact with an
a infected bat.
b It can th
herefore be a
reasssuring thoug
ght that baseed on the reesults of thiss study racccoons are highly refracttory
to EB
BLV-1 virus infection. The highesst risk that thhe raccoon population will becom
me
infeccted with rabbies is by ree-emergence of this dissease by acccidental impportation off dog
rabiees. Althoughh the immeddiate risk may
m not seem
m high it muust be stresssed that no
experience and no
n tools aree readily avaailable to coontrol a rabiies outbreakk among
many.
raccooons in Germ
c
as part of thee Lyssavirus Research Network annd sponsoreed by
The sstudy was conducted
the G
German Fedderal Ministrry of Educaation and Reesearch (graant nr. 01KII1016A).
CO.555
mes in the Balkan
B
region: achieveements and
d perspectivves
Rabiies control programm
in M
Macedonia
Cliquuet F1, Mrennoski S2, Daavcheva K3, Kirandjiskki T4, Nakov
va E4 - 1Nanncy laboratoory
2
for raabies and wildlife,
w
Frannce, Facultty of Veterinnary Mediccine in Skop
pje - Macedoonia,
3
4
Agrriconsulting Europe SA
A - Macedonnia, Food & Veterinary
y Agency - Macedoniaa
Throough the Insstrument andd Pre-Accesssion Assisttance, Europpean Unionn provides
fundiing to suppoort the conttrol and erad
dication of cclassical sw
wine fever an
nd rabies inn
sevenn candidate or potentiaal candidate countries of
o the Westeern Balkanss, where the
mainn reservoir and
a vector of
o rabies is the
t red fox (Vulpes
(
vulp
pes). Most of
o those
counntries have reinforced
r
m
measures
aim
ming to conntrol the diseease and haave initiatedd oral
vacciination proggrammes aggainst rabiess which starrted in Kosoovo in sprinng 2010. The
projeect was initiiated in Maccedonia in August
A
20100 and the first objective was to
imprrove passivee surveillancce and reporting througgh enhancinng public aw
wareness about
the riisks of the disease
d
and training forr stakeholdeers. Macedoonia is a 25,,713 km2
counntry bordered by four innfected coun
ntries. In Macedonia,
M
th
here had beeen no casess
reporrted since 2000. The firrst oral vacccination cam
mpaigns weere implemeented in spriing
and aautumn 2011 in the whhole country
y using SAD
DB19 vaccin
nes and aeriial distribution.
The ffirst case off rabies wass detected in
n July 2011 on a fox in the centre of
o the counttry.
Eighht other casees have beenn reported inn the centree, in the eastt and in the south parts of
Maceedonia. Sevveral cases have
h
been issolated at cllose proxim
mity with Buulgaria and
Greeece, requirin
ng the impleementation of
o addequatte measuress in those coountries. Thhe last
case was recorded in a cat in
i March 20012. The nuumber of sam
mples and of
o species
mpared to thaat of 2009 and
a 2010. Thhis
analyysed in 2011 was signifficantly incrreased com
refleccts a higherr level of aw
wareness of rabies amonng hunters, general pubblic and
profeessionals inv
volved in raabies controol activities as well as increased
i
qu
uality of thee
rabiees surveillan
nce and monnitoring. Thhe efficacy of
o the camppaigns was assessed
a
thrrough
moniitoring healthy foxes (w
with active participatio
p
on of hunters) in all vacccinated areas
and rrevealed a very
v
high peercentage off bait uptakee (estimatedd at 91%) annd an rabiess
antibbody responnse of foxes at 59%. Thhe genetic chharacterizattion of eightt strains isolated
in M
Macedonia haas been unddertaken to identify
i
rabies variantss circulatingg in the counntry.
The ttested isolattes were ressolved in the East Euroopean groupp with a highh nucleotidee
identtity of the nucleoprotei
n
gesting wild
dlife movem
ments
n gene founnd for all isoolates, sugg
of rabbies in the region.
r
The multi
m
annuaal rabies conntrol prograamme will be
b shortly deescribed (raabies
surveeillance netw
work, plannning, organiisation, impplementationn and evaluaation of the
camppaigns, labo
oratory inveestigations) as well as aachievementts done in thhe Balkan
regioon. Perspecttives for streengthening collaboratioon with the neighbouring countriees for
the nnext years will
w also be discussed.
d
CO.556
SULTS OF THE FIRS
ST ONRAB
B® SAFETY AND IM
MMUNOGE
ENICITY
RES
FIEL
LD TRIAL
L IN RACC
COONS IN THE U.S.
Chippman R1, Veercauteren K2, Nelson K1, Algeo T1, Slate D1 - 1USDA, APHIS
A
Wilddlife Servicees, 2USDA, APHIS, WS
W - Nationaal Wildlife Research
R
Ceenter
mmunogeniciity field triaal to evaluatte a live reccombinant human
h
A saffety and im
adennovirus (sero
otype 5)-rabbies glycoprrotein vacciine (ONRAB
B®) in racco
oons and skkunks
was cconducted in
i the U.S. in
i 2011. Ap
pproximatelly 80,000 Ultralite
U
baitts (Artemis
Techhnologies, Guelph,
G
ON, CAN) werre distributeed at 75 baitts/km2 alongg 750m fligght
lines in 4, 127 km
k 2 study arreas in south
heastern Weest Virginiaa, U.S. The bait was
s
blisterr pack that contained
c
thhe ONRAB® vaccine with
w a waxyy
compposed of a small
coatiing matrix of
o attractantts impregnatted with tetrracycline biiomarker, annd camoufllaged
by a green dye. No phone calls
c
from human
h
or peet bait contaacts were reeported through a
n
provvided on eaach bait. Loow human population
p
d
density
mayy
toll-ffree phone number
largeely account for no reported bait contacts. No tissue abnoormalities were
w observeed in
captiive cottontaail rabbits, opossums,
o
foox squirrelss, eastern wiild turkeys, and woodraats at
®
a 10xx ONRAB dose, and field
f
histopaathology results shouldd be availabble in Decem
mber
20122. Rabies viirus neutraliizing antiboody (RVNA
A) was higheer among raaccoons
(P<00.05) in postt-ONRAB® samples (4
49.4%, n=2996) than in naïve
n
pre-O
ORV samplees
(9.6%
%, n=395). Biomarkerr was higherr (P<0.05), among posst-ONRAB® raccoons
samppled, an indiication of vaccine-indu
v
uced RVNA
A’s. The 49.4% RVNA
A populationn
levell in raccoon
ns is the highhest observeed in the U..S. for a firsst time oral rabies
r
vaccine
distriibution even
nt. Skunk sample
s
size was inadeqquate to asseess ONRAB
B® effects. Field
F
trial rresults warrranted replication and expansion
e
inn 2012 to assess raccooon populatioon
immuunity from a second ON
NRAB® triaal in four more
m
states, including
i
O
Ohio
urbansubuurban habitaats. These collaborativee trials, whiich will conntinue to brin
ng togetherr
multiiple discipliines from coounty, statee, federal annd internatioonal jurisdicctions in thee
®
spiritt of One Heealth, shouldd provide a basis to dettermine if ONRAB
O
is suited to
achieeve raccoon
n rabies mannagement gooals.
CO.557
EFERENCE
ES OF SEL
LECT ATT
TRACTANT
TS IN THE
E COATIN
NG OF ONR
RAB
PRE
VAC
CCINE BAIITS BY RA
ABIES RES
SERVOIR SPECIES
Johnnson SR1, Beerentsen AR
R1, Leland B2, Oertli E3, VerCauteeren K1 - 1USDA-APH
U
HISWS-N
National Wildlife
W
Reseearch Centeer, 2USDA-A
APHIS-Willdlife Servicces, 3Texas
Depaartment of State
S
Healthh Services
m
annd researcheers in the United Statess are assessiing how thee
Rabies control managers
t United States
S
oral
Canaadian vaccinne ONRAB® may perfform if integgrated into the
rabiees vaccinatio
on (ORV) program.
p
A measuremeent of success of any ORV
O
program
m is
bait uptake
u
by taarget speciees. The attraactant used in
i the bait matrix
m
surroounding a
vacciine influencces bait uptaake and vacccination ratte. Our objeective is to determine
d
w
which
flavoor of attractaant in the ONRAB®
O
cooating is thee most prefeerred by rabbies reservoir
speciies in the fieeld. In Texaas (TX) we are evaluatiing four attrractants (sw
weet, fish, eggg,
and ccheese) in areas
a
inhabitted by raccooons (Procyyon lotor), skunks
s
(Meppthis mephiitis),
foxess (Urocyon cinereoarggenteus), andd coyotes (C
Canis latran
ns). In Puerrto Rico (PR
R),
we arre comparin
ng the prefeerence of moongoose (H
Herpestes au
uropuctatus)) for cheesee,
coconut, and fish attractants. We moniitored bait stations
s
with
h animal-acttivated cam
meras
s
(untouuched, distuurbed, and removed).
r
I TX, we
In
and rregular checcks of bait status
offerred 540 baitts of which 102 were reemoved, witth cheese an
nd fish mosst often remooved
(bothh 25%) follo
owed by eggg (21%) andd then sweeet (15%) andd unflavored controls
(14%
%). Image sccoring from
m camera datta is underw
way. In PR,, mongoose removed baits
on 388 of 343 occcasions. Though all data
d are not yyet fully an
nalyzed, it apppears
monggoose prefeer cheese, foollowed clossely by fishh. Findings in both TX
X and PR aree
sugggesting that sweet
s
flavors are least attractive too rabies reseervoir speciies. To
confi
fidently statee which attrractants willl likely perfform the besst, we need to completee the
analyyses of these data and do
d more exttensive trialls, especially
y in raccoonn habitat in the
easteern United States.
S
CO.558
ALUATION
N OF NON
N-TARGET
T ANIMAL
L EXPOSUR
RE TO HU
UMAN
EVA
ADE
ENOVIRUS
S RECOMB
BINANT ORAL
O
RAB
BIES VACC
CINE- OH
HIO 2012
Walllace R1, Blaanton J1, Voora N1, Dyerr J1, Smith K1, Chipmaan R2, Rupprrecht C1 - 1US
U
Centters for Diseease Control and Preveention, 2US Centers for Disease Coontrol and
Preveention
ORV) is the primary maanagement practice
p
forr controllingg
Oral Rabies Vacccination (O
mong raccooons and foxxes. Two OR
RV
wildllife rabies inn the Unitedd States, parrticularly am
bait designs
d
are primarily utilized
u
for the
t distributtion of vacccinia rabies glycoproteiin
(VRG
G) vaccine: a fish meall polymer block and a coated
c
sachhet. A primaary public heealth
concern related to ORV baiit distributioon is non-taarget contact between thhe ORV andd
mestic pets. The VRG virus
v
strain used in OR
RV is attenuated in micee, but
humaans and dom
humaan percutan
neous exposure to ruptuured sachetss has resulteed in localizzed vacciniaa
viruss infection in
n very rare cases. Receently, a new
w recombinaant human adenovirus
a
O
ORV
(AdR
RG) has beeen developeed. This vacccine is incoorporated inn ultralight bait
b which has
h
not ppreviously been
b
used inn the United
d States. Surrveillance foor human coontact is
impoortant, particcularly amoong young children
c
thatt may have contact witth the bait, due
d
to theeir lower prrevalence off prior expo
osure and im
mmunity to human
h
adennoviruses.
To evvaluate poteential differrences in co
ontact rates between
b
thee VRG and AdRG bait
typess CDC, the Ohio Deparrtment of Health,
H
and USDA/WS
U
will conducct an
invesstigation duuring ORV baiting
b
in Northeastern
N
n Ohio in Auugust 2012. The focus of
this iinvestigation will be too ensure thaat public heaalth program
ms are in place to captuure
evennts of humann and domesstic animal bait contactt, ensure apppropriate prrotocols aree in
placee in case of a severe addverse eventt from a baitt contact, an
nd evaluate whether the
AdR
RG vaccine bait
b matrix is
i associated with a diffferent human detectionn rate comppared
to baait types useed for distribbuting VRG
G. Updated gguidelines related
r
to ap
ppropriate
manaagement of potential coontacts withh AdRG baiits during ORV
O
activities may be
developed basedd on findinggs from this investigatioon.
CO.559
a Pathogenesis of Bat Rabies
Multtidisciplinaary Approaach to Epizoootiology and
Viru
uses in the United
U
Stattes
Ellisoon JA1, Johhnson SR2, Kuzmina
K
NK
K3, Gilbert AT3, Carso
on WC3, Blaanton JD3,
2
3 1
VerC
Cauteren K , Rupprechtt C - Centters for Diseease Controol and Preveention, Atlannta,
Georrgia, USA - Division off High-Con
nsequence Pathogens an
nd Pathologgy, 2Nationaal
Wilddlife Researcch Center, United
U
Statees Department of Agricculture - Wiildlife Serviices,
Fort Collins, Co
olorado, USA
A, 3CDC
Zoonnotic diseasee surveillannce is typicaally initiatedd after an annimal pathogen has cauused
diseaase in humaans. Early deetection of potentially
p
h
high-risk
paathogens wiithin animall
hostss may facilittate medicaal interventio
ons to copee with an em
merging diseease. To
effecctively spillo
over to a noovel host, a pathogen may
m undergoo genetic chhanges resullting
in vaarying transm
mission pottential in thee new host and
a potentiaally to humans. Rabiess
viruss (RABV) iss one modell pathogen to
t consider for studying the dynam
mics of emeerging
infecctious diseasses under booth laboratoory and fieldd conditionss. The evoluutionary hisstory
of RA
ABV is chaaracterized by
b regularlyy documenteed spilloverr infections and a seriess of
notabble host-shifts. Within this contextt, enhancedd field surveeillance to im
mprove
detecction of spilllover infecttions will reequire validdated techniqques to non
n-invasively
differentiate infeected from non-infecte
n
ed individuaals. In this sttudy we evaaluate the use of
graphy to detect thermaal changes associated
a
w experim
with
mental RAB
BV
infraared thermog
infecction in big brown bats (Eptesicus fuscus)
f
in a captive coolony. Our results indiccated
that 62%
6
of rabiid bats had detectable
d
f
facial
tempeerature decrreases (-4.6°°C, SD ±2.55),
comppared to pree-inoculation baseline values.
v
Thesse data sugggest potential utility forr
discrriminating rabid
r
bats inn natural field settings. In addition, focusing upon
u
RABV
V
circuulating in the United Sttates betweeen 2008 - 20012, we connfirmed spillover eventts of
bat R
RABV amon
ng carnivores and identtified cross--species tran
nsmission events
e
causeed by
four lineages off RABV assoociated with
h insectivorrous bats. Thhis study prrovides a
ABV pathobbiology and spillover dyynamics am
mong and beetween bats and
glimppse into RA
a varriety of messocarnivoress.
CO.660
OWLEDGE
E, ATTITU
UDES AND
D PRACTIC
CES AMON
NG POPUL
LATIONS
KNO
EXP
POSED TO
O BATS IN SOUTHER
RN NIGER
RIA
Osinnubi MOV1, Recuenco S1, Kuzminn I1, Haberling DL1, Blaau DM1, Daavis LB1,
Ehim
miyein AM2, Ogunkoyaa AB2, Ruppprecht C1 - 1Centers forr Disease Control
C
and
2
Preveention - Divvision for High-Conseq
H
quence Pathhogens and Pathology,
P
Ahmadu Bello
B
Univversity, Zariia - Veterinaary Medicin
ne
wide, all butt one
Amoong the currently recognnized speciees in the Lyyssavirus geenus worldw
(Mokkola virus) has
h been ideentified in bats.
b
Recentt discovery of emerging pathogenss of
zoonnotic importtance in batss raises concerns aboutt health riskks of populaations that
direcctly or indireectly come in contact with
w these diverse
d
mam
mmals. Idanrre, an ancieent
city ssituated in Southwest
S
N
Nigeria,
connducts a uniique bat festtival twice annually,
a
w
where
the ppopulations have traditiional practicces that invoolve capturiing and connsuming batts.
Thesse activities bring them
m into direct contact witth bats that are
a importaant not only
becauuse of potenntial exposuure to lyssavviruses, but also to otheer emerging
g pathogens. A
know
wledge, attittude and praactices survvey was devveloped and administereed to investtigate
the ppotential risk
k of exposuure to emerg
ging diseasees among these populattions living
arounnd bat caves, as well ass those who
o participatee in the bat festival.
f
Serrum samplees
weree collected from
f
humanns and bats in
i the studyy area, and were
w analyzed for
lyssaaviruses andd other potenntial microoorganisms.
Participants (n=142) were recruited
r
froom 90 households in 5 different co
ommunitiess (4
%)
rurall and 1 urbaan). Participaant ages rannged betweeen 9 – 83 yeears. Twentty-one (28%
particcipants claimed to have participateed in the baat festival annd 15 (71%) participateed
twicee a year. Off those that participatedd in the festtival, 14 (677%) were in
nvolved in
multiiple activities: 14 (67%
%) reported involvemennt in bat preeparation/coonsumption,, 12
(75%
%) hunted baats, 4 (19%)) sold bats, and 5 (24%
%) watched the
t events. Thirty
T
(26%
%)
studyy participan
nts claimed to
t have enteered a bat caave and 53 (50%) touched a live bat.
b
Eighhteen (18%) participants who reporrted direct contact
c
with
h bats also reported
r
scrattches and 144 (15%) repported bat biites. Ninety-three particcipants (92%
%) made noo
attem
mpt to protect their fam
milies from bat
b bites, whhile 5 (5%) prevented bats
b from
enterring the hom
me, and 1 (11%) reportedd destroyingg bats enterring the hom
me. Only 100
(15%
%) participan
nts claimed to have an extensive knowledge
k
o rabies. However,
of
H
33
(53%
%) would doo nothing if bitten or sccratched by a bat. A total of 102 bllood samplees
weree collected from
f
humanns. Neutralizzation againnst rabies viirus was dettected in 3 (3%).
(
Therre was no ev
vidence of neutralizatio
n
on against non-rabies
n
lyyssaviruses.. Among thee 145
bat sera, seropreevalence of Lagos Bat Virus
V
(LBV
V) antibodiees was obserrved in
Roussettus aegypptiacus (51%
%) and Eidoolon helvum
m (24%) species. Some of the LBV
Vposittive sampless additionallly neutralizzed Shimonii bat virus (SHIBV).
Our results
r
indiccate an insuufficient knoowledge aboout rabies among the sttudy populaation
and aan increased
d exposure through
t
praactices and attitudes
a
tow
wards bats, that maybe
harbooring unkno
own zoonottic pathogenns. This callls for plans to
t provide continuous
c
surveeillance of important
i
pathogens off risk to hum
man health, and the neeed to have
preveentive meassures and reesponse straategies in plaace to safe-guard humaan health.
CO.661
ACE-TIME DYNAMIC
CS OF ATTACKS BY
Y HEMAT
TOPHAGO
OUS BATS AND
SPA
GEO
OGRAPHIC
C ACCESS
S TO HEAL
LTH CARE IN A RE
EGION OF MEXICO.
Zaldiivar-Gomezz A1, Arteagga-Troncoso
o G2, Velazzquez-Quiro
oz IR3, Delggado-Urbinaa C1,
1
1 1
Ram
mirez-Hernan
ndez MD , Jimenez-Es
J
strada JM - Laboratorrio Estatal de
d Salud Púbblica
2
del E
Estado de México-ISEM
M
M - Laborattorio de Rabbia, Institutto Nacional de
Perinnatología, 3Laboratorio
L
o Estatal de Salud Públiica del Estado de Méxiico-ISEM Labooratorio de Biologia
B
Moolecular
The oobjective off this study was to estim
mate the spaatial patternns in rates of aggressionn by
vamppire bats (D
Desmodus rootundus) andd identify human
h
popu
ulations withh limited
geoggraphic acceess to mediccal service with
w post-exxposure propphylaxis.
THOD: A seet of points georeferencced with case reports of
o aggression
n by bats inn
MET
humaans was useed to model the rates-addjusted of aggression
a
a
against
residdents of locaal
comm
munities in the south of the State of
o Mexico. A continuouus predictioon area was
consttructed baseed on the raates of aggreession from
m events duriing the yearr 2000 (outbbreak
of rabbies cases in animals, and
a attacks to humans)) and during
g the period 2001 to 2009
(stabbility in the frequency
f
o cases) usiing a spatial interpolatiion method (Poissonof
Krigiing). The cooverage areas of health
h services w
were calculatted throughh a network
analyysis, estimatting travel times
t
(30 to
o 60 minutess) to the neaarest hospitaal unit. Bothh
estim
mates were integrated
i
w risk maaps.
with
ULTS: Twoo thousand one hundred ninety peoople of diffe
ferent sexes and ages w
were
RESU
attacked by vam
mpire bats inn the region; 94.8% of attacks
a
were recorded in
i the
o Tejupilco and Luvian
nos. In 20000, there wass an outbreaak of rabies in
muniicipalities of
anim
mals, with an
n increase of 74.4% in the numberr of attacks (1629
(
recorrded). Durinng
20011-2009, the rate
r of aggrression by gender
g
show
wed a signifiicant increaase in womeen
(58.66%) in contrrast to men (41.4%). By age groupp, the highest percentagge of attackks
was ffrom ages 1 to 19. By anatomical
a
region more attacks weere recordedd in the uppper
extreemities with
h 784 cases. Seasonal aggression
a
w observeed, with greaater frequenncy in
was
the months
m
of April
A
to Auggust. Mappinng was deveeloped for both
b
Poissonn-Kriging model
m
desiggns, a trend toward a hiigher rate of aggressionns being observed in bo
oth models in
the nnorth-west, in
i the municipality of Luvianos.
L
T distribuution and seasonality iss
This
relateed to severaal factors suuch as: high productionn and marketing of livesstock in thee
muniicipality, the physical and
a geograp
phical condiitions in thee region thatt favor the
surviival of hemaatophagous bat, and soocio-econom
mic conditioons with hig
gh level of social
vulneerability andd limited acccess to heaalth servicess of basic lev
vels.
NCLUSION
N: The Geoggraphic Infoormation Systems in Heealth providde a tool for geoCON
statisstical analyssis, manageement and planning which is essenntial becausee it is possibble to
addreess the risk of aggression toward humans
h
by vampire
v
batts from a peerspective which
w
is spaatial, system
mic, multi-ccausal and in
nterdisciplinnary.
Acknnowledgements
The aauthors wissh to acknow
wledge to María
M
Eugennia Jaimes from
fr
Comitee de Fomentto y
Proteección Pecu
uaria del Esttado de Méx
xico. Faculttad de Geog
grafía-UAEM
Mex for hiss
assistance with the creationn of cartogrraphy. MD Gabriel
G
O'sh
hea Cuevass from Instittuto
de Saalud del Esttado de Méxxico.
CO.662
BIES VIRU
US MONITORING IN
N BATS FR
ROM THE DIRECT AREA
A
OF
RAB
INFL
LUENCE OF
O HYDRO
OELECTR
RIC POWE
ER PLANT
T IN JIRAU
U, RONDÔ
ÔNIA,
BRA
AZIL
Almeeida MF1, Rosa
R
AR1, Martorelli
M
L 1, Kataooka APAG1, Aires CC2 - 1Centro de
LFA
d
2
Conttrole de Zoo
onoses-COV
VISA-PMSP
P, Museu de
d Zoologiaa da Universsidade de Sãão
Pauloo
In Roondônia Staate, North reegion of Braazil is beingg built at Maadeira Riveer, the
hydroelectric poower plant of
o Jirau. Thee implemenntation of hu
uge enterprises such as the
mands a seriies of alteraations in thee environmeent, such as vegetation
hydroeletric dem
d movementt of land in large
l
extenssion along with
w other antropic
a
suppression and
h environmeental impacct. These acttivities can destroy the artificial annd
activvities of high
naturral shelters for bats. Seeveral studiees have poinnted an assoociation betw
ween these
envirronmental changes
c
andd outbreaks of rabies. Monitoring
M
J
Jirau´s
bat population
p
b
began
in 20010 and it was
w made ovver a period of three years, with regular visits lasting 10 to
t 12
days, twice a yeear, totalizinng six campaigns. First,, the local population
p
w intervieewed
was
o
aiminng at elicitinng what theey knew aboout rabies annd bats and to locate thhe shelters of
thesee animals. Inn the periodd of three yeears, 158 peeople were interviewed
i
d, 4.387 batss
weree captured by mist net or
o active seaarch and 3.8852 of them
m were loosee after this
proceeeding. Reggarding to raabies, 535 bats
b and 4866 sera were sent for diaagnosis and
antibbodies dosagge respectivvely at CCZ
Z-SP by Direect Fluoresccence and Simplified
S
Fluorrescence Innhibition Miicrotest. All bats were negative
n
to rabies. The global
prevaalence of anntibodies waas 15.4% ussing 0.5IU/m
ml as a cut-off (0% in the
t first
camppaign, 2.8%
%, 6.4%, 19.22%, 14.8% and 36.3% in the subsequent cam
mpaigns). In the
intervview 93.5%
% declared too know the disease, hoowever rabiees was assocciate to doggs
and jjust 52.5% knew
k
that bats
b could trransmit rabies. Regardiing the type of houses,
93.5%
% were vulnnerable to bats
b presencce and bats were
w roostinng in 70% of
o these houuses.
The rresults for rabies
r
antiboodies dosagge showed a progressive increase for
f every
subseequent cam
mpaign. As thhe first cam
mpaign was performed
p
before
b
the demolition
d
o the
of
houses and vegeetation supppression in the
t direct arrea of influeence of the hydroelectri
h
ic,
thesee results couuld be refleccting the perturbation inflected to the populattion of bats such
as the stress of the
t forced migration
m
to search for new
n shelterrs and forag
ging areas ass well
f space annd food with
h the bats poopulations already
a
estaablished in thhe
as the disputes for
ment when th
here is the reestablishin
r
ng of the coolonies.
new shelters up to the mom
he contact with
w the viruus,
Conssidering justt the presennce of antiboodies as inddicative of th
the aaverage of tiiters in the first
f
and seccond campaaigns was 0,,03UI/ml annd 0,04UI/m
ml
whilee in subsequ
uent campaiigns this avverage was 00,21UI/ml, 0,49UI/ml,
0
0,24UI/ml and
0,38U
UI/ml, respeectively. When
W
statisticcally analyzzed by Krusskal-Wallis (H=1.611,9942;
p<0,00001) and Dunn
D
(R1=
=44.330) theese tests connfirmed thatt there is sig
gnificant
difference in thee results obsserved amonng the camppaigns and the
t differennce observedd
betw
ween the resu
ults of first campaign and
a subsequuent campaigns is significant. Thesse
data indicate thee increase of rabies viruus circulatioon among th
hese bat poppulations annd
r
outbreak that shoould be mon
nitored.
sugggest a potenttial risk of rabies
CO.663
LIMITAÇÃ
ÃO DE ÁRE
EAS DE CONTROLE
E NO ESTA
ADO DE SÃO
S
PAUL
LO
DEL
PAR
RA A RAIV
VA DOS HE
ERBÍVOROS NAS REGIÕES
R
D SERRA
DA
A DA
MAN
NTIQUEIR
RA E CANA
ASTRA
Cavaalcanti MP1, Nogueira VS
V 1, Gomes MN1 - 1CO
OORDENA
ADORIA DE DEFESA
A
AGR
ROPECUAR
RIA - SECR
RETARIA DE
D AGRICU
ULTURA E ABASTE
ECIMENTO
O
os focos de raiva
r
em heerbívoros esstá associada às localizaações dos
A proogressão do
abriggos de Desm
modus rotunndus que por sua vez, estão relacio
onados às prrincipais feiições
geoggráficas de uma
u região. Muitos estuudos considderaram elevvadas altitudes, rios ouu
deterrminado tipoo de cobertuura da terra como fatorr protetor ouu causal parra difusão de
d
epideemias. Geottecnologias foram utilizzadas por Gomes
G
et al (2011) paraa gerar um
modeelo descritivvo com três camadas so
obrepostas: a enfermid
dade caracterizada
por uuma funçãoo kernel doos focos de raiva
r
entre 1992 e 2003
3, os tipos de uso e
cobertura da teerra obtidoss por classiificação dee imagens de
d satélites e a altitudde
a
a ennfermidade esteve forteemente molddada
oriunnda do radarr SRTM. Seegundo os autores,
pelass áreas de veegetação rasteira (pastaagens). O reelevo formo
ou os mosaicos de uso e
cobertura da terrra, o qual deeterminou os
o locais de grassamentto ou não daa enfermidaade.
S
da Maantiqueira nna
Consstataram-se regiões perrmeáveis dee menores alltitudes da Serra
divissa de estado com Minass Gerais quee eram carreeadores de casos,
c
assim
m como, áreeas
delim
mitadas pelaa serra da Quebra
Q
Canggalha que diivide horizo
ontalmente o Vale do
Paraííba. A análiise temporall demonstro
ou que os foocos tinham
m sentido do estado de
Minaas Gerais paara o de Sãoo Paulo. Freente às consttatações, a Coordenado
C
oria de Defeesa
Agroopecuária doo estado de São Paulo (CDA)
(
prioorizou uma faixa
f
na divvisa estadual
para realização de
d trabalhoss de controlle da enferm
midade. Os locais
l
relataados como
r
s à introduçção de novoos casos. Dee certa formaa,
carreeadores tambbém estão relacionado
validdando o prim
meiro modeelo descritivvo. Focos, aggora identifficados com
mo pontos dee
coorddenadas adqquiridos porr receptor de
d GPS entree os anos dee 2008 e 2012, demonsstram
que o vale do rio
o Sapucaia e Sapucaia--Mirim propporcionam faixas
f
de menores altituudes
na Seerra da Man
ntiqueira parra introduçãão de raiva na Região de Bragançça Paulista e São
José dos Campo
os. A Serra da
d Mantiqueira, na região de Cacoonde e Cruzzeiro, tambéém
q possibiliitam maior permeabilid
p
dade de focoos.
possuuem cortes de menoress altitudes que
Por ffim, na regiãão da Serra da Canastraa também ocorre
o
tais formações
f
de
d baixa altiitude.
Assim
m, estas anáálises descriitivas sugerrem que a CDA
C
possa definir
d
menoores áreas de
d
priorridade de co
ontrole da ennfermidade, que delimiitariam áreaas específicaas sentinelaas e
de coontrole de Desmodus
D
rootndus nas faixas
f
de menores
m
altituudes nas serrras aqui
estuddadas. Trabaalhos conjuuntos com e estado minneiro deveriaam ser fomeentados.
Agraadecimentoss a Coordennadoria de Defesa
D
Agroopecuária e FAPESP prrojeto Nº 033123119-0
mes MN, Moonteiro AMV
V, Escada MIS.
M Raiva bovina seguundo os moosaicos de uso
u e
Gom
cobertura da terrra no estadoo de São Pauulo entre 19992 e 2003. Arq. Bras. Med. Vet.
Zooteec., v. 63, p.
p 287, 20111.
CO.664
LICACIÓN
N DEL MA
ANEJO INT
TEGRADO
O DE PLAG
GAS (M.I.P
P.) EN EL
APL
CON
NTROL DE
EL “DESM
MODUS RO
OTUNDUS
S” EN COS
STA RICA
Sanccho VH1 - 1Senasa,
S
Cossta Rica - Raabia
mentan añoo con año poor causa dell
Las ppérdidas ecoonómicas enn nuestra gaanadería aum
Desm
modus rotun
ndus (murcciélago hemaatófago) esttimándose en
e $800.0000.00 por añoo.
La prrincipal enffermedad quue transmiteen es la rabiia paralítica bovina y laa ecología de
d
nuesttro país hacce propicia la
l multiplicaación de estte vector.
De laas 110 especcies de murrciélagos qu
ue existen enn Costa Ricca, solamentte tres son
hemaatófagas y generalment
g
te los ganadderos no disstinguen esttás de las ottras especies que
tieneen una gran importanciaa ecológica, por lo quee eliminan ecosistemas enteros
utilizzando métod
dos equivoccados en cueevas, troncoos huecos y otros refug
gios, ya que por
vivirr en grupos numerosos
n
son los aniimales más susceptiblees a la extincción.
El P
Programa Naacional de Rabia
R
Paraliitica Bovinaa del Minissterio de Ag
gricultura y
Ganaadería llevaa a cabo el control
c
de este vector, tomando
t
en
n cuenta el concepto
c
de M
MIP (Manejo Integradoo de Plagas), el cual se concibe coomo un sisteema de conttrol
que cconsidera ell medio ambbiente en el que vive laa plaga, su dinámica
d
pooblacional,
previiendo conseecuencias eccológicas y económicas, seleccion
nando métoddos de contrrol
para reducir las poblaciones por debajjo del daño económico, de salud an
nimal y saluud
c
dee Educaciónn Sanitaria que
q comprennde mil
públiica. Por meddio de una campaña
quiniientas charlas y demosttraciones dee métodos a más de quuince mil gaanaderos y
estuddiantes se haa logrado cooncientizar al productoor ganadero para que noo elimine laas
especcie benéficaas en sus finncas.
CO.665
o Myotis lu
ucifugus too heterologoous and hom
mologous rabies
r
virusses.
Suscceptibility of
Daviis AD1, Jarvvis J1, Poulioott C1, Morgan S2, Ruddd RJ1 - 1Wadsworth
W
C
Center
- Division
of Innfectious Diseases, 2Staate Universiity of New York,
Y
Albanny - Departm
ment of Ecoology
and E
Evolutionarry Biology,
R
Rabies virus (RV) mainttenance in bats
b is not w
well understtood. Eptesiicus fuscus,
Myottis lucifuguss, and Tadaarida brasiliiensis are thhe most com
mmon bats species
s
in thhe
US. T
These colon
nial bat speccies also havve the mostt frequent coontact with humans and
domeestic animalls. Howeverr, the Lasionycteris nocctivagans/P
Perimyotis subflavus
s
(Ln/P
Ps) RV is asssociated with
w the majoority of hum
man rabies virus
v
infectiions in the
Uniteed States an
nd Canada. This
T is of in
nterest becaause the L. noctivagans
n
s and P.
subfllavus bat species are m
more solitaryy bats with less
l frequennt human intteraction.
O
Our interest was
w to deterrmine the liikelihood off a colonial bat species becoming
infeccted with annd transmittiing a heteroologous RV. To determ
mine the poteential of
heterrologous RV
V infection in colonial bat species,, M. lucifug
gus bats werre inoculatedd
with a homologoous or one of
o two heterrologous (E
E. fuscus andd L. noctivaagans) RV.
d the diseasse process, bats
b
Addiitionally, to determine if the route of exposuree influenced
weree inoculatedd either intraamuscularlyy (i.m.) or suubcutaneoussly (s.c.) wiith a
homoologous or heterologou
h
us RV. Batss were obserrved for 6 months.
m
Survvivors weree
challlenged i.m. with a hom
mologous RV
V and obserrved for an additional
a
6 months.
O
Our results demonstrate
d
intramuscuular inoculattion results in a more rapid
r
progreession
of disease onset as compareed to a signiificantly lonnger incubaation time inn bats inocuulated
Additionally
y, cross prottection was not consisttently achiev
ved in bats previously
s.c. A
inocuulated with a heterologgous RV following a sixx month chaallenge withh a homologgous
RV. Finally, batts that develloped rabiess following s.c. inoculaation were significantly
s
y
moree likely to shhed virus inn their salivaa and demonstrated inccreased viraal tissue troppism.
In suummary, batts inoculated via the s.cc. route are more likely
y to shed virrus thus
increeasing the potential for transmissioon.
CO.666
C’S GLOBA
AL DISEA
ASE DETEC
CTION PR
ROGRAM AND
A
THE
CDC
INTE
ERNATIO
ONAL HEA
ALTH REG
GULATION
NS: PROVIIDING EA
ARLY
WAR
RNING TO
O CDC FOR
R HUMAN
N RABIES OUTBREA
AKS
Chrisstian KA1, Arthur
A
RR1 - 1Centers for
f Disease Control and Preventioon - Global
Diseaase Detectio
on and Emeergency Ressponse
RS alerted puublic healthh leaders thaat novel pathhogens could be
In 20003, the spreead of SAR
transsmitted alon
ng internatioonal travel routes
r
with unpreceden
u
nted speed. With
W the
realizzation that an
a outbreakk anywhere in the worldd was a poteential threatt to virtuallyy all
counntries, the United Statess Congress in
i 2004 authhorized the appropriatiion of fundss to
estabblish a Globbal Disease Detection
D
(G
GDD) proggram, based at the CDC
C, with the aim
a
of prromptly deteecting and mitigating
m
thhe consequeences of em
merging threeats. The GD
DD
progrram providees a platform
m to develoop and strenngthen globaal capacity to
t rapidly detect,
identtify, and con
ntain emergging infectioous disease and bioterro
orist threatss in line withh the
Interrnational Heealth Regulaations (IHR
R), which enntered into force
fo in Junee 2007 and
legallly requires all signatorry nations too establish ssystems to detect
d
and reespond to new
n
diseaase threats. The
T GDD program
p
waas subsequenntly selectedd by WHO as a key parrtner
to heelp implemeent the IHR (2005) for its 194 mem
mber states and in 2009
9 was designnated
a WH
HO Collabo
orating Centter for Implementation of IHR Nattional Surveeillance andd
Respponse Capaccity. A signiificant compponent of GDD
G
is the GDD
G
Operaations Centeer
(GDD
DOC), an epidemic inttelligence unnit which uses novel, event‐based
e
surveillancce
technniques to prrovide CDC
C programs with
w a singlle source off reliable, coomprehensivve,
and hhigh quality
y informatioon on international diseease outbreaaks, and proovides logisttical
and ffinancial sup
pport to CD
DC program
ms for emerggency deplooyments to internationaal
outbrreaks. Tech
hnological advances havve revolutioonized the way
w informaation is
accesssed, and evvent-based surveillance
s
e provides a mechanism
m for the org
ganized andd
rapidd collection and verification of info
formation abbout events that are a riisk to publicc
healtth, particulaarly with reggard to emerging zoonooses, which
h countries sometimes
s
cannnot or do nott report to thhe global puublic healthh communityy. A re-emeerging, globbal
zoonnosis that thee GDDOC actively
a
moonitors is rabbies in bothh animals annd humans.
Sincee 2009, the GDDOC haas supported
d provided epidemioloogic, logisticcal, or finanncial
support to CDC’s Rabies Program for emergency deploymen
nts to the Doominican
o mitigate outbreaks
o
off human rabbies associatted
Repuublic, Peru, Ecuador, annd Kenya to
with canine and vampire baat rabies. Beecause of thhe GDDOC’’s work to actively
a
idenntify
s
e data to CD
DC’s Rabiess Program, CDC
C
and rreport rabies-related evvent-based surveillance
is bettter position
ned to respoond to a requuest for techhnical assisstance by the affected
counntry and estaablish core capacities
c
in
n compliancce with IHR
R.
CO.667
SULTS WE
EBSITE FO
OR RABIES
S DIAGNO
OSTIC CON
NSULTAT
TION INTO
O
RES
THE
E HEALTH
H SERVICE
ES OF COA
AHUILA, MEXICO.
Fernaandez MM1, Solis MEP
P2, Romeroo MAR2, Agguilar AMB
B1 - 1Laborattory of Publlic
2
Heallth Saltillo, Coahuila,
C
M
México,
Heealth Services of Coahu
uila
kground: Health
H
Serviices of Coahhuila had noot a rabies diagnostic
d
laaboratory. Itt was
Back
needded to send samples
s
to Nuevo
N
Leonn and InDR
RE to solve this
t limitatioon. On Apriil,
20100, the State Authorities
A
establishedd this laboraatory that is placed insidde the faciliities
of the State Laboratory of Public
P
Healtth. On July,, 2010 the laaboratory foormalized
operaations with the InDRE.. The laboraatory has 2 eemployees: a professioonal diagnosstics
speciialist and a laboratory technician.
t
mple shipm
ments for rab
bies virus monitoring
m
too other statees. Extempoorary
Challlenges: Sam
recepption of other states ressults. No cleear idea aboout rabies viirus circulattion and sannitary
risk sstatus in thee State. Exppensive operrational costts. Several criteria
c
of raabies PEP based
b
on laab results. The
T laboratoory had not a working algorithm
a
fo
or technical and
epideemiologicall reports as well
w as for laboratory
l
r
results.
The lab had nott a standardd
protoocol to estab
blish a timely diagnostiic.
T develop a feasibilityy study for the
t operatioon of a rabiees
Alterrnatives off Solution: To
diagnnostic laborratory and too identify thhe mechanissms and sup
pport elemeents to estabblish
this llaboratory. Review
R
the operative and
a financiaal plans and
d rabies vacccines availaability
for P
PEP. Websitte design for electronicc consultatioon via Intern
net, to get th
he timely
diagnnostic accorrding with the
t needs an
nd request fr
from the opeerative unitss.
Operration: Thee Epidemiollogical survveillance of rabies in thee State is deeveloped in
threee steps: the pre-analytic
p
c step is for the data collection andd integrationn of the file with
the suupport of th
he sanitary regions
r
thatt send the saamples that could be cooming from
m
bittenn animals, for
f monitoriing purposees and from wildlife. Thhe Analyticcal step for the
t
laborratory resultt and capturre of the epiidemiologiccal info as well
w as the laaboratory reesult
in thee website. Each
E
case iss identified with
w an ID key that couuld be revieewed by inteernet
for each sanitary
y region. Thhe post-anallytic step is to follow up
u the resultts. This is
he sanitary regions
r
to deevelop the recommend
r
ded preventiion and conttrol
developed by th
A users haave an ID keey access thhat is individdual and noon-transferabble,
meassurements. All
once it is approv
ved by the State
S
Authorities. The website
w
has different acccess levelss and
f capturinng the initiall
just tthe rabies laaboratory peersonnel havve administtrator level for
info, modificatioons and corrections of each case. All
A the otheer users, adm
ministrative
o
peersonnel of the
t sanitaryy regions, haave
persoonnel of thee central levvel and the operative
an ID
D key to con
nsult the labb results. Th
he author waants to thannk to the Staate Authoritties
and tthe workforrce of the Health Servicces in Coahuuila for the support recceived for thhe
elabooration and presentation of this papper.
CO.668
MAL BITE
ED AND HUMANE
H
R
RABIES
SU
URVEILLA
ANCE IN IRAN
I
ANIM
Shirzzadi MR1 - 1ministery of
o health iraan - zoonosees
man rabies inn Iran is woorking with the aim of having
h
no
Survveillance sysstem of hum
humaan rabies an
nd 100% acccess to heallth services for all the community.
c
. Further moore,
coopperation with
h veterinaryy organization and enviironment prrotection orgganization aabout
vacciination of dogs
d
and catts, samplingg of suspicioous animalss and also trraining the staff
s
and ccommunity and attendiing rabies national com
mmittee meeetings is perrforming duuring
the yyear.
Iran hhas 31 provvinces and 386
3 districtss with a diffferent weathher conditio
on as
Mediiterranean weather
w
in thhe north of Iran and hoot and dry weather
w
in thhe south. Ass a
resullt the domin
nant carnivoores in the north
n
and Noorth West of
o Iran are wolves
w
and in
i
otherr areas of th
he country itt's fox. How
wever, in thee whole cou
untry jackals are found
frequuently and have
h
been coonsidered as
a the reservvoir of rabiees in the natuure.
The hhealth centeer in villagees includes health
h
housees with trainned staff annd in the nexxt
levell there are ru
ural health centers
c
with
h a family physician.
p
Inn the cities there
t
are heealth
postss in the firstt level and urban
u
health
h centers in the next lev
vel which haave physicians;
and aalso there arre hospitals in the distrricts. The coomplicated disease
d
are referred froom
healtth houses annd health poosts to rural or urban heealth centers and in neccessary to
hospitals.
nd rabies coontrol progrram educatioonal measurres in contrrol and
In annimal bite an
preveention has been providded in all levvels. In eacch district th
here is faciliitated prevention
and ccontrol centter for humaan rabies in one of the urban
u
or rurral health ceenters or
hospitals. More than 700 ceenters for coontrol and prevention
p
o human raabies are
of
t
abou
ut preventinng
perfoorming in 3886 districts with 24-houur staff thatt had been trained
treatm
ment methoods. If the caases of anim
mal bite refeer to health houses, heaalth posts orr
healtth centers th
he lesion wiill be washeed with wateer and soap for 15-20 minutes
m
afteer
regisstration of thhe patients and
a they wiill be referreed to preven
nting treatm
ment center for
f
humaan rabies to start vaccinnation and serum
s
theraapy if needed. Further more
m
the
patieents' informaation will be sent to thee district heealth center to be declarred to the
preveenting treatm
ment centerr for the folllow-up. If thhe cases of animal bitee don't visit the
preveenting treatm
ment centerr for rabies they
t
are acttively follow
wed-up. In the
t district
healtth center thee informatioon of the paatients will be
b registeredd in the onliine portal
reporrting system
m. The onlinne informatiion can imm
mediately bee seen after registrationn by
the pprovince or national
n
levvel.
In the national leevel the nattional guideeline for rabbies surveillaance had beeen publisheed
n of nationaal rabies com
mmittee andd they are distributed
d
too all the heaalth
with cooperation
venting treattment centeers for rabiees in Iran.
centeers and prev
Conssidering the national raabies commiittee decisioons two typees of vaccin
nes includinng
VER
RO and PCE
EC and speccific human rabies serum
m are proviided and disstributed to all
the ppreventing centers
c
for rabies.
r
Baseed on the tabble of inform
mation received from preventing
p
t
treatment
ceenters for raabies,
in thee year 2011 about 1350095 cases of animal bitte and 7 casses of rabiedd had been
regisstered.
Baseed on the naational guideeline of rabiies two typees of prevennting treatm
ment measures
are cconducted affter animal bite which include 3-ddose and 5-d
dose treatmeent. The rabbies
contrrol program
m in Iran is successfully
s
y performingg due to 0n
n time measu
ures.
CO.669
wards huma
an rabies ellimination in
i Eastern Europe an
nd Middle East:
E
curreent
Tow
statu
us and prosspects
Vrannjes N1 - 1PA
ASTEUR IN
NSTITUTE
E NOVI SAD
D, SERBIA
A, EUROPE
E - Epidemioology
Depaartment
xpert Bureauu (MEEREB
B)
Folloowing the 1st Middle Eaast and Easttern Europee Rabies Ex
meetting held in Turkey in 2010,
2
the grroup of rabiies experts from
f
8 partiicipating
counntries met inn June 2012 in Paris, Frrance for thee 2nd meetin
ng, togetherr with
repreesentatives from
f
the Insstitute Pasteeur and the Global Alliiance for Raabies Controol to
discuuss the rabiees situation in their resppective couuntries, its evvolution, thhe problems
encountered and
d the solutioons.
man rabies, number
n
of animal
a
bites and post-exxposure pro
ophylaxis (P
PEP)
Hum
admiinistered aree notifiable in all MEE
EREB countrries howeveer reliable rabies data in the
regioon still remaain scarce esspecially reegarding hum
man rabies reporting.
r
T is evenn
This
moree pronounceed in countrries that are not part of the Rabies Bulletin Eu
urope. Thankks
howeever to MEE
EREB mem
mbers’ activee participatiion, a cleareer image off the rabies
situaation in the region
r
is beecoming moore apparentt.
REB countriees (Croatia and Serbia)) have not reecorded hum
man deaths from
Somee of MEER
rabiees for more than 30 yeaars. Howeveer, persistennce of rabiess in wildlifee (foxes)
requiires constannt surveillannce and maintenance off a high leveel of vigilannce and pubblic
healtth interventiions in ordeer to preventt human rabbies cases. In
I other ME
EEREB counntries
rabiees is presentt not only inn wildlife buut also in doogs, which increases
i
thhe risk of
transsmission to humans.
h
In fact, humann cases stilll do occur inn these counntries.
In coountries wheere the inciddence of huuman rabies did not excceed 0.15 peer million
inhabbitants durinng the last 3 years, the incidence of
o reported bites
b
and PE
EP remained
stablle. In the tw
wo countries where the incidence
i
of human rab
bies is the highest,
h
nd 0.67-1.377, respectiveely), the meean PEP
Kazaakhstan andd Georgia (00.38-0.52 an
inciddence increaase rate wass of over 15%
%. In 2011,, 0.4% of Kazakh
K
popuulation receiived
PEP.. In Georgiaa, PEP inciddence reacheed 0.9% - thhe highest known
k
PEP incidence in
i the
worldd, and 2-50 fold higherr than in othher MEERE
EB countriess. All MEER
REB countrries
use ccell culture vaccines
v
thrrough intram
muscular addministratio
on. Rabies im
mmunoglobbulin
(RIG
G) is locally produced inn Croatia, Serbia,
S
and Ukraine.
U
EREB particcipants stateed that hum
man rabies transmitted by
b dogs can be and shouuld
MEE
be elliminated from the regiion and disccussed requiirements to conduct a successful
s
rabies
elimiination prog
gram. Althoough in all MEEREB
M
coountries a leegal framew
work for rabbies
contrrol exists, most
m often raabies eliminnation is nott consideredd as a publicc health prioority.
In adddition, although some rabies conttrol measurees are alreaddy in place, there is stilll a
signiificant dispaarity amongg the MEER
REB countries as to eacch country’ss response too its
rabiees situation. Furthermorre, some off the conditions for succcessful rabies eliminatiion
progrrams are no
ot met. MEE
EREB mem
mbers agreedd that reinforced interseectional
collaaboration an
nd “One Heaalth approacch” are the keys for succcessful rabbies control..
CO.770
MPARATIV
VO DE LA
A INVERSIIÓN EN BIIOLÓGICO
OS ANTIR
RRÁBICOS
S
COM
PAR
RA LA PRE
EVENCIÓN
N DE LA RABIA
R
HU
UMANA TR
RANSMITIIDA POR EL
E
PER
RRO EN MÉXICO
Varggas FP1, Guttiérrez VC1, Chávez IA
AF1, Lezanaa MAF1 - 1Secretaría
S
dee Salud/Méxxico CEN
NAPRECE
ón de la rabiia humana transmitida
t
por el perro
o en México
o, los biológgicos
En laa eliminació
antirrrábicos han
n sido elemeento básico para
p alcanzzarlo, por elllo el gobiernno federal ha
h
destinado presup
puesto para adquirirloss, gasto perm
manente dessde 1990 a la
l fecha.
A coontinuación se establecee comparatiivo de la déccada de los 90’s con ell período deel
20000, se utilizan
n indicadorees de resultaado (caso dee rabia hum
mana transmitida por el perro
y casso de rabia canina)
c
y dee proceso (ppersonas agrredidas, trattamientos inniciados y perros
p
vacuunados contrra la rabia), las cantidad
des de los biológicos
b
antirrábicos
a
adquiridos y
utilizzados (vacuuna antirrábiica humana,, inmunogloobulina antiirrábica hum
mana y vacuuna
antirrrábica caninna) en dólarres americannos y el efecto logradoo con los sigguientes
resulltados:
0’s se registtraron
Casoos de rabia humana trransmitida por perro,, en la décadda de los 90
240 ccasos en 20 entidades federativas
f
de
d las cualees cuatro aportaron el 49%
4
del totaal, en
contrraste en el período
p
del 2000
2
se nottificaron cuaatro casos en
e sólo dos entidades,
e
s
se
dejarron de preseentar a partiir de 2006.
d los 90’s se
s acumularron 13 mil casos
c
en 31 de
Casoos de rabia canina, en la década de
las 32 entidades, en contrasste se reducee para el peeríodo del 20000 a 921 casos
c
en 23
9
entiddades (decreemento del 93.3%).
didas, en la década de los
l 90’s se acumularon
a
n 947 mil peersonas
Personas agred
agreddidas por annimal sospechoso de raabia con unaa tasa promeedio de 1066.2 agredidoos por
cada 100 mil hab
abitantes, miientras que para el perííodo del 20000 suman 1.2 millones de
c una tasaa de 96.4 poor cada 100 mil habitanntes.
estass personas, con
Personas que see indica vacunar, en la década dee los 90’s see acumularoon 334 mil
ntras que parra el períoddo del
persoonas que iniiciaron trataamiento antiirrábico (355.3%), mien
20000 lo iniciaron
n 378 mil (331.3%).
una antirrá
ábica canin
na aplicada, en la décaada de los 900’s se aplicaaron 101.3
Vacu
milloones de dosiis a igual núúmero de annimales de compañía,
c
m
mientras
que para el peeríodo
del 22000 se incrrementó a 1996.2 millon
nes de dosis aplicadas (93.7% de au
umento).
una antirrá
ábica humaana aplicad
da, en la déccada de los 90’s se utiliizaron 1.4
Vacu
milloones de dosiis que correespondió en tratamientoos completo
os (cinco doosis) al 56.4% y
el ressto (44.6%)) en incompletos (tres dosis
d
en proomedio), misma cifra ap
plicada en el
e
períoodo del 20000 pero se innvierte el co
onsumo en trratamientoss completos 46.1% y enn
incom
mpletos el 54.9%.
5
unoglobulina antirráb
bica human
na aplicadaa, inicia su uso
u a partir de 1998
Inmu
acum
mulando hassta el año paasado 385 mil
m frascos aplicados
a
enn 97 mil perrsonas con
agressión de riesggo grave.
La innversión quee hizo la Seecretaría de Salud de 19990 a 2011 en estos bioológicos fuee de
$1055.4 milloness de USD, que
q comparaado con el gasto
g
que see dejó de apllicar en meddidas
de coontrol de rab
bia canina en
e los casoss que se evittaron en estee período (9
92,028)
estim
mado en $22
25 millones de USD, reepresentó ecconomías paara el país por
p $120
milloones de USD
D, lo cual juustifica estaa inversión en
e dichos biiológicos.
CO.771
k model to estimate boovine rabiees occurren
nce in Brazil
Quallitative risk
Bragga GB1, Leitte BM2, Sanntos EFS3, Dias
D RA4 - 1FMVZ-US
SP - Laborattório de
Epiddemiologia e Bioestatísttica, 2Brazillian Ministrry of Agricuulture, Livesstock and Food
F
Suppply - Animaal Health Deepartment, 3Brazilian Ministry
M
of Agriculture,
A
, Livestock and
Foodd Supply - Animal
A
Health Departm
ment, 4Facullty of Veteriinary Mediccine, Univeersity
of Saao Paulo - Department
D
of Preventive Veterinaary Medicin
ne and Anim
mal Health
mic in Braziil and despitte the controol efforts, thhe
Boviine rabies iss still considdered endem
diseaase still spreeads in an innsidious waay and the main
m vector is the vamppire bat,
Desm
modus rotun
ndus. This project
p
aimeed to create a predictivee model to estimate
e
thee
probaability of bo
ovine rabiess outbreaks in each muunicipality of
o 21 out of 27 Braziliaan
Statees. The risk was estimaated using co
oncepts of rreceptivity and
a vulneraability.
Quesstionnaires were
w sent too the Local Veterinary Units of each State and
d covered a
numbber of questtions relatedd to the surv
veillance off possible rissks, such ass: bovine
outbrreaks, active roosts, baats positivityy and spatiaal changes. The
T bovine density andd
geom
morphologicc features were
w obtaineed from national registrries and geoographic
inforrmation systtems. The riisk results where
w
comppared with thhe 417 boviine outbreakks in
20100 distributedd throughouut the municcipalities. Ouut of 5016 municipaliti
m
ies accessedd,
217 ((4,3%) weree rated as haaving a highh risk for thhe virus spreead, 1277 (2
25,5%) as
mediium risk, 20045 (40,8%)) as low risk
k, and 544 (10,8%)
(
as negligible
n
riisk. In 933
(18,66%) cases thhe risk was unable to be determineed because the
t lack of information
i
n.
From
m 417 municcipalities prresenting heerbivores ouutbreaks in 2010,
2
183 (4
43,9%) werre
ratedd as high risk, 196 (47,00%) as meddium, 23 (5,5%) as low
w and in 15 (3,6%)
(
the risk
r
was nnot determiined. The reesults showeed that placees with outbbreaks weree skewed
towaards areas with
w higher risk
r for the virus
v
spreadd. In the futu
ure, these models
m
couldd
allow
w the targetiing of effortts, adoption
n of control measures directed
d
to certain locattions,
optim
mization of the control team’s trannsit and a beetter understtanding of rabies
r
spreaad.
Addiitionally, effforts need to be made to
t stimulatee the continuuous surveilllance of rissk
and rreduce areass with lack of informattion.
CO.772
TIMIZATIO
ON OF SPA
ATIAL AC
CCESSIBIL
LITY TO VACCINAT
V
TION POIINTS
OPT
OF 22009 RABIIES VACCIINATION CAMPAIG
GN IN SAO
O PAULO CITY,
C
BRA
AZIL
Dias RA1, Polo G1, Guillouux AGA1, Ferreira F1, Amaku
A
M1 - 1Universiddade de Sãoo
mento de Medicina
M
Veeterinária Prreventiva e Saúde Anim
mal
Pauloo - Departam
Paulo city has
h more thaan 2.5 million dogs andd 560 thousand cats. Th
hese populaations
São P
are ddistributed irrregularly thhrough the territory,
t
m
making
it diffficult to allo
ocate healthh
serviices. One off the most im
mportant heealth interveentions overr the animall populationn is
the vvaccination against rabiies, presumaably responnsible for thee ending off the rabies
epideemics in dogs and cats in 1983. Th
he aim of thhis study is to
t evaluate the accessibbility
to vaaccination points duringg the 2009 mass
m vaccinnation camppaign. To acchieve this, we
used a two-step floating cattchment areea (2SFCA) method im
mplemented in a geograpphic
inforrmation systtem environnment, undeer the hypotthesis of opttimal distribbution of
vacciination poin
nts to assuree the potential use of thhe service by 100% of the
t target
popuulation. Areaas with low
w accessibilitty were obsserved, espeecially in deense populatted
areass, for both dogs
d
and catts. An algorrithm to opttimize the vaccination point
p
locatioon
was ccreated in order
o
to impprove accesssibility, andd scenarios with
w lower number
n
of
vacciination poin
nts were creeated. This methodolog
m
gy can be ussed to offer vaccinationn
againnst rabies ev
venly to thee entire popuulation of annimals, espeecially afterr the halt off this
contrrol strategy from 2010 and 2011. The
T model considered
c
the same loocation of
vacciination poin
nts for dogs and cats, when
w
approppriate.
CO.773
D COLLAR
R CAMPAIIGN: HELP
PING TO CONTROL
C
L RABIES IN A
RED
HUM
MANE, EFFECTIVE AND SUSTAINABL
LE WAY.
Gebaara RR1, Gaamboa M2, Quintero
Q
RM
M3, Hiby E4, Russell E4 - 1WSPA-- World Socciety
for thhe Protectio
on of Animaals - WSPA
A Brasil, 2WSPA- World
d Society fo
or the Protection
3
of Annimals - WS
SPA Mesoaamérica, WSPAW
Worlld Society for
f the Proteection of
Anim
mals - WSPA
A Suramériica, 4WSPA
A- World Soociety for the Protection
n of Animalls WSP
PA Internatiional
e
55
5,000 peoplee die of rabies every yeear1, and 99
9% of these are caused by
An estimated
bites from rabid
d dogs.2 Thee global ecoonomic burdden of rabies is estimateed to be 4,0000M
D/year3. Masss dog culls are organizzed by goveernments in misguided attempts to
USD
contrrol the diseaase and limiit outbreakss however thhis approachh has repeattedly been
show
wn to be inefffective for rabies conttrol4. Thesee culls incluude gassing, shooting,
electtrocution, beeating, and poisoning, cause the neeedless death of millions of dogs each
e
year,, furthermorre such inhuumane meth
hods are conndemned internationallly5. Converrsely,
evideence shows that vaccinnating at least 70% of a dog populaation annually creates a
barrier of healthhy immune dogs
d
that sto
ops the diseease from sppreading.2
W
Rabiees Day 20111, WSPA laaunched the Red Collarr Campaign (RCC), whhich
On World
is a gglobal camppaign to endd the inhumaane culling of dogs in the
t fight against rabies. Our
goal is to convin
nce and assiist governm
ments to conttrol rabies in
i an effectiive, humanee, and
mane cullingg of dogs.
sustaainable way through maass vaccinattion withouut the inhum
(
is an example of best
b practicee in rabies control
c
as itt has been
Latinn America (LA)
comm
mitted to do
og vaccinatiion since thee 80´s. Usinng this apprroach, dog rabies
r
casess in
LA ddeclined fro
om 25,000 inn 1977 to 1996 in 2011, a decrease of 99% andd subsequenntly ,
humaan rabies also fell to onnly 15 in 20011, a decreaase of 96%..6 Thus, mass vaccinatiion
of doogs has becoome an esseential compo
onent in rabbies control by halting disease spreead,
alonggside pre an
nd post-expoosure prophhylaxis, survveillance annd informatiional campaaigns.
This commendaable work reeveals the im
mportance of
o creating political
p
willl and providdes
her regions to change their
t
policy and practicce.
inspiiration to oth
o
in the previouusly rabies-ffree island of
o Bali,
In 20008, a rabiess outbreak occurred
Indonnesia, leadin
ng to a widespread dog
g cull. In 20010, in respo
onse to this unnecessarry
killinng, WSPA worked
w
togeether with thhe Bali Aniimal Welfarre Association (BAWA
A)
and cconvinced Bali’s
B
goverrnment to sttop killing dogs
d
and im
mplement thee first islanddwidee mass dog vaccination
v
, reaching 70%
7
coveragge and savin
ng over 3000,000 dogs from
f
a neeedless death
h. During thhis vaccinatiion period, human
h
rabiees cases deccreased by 335%,
and ddog cases byy 76%.6 The Disease Innvestigationn Centre in Bali reportss that people in
Indonnesia now believe
b
in thhe power off vaccinationn and the syystems and procedures
p
developed durin
ng WSPA annd BAWA´s work in Bali
B are beinng adopted as
a national
guideelines.
011, WSPA supported the
t Bangladdeshi govern
nment to caarry out a piilot
In Noovember 20
dog vvaccinationn project in Cox’s
C
Bazarr, vaccinatinng more thaan 70% of th
he dog
popuulation in tw
wo weeks. WSPA
W
is now
w working with the Goovernment to
t develop a
natioonal action plan
p for elim
minating rabbies throughh mass dog vaccination
n.
PA’s RCC has
h continueed to gather global mom
mentum since these succcesses,
WSP
proviiding furtheer evidence for the conttribution off positive an
nimal welfarre to humann
healtth.
1. Knobel, D.L. et al. Re-evaluating the burdenn of rabies inn Africa andd Asia. Bullletin
3
(20005)
of the World Heealth Organiization 83, 360-368
pdated Sept 2010. Acceessed
2. World Heaalth Organizzation Rabiees Fact sheeet No.99, up
who.int/mediacentre/faactsheets/fs0099/en/
July 2012www.w
o the Globaal Burden of Canine Raabies: Humaan &
3. Hampson K et al. Reassessment of
mal Costs. Presented
P
byy S Cleavelaand to OIE Global Connference on Rabies Conntrol:
Anim
Towaards sustainnable prevenntion at souurce, 7-9 Sepptember 2011. Accesseed July 20122
www
w.oie.int/eng
g/A_RABIE
ES/presentaations_rage/S31%200Socioecon
nomicBurdeen_DrHamppson.pdf
4. WHO (200
05) Expert consultation
c
n on rabies: First
F
report. Technical Report Serries
Geneva: WH
HO
931 G
5. World Organisation foor Animal Health
H
(OIE), Terrestriaal Animal Health
H
Codee
chapter 7.7 Strayy dog contrrol. Accesseed July 20122
&L=0&htmffile=chapitrre_1.7.7.htm
m
http:///www.oie.iint/index.phhp?id=169&
6. Tamayo HS
S. Eliminatiion of humaan rabies traansmitted by
y dogs in thhe Americass:
P
n to OIE Gllobal Conferrence on Raabies Controol: Towardss
Achiievements. Presentation
sustaainable prev
vention at soource, 7-9 September
S
22011. Accesssed July 20012
www
w.oie.int/eng
g/A_RABIE
ES/presentaations_rage/S15%200CaseReport%20Amerrica_DrTam
mayo.pdf
7. World Sociiety for Prootection of Animals
A
(W
WSPA). Casee study- Rab
bies controll:
d disease prrevention, 2012.
2
Accesssed July 20
012 www.w
wspaDog welfare and
g/red-collar//case-studiees/default.asspx
internnational.org
PO
OST
TER
RES/// PO
OST
TERS
S
PT.0001
VITRO AN
ND IN VIVO
O INHIBIT
TION OF RABIES
R
VIRUS REP
PLICATION
IN V
BY R
RNA INTE
ERFERENC
CE.
Ono EAD1, Iam
mamoto K2, Castilho
C
JG
G2, Carnieli Jr
J P2, Oliveeira RN2, Acchkar SM2,
2
2
1 1
K
I , Braandão PE - Universiddade de São Paulo - Meedicina
Carriieri ML , Kotait
Preveentiva e Saú
úde Animall, 2Instituto Pasteur de São
S Paulo
mals and leaads to more than 55,0000
Rabies is a zoonnotic diseasee that affectts all mamm
r
virus (RABV) (M
Mononegaviirales:
humaan deaths evvery year3, caused by rabies
Rhabbdoviridae: Lyssavirus)). The searcch for antiviirals againstt rabies is on
ne of the
fronttiers in the field
f
but, deespite a prottocol (the Milwaukee
M
P
Protocol)
baased on ketaamin,
ribavvirin, midazzolam and am
mantadin was
w successfful after the treatment of
o a human
patieent,4 it was shown
s
as noot reproduciible. RNA interference
i
e is an altern
native as
antivviral technollogy against RABV alrready shownn as effectivve in vitro in
n cell culturres
1,2
, buut no reportts on its in vivo
v
use ex
xist hitherto.. The aim of
o this study
y was to asssess
the ddecrease in the
t titer of rabies
r
virus both in vitrro and in vivvo using short-interferiing
RNA
As. To this end,
e three siiRNAs weree used with antisense strands compplementary to
rabiees virus nucleoprotein (N)
( mRNA.. BHK-21 cells monolaayers were infected
i
witth
1,0000 to 0.1 TCIID50 of PV
V and after 2 hours the cells
c
were trransfected with
w each of tree
RNA
As in separatte using Lippofectaminee-2000™. All
A three siR
RNAs reduced the titer of
PV sstrain in a leeast 0.72 loggTCID50/m
ml and no cyytotoxic effeect was obseerved in thee
monoolayers treaated with Lippofectamine-2000™. Swiss
S
albino
o mice infeccted with 100.000
to 1L
LD of PV sttrain by the intracerebraal route werre also transsfected afterr two hours of
infecction with a pool 3 siRN
NAs with Lipofectamin
L
ne-2000™ by
b the intraccerebral rouute,
resullting in a suurvival rate of
o 30% in mice
m inoculaated with 1000 LD50, while
w
the sam
me
dose led to 100%
% mortality in untreated animals. Lipofectam
L
mine-2000™
™ showed noo
ontrol mice. These resuults suggestt that intraceerebral adm
ministration of
o
toxicc effect in co
siRN
NAs might be
b an effectiive antivirall strategy foor rabies.
The aauthors are grateful to FAPESP (F
Fundação dee Amparo à Pesquisa do
d Estado dee São
Pauloo), Grant # 2008/515199-8, and Passteur Instituute of São Paulo for thee financial
support.
ERENCES
REFE
1.Braandão PE ett al. Short-innterfering RNAs
R
as anttivirals agaiinst rabies. BJID, v. 111, p.
224, 2007.
m
artifficial
2. Isrrasena N et al. Inhibitioon of rabiess virus repliccation by multiple
microoRNAs. A Res; v. 84, n. 1, p. 76, 2009.
WHO Expertt consultatioon on rabiess, p.2, 2005.
3. W
Willoughby Jr
J et al. Surrvival after treatment
t
off rabies withh induction of coma. New
N
4. W
Englland Journall of Medicinne, v. 352. n.
n 24, p. 25008, 2005
PT.0002
RESSÕES DE
D Desmod
dus rotund
dus (E. Geooffroy) EM ANIMAIS
S
AGR
DOM
MÉSTICOS
S PRÓXIM
MOS A ÁRE
EA URBAN
NA DA ILH
HA DE SÃ
ÃO
LUÍS
S/MARAN
NHÃO/BRA
ASIL
Arruuda RCN1,2,33,4 - 1MAPA
A - SFA-MA
A, 2Santos EFS
E - Coorddenação MA
APA, 3Falcãão
4
MA, - AGED-M
MA, Gualteer PP - AGE
ED-MA
umo
Resu
Agreessões de Deesmodus rotundus em animais
a
sãoo acontecim
mentos comu
uns no
Maraanhão, comoo no Brasil,, com a difeerença que nesse
n
Estado
o ainda há agressões
a
em
m
humaanos e houv
ve transmisssão do víruss da raiva, ccomo ocorrido em 24 pessoas no ano
a
de 20005. O objeetivo do pressente estudoo foi chamaar a atenção à presença de D. rotunndus
próxiimo a humaanos em áreea urbana dee São Luís/M
MA, mostraar opção de monitorameento
de árrea de forrag
geamento attravés anilhhamento de morcegos, como tambéém, identifiicar
caraccterísticas im
mportantes nos indivíd
duos para avvaliações e reavaliações
r
s. Executouu-se a
armaação de redees em currall, caracterizzado o sentiddo Leste-Oeeste em relaação nascim
mento
e pôrr-do-sol num
m croqui o que
q facilitarria no direciionamento de
d possíveiss abrigos doos D.
rotunndus. Dos morcegos
m
caapturados, mediu-se
m
em
m milímetross o antebraçço direito coom
paquuímetro de plástico,
p
o peso em gram
ma foi deterrminado por um dinam
mômetro (1000g).
As annilhas de 4m
mm de diâm
metro numerradas foram
m colocadas no antebraçço do morceego
menssurado com
m objetivo dee novos monnitoramentoos. Observoou-se a ossifficação das
epífises das falaanges das assas, que nos indivíduos jovens são mais maleááveis, e nos
m
calcifficados. O estado
e
repro
odutivo foi avaliado
a
de
adulttos os ossoss estavam melhor
form
ma visual e leeve palpaçãão, nas fêmeeas com um
m volume maaior no abdô
ômen e nos
machhos quando os testículoos estavam na
n bolsa esccrotal. No controle dos espécimes,,
efetuuou-se a aplicação da pasta vampirricida (warffarina a 2%)) no dorso dos
d morcegoos,
por vvolta das 244 h quando havia
h
se enccerrado as capturas
c
e reecolhidas todas rede em
m
voltaa do curral. Foram
F
captturados 5 indivíduos, ssendo todos da família Phyllostom
midae,
mas de espécies diferentes, um frugívooro (Artibeuus), três hem
matófagos (D
Desmodus) e um
ossophaga),, este últimoo não foi coolocado anell, já que nãoo era já
nectaarívoro (Glo
especcífico para o seu tamannho diminutto. Observou-se que os D. rotunduus foram peggos
depoois da 21:17h
h, indicandoo que onde há a presennça humana a alimentaçção dos
indivvíduos podee acontecer mais
m tardiam
mente, deviido ao ambiente mais calmo. As
cheggadas ou enttradas dos morcegos
m
heematófagos aconteceram
m pelo Nortte, essas ‘rootas
de vôôo dos vamppiros’ indiccam possiveelmente as loocalizações de abrigos. A presença de
uma fêmea de D.
D rotundus prenha e um
m macho esscrotado inddicava que a colônias esstão
em pplena atividaade sexual. Com o pressente trabalhho foi possíível concluirr que a pressença
de D
D. rotundus próximo
p
a humanos
h
é um
u risco quue deve ser mitigado.
m
O anilhamennto é
uma forma de gaarantir o moonitoramentto de morceegos em várrios estágioss da vida, seendo
mportante anotar
a
todoss os dados em
e fichas appropriadas para
p saber a
extreemamente im
evoluução da eficciência do trratamento efetuado,
e
beem como o croqui
c
da árrea, para
conhhecer possívveis abrigos através da captura
c
na fonte
f
de alim
mentação, e os dados
biolóógicos dos morcegos,
m
p identificar e/ou acoompanhar in
para
ndivíduos, reprodução,
r
,
testes laboratoriais, e observar a evoluçção de anticcorpos da raaiva, de tem
mpo em temppo,
para melhor avaaliar riscos.
Palaavras-chaves: anilhas, encefalite, raiva, herbíívoros, mitiigação de risco e
quiróópteros.
___________________________________
1
Fisccal Federal Agropecuáário do Miniistério da Agricultura, Pecuária
P
e Abastecime
A
ento
(MA
APA)
2
Fisccal Estaduall Agropecuáário da Ageencia Estaduual de Defessa Agropecu
uária do Esttado
do M
Maranhão (A
AGED-MA /BRASIL).
PT.0003
CO SITUAC
CIONAL DA
D RAIVA
A ANIMAL
L NO ESTA
ADO DO RIO
R
DIAGNÓSTIC
DE JJANEIRO
Meneeguete PS1 - 1Secretaria de Estadoo de Saúde do
d Rio de Jaaneiro - Coo
ordenação de
d
Vigillância Amb
biental em Saúde
S
e Saúdde do Trabaalhador
o de Janeiroo está localizzado na Reggião Sudestte, com umaa populaçãoo de
O Esstado do Rio
15.8556.915 habiitantes, seguundo estimaativa do IBG
GE, com um
ma densidadde demográffica
de 433.696,054 KM².
K
Possuui 92 municcípios distriibuídos em 9 regiões (M
Metropolitanna I, Metroppolitana II,
Baixxada Litorân
nea, Médio Paraiba,
P
Centro Sul, Seerrana, Baíaa de Ilha Graande, Nortee e
Norooeste Flumin
nense), limiitando-se aoo Norte com
m o Estado de
d Minas Geerais, ao Suul São
Pauloo e a Noroeeste com o Espírito
E
Sannto.
Em
m nosso Esttado o Progrrama Estaduual de Preveenção da Raaiva iniciou
u a campanhha de
vacinnação animaal no ano dee 1983, o qu
ue determinnou à diminu
uição signifficativa dos
casoss de raiva em
m cães e gaatos e como conseqüênccia a reduçãão de casos em humanoos.
Apóss 21 anos seem ocorrênccia de caso de
d raiva hum
mana no Esstado do Rioo de Janeiroo, foi
confi
firmado um caso em dezembro de 2006
2
no muunicípio de São José doo Vale do Rio
R
Pretoo devido a acidente
a
com
m morcego.
As açções de con
ntrole repressentadas pella coberturaa vacinal, attingindo índ
dices maiorees do
que 990% da pop
pulação estim
mada canina e felina enntre 1990 e 2009 contriibuíram parra a
obtennção de resu
ultados satissfatórios. No
N ano de 2010, houve uma estimaativa de vaccinar
aproxximadamennte 2.500.0000 animais. Porém,
P
apóós a ocorrênccia de eventtos adversos nos
estaddos de São Paulo
P
e Rio de Janeiro,, o Ministério da Saúdee, baseado nos
n resultado
encontrado nas avaliações
a
l
laboratoriai
s de algumaas a partidass e dos estuudos
epideemiológicoss efetuados,, decidiu pela suspensãão definitivaa do seu usoo na rede púública
de Saaúde, ficand
do o Estado sem coberttura vacinall desde entãão.
Nos últim
mos anos a ocorrência de raiva no Estado do RJ está resttrita a animaais da
mos ocorrênccia de raiva em cães e gatos
g
área rural: bovinnos, eqüinoss e morcegoos. Não tem
c
e gatoss no
desdee 2002, indiicativos quee as campannhas de vaciinação antirrrábica em cães
Estaddo do Rio de
d Janeiro têêm atingindo o objetivoo proposto.
No Estaddo do Rio e Janeiro, enntre os anos de 2007 e 2010
2
foram confirmadoos
e morcegoos não
170 ccasos de raiiva positivos em bovinoos, 40 em eqüinos, 14 em
hemaatófagos e 03
0 em morcegos hemattófagos e 044 outros.
Dos 98 casos co
onfirmados no
n Estado em
e 2011, 133 % ocorreraam no muniicípio do Rio
R de
P
resp
pectivamentte).
Janeiiro e 12% em Valença (Região Meetropolitanaa e Médio Paraíba,
Os quue casos po
ositivos vêm
m aumentand
do ano a ano, notadameente em aniimais que nãão
bovinnos, e que apenas
a
uma pequena paarcela é captada pelos serviços
s
de vigilância.
PT.0004
MPLETE GENOME
G
S
SEQUENC
CE ANALY
YSIS OF FIIVE RUSSIIAN RABIES
COM
VIRU
US FIELD
D ISOLATE
ES
Chuppin S1, Cherrnyshova E1, Metlin A1 - 1Federal Centre for Animal Heaalth
g
of Russian rab
bies virus fieeld isolates have been sequenced.
Five complete genomes
w collecteed from diff
fferent regioons of Europ
pean part off Russia. Thhe
Thesse isolates were
genoome lengths were 119233 nt (3 isolaates), 119277 nt (1 isolatte) and 11914 nt (1 isolate).
All ggenomes rettained basic genetic feaatures charaacteristic forr genotype 1 lyssavirusses.
Fourr isolates weere geneticaally close to European lineage
l
of raabies virus and shared 93,495,1%
% nucleotid
de identity with
w strain 9147FRA
9
frrom France comparing complete
genoomes. One issolate from reindeer was geneticallly close to Arctic lineaage of rabiees
viruss and shared
d 99,7% nuccleotide iden
ntity with sttrain A09066 from Alasska compariing
gene N sequences. This is the
t first sequ
uenced com
mplete genom
me of Arctiic lineage raabies
k
th
his is the firrst proven case
c
of Arcttic lineage rabies
r
viruss. Furthermoore to our knowledge
viruss distribution in Europee.
PT.0005
NESS OF PA
ARENTER
RAL VACC
CINATION
N OF STRA
AY DOGS
EFFECTIVEN
AINST RAB
BIES TO REDUCE
R
T
THE
INCID
DENCE OF
F HUMAN
N RABIES
AGA
DEA
ATHS DUE
E TO STRA
AY DOG BIITES.
Jayassundara VK
K1 - 1Dehiwaala Mt. Lavvinia Municiipal councill Sri Lanka - Health
Rabies is one off the notifiabble zoonosees in Sri Lannka. 96% off human rabbies deaths occur
o
r
annd the transm
mitter of thee disease in the
due tto dog bites. The dog iss the main reservoir
comm
munity.In Sri
S Lanka, vaccination as
a a prophylactic measuure to contrrol canine raabies
was ppracticed on
nly for dom
mestic dogs till
t 2006. Ellimination was
w the main control
meassure for straay dogs duriing this periiod.The new
w strategy of
o mass pareenteral
vacciination of sttray dogs inn addition to
o the domesstic dogs, innstead of stray dog
elimiination, has been impleemented by the health aauthorities of
o Sri Lankaa after 20077.
Aparrt from that,, chemical and
a surgicall birth contrrol methods also were introduced
i
t
to
contrrol the strayy dog populaation densitty since 20007. An Autooplunger, a newly
n
invennted
instruument in Srri lanka wass utilized forr intramuscular vaccinaation of straay dogs.
m objecttive of this study
s
was too find out thhe effectiveeness of masss vaccinatiion of
The main
strayy dogs again
nst rabies ass a prophylaactic measurre to reducee the incidennce of humaan
rabiees deaths duue to stray dogs
d
in Sri Lanka.
L
d
of epidemiology and the
t
The ddata obtaineed from quaarterly bulleetins of the department
depaartment of thhe public heealth and veeterinary serrvices of thee ministry of
o health
betw
ween 2007 annd 2010 weere analyzed
d.
man deaths due to strayy dog bites has
h increaseed
The ddata revealeed the incidence of hum
from
m 16 in 2001 to 28 in 20006 whereass the incidennce has deccreased from
m 21 in 20077 to
18 inn 2010.The total
t
numbeer of femalee stray dogs subjected to
t chemical birth controol
weree 90380,853339,53931 and
a 39888 inn 2007, 20008, 2009 andd 2010 respectively. Tootal
numbber of femaale stray doggs surgicallyy sterilized were 1848, 119546, 2220280 and
907664 for these correspondding years. It
I has been reported
r
thaat Some proovinces havee
not ccarred out proper
p
vacciination and population control measures on sttray dogs duue to
varioous issues inn 2010 comp
mpaired to 20
009.
The sstatistical annalysis reveealed a weakk negative correlation
c
(r=
( 0.27,
R²=00.072) betw
ween the maass vaccinattion of strayy dogs againnst Rabies and
a the
inciddence of hum
man rabies deaths
d
due to
t stray dogg bites.
wever, the mass
m vaccinaation of stray
y dogs agaiinst rabies helps
h
to estaablish herd
How
immuunity in straay dog reserrvoir and itss feedback eeffect helps to arrest thhe transmisssion
chainn of virus am
mong stray dogs. Thereefore in connclusion, maass vaccinattion of strayy
dogs makes the public less susceptiblee for rabies ddue to strayy dogs.
v
of stray dogs and popuulation conttrol
Furthher strengthhening of annnual mass vaccination
meassures to a grreater extennt in a methoodical way aat provinciaal level in fuuture is
recom
mmended.
n
vacccination proogramme is funded by Health Minnistry of sri
This on-going national
y was not fuunded.
lankaa. My study
man Rabiess, Mass vacccination
Key words: Straay Dog, Hum
PT.0006
NTROLE DE
D Desmod
dus rotundu
us E EDUC
CAÇÃO SA
ANITÁRIA
A EM
CON
PEQ
QUENAS COMUNIDA
C
ADES DA BAIXADA
A MARANH
HENSE – BRASIL
B
Santaana SS1 - 1Agência
A
Esttadual de Deefesa Agroppecuária do Maranhão--AGED-MA
AProggrama Estaddual de Conttrole da Raiiva dos Herbbívoros e Outras
O
Enceffalopatias
e pessoas e
Este trabalho tevve o objetivvo de traçar o perfil doss ataques dee vampiros em
B
Maaranhense (rregião alagaada), controlar a populaação de
semooventes na Baixada
Desm
modus rotun
ndus, bem como,
c
mostrrar que a edducação saniitária é umaa possibilidaade
de traansformaçãão de compoortamento dos
d indivíduuos avaliado
os. Na regiãoo estudada, as
áreass mais altas onde os aniimais se alim
mentam durrante o perííodo das cheeias, são
denoominadas peelos popularres de ‘Teso
o’, no períoddo das secass os servidoores da Agênncia
Estaddual de Deffesa Agropeecuária do Maranhão
M
(A
AGED-MA)), atenderam
m demanda de
espolliações em animais e humanos
h
em
m dois povoaados a 2 km
m entre si. No
N total foram
m
visitaadas 48 casaas, sendo 35
3 em Fradees e 13 no povoado
p
Teso da Taperra. Há histórico
de abbertura de novas
n
áreas dentro
d
de reemanencenttes de matass que estariaam próximaas aos
povooados em estudo. Nas comunidade
c
es agredidas foram realiizadas palesstras sobre
Raivaa Humana e Animal e Bioecologia
B
a de Morceggos Hematóófagos para 86 pessoas em 3
ocasiiões distintaas. Realizouu-se um queestionário paara avaliar o sistema dee criação dee
anim
mais e mordeeduras do Desmodus
D
rootundus. Pella análise dos resultado
os, observoou-se
que 113 humanoss foram agreedidos, denttro de uma população de
d 198 pesssoas,
uma criança (0,,50%) e 12 outras adulttas (6,06%), perfazenddo um total de
d 7% da
mpre proteggidas com mosquiteiro
m
s. Da
popuulação exposta. As criannças dormeem quase sem
locallização das espoliaçõess nos humannos, 92,30%
% foram nos pés, indicaando que as
pessooas que dorm
mem em redes , ficam com as extrremidades inferiores
i
esstão mais
vulneeráveis a ataaque de vam
mpiros e 81%
% das pessooas entrevisstadas não tiinham
esclaarecimentos necessários a respeito da raiva. A maior freqquência de mordeduras
m
em
pessooas aconteceram num período dee 4 meses (992,30%), enntre os mesees de agostoo e
outubbro e o restaante a no máximo
m
1 anno. Indicanddo que as ag
gressões poddem ter ocorrrido
em fu
função do deesmatamentto de áreas próximas
p
aoos povoadoss e no períoodo do
desloocamento dee animais para outras áreas.
á
Todass as pessoass agredidas, só procuram
mo
serviiço de saúdee para tratam
mento preveentivo após a primeira palestra, deemonstrandoo a
impoortância da educação
e
saanitária em áreas de risco para raivva. Pela ausência de casos
clíniccos, observaa-se que a população
p
d morcegoss hematófaggos não estaava enfermaa com
de
víruss rábico. Daas pessoas ennvolvidas em
e entrevistta, (20,87%)) delas desccreveram coomo
abriggos, ocos dee árvore e buueiros na Feerrovia Caraajás. Verificcou-se que dentre
d
47
criaddores, 51,6%
% destes deiixavam os seus
s
animaiss soltos no período,
p
e os
o que ficavam
próxiimos as resiidências serriam um atrativo aos morcegos.
m
No período annalisado,
obserrvou-se quee apesar de haver
h
animaais nas redoondezas, os D. rotunduss, tinham
prefeerências porr alimentar-se de sangu
ue humano. Com base nas
n elucidaçções de cam
mpo,
sistem
ma de criaçção e caracteerísticas da área, concluui-se que háá muitos abrrigos ao lonngo
da Feerrovia Caraajás que corrta os municcípios do Esstado e poucos nos cam
mpos alagaddos;
as moradias messmo sendo de
d alvenariaa, não possuuem forros ou outro tippo de proteçção
impeedindo a enttrada dos heematófagos;; no regime de criação dos animaiss na baixadaa, na
estaçção chuvosaa os médios e/ou grandee animais estão próxim
mo às residên
ncias e no
períoodo mais secco são criaddos soltos, os
o morcegoss não acomppanham esssa
moviimentação; o controle de
d morcegoos hematófaggos e a educcação sanitáária são meddidas
de fáácil visualização e convvencimento em comuniidades; facillmente o hoomem pode se
tornaar uma fontee de alimentação alternnativa em árreas onde hoouve alteraçções de
ambiiente, movim
mentação ouu morte de animais e não
n há circulação viral na
n área já que
q o
resulltado do diaagnóstico labboratorial foi
fo negativo para raiva e não houv
ve casos da
enferrmidade.
Palaavras-Chave: Raiva, Edducação San
nitária, Dessmodus rotuundus e Baix
xada
Maraanhense.
___________________________________
1
Agêência Estaduual de Defessa Agropecu
uária do Maaranhão - AGED/MA.S
A
São LuísMA//Brasil
2
Minnistério da Agricultura,
A
Pecuária e Abastecimeento.São Lu
uís-MA/Braasil
PT.0007
YLOGENETIC RELA
ATION AM
MONG BOV
VINE RAB
BIES CASE
ES FROM
PHY
MIN
NAS GERA
AIS AND SA
AO PAULO
O STATES
S, BRAZIL
L: A POSSIBLE
COM
MMON OR
RIGIN
Garccia AIE1, Sillva SOS1, Peixoto
P
H1, Brandão
B
PE
E1, Cunha EM
E 2, Richtzeenhain L1 1
2
FMV
VZ USP - Veterinary
V
P
Preventive
M
Medicine
annd Animal Health,
H
Instituto Bioloogico
de Sãão Paulo - Laboratory
L
o Rabies annd Viral Enncephalitis
of
b hematophhagous bat Desmodus
D
r
rotundus
haas representted a
Boviine rabies trransmitted by
publiic health ch
hallenge andd an econom
mical burdenn to livestocck industry of Brazil sinnce
19111, when the first rabies outbreak inn this speciees was officiially docum
mented in thee
southh of the cou
untry. Severral studies revealed
r
tem
mporal and spatial
s
patteerns of endeemic
preseentation alteernating witth epidemicc periods. Laast registereed epizooticcs occurred
betw
ween 1997 annd 2002, in Sao Paulo affecting boovines and equines
e
neaar the borderr of
Minaas Gerais. The
T objectivve of the preesent researcch is to perfform a phylogenetic
analyysis of rabiees cases in livestock
l
fro
om Minas Gerais
G
muniicipalities neext to Sao
Pauloo´s epizootiic area in thhe subsequennt period (fr
from 2000 to
o 2009) andd try to assoociate
moleecular data to
t geographhical and eco
ological facctors. 32 parrtial DNA sequences were
w
obtaiined corresp
ponding to rabies
r
exterrnal glycoprrotein (540 nucleotides
n
of length). To
inferr genetic relations amonng virus iso
olates were performed
p
analysis
a
app
plying NeighhborJoiniing algorithm
m and Kimuura 2 param
meter evoluttion model with
w 1000 bootstrap
b
repettitions. The proposed phylogeny
p
p
presented
heere is in agreeement withh a previouss
workk (Carnieli, virus researrch,v 144, p 215, 2009)) which prompts the prresence of thhree
mainn sub lineages in the Saao Paulo´s epizootic
e
areea, near to the frontier with
w Minas
Geraais: “Old Strrain”, RD1 and RD2/R
RD3. The olld Strain is composed
c
b isolates
by
previious to 1998
8. The strainn RD1 is inntegrated byy viruses isolated after 1998
1
and haas a
different geneticc origin. Linneage RD2//RD3 has suubtle geneticc differencee when
D1 even thouugh they em
merged in thhe same period of time and also haave
comppared to RD
an ovverlapping pattern
p
of geographic distribution.
d
. All samplees from Minnas Gerais
analyysed in this research weere classifieed as RD2/R
RD3.
This genetic sub
b-lineage is located alonng the bordder between the two staates, mainly over
g
sub-lineage is
the loowest portioons of Serraa da Mantiqqueira mounntain. This genetic
slighhtly divergen
nt from rabiies isolates from the rest of the couuntry and iss possible too
differentiate it among
a
rabiees viruses from inner municipalitie
m
es of São Paaulo (far froom
o
isolatees involvedd in previouss epidemic waves
w
as thhe old
epizoootic area), even from older
and R
RD1 Strain.. Molecularr analysis suuggests a poossible invollvement of Artibeus
A
lituraatus in the rabies
r
cyclee from innerr municipaliities of São Paulo state. Geographiic
analyysis showedd that the caases were mainly
m
distribbuted in thee Mata Altlaantica biomee and
alongg the rivers:: Pardo, Moogi-Guaçu, Piracicaba/J
P
Jaguarí and Grande. Molecular
M
analyysis suggestts that rabies virus circuulating in bovines from
m south of Minas
M
Geraais betweenn 2000 to 20009 is genetically indisttinguishablee from one of the genettic
strainns involvedd in the last epizootics
e
r
registered
inn Sao Paulo from 1997 to 2002, caalled
here as RD2/RD
D3.
ES, FAPESP
P process nuumber 20111/06236-0
Acknnowledgements: CAPE
PT.0008
ANCE DE LA
L RABIA
A BOVINA EN EL ES
STADO SA
AN LUIS POTOSI
P
AVA
(ME
EXICO)
Cam
macho J1 - 1SERVICIOS
S
S DE SALU
UD DE SAN
N LUIS POT
TOSI - ZOO
ONOSIS
nzaron en el
e año de 1992, hasta 1999 cuando
o aun no exiistía en el país
p la
Los ccasos comen
caraccterización antigénica.
a
Se analizarron 30 muesstras resultaando 17 possitivas con 11
muniicipios afectados, 8 ceercanos a laa capital del estado inclluyendo La misma y soolo 3
fueroon de la zon
na huasteca, se infiere que
q los 8 caasos de los municipios
m
cercanos
c
a la
l
capittal del edo. Fueron causados por caninos ya que
q se presen
ntaban en promedio
p
400
casoss anuales enn perros . essto reforzado por que ya
y no se pressentaron máás casos al
contrrolarse la raabia canina en
e 1996.
Los 3 de la zonna huasteca se sospechaa que ya inniciaba el prroblema de los
l casos
M
Vampiro.
originados por Murciélago
000-2011 see procesaron
n 1039 mueestras con 6225 positivass ya
En ell periodo 20
caraccterizándosee 471 de ellas.
inicioo un brote en
e la zona huasteca:
h
dee 3 municipios afectado
os se increm
mento en
prom
medio anual 2.6 los muunicipios afeectados, paara hacer un total en el estado
e
de
41 ddesde 1992.
obserrvándose ell avance connstante de lo
os casos haccia el centroo del estadoo y muy
probaablemente a los estados vecinos, por
p falta de medidas prreventivas como la
vacuunación al gaanado, observándose ya afectacioones a otross animales de
d interés
Econnómico com
mo Equinos, Caprinos, ovinos y a fauna Silveestre como mustélidos.
m
Existtiendo la lattencia de unn probable ataque
a
y trannsmisión a los seres huumanos.
La prrincipal varriante antigéénica que see ha encontrrado es la V11
V en meno
or proporcióón es
la V33 y V5.
PT.0009
M
DE NORM
MAS TÉCN
NICAS DO TRATAM
MENTO
ESTUDO DO MANUAL
OFILÁTICO
O ANTIRR
RÁBICO HUMANO
H
P
PARA
PRO
OFISSION
NAIS DA 155ª
PRO
CRE
ES – CRAT
TEÚS/CE, 2010.
2
Dennnis DFCM1 - 115ª Coorrdenadoria Regional
R
dee Saúde Craateús
A raiva é uma doença inffecciosa aguda, cara
acterizada por um qu
uadro
p
óbito em poucoss dias. Aprresenta do
ois ciclos
neurrológico, que evolui para
básicos de tran
nsmissão: o urbano, cujos principais rese
ervatórios são
s os cãe
es e
os, e o silve
estre, que ocorre prin
ncipalmentte entre mo
orcegos, macacos
m
e
gato
rapo
osas, com destaque
d
p
para
o sag
gui no Esta
ado do Cea
ará. Essa doença
d
é de
d
extre
ema imporrtância para
a saúde pú
ública, devvido a sua letalidade de
apro
oximadame
ente 100%. O diagnó
óstico laborratorial da raiva é de
e fundamen
ntal
impo
ortância pa
ara o tratam
mento proffilático hum
mano pós-e
exposição, mediante a
apliccação de im
munobiológ
gicos espe
ecíficos, e para a ado
oção de me
edidas que
e
visam
m o contro
ole da doen
nça. O obje
etivo deste
e trabalho foi
f realizarr estudo do
o
man
nual de norrmas técniccas do trattamento prrofilático an
ntirrábico humano
h
co
om
profiissionais de saúde no
o universo
o dos 11 municípios que
q compõ
õem a 15ª
CRE
ES - Crateú
ús. A partirr desta açã
ão foi realizzado estud
do quantita
ativo descritivo,
base
eado no Arrt.6º, inciso
o III da lei Nº
N 8.080/9
90, que incllui no campo de atua
ação
do S
SUS a orde
enação da formação de recurso
os humano
os na área de saúde..
Dura
ante os me
eses de outubro e no
ovembro do
o ano de 2010, foram
m realizada
as 05
oficin
nas, sendo
o dois méd
dicos veterinários do NUVET/SESA e um da 15ª CR
RES
Cratteús os faccilitadores, e foram ca
apacitadoss 42 enferm
meiros,12 médicos,
m
0
05
agen
ntes de imunização, 03 coordenadores de
e vigilância
a sanitária, 08
veterinários e 09
0 agentess de contro
ole de endemias. Tra
atar pessoa
as agredidas
animais é atividade
a
r
rotineira
da
as equipess de Atençã
ão Básica / Estratégia
por a
Saúd
de da Fam
mília e da Vigilância
V
e Saúde que
em
q devem
m ser dese
envolvidas
visan
ndo tornarr viáveis oss princípioss e diretrize
es de acessso universsal e da
integ
gralidade do
d cuidado
o conforme a necessidade de saúde utilizando a
epidemiologia para o esttabelecime
ento de prio
oridades. Concluímo
C
os que a
ormações relativas
r
ao
o processo
o saúde-do
oença, as doenças
d
e
difussão de info
suass formas de
e prevençã
ão e contro
ole proporccionam um
m melhor co
onhecimen
nto
sobrre os fatore
es condicio
onantes e a situação de saúde. Capacitar profission
nais
que trabalham em diferentes setore
es da saúd
de favorece a definiç
ção de
o e na man
nutenção das
d ações de controle de agravvos a
priorridades, a elaboração
sere
em desenvo
olvidas de acordo co
om as suass especificiidades.
PT.0010
DEMIOLÓ
ÓGICO NA
A PROFILA
AXIA DA RAIVA
R
NA
A 15ª CRES
S–
ESTUDO EPID
ATEÚS/CE
E, NOS ANO
OS DE 200
09, 2010 e 2011.
2
CRA
Dennnis DFCM1 - 115ª Coorrdenadoria Regional
R
dee Saúde Craateús
As m
mordedurass causadas por anim
mais são motivo de grrande preo
ocupação
devid
do a possibilidade de
e transmissão de zoo
onoses, prrincipalmen
nte raiva. Este
E
estudo descritiivo quantita
ativo foi re
ealizado na
a 15ª CRES
S – Coorde
enadoria
aúde – Cra
ateús, form
mada por onze
o
munic
cípios: Ararrendá,
Regional de Sa
pendência, Ipaporang
ga, Ipueira
as, Monsen
nhor Tabos
sa, Nova
Cratteús, Indep
Russsas, Novo Oriente, Poranga,
P
Quiterianóp
Q
polis e Tam
mboril. Os dados
d
foram
colettados atravvés da fich
ha mensal de profilaxxia da raiva
a , enviada
a pelos
mun
nicípios a CRES
C
até o quinto dia
a útil de ca
ada mês co
om dados referentess ao
mês anterior. No
N ano de 2009, fora
am atendid
das 577 pessoas, dass quais 87,,9%
eberam trattamento, no
n ano de 2010,
2
foram
m 587 pessoas atend
didas com
rece
89,6
6% de trata
amentos, já
á em 2011, foram 812 atendime
entos com 93,3% de
e
trata
amentos. Quanto
Q
as mordedura
m
as, no ano de 2009, os
o cães foram
resp
ponsáveis por
p 69,2% do total e os gatos por
p 19,2%, no ano de
e 2010, cãe
es
foram
m 71% e os
o gatos 19
9,2%,em 2011 os cãe
es foram 72,4%
7
e ga
atos 21,4%
%,
send
do que as demais
d
mo
ordeduras foram causadas por primatas, morcegoss,
rapo
osas, bovin
nos e equin
nos. A quantidade de
e cães e ga
atos obserrvados pela
a
quan
ntidade destes anima
ais agresso
ores foi de
e 40,2%, 44
4,5% e 46,,3%,
resp
pectivamen
nte nos ano
os estudad
dos. O núm
mero de ate
endimentos nos anoss é
basttante eleva
ada e vem apresentando aumento. Os da
ados també
ém nos
reve
elam que os cães são
o os princip
pais agresssores, seg
guido peloss gatos, com
quan
ntidade de animais agressores
a
observado
os ascendente. O fatto de anim
mais
dom
mésticos serem os agressores mais
m
signifiicativos nã
ão justifica o alto
perccentual de tratamento
os anti-rábico human
no, mostran
ndo a nece
essidade de
capa
acitação de
e médicos e enferme
eiros em attendimento
o profilático
o, bem com
mo
uma
a maior inte
egração en
ntre ESF e os profisssionais de controle
c
de
e zoonosess na
indiccação de trratamento..
PT.0011
TITU
ULAÇÃO DE
D ANTICO
ORPOS NEUTRALIZ
ZANTES DE
D VÍRUS DA RAIVA
A
DOS
S DOCENT
TES DO CURSO DE MEDICINA VETERIINÁRIA DA
A
UNIV
VERSIDAD
DE FEDER
RAL DE GOIÁS (UFG
G), CAMPUS JATAÍÍ.
Meirrelles-Bartoli RB1, Cruzz CA1, Sousa DB1, Asssis LN2, Coosta KD3, Crruz EG3,
Rezeende Júnior SA4, Barceelos AA5 - 1Docente
D
da Universidaade Federal de Goiás /
Cam
mpus Jataí - Laboratório
L
o de Sanidadde Animal, 2Aluna do curso
c
de Meedicina
Veterinária da Universidad
U
e Federal dee Goiás/Cam
mpus Jataí, 3Aluna do curso de
Campus Jataaí, 4Técnico do Laborattório
Enfermagem daa Universidaade Federal de Goiás/C
Análise Clíniica Veterináária da Univversidade Feederal de Goiás/Campu
G
us Jataí,
de A
5
Técnnico de Enffermagem da
d Universiddade Federaal de Goiás. Campus Jaataí
A pro
ofilaxia da raiva hum
mana pode ser feita pré
p ou pós--exposição
o ao vírus. A
profiilaxia pré-e
exposição, realizada com vacin
nas, é indic
cada para as
a pessoas
que, devido à atividade
a
p
profissiona
al, correm o risco de exposição
e
ao vírus,
uisadores, professore
es e alunos que trabalham com
m
como veterinárrios, pesqu
mais potenc
cialmente infectados com o vírus da raiva
a. A profila
axia pósanim
expo
osição é indicada parra as pessoas que accidentalme
ente se exp
puseram ao
a
víruss; combina
a a limpeza
a da lesão e a administração da
a vacina, is
soladamen
nte
ou e
em associa
ação com o soro ou a imunoglo
obulina hum
mana anti-rrábica. O
objetivo do trabalho foi avaliar
a
o pe
erfil sorológ
gico de antticorpos ne
eutralizante
es
M
V
Veterinária
da
de vvírus da raiva dos doccentes do curso de Medicina
Univversidade Federal
F
de
e Goiás, Ca
ampus Jata
aí. Foram coletados 5 mL de
sang
gue de 17 professore
es e encam
minhado pa
ara o Institu
uto Pasteu
ur 2 mL de soro
cong
gelado de cada
c
amosstra em microtubos acondicion
a
ados em caixa
c
de iso
opor
com gelo recicclável manttendo uma
a temperatu
ura entre 4º
4 a 8º grau
us. Todas as
ostras estavvam devidamente ide
entificadass e acompa
anhadas de ficha de
amo
requ
uisição. O Microteste
M
Simplificado de Inibição de Flu
uorescência foi utiliza
ado
pelo Instituto Pasteur
P
para a titulaçção dos anticorpos. Das
D 17 amo
ostras, 07
2%) aprese
entaram tittulação sattisfatória (≥
≥ 0,5 UI/mL) e 10 (58
8,8%)
(41,2
insattisfatória (˂
˂ 0,5 UI/mL). Por me
eio da ficha
a epidemio
ológica fora
am observa
adas
algumas caraccterísticas em
e relação
o à profilaxxia da raiva
a recebida
a por estes
essores. Dos
D 17 doccentes, 12 (70,6%)
(
re
eceberam profilaxia
p
p
pré-exposiç
ção
profe
dura
ante o curs
so de Mediccina Veterinária e 05
5 (29,4%) já haviam recebido
r
profiilaxia pós-e
exposição antes da faculdade.
f
Dos 12 prrofessores que
rece
eberam a profilaxia
p
prré-exposiç
ção, 01 (8,3
3%) relatou
u realizar exames
e
de
e
soro
ológicos an
nualmente, 04 (33,4%
%) confirma
aram ter re
ecebido do
ose reforço sem
a rea
alização de
e sorologia
a prévia e os
o outros 07
0 (58,3%)) disseram
m nunca terr
rece
ebido uma dose reforrço e nem ter
t realizad
do sorologia. Os 05 docentes
d
q
que
realizaram pro
ofilaxia pós-exposição
o recebera
am soro-va
acinação, sendo
s
que, 02
%) também
m receberam o esque
ema de pro
ofilaxia pré--exposição
o no mome
ento
(40%
que entraram na
n faculdade, sem re
ealização de
d sorologiia prévia; 01
0 (20%) disse
d
m tomado dose reforrço; 01 (20%) comentou
nuncca ter realizado sorollogia e nem
ter fe
eito um no
ovo tratame
ento pós-exxposição (soro-vacin
(
nação), ma
as nunca
realizado um exame
e
soro
ológico; e 01
0 (20%) relatou
r
que
e assim qu
ue entrou no
n
municou já ter
t realizad
do profilaxiia póscurso de Mediccina Veterinária com
osição, e que
q faz monitoramentto sorológiico periodicamente, recebendo
r
o
expo
dose
e reforço somente qu
uando apre
esenta titulação insattisfatória. Desta
D
form
ma,
pode
emos conccluir que exxiste a neccessidade de
d alertar e estimular a realizaçção
de sorologias periódicas
p
e prévias à dose refforço para que estes profissionais
postos a re
eações advversas pelo
o recebime
ento de trattamentos
não sejam exp
necessário
os.
desn
Agra
adecimento
o ao Institu
uto Pasteur pela realização dass titulaçõess.
PT.0012
MPARATIV
VO DA OC
CORRÊNC
CIA DE RA
AIVA EM HERBÍVOR
H
ROS
ESTUDO COM
TENDIME
ENTOS ANTIRRÁBIC
COS HUM
MANOS NA
A REGIÃO DE SÃO JOSÉ
J
E AT
DO R
RIO PRET
TO NO PER
RÍODO DE
E 1999 A 20011
Boccchi MR1,2, Paulino
P
FS1, Tomé MFA
A1 - 1Fatec Rio Preto - Faculdade de Tecnoloogia
2
em A
Agronegócioo, Grupo de
d Vigilânciaa Epidemiológica 29 - SJRP
A raiiva, que noss bovinos see manifesta de forma paralítica, accarreta granddes prejuízoos
aos ppecuaristas. No homem
m pode caraccteriza-se poor uma enceefalite de allta letalidade. O
objettivo deste trrabalho foi estudar
e
a distribuição temporal
t
e geográfica
g
d raiva em
da
herbíívoros e doss atendimenntos antirrábbicos humannos na regiãão de São Jo
osé do Rio
Pretoo. As fontess de dados foram
fo
Grupoo de Vigilânncia Epidem
miológica e Escritórios de
Defeesa Agropeccuária, a tabbulação foi feita
f
utilizanndo Excel e o mapeamento utilizanndo
progrramas TrackMaker e Google
G
Eartth. Os resulttados mostrraram que o número de
AAR
RHs (Atendiimentos Anntirrábicos Humanos)
H
p contato com
por
c
herbívo
oros seguem
m
padrõões mensaiss semelhanttes ao longo
o dos anos, ccom aumen
nto em feverreiro/abril e
setem
mbro/outubrro e o mesm
mo padrão ocorre
o
com nos
n focos em
m herbívoro
os. A notificcação
de raaiva em herbbívoros ocoorreu em 19999, 2000, 20003, 2006, 2007,
2
2008, 2010 e 2011,
próxiima a cursoos d’água e área
á antropiizada. Foram
m encontraddos 25 proppriedades coom
focoss de raiva (bbovinos/equuinos) totaliizando 57 annimais em 14
1 municípiios. Evidencciase coorrelação enntre aumentoo de casos de
d raiva em herbívoros seguido dee aumento do
d
númeero de pesso
oas tratadass. Estes aum
mentos podem coincidirr com períoddos de aumento
de plluviosidade, cheias de rios
r e melho
oria de pasttagem. O co
oeficiente dee incidênciaa
demoonstrou umaa grande vaariabilidade,, dados quee nos direcio
onam a novas pesquisaas,
assim
m como a faalta do dimeensionamento dos prejuuízos econômicos e subbnotificaçãoo de
casoss. Essa dificculdade em contabilizaar os casos, somados àss várias fonttes de
inforrmação dificculta a obtennção de daddos. Concluui-se que a raiva causa perdas
p
econnômicas e rissco à saúde pública, po
orém não esstimados. A ocorrência de raiva em
m
herbíívoros e ateendimentos antirrábicoss humanos coincide
c
e apresenta
a
auumentos noss
períoodos de feveereiro a marrço e setembbro a outubrro. Há relaçção dos caso
os com o maapa
hidroográfico da região, direecionando a pesquisas a respeito do
os fatores condicionan
c
tes.
É diffícil padroniizar dados de
d diversas fontes, sugeere-se a criaação de bannco de dadoss que
perm
mita comparttilhar as infformações entre
e
as instiituições.
PT.0013
ALIAÇÃO DE QUIRÓ
ÓPTEROS
S POSITIV
VOS PARA RAIVA E SITUAÇÃ
ÃO
AVA
VAC
CINAL DO
OS GATOS NO BLOQ
QUEIO DE FOCO DE
E BOTUCA
ATU-SP, EM
M
UM PERÍODO
O DE 5 ANO
OS
Carvvalho VM1,2,3,4,5 - 1Facuuldade de Medicina
M
Vetterinária e Zootecnia
Z
- UNESP
2
Cam
mpus Botucaatu - Depto de
d Higiene Veterinária
V
e Saúde Pú
ública, Proff. Dr. Cassiaano
3
Victóória - Deptoo de Higienee Veterináriia e Saúde Pública,
P
Prrof. Dr. Carllos Roberto
Padoovanni - Dep
pto de Bioestátistica, 4Gabriella
G
K
Koppany
González - Eqquipe de
Vigillância Amb
biental em Saúde,
S
Prefeeitura de Bootucatu, 5Vaaldinei Moraaes Campannucci
da Siilva - Equip
pe de Vigilâância Ambieental em Saúúde, Prefeittura de Botuucatu
os - Raiva
Palaavras Chavee: Quirópteros – Felino
1. Introduçãoo
O preesente trabaalho avaliouu a situação do risco dee transmissãão da Raiva em felinos
levanndo-se em consideração
c
o a incidênccia de quiróópteros posittivos para a Raiva e o
perceentual da co
obertura vaccinal em gattos nas áreaas de Bloqueeio de Foco
o no municíppio
de Botucatu-SP,, através de um estudo retrospectivvo no períoddo de 2005 à 2009. Parra
d bloqueio de foco forrnecidos pella Equipe dee
análiise foram uttilizados os relatórios de
Vigillância Amb
biental em Saúde
S
(EVA
AS) da Prefeeitura Municcipal de Bottucatu.
2. Resultadoss e Discussãão
No período
p
avalliado, constaatou-se 9 caasos de quirrópteros possitivos para Raiva no
muniicípio, de 4 espécies diiferentes. Duurante os 5 anos de Blooqueio de Foco,
F
10.6255
residdências foram
m visitadas e avaliado a situação vacinal
v
e idade de 5.1003 cães e 980
gatoss.
Posteeriormente os
o dados levvantados peelo Bloqueioo de Foco foram
fo
comparados com
m os
resulltados obtiddos na Camppanha de Vaacinação Anntirrábica noo mesmo peeríodo, paraa
avaliiar se os feliinos domiciiliados no perímetro doo Foco estavvam corretaamente
imunnizados conttra Raiva e se há risco real de trannsmissão da doença.
3. Conclusãoo
Conssiderando qu
ue a média da coberturra vacinal naas Campanhhas Antirráb
bicas no perríodo
de 20005 à 2009 foi 52,65%, índice abaaixo do precconizado pela Organizaação Mundial da
Saúdde (OMS) e que na avalliação do esstado vacinaal dos gatoss conferidoss pelo Bloquueio
de Fooco indicam
m que 92% dos
d felinos não estavam
m previamennte imunizaados, concluuindo
que hhá risco de reintroduçã
r
o da Raiva em humanoos através do ciclo: quirópteros –
felinoos – humannos, caso as Campanhass de Vacinaação não atin
njam a Metta Vacinal para
p
gatoss no Municíípio de Botuucatu.
mentos
4. Agradecim
D
de Higiene e Saúúde Pública da FMVZ – Unesp Caampus Botuccatu, a Equiipe
Ao Depto
de V
Vigilância Am
mbiental em
m Saúde da Prefeitura de
d Botucatuu – SP e a ag
gência FAP
PESP
pelo financiameento da pesqquisa.
PT.0014
NITORAM
MENTO DE
E MORCEG
GOS (QUIROPTERA
A) COMO ESTRATÉ
ÉGIA
MON
DE V
VIGILÂNC
CIA DA CIIRCULAÇÃ
ÃO DO VÍRUS DA RAIVA
R
NO RIO
GRA
ANDE DO SUL
Witt AA1, Doninni MAW, Predebon
P
J, Diedrich G, Prato R - 1Centro Estaadual de
S
- Programa
P
E
Estadual
de Controle
C
e Profilaxia
P
d Raiva
da
Vigillância em Saúde/RS
Morccegos são annimais com
muns em áreaas urbanas no
n Rio Gran
nde do Sul, principalm
mente
em ggrandes cidaades. Atualm
mente, dentrre os animaais sinantróp
picos observvados nessaas
áreass os morceggos são, provvavelmentee, os que cauusam maiorr preocupaçãão por partee das
autorridades de saúde
s
públicca. Segundo
o o Ministérrio da Saúdee, desde 20004, os morccegos
são oos principaiss agentes naa disseminaação do víruus da raiva no
n Brasil. Diante
D
desse
cenárrio, a Secretaria Estaduual de Saúdee através doo Centro Estadual de Vigilância
V
em
m
Saúdde (CEVS), instituiu o Programa
P
d Monitoram
de
mento de Morcegos,
M
co
om o objetivvo de
estuddar a importtância dos quirópteros
q
n ciclo urbbano da raivva. O monittoramento de
no
d
raivaa através de amostras de
d morcegoss era realizaado até o ano
o de 2011, de
d forma
passiiva, onde oss morcegos encontrado
os em situações não habbituais (caíddos no chão,
dentrro de casa, etc.),
e
eram eenviados paara investigaação laborattorial sem terem sido
identtificadas e catalogadas
c
as espéciess envolvidass neste processo. Sendoo assim, o CEVS
C
passoou então a identificar e catalogar os
o animais enviados
e
pela populaçãão, com o
objettivo de traçaar estratégiaas para o maanejo de moorcegos em áreas urbannas no Estaddo.
Além
m disto, foi estabelecida
e
a rotina para coleta de morcegos, para
p obtençção de amosstras
de saaliva, sanguue e tecido cerebral
c
de indivíduos
i
eem colôniass de diversas regiões doo
Estaddo. A maiorr parte dos morcegos
m
é anilhada e solta para verificar
v
desslocamentoss
entree cidades e regiões
r
posiitivas para raiva.
r
No anno de 2011 foram
f
enviaadas para annálise
268 aamostras dee quirópteroos, das quaiss apenas seiis indivíduos de morceg
gos nãohemaatófagos ressultaram positivos. A maioria
m
das amostras dee morcegos pertence à
famíllia Molossid
dae, onde see observam
m espécies beem adaptadas à vida naas cidades. Com
C
base neste moniitoramento o Rio Grandde do Sul esstá investind
do na preveenção e
mente, o
invesstigação da circulação do vírus rábbico nas áreeas urbanas, onde atualm
morccego é consiiderado o prrincipal ageente transmiissor.
PT.0015
TECTION OF
O B LYM
MPHOCYT
TES IN THE
E CENTRA
AL NERVO
OUS SYST
TEM
DET
OF C
CATTLE AND
A
HORS
SES NATU
URALLY IN
NFECTED
D WITH RA
ABIES VIR
RUS.
Achkkar SM1, Feernandes ER
R1, Carrieri ML1, DUARTE MIS2 - 1Instituto Pasteur Labooratório de Diagnóstico
D
o da Raiva, 2Faculdade de Medicinna da Univeersidade de São
S
Pauloo - Departam
mento de Paatologia
moral immun
nity has an essential
e
prootective funnction in thee course of rabies
r
viruss
Hum
infecction. Virus--neutralizinng antibodiees, under thee control of T helper ceells, play a
criticcal role in im
mmunoprotection. Therrefore, our aim was to detect the presence
p
of B
lympphocytes in samples of central nerv
vous system
m (CNS) fro
om cattle annd horses annd
comppare the finddings betweeen species.. For this weere selectedd four samplles of cattlee and
four samples of horses. Fraagments CN
NS (cortex, hippocampu
h
us, cerebelluum and braiin
m) were analy
yzed by imm
munohistocchemical reaaction. All immunostaained cells were
w
stem
quanntified with the
t aid of thhe graticule of 1 cm2 cooupled in ligght microsccope with 100x
eyepiece and 400 x objectivee, were quanntified 40 fiields in everry fragmentt of the CNS
S.
pressed in number
n
of cells
c
per mm
m2. When coomparing thhe populatioon of
Resuults were exp
B lym
mphocytes in
i cattle andd horses, wee found a sttatistically significant
s
in
ncrease of these
t
cells in cattle in relation to horses (p = 0.0006). By analyzing
g the differeent brain areeas in
nificant diffferences
sampples obtaineed from bovvine not wass found statiistically sign
betw
ween areas, but
b in samplles of horsees was foundd a greater presence
p
off these cells in
the bbrain stem (p
p = 0.0266)). We could also observve that the meningeal
m
a perivasccular
and
envirronments were
w where we
w find morre immunosstaining for B lymphocyytes. The
detecction of B lyymphocytess in CNS saamples of caattle and horrses was mo
ore pronounnced
in meeningeal annd perivascuular environnments, sugggesting that these cells would be
enterring the CN
NS by breakiing the bloo
od brain barrrier, howevver, the induuction of speecific
antibbodies for viral
v
clearannce is delayeed, occurrinng only wheen the animaal already have
h
in sevvere neuronnal damage.. Although we
w have dettected B cellls in situ inn the samplees
studiied, these ceells were in small amouunts mainlyy in sampless of horses. The
collaaboration inttercellular between
b
CD
D4 + T lympphocytes an
nd B lymphoocytes for
activvation of theese cells andd consequenntly inductioon of speciffic antibodiees to the virrus
may be impairedd because CD4
C + T lym
mphocytes when
w
enterinng the CNS
S may underrgo
gh its associiation with infected neuurons that up
u expressin
ng FASL annd
apopptosis throug
bind to CD4 + T cells expreessing FAS occurring so
s the deathh of these im
mmune cellss
v
Thesee findings arre importan
nt for
essenntial for prootection agaiinst rabies virus.
undeerstanding how
h the imm
mune responnse is maniffested in theese animal species
s
and also
to im
mprove undeerstanding of
o the pathogenesis of rrabies in catttle and horsses.
TUTO PAS
STEUR/FAC
CULDADE
E DE MEDIICINA DA USP
U
Finaancial support: INSTIT
PT.0016
A
us Expressin
ng G protein of Rabiees in mice
Efficcacy of Reccombinant Adenoviru
Yangg DK1, Kim
m HH1, Songg JY1 - 1QIA
A - Viral dissease
kground: Since rabies case occurrred again inn 1993, a number of aniimal rabies had
Back
beenn reported up
p to 2011. Even
E
thoughh animal rabbies cases seeemed to bee decreased, the
contiinuous outbbreak was iddentified at some
s
countties of Ganggwon Provinnces of Korrea.
Althoough nationnal mass vacccination prrogram withh live and innactivated vaccines
v
to
domeestic and peet animals has
h blocked dog-to-dogg transmissio
on, most off rabies casees are
relateed with to animal
a
bittenn by rabid raccoon
r
doggs and rabiees in wild an
nimals are not
n
eradiicated. A saafe and effecctive vaccinne is neededd for the imm
munization of wild anim
mals
and ddogs. Humaan adenoviruuses have been
b
studiedd as viral vector. In thiss study, we
consttructed threee kinds of recombinan
r
nt adenoviruus expressing rabies prooteins and
checkked efficacy
y of the connstructs in mice.
m
M
Raabies virus (RABV) cirrculating in
n Korea wass isolated ussing
Mateerial and Methods:
neurooblastoma cell
c (NG1088-15) in 20009. The RAB
BV designaated as KRV
VB0910 straain
was ppropagated in the NG108-15 cellss for the clonning of genes. In orderr to analysiss the
glycooprotein (G) and nucleoocapsid (N)) genes of thhe strain, thhe G and N genes
g
were
ampllified with three
t
kinds of
o primers and
a cloned into pENTR
R/D-TOPO cloning vecctor
respeectively. Affter cloning three geness (Nfull, Gfu
full,G-TMCD
D), each plaasmids
contaaining the genes
g
were transfected
t
into TOP100 competentt cells. The purified
plasm
mids were mixed
m
with pAd/CMV/
p
/V5-DEST gateway
g
vecctor and thee mixtures had
h
reacttion with LR
R Clonase II enzyme too catalyze thhe LR recom
mbination reeaction. Aftter
confi
firming the expression
e
c
clones,
the clones
c
weree digested with
w Pac I to
o expose thee
ITRss and transfeected into thhe 293A celll lines to coonstruct reccombinant adenovirus
a
(reAddV) expresssing N and G genes of RABV. Thhe 293A cellls transfecteed with the
clonees showed specific
s
cytoophatic effeect. For 6 daays after inooculation, thhe cells werre
stainned with monoclonal anntibodies annd FITC connjugated goaat anti humaan IgG+IgM
M and
exam
mined by fluuorescent microscopy.
m
T check effficacy of thhree kinds of
To
o reAdVs, tthe
reAddVs containiing 108.0 TC
CIID50/ml was
w inoculatted into 4 weeks
w
old Baalb/C mice.
Survvival rate and change off body weig
ght of the mice
m were chhecked for 17 days afterr
challlenge.
ults: We succcessfully reeconstructed three kindds of reAdV
Vs (Nfull, Gfull,
G
G-TM
MCD)
Resu
in 2993A cells. The
T titer of reAdVs
r
rangged from 1007.7 to 108.0 TCID50/ml.. Four groupps of
micee (Gfull, G-T
TMCD, Nfuull+Gfull, Nfull+G-TM
N
MCD) were inoculated with
w 0.2 mll
reAddV and half of mice in each
e
group were challeenged with CVSN2c sttrain
intram
muscularly 21 days aftter inoculatiion. All micce did not shhow any typ
pical rabies
sympptoms and showed
s
com
mplete protection. On thhe other hannd, half of mice
m in threee
groupps (Gfull, G-TMCD,
G
G
G-TMCD+N
Nfull) did noot show com
mplete proteection againnst
challlenge by inttracranial (IC) route. However, thee one group inoculated with
Nfulll+Gfull reA
AdVs revealed 100% su
urvival rate. These dataa demonstraated the poteential
of the reAdV as a safe rabiees vaccine.
Vs in 293A cells. The combination
c
n of
Concclusion: Wee constructeed three kinnds of reAdV
two kkinds of reA
AdVs (Nfulll+Gfull) maay be a usefful tool in seearch of rabbies vaccinee
canddidate for annimals and further
f
study
y related to oral vaccinnation of doggs and raccooon
dogs is needed in the near future.
PT.0017
R
Viru
us Glycoprrotein Fragments with
h the Nicotiinic
Interraction of Rabies
Acettylcholine Receptor
R
Rideout SA1, Haarris MB2, Hueffer
H
K3, Schulte MK
K2 - 1Univeeristy of Alaaska Fairbannks,
2
3
Univversity of Alaska
A
Fairbbanks, Univ
versity of Alaska
A
- Vetterinary Medicine
The rrabies viruss glycoproteein (RVG) innteracts witth Torpedo and musclee nicotinic
acetyylcholine receptors (nA
AChR). The field of Liggand Gated Ion Channeels, such as the
nicottinic receptoors, has bennefited greaatly over thee last decadee due to thee discovery of
non-m
membrane bound
b
Acettylcholine Binding
B
Protteins (AChB
BP). Since nicotinic
n
acetyylcholine receptors andd the ACHB
BP share signnificant seqquence and structural
s
homoology in thee neurotoxinn binding do
omain, the A
AChBP cou
uld provide a useful moodel
for sttudying the molecular basis
b
of thee RVG/nAC
ChR interacttion.
In this study we investigateed the interaaction betweeen RVG neeurotoxin like peptide
ments and thhe AChBP. Surface Plaasmon resonnance (SPR
R) was used to assess
fragm
bindiing kineticss to the AChhPB. Electroophysiologyy experimennts were useed to compaare
thesee results to interactions
i
between thhese RVG fragments
fr
an
nd human nicotinic
n
acetyylcholine receptor subtyypes. RVG
G fragments were shown to bind with
w micromolar
affinnity to the Lyymnaea AC
ChBP. SPR
R permits detterminationn of on and off
o rates forr
bindiing of all 6 fragments. Our data shhow slow on
o rates (ka=
= 100-300 1/M•s)
1
with off
rates (kd = 0.01--0.004 1/ M•s)
M corresponding to binding
b
withh a dissociatte rate (Kd of
omolar). Vooltage clamp
p electrophyysiology daata obtained using Xenoopus
25.4--60.3 micro
oocyytes shows similar
s
Ki values for innhibition of acetylcholinne induced responses on
o
alphaa4/beta2 nA
AChR.
PT.0018
Rabiies in Iran
Baghhaipour MR
R1 - 1Milad Hospital
H
b a virus, Lyssavirus
L
r
rabies
that affects
a
the nervous
n
systtem
Rabies is a diseaase caused by
ults in deathh unless treaated quicklyy. Rabies is found in maammals in all
a
and uusually resu
regioons of the world.
w
The disease
d
infeccts domesticc and wild animals,
a
andd is spread to
t
peopple through close
c
contacct with infected saliva via bites or scratches. Dogs
D
are thhe
mainn host and trransmitter of
o rabies butt bats, foxess, raccoons, coyotes, wolves
w
may
transsmit the diseease as welll. More thann 50 000 people die of rabies everyy year.
The vvirus spread
ds through the
t central nervous
n
system and proogressive, fatal
fa
inflam
mmation off the brain and
a spinal co
ord developps. After an incubation period of 1–3
montths (days to years) the initial
i
sympptoms start with
w fever and
a often paain or
paraeesthesia at the
t wound site.
s
In abouut 35% of thhe patients, Rabies
R
has a longer course
.The muscles grradually beccome paralyyzed, startinng at the sitee of the bite or scratch. A
med the paraalytic form and
a
comaa slowly devvelops, andd eventually death occuurs. It is nam
its diiagnosis is difficult.
d
Thhe majority of patients show a furious form annd exhibit siigns
of hyyperactivity, excited beehavior, hyd
drophobia annd sometim
mes aerophobbia. After a few
days, death occu
urs by cardiio-respiratorry arrest.
No teests are avaailable to diaagnose rabiees infectionn in humans before the onset of clinnical
diseaase. For susp
pected anim
mals, taking tissue sampples (often brain
b
tissue) and using
immuunofluoresccence or imm
munologicaal techniquees to detect the
t virus is the mainstaay of
diagnnosis.A persson exposedd to rabies should
s
ideallly be treateed within 12
2 hours of thhe
expoosure and deefinitely witthin 48 hourrs for the beest outcomee. They are usually
u
treaated
with rabies imm
mune globuliin and rabiees vaccine innitially and then get sevveral additiional
vacciine injectionns.
The nnumber of infected
i
perrson has beeen decreasedd during paast decade inn Iran. Mostt of
them
m are males and
a under ten years oldd. More thann 250 centeers provide post
p exposuure
treatm
ment for Raabies and Paasture Instittute have prroduced IG and Vaccinne of Rabies
sincee 85 years ago.
a
PT.0019
E
e Saúde - Alcance, eestratégias, metodologgia.
em
Proggramas de Educação
Reichhmann MLB
B1, Nunes VFP
V 2, Santoos MB3, Viaaro O4 - 1Insstituto Pasteeur - Assisteente
Técnnico, 2Prefeiitura do Muunicípio de Jundiaí
J
- Jarrdim Botâniico, 3Ministtério do Meio
4
Ambbiente - Instiituto Chico Mendes, Secretaria
S
M
Municipal
dee Saúde de São
S Paulo Centtro de Contrrole de Zoonnoses
Introodução:
A Edducação em Saúde devee ser estrutuurada em açções program
máticas. Os profissionaais da
área devem conhhecer a reallidade local a ser traballhada, estrattégias especcíficas que
d princípio
os de promooção da saúd
de, preservaação do meiio
favorreçam a inccorporação de
ambiiente, a interação saudáável com an
nimais de esstimação, effetivando a guarda
g
respoonsável. Os interlocutoores devem ser
s estimulaados a comppreender o assunto,
a
anaalisar
as informações, aplicar os conteúdos
c
aprendidos
a
e experimenntar mudançças em seus
ortamentos, transferinddo e replicanndo os conceitos adquirridos e
hábittos e compo
incorrporando-oss a sua cultuura e a seus valores.
Objeetivos:
Divuulgar os trab
balhos de edducação em saúde do Estado
E
de Sãão Paulo – SP,
S em 20111.
Mateeriais e Métodos:
Pesqquisa docum
mental nos prrogramas de educação e promoção
o da saúde nos
n municíppios
que uutilizam meetodologia compatível
c
c
com
as pesqquisas cientííficas, geran
ndo um roteeiro,
descrrevendo ativ
vidades e reesultados obbtidos.
Resuultados:
O rotteiro utilizaado incluiu resumidame
r
ente: conheccer e visitarr o local habbitado peloss
interllocutores, iddentificar problemas na interação com animaais, estabeleccer objetivoos e
estraatégias comppatíveis com
m aquilo quee deve ser trabalhado,
t
avaliar periodicamentee os
resulltados e corrrigir vieses.. As ações programátic
p
cas geraram instrumentos que serviram
de noormas e direetrizes para informar, nortear
n
e eduucar interloocutores de diferentes
d
classses socioecoonômicas e educacionais, fortaleceendo vínculo
os para umaa melhor
qualiidade de vidda.
Concclusão:
A inccorporação de program
mas de educaação em saúúde por proffissionais daas áreas de
Saúdde, Meio Am
mbiente e Saaúde Animaal, desde o iinicio de suas carreirass, utilizandoo
metoodologia com
mpatível coom suas atribbuições proopicia maiorr efetividadee nos resultaados.
PT.0020
ALIAÇÃO DOS ESTR
RATOS DA
A POPULA
AÇÃO CAN
NINA DE MUNICÍPI
M
IOS
AVA
DO IINTERIOR
R DO ESTA
ADO DE SÃO
S
PAUL
LO – BREV
VE AVALIA
AÇÃO DA
LITE
ERATURA
A.
2
Reichhmann MLB
B1, Alves MCGP
M
, Maatos MR3, Dominguez
D
M 4, Dom
MHS
minguez MH
HS4 1
2
Instiituto Pasteuur - Assistennte Técnico, Secretariaa de Estado da Saúde de
d São Pauloo Instittuto de Saúdde, 3Secretaaria de Estaddo da Saúdee de São Paaulo - Superintendênciaa de
Conttrole de End
demias, 4Proofissional au
utônoma
Introodução: Um
ma das princiipais questõ
ões relacionaadas ao cuiddado e proteeção da saúúde
humaana e animaal consiste na
n determinação do núm
mero de annimais e de suas
s
demaiss
caraccterísticas demográfica
d
as. Esta avalliação semppre foi possíível em um dos estratoss, a
de cãães com pro
oprietários, uma
u vez qu
ue a presença nos domiccílios permiite sua
contaagem. A Téécnica Pasteeur São Paullo (TPSP) foi
f um modeelo de avaliação de
popuulações caniinas de áreaas urbanas, desenvolvid
d
do em 2001,, que permittiu a estimaação
da poopulação seegundo os estratos
e
de restrição
r
e dependência
d
a.
Metoodologia: Em
m 2002, a TPSP
T
foi aplicada nos municípios
m
do interior do Estado dde
São P
Paulo para estimar
e
o núúmero de cãães existentes segundo estratos e conhecer
c
aspecctos relacionados à vaccinação e diinâmica poppulacional. A amostra foi
f compostta de
100 ssetores censsitários, disttribuídos em
m 41 municíípios.
Resuultados: Foraam obtidos dados que se
s situavam
m acima da relação
r
até então
e
dispooníveis, indicando umaa proporção de um cão para cada quatro
q
habitantes. Salieentase a bbaixa idade dos cães, em
e média, de
d quatro annos, indicativva da velocidade da
repossição populacional. A maior
m
conceentração (599%) de cãess foi de até três
t anos,
geranndo preocuppações com
m a introduçãão continuaa de animaiss susceptíveis e os posssíveis
prejuuízos no conntrole da leiishmaniose e demais zooonoses. Estas dificulddades ficam
preseentes quand
do se observva a alta percentagem (220%) de pro
oprietários que
q
descoonheciam see seus anim
mais foram vacinados
v
coontra a raivaa nos 12 meeses anteriores e
quanntas crias forram geradass nos partoss ocorridos nno mesmo prazo
p
de tem
mpo.
Concclusões: Meesmo para cães com prooprietário, parcela
p
teoriicamente mais
m protegidda, a
convvivência com
m animais de
d estimação
o não tem see revestido da responsaabilidade leggal
para prevenção de doenças que envolvvam as pessooas e os aniimais. Recomenda-se a
inserrção de proggramas supllementares, como os dee educação, de registro e concessãão de
licennças e a interação com profissionai
p
is de diversaas especialiidades para obtenção dee
resulltados concrretos no equuilíbrio ecollógico de cãães mantidoos em domiccílios.
PT.0021
LIZAÇÃO
O DO GEOP
PROCESSA
AMENTO COMO FE
ERRAMEN
NTA NO
UTIL
MON
NITORAM
MENTO E CONTROL
C
LE POPUL
LACIONAL
L DOS MO
ORCEGOS
HEM
MATÓFAG
GOS NAS ILHAS
I
FLU
UVIAIS DO
O MUNICIIPIO DE ANANINDE
A
EUAPAR
RÁ/BRASIL
L.
Ram
mos OS, Barrreiros MA, Souza EM, Souza SMF
F, Melo TIS
S
RES
SUMO
mportância do
d ciclo silv
vestre envolvendo os morcegos
m
hem
matófagos na
n
É creescente a im
regiãão, tem sidoo observada uma mudannça no perfiil epidemiollógico da raaiva humanaa no
estaddo do Pará, tornando
t
oss morcegos os principaais responsávveis pela dooença.
O preesente trabaalho visa forrnecer subsídios possibbilitando um
m banco de dados
d
geoggráficos dos principais eventos,
e
parra gerar mapas de áreas propicias para o ataquue
espolliativo de morcegos
m
heematófagos, fazendo o ccontrole poppulacional dos
d quirópteros,
cadastrando e geeorreferenciiando abrigos e as proppriedades doos moradorees ribeirinhoos
P
Santta Rosa e Viiçosa onde os
o animais (suínos, bovvinos
das ilhas Sasuneema, João Pilatos,
e Avves) são ataccados.
Diannte dos fatoss, o sistema de Vigilânccia em Saúdde do municcípio de Ananindeua
progrramou as açções em 20112 usando as
a técnicas do
d geoproceessamento. Os
O mapas fo
foram
elaboorados utilizzando-se o sistema
s
AR
RC.GIS 9.3, as bases caartográficas do Municíppio
de A
Ananindeua e o equipam
mento GPS MAP
M
60 CS
SX Garmin..
Foram
m atendidass, 05 propriedades na Ilha João Pillatos, 01 naa llha viçosaa e 01 na Ilhha
Santaa Rosa. Sen
ndo capturaddos 35 morccegos hemaatófagos geoorreferenciaados e
cadastrados 06 abrigos
a
artifficiais e 03 abrigos natuurais e 07 pontos
p
de caaptura geranndo
mapaa para fazerr uma avaliaação espaciaal. Certas caaracterísticaas de uma reegião
deterrminam altaas populaçõees de D.rottundus em áreas
á
próxim
mas aos rioss, geralmentte
favorrecem a exiistência de um
u numero maior de abbrigos Lordd (1988).
Os abbrigos artifiiais georrefe
ferenciados, a maioria são:
s Caeirass desativadaas e casas dee
taipaa abandonaddas foram enncontrados morcegos em
e ocos de árvores
á
dennominadas
siriúbba (Avicennnia nítida) ao
a longo doss rios. Invesstigação epiidemiológicca sobre o caaso
de raaiva em Aug
gusto Corrêa relata a exxistência dee grande poppulação de morcegos
m
q
que
habittam os manguezais e abbrigam-se no
n oco da sirriubeira Ministério da Agricultura
A
a
Pecuuária e Abasstecimento (2005).
(
Ao se inter-relaccionar as baases de dado
os georreferrenciados e espresa-las visualmentte
a
e ass propriedaddes atacadass por
atravvés do mapaa digital, pode-se obserrvar que os abrigos
morccegos localizam-se as proximidade
p
es dos rios, possivelmeente os mesm
mos deslocaamse atrravés dos manguezais
m
q margeiaam as ilhas..
que
O coontrole da poopulação doos morcegoss hematófaggos conform
me o mapa, ajuda
a
na
caraccterização do
d transmisssor, identificcando as coomunidades mais afetad
das e defininndo a
situaação de risco
o, estabelecendo uma ação
a
estratéggica especiffica de vigillância a saúúde.
go hematófaago, mapa
Palaavras-Chave: Ananindeua, georrefferenciamennto, morceg
G
Vazz Secretáriaa de Saúde de
d Ananindeeua pelo apooio, e
Agraadecemos a Dra Ivete Gadelha
os aggentes de enndemias Arttemis, J.Theeodósio, Avviz, Augustoo, Erivaldo e Edmilsonn por
suas dedicações na captura..
PT.0022
YLOGEOG
GRAPHY OF
O THE RA
ABIES VIR
RUS ISOLA
ATED FRO
OM CANID
DS
PHY
IN T
THE NORT
TH AND NO
ORTHEAS
ST OF BRA
AZIL
Carnnieli Jr P1, Oliveira
O
RN1, Castilho JG
J 1, Vieira LFP2 - 1Insstituto Pasteur, Brasil 2
Diaggnóstico, Innstituto de Defesa
D
Agro
opecuária e Florestal doo Espírito Santo,
S
Espíriito
Santoo, Brazil
R
Viru
us (RABV), typically foound in doggs in Brazil, still
Antiggenic variannt 2 of the Rabies
circuulates in the North and Northeast of
o Brazil in spite of mass vaccinatiion campaiggns
againnst rabies in
n domestic animals.
a
Preevious studiies have shoown that thiis variant is
madee up of two main genettic lineages:: one found in dogs (Ca
anis familia
aris) and anoother
typiccally found in wild caniids, particullarly the craab-eating foox (Cerdocyyon thous),
whicch is known in Brazil ass cachorro do mato. C.. thous is cuurrently the only wild land
speciies in whichh RABV is continually isolated, with
w an averaage of 20 caases/year.
Althoough previo
ous studies and
a phylogeenetic analyyses of the N and G gen
nes and
interggenic G-L region
r
all coonfirmed th
he existence of these tw
wo genetic liineages, thee way
in whhich the lineeages had dispersed
d
tem
mporally annd spatially remained too be elucidaated.
We ssequenced a 1388 nucleeotide-long region of thhe RABV N gene from
m 53 typical
canidd samples. The
T phylogeeography off the datasett obtained was
w determiined using thhe
BEA
AST (Bayesiian Evolutioonary Analyysis Samplinng Trees) program withh the follow
wing
data for the scrip
pt: Evolutioonary Modeel K3Puf+G4; Partitioniing into coddon positionns
r
uncoorrelated lognormal; Demographic
D
c model
(1+2)+3; Molecular clock relaxed
wth; Disperrsion model relaxed ranndon walkin
ng (RRW). The
T
coaleescent exponential grow
resullts can be su
ummarized as follows. The commoon ancestorr of the RAB
BV in the
sampples analyzeed became established
e
around
a
1900 in the borrder region between
b
thee
states of Paraíbaa and Pernam
mbuco and diversified into the lin
neages typically found iin C.
familliaris and C.
C thous. Aroound 1910 the
t original C. thous linneage diverrsified into ttwo
mainn sublineagees in the sam
me area, onee of which migrated
m
to the south and
a the otherr to
the nnorth. Then, around 19330 the southhern sublineeage diversiified even fuurther and
moveed in two diirections: tooward the sttate of Bahia and towarrd the state of Pernambbuco.
In appproximately 1945 the northern su
ublineage off C. thous allso diversifiied, resultinng in
a subblineage thaat remained in the state of Pernambbuco and an
nother that diversified
d
e
even
furthher and movved toward the
t state of Piauí. The lineage
l
typiically found
d in C. familliaris
diverrsified aroun
nd 1945 andd moved tow
ward the noorth and souuth. Howeveer, around 1970
1
it undderwent a major
m
diverssification when one of the
t 1945 su
ublineages divided
d
into two
subliineages, onee of which migrated
m
to the states oof Pernambu
uco, Sergipee and Alagooas
and tthe other to the states of
o Pará and Piauí.
P
Analyysis with thhe RRW moodel allowedd us
to inffer not only
y the movem
ment of the virus
v
lineagges isolated from C. fam
milaris and C.
thouss among thee municipallities sampleed, but also the probable location where
w
dispeersion of thee lineages occurred.
o
Fuurther studiees covering a longer peeriod and ussing
moree samples arre required to confirm the
t pattern of movement of the virrus describeed
here so that its geographic
g
s
spread
can be
b predictedd and blockked.
B
Finanncial Suppoort: Institutoo Pasteur, Sãão Paulo, Brazil
PT.0023
RELIMINA
ARY ANAL
LYSIS OF THE GLY
YCOSYLAT
TION PAT
TTERN OF
F
A PR
SER
RUM IgG FOLLOWIN
F
NG PRE-E
EXPOSURE
E SCHEME
E FOR HU
UMAN RAB
BIES
VAC
CCINE
Silvaa SR1, Cunhha Neto RS2, Chaves LB
B1, Faquim
m-Mauro EL
L3, Silva AC
CR1 - 1Instituuto
2
Pasteeur - Seção de Diagnósstico, Instittuto Butantaan - Laborattório de Biooquímica e
Biofí
física, 3Instittuto Butantaan - Laborattório de Imuunopatologiia
T currenttly recommeended by WHO
W
for preevention inddividuals
Intrroduction: The
expoosed to rabiees virus (RV
V) is pre-expposure propphylaxis (PE
EP) through
h the
admiinistration of
o three dosees of rabies vaccine byy intradermaal (ID) or in
ntramuscularr
(IM) route. The previous sttudy showedd that the peersistence of
o protectivee antibodies titer
ule is more long-lived, suggestingg a different clearance mechanism
m
in thee IM schedu
of
antibbody. It known that carbbohydrate modification
m
n of antiboddies could innduce
accellerated cleaarance or moodulate of antibody
a
efffector functiions. All enddogenous IggG
antibbodies contaain an N-linkked glycan in the Fc reegion and abbout 20% coontain glycaans
elsew
where on thee molecule to maintainn the two heeavy chains of the Fc inn an open
confo
formation reequired for interactions
i
with activaating Fcγ receptors (FcγγRs). Objecctive:
Baseed on these data,
d
we com
mpared the glycosylatiion patterns of serum IggG of
indivviduals whoo received thhe PEP scheedule by ID or IM routee. Methodss: The Puriffied
Veroo Cell Vacciine (PVCV)) (Sanofi /Pasteur Labooratory) wass utilized, with
w a minim
mum
of 2.5 IU per doose. Sera of individuals who previoously receivved the PEP
P by the IM (n=
v
dosse) or by thee ID (n= 19;; 0.1 mL /doose) route were
w collecteed to
15; 00.5 mL per vaccine
IgG detection
d
an
nd its subclaasses (IgG1, IgG2, IgG
G3 and IgG44) by antigeen-specific
ELIS
SA and to measured
m
RV
V neutralizinng antibodiies by rapid fluorescentt focus
inhibbition test (R
RFFIT). Thee N-linked oligosaccha
o
aride chain of
o purified IgG
I antiboddies
from
m serum wass evaluated by
b a lectin enzyme-link
e
ked immuno
osorbent assay. Resultts:
All 334 persons tested
t
developed titers of antibodyy ≥ 0.5 IU/m
mL, with a maximum
m
geom
metric mean
n titer of 18.0 IU/mL. There
T
was noo statisticallly significan
nt relation
betw
ween neutralizing antiboody titer andd route of addministratioon. Similarlly, the majoority
of the anti-rabiees humoral responses
r
w restricteed to the IgG1 subclasss in both grooups.
were
wever, the lectin-bindingg assay showed differeence on the monosacch
m
aride contennt,
How
mainnly of galacttose, neuram
minic acid, mannose
m
annd fucose beetween seruum IgG1
indivviduals whoo received thhe PVCV by
y ID or IM route. Concclusion: Thhese results
sugggest that IgG
G1 serum off individualss vaccinatedd intramuscuulary with PVCV
P
are more
m
glycoosylated thaan IgG1 seruum of subjeects who recceived the vaccine
v
intraadermally. This
T
differential glyccosylation patterns betw
ween antiboodies is encoouraging and warrants
furthher examinaation.
PT.0024
ÁLISE EPIDEMIOLÓ
ÓGICA DA
AS AGRES
SSÕES POR
R ANIMAIIS
ANÁ
SILV
VESTRES NA 10ª RE
EGIÃO DE SAÚDE/L
LIMOEIRO
O DO NOR
RTE – CE/ 2007
2
A 20011.
LIMA
A FMG1, CHAVES
C
CS
S1, RODRIG
GUES VC2 - 1Secretaria de Saudee do Estado do
Cearrá - 10ª Coordenadoria Regional dee Saúde de Limoeiro do
d Norte, 2Secretaria dee
Saúdde do Estado
o do Ceará - 10ª Coorddenadoria Reegional de Saúde
S
de Liimoeiro do
Nortee
INTR
RODUÇÃO
O: no Brasil, a partir de 2004, aum
mentou o núm
mero de cassos humanoss de
raivaa transmitidoos por anim
mais silvestres(MS, 2011). O Cearáá, em 2008 e 2010 tevee dois
óbitoos transmitid
dos por saguui. Apesar de
d não regisstrar casos humanos,
h
auumentaram as
agresssões por annimais silveestres na 10ªª Coordenaddoria Regional de Saúd
de, de 29 caasos
em 22007 para 355 em 2011. Nesse perío
odo 12,5% das
d amostraas foram positivas, um
caníddeo silvestree e um morccego hemattófago(Desm
modus rotun
ndus). OBJE
ETIVO:
analiisar a epidem
miologia daas agressõess por primatta não humaano, quirópttero e caníddeo
silvestre na 10ª Região
R
de Saúde
S
- Lim
moeiro do Norte - CE, de
d 2007 a 20011, enfocaando
ológico paraa a raiva a que
q sua população está submetida.
o riscco epidemio
MET
TODOLOG
GIA: o estudo trata-se de
d uma pesquisa docum
mental e bibbliográfica dos
d
relatóórios do Sisstema Nacioonal de Agravos de Notificação(SIINANNET,, 2012), da Ficha
F
Epiddemiológica de Profilaxxia da Raivaa Humana(110ª CRES) e artigos reccentes.
RES
SULTADOS
S: na Regiãão de Saúde predominaaram as agreessões por primatas
p
nãoo
humaanos (71,1%
%), seguidoss por quiróppteros(18,4%
%), e canídeeos silvestrees(10,4%). O
sexo masculino foi o mais agredido
a
po
or quirópteroo(62,1%) e canídeo silvvestre(75%). O
% nas agresssões por prrimatas não humanos, com
c
um
feminnino se expôs em 55,3%
aumeento de 17 agressões
a
em
m 2007, parra 25 em 20011. Mãos e pés tiveram
m 42,98% das
d
lesõees, membross inferiores 24,1%, membros supeeriores 21,499%, tronco 5,26%,
cabeçça/pescoço 4,38%, mucosa 1,75%
%. Registraraam-se 67% dos ferimenntos como
únicoo, e 33% múúltiplo. Em 47,7% dos acidentes o ferimento foi profund
do, 47,2%
superrficial, e 4,9
9% dilaceraante. O soro antirrábicoo foi adminiistrado em 60,2%
6
dos
pacieentes, revelaando que occorreu um maior
m
númerro de acidenntes graves..
CON
NCLUSÃO
O: embora nãão haja histó
órico de raiiva humana,, o registro de animais
silvestres positivvos indica a circulação do vírus naa região, que aliado ao alto percenntual
de leesões gravess, potencialiiza o risco de
d transmisssão caso a vigilância
v
doo agravo sejja
negliigenciada.
PT.0025
MORAL IM
MMUNE RESPONSE
R
E IN DOGS
S AND CAT
TS VACCIINATED
HUM
AGA
AINST RAB
BIES IN TH
HE MUNICIPALITIIES OF DR
RACENA AND
A
PRE
ESIDENTE PRUDENTE, SP, BR
RAZIL.
Albaas A1, PÍCO
OL0 MR, SO
OARES CN, BACHEG
GA HVH, TA
ARUMOTO
O MH - 1AP
PTA
- PÓLO ALTA SOROCAB
BANA
n Brazil, haas been mad
de campaignns to vaccin
nate dogs an
nd cats once a
Introoduction: In
year in almost all
a municipaalities and thhese animals are among
g the main transmitters
t
s of
rabiees to humanns. The preseence of rabiies antibodiies in animaals is likely a good indicator
that they
t
are imm
munized annd protectedd this zoonosis.Objectiv
ve: This exxperiment aiimed
to invvestigate thhe humoral immune
i
ressponse in doogs and catss vaccinated
d against rabbies
in thee Municipalities of Preesidente Pruudente and Dracena
D
12 months beffore the masss
vacciination cam
mpaign condducted in 20009. Method
ds: In this paper
p
are repported the
resullts of 834 seera from doggs and cats involving these
t
animaals. It was used
u
for this
purpoose, the techhnique of Rapid
R
Fluoreescent Focuus Inhibitionn Test (RFF
FIT) and was
consiidered as reeactant sera with valuess ≥ 0.5 IU/m
ml. Results:: Thus, Pressidente
Pruddente had 1553 (51.0%) samples
s
reaactants for dogs
d
and 599 (32.6%) reeactants forr
cats, while the Municipality
M
y of Dracenna had 110 (52.1%)
(
sam
mples reactaants for dogs
and 771 (50.0%) for cats. Coonclussion:: In this papper, is discusssed the
vacciinal coverag
ge of the annimals invollved in this experimentt. It was obsserved low
perceentages of titres ≥ 0.5 IU/ml,
I
especciallity in cats of Presiddente Prudeente.
P, process 08/54266-3.
0
Finanncial Suppoort: FAPESP
PT.0026
VELOPME
ENT OF RE
EAL TIME
E RT-PCR (TAQMAN
(
N) FOR DE
ETECTION
N
DEV
AND
D GENETIC
C CHARA
ACTERIZA
ATION OF ANTE-MO
ORTEM HUMAN
H
RAB
BIES
Morii E1, Scheffe
fer KC1, Fahhl WO1, Oliiveira RN1, Asano KM1, Carnieli Jr
J P1, Carrieeri
1
1 1
ML , Kotait I - Instituto Pasteur
P
de São
S Paulo - D
Diagnósticoo
man rabies is still an im
mportant pubblic health problem
p
in some
s
Brazillian regionss.
Hum
Usuaally, ante-m
mortem diagnnosis of rabbies is made by demonsstration of virus
v
antigenn by
direcct immunoflluorescencee in corneal or conjuncttival smearss and skin biopsies;
b
howeever, this technique hass a low senssitivity. Reccently, moleecular techn
niques such as
the reeverse transscriptase poolymerase chhain reactioon (RT-PCR
R) and nucleeic acid
sequeence based amplificatioon assay (N
NASBA) havve been dev
veloped to im
mprove the
sensiitivity and specificity
s
o ante as well as post-m
of
mortem diaggnosis of raabies. Rapidd and
accurrate diagnosis of ante-m
mortem hum
man rabies iis essential for effective medical
manaagement andd to ensure appropriatee post-expossure prophyylaxis of pottential contaacts
with the patient.. The presennt study wass carried ouut to evaluatte the sensittivity and
man) in com
mparison witth RT-PCR and DNA
speciificity of reaal time RT--PCR (Taqm
sequeencing for the
t diagnosiis of rabies.. From Junee through Ju
uly 2012, ninne specimenns
from
m three patieents with rabbies were suubmitted to the Pasteurr Institute foor rabies
diagnnosis. Five saliva (2551 and 2613 to 2616) annd two hair follicles (25552 and 26112)
speciimens were collected serially from
m patient susspected of having
h
rabiees from Matto
Grossso (MT) staate. Saliva (3550
(
and 4109)
4
specim
mens were collected
c
froom patients
rabiees suspectedd from Minaas Gerais (M
MG) and Maranhao (M
MA) states, respectively
r
y. The
posittive rabies results
r
weree confirmed by RT-PCR
R using prim
mers targeteed to
nucleeoprotein (N
N) gene andd all of specimens weree identified as compatib
ble with
hemaatophagous bat lineagee (variant 3) by DNA seequencing, with
w the excception of the
t
salivva collected from MA patient,
p
whicch was genootyped as caanid lineagee (variant 2)). A
real time
t
RT-PC
CR (Taqmann), with two
o primers annd probe setts targeting to N, has beeen
descrribed in ordder to validaate an alternnative methood for rabies diagnosis in ante-morrtem
sampples. This method
m
was capable of accurately
a
i
identifying
t variant 3 in saliva
the
speciimens collected from MT
M patient previously
p
ggenotyped as
a hematophhagous bat
lineaage. The haiir follicle annd saliva speecimens froom MT and MG patientts, respectivvely,
yieldded high Ct (threshold cycle)
c
valuees (betweenn 35 and 38)), suggestingg low viral load.
This assay failedd to detect amplificatio
a
on in the chaallenge viruus standard (CVS)
(
straiin
M patient (genotyped
(
as variant 2)
2 due to miismatches
and ssaliva colleccted from MA
betw
ween the prim
mers/probe sets and thee target N ggene. Thus, our
o results showed
s
the
usefuulness of reaal time RT--PCR as a raapid alternaative to DNA
A sequencinng (at least four
f
timess faster) forr the confirm
mation of raabies diagnoosis.
PT.0027
R
A
ANTIBODIE
ES IN DOG
GS PRIME
E VACCINATED
KINETIC OF RABIES
C
HF2, Martorellii LFA3, Kattaoka APAG
G3, VICTOR
RIA C1,
Babbboni SD1, COSTA
4
1 1
PAD
DOVANI CR
R , Modolo JR - UNE
ESP -Botucaatu - Departtamento de Higiene
Veterinária e Saaúde Públicaa, 2UNESP-- Araçatubaa - Departam
mento de apo
oio, produçção e
3
C
de Controle
C
de Zoonoses
Z
- Laboratórioo de Zoonosses e Doençças
saúdee animal, Centro
Trannsmitidas po
or Vetores, 4UNESP -B
Botucatu - Departament
D
to de Bioesttatística,
Instittuto de Biocciências
w
populaation (3.5 biillion people) occupies areas wherre
Introoduccion: Half of the world
theree is an increase of dogs, cats and ro
odents, and zoonosis trransmitted by
b these aniimals,
wherre rabies is the
t most woorrisome. Thhe protectioon of humanns against urban
u
rabies is
achieeved mainly
y by the proophylaxis off dog and caat with vacccines that ind
duce a
minim
mum of anttibody titerss (≥ 0.5 IU/m
mL). This study aimed
d to evaluatee the kineticc of
antibbodies in do
ogs that weree prime vacccinated witth Fuenzalid
da&Palácios modified
vacciine during 12
1 months. Materials and
a methodss: The study
y was condu
ucted in 20009
durinng the 41st Annual
A
vaccination cam
mpaign agaainst rabies in
i dogs andd cats in the
urbann area of Bootucatu, Sãoo Paulo, Braazil. It was considered to determinne the sample
size aat 95% conffidence alonng an estim
mation error of the orderr of 10% ran
ndom
contrribution associated withh the casuall participation of the an
nimal. Thuss, the study
invollved 576 doogs over thee age of threee months, never
n
vaccin
nated against rabies before,
regarrdless of breeed, age or sex random
mly selected for blood collection.
c
S
Samples
werre
colleected during
g five momeents at the sttudy periodd, which are: 0 (first dose of vaccinne), 1
[30 ddays after first vaccinattion (dafv) and
a the secoond dose off vaccine], 2 (60 dafv), 3
(180 dafv) and 4 (360 dafv)). Thirty day
ys after the first vaccinnation (timee 1) dogs
t same typpe of vaccin
ne (Fuenzallida&Paláciios
receiived the boooster vaccinnation with the
modiified vaccin
ne- Institute of Technollogy of Paraaná - TECPA
AR®). Seru
um neutralizzing
antibbodies to rabbies was helld at the Ceenter for Zooonosis Conttrol of São Paulo
P
trouggh the
rapidd fluorescen
nt focus inhiibition test (RFFIT).
(
Reesults: Totaal (urban and
d rural) of rrabies
vacciination coveerage reachhed 81.36% in Botucatuu / SP, and 3229
3
(16.97
7%) of dogss in
urbann were first time vaccinnated. At 3660 days afteer first vacciination, onlly 51.5% off the
dogs had protecttive titers thhroughout over
o
all period of the stuudy (over 5 moments)..
b the Conttrol Program
m, the caninne rabies
Discuussion and conclusion:: In Brazil, by
vacciination is annnual and mandatory
m
a
after
three m
months of ag
ge. The low concentratiion
of anntibodies aftter vaccinattion of dogss against rabbies has been reported, also
the iimmune ressponse of doogs with priime vaccinaation (Fuenzzalida&Paláácios modifi
fied
vacciine) withouut booster was
w evaluated
d, where thee results observed rapid
d decline off
antibbody titers, suggesting
s
t many animals
that
a
cannnot be proteected betweeen the
camppaigns. Acccording to thhe Institute Pasteur
P
in 22000, titers below
b
0.5 IU/mL do noot
confe
fer protection against raabies. In add
dition, rabiees can infectt the dogs at
a any periodd
durinng the camp
paigns. The coverage of vaccinatioon shows weeak protectiion, since onnly
51.5%
% of dogs had
h protectivve titers at all
a time of m
moments annalyzed. Furrthermore, it
i is
also kknown that half of the dogs primee vaccinatedd are not imm
munized ag
gainst rabiess
viruss after 12 months,
m
the results allow
w us to inferr a new vacccination straategy in doggs
prim
me vaccinated should bee adopted.
nowledgem
ments
Ackn
This work was supported
s
by the Grantt of Fundaçãão de Ampaaro a Pesquiisa do Estaddo de
São P
Paulo-FAPE
ESP (2009-09098-8). Municipalityy of Botucaatu, Environnmental Heaalth
Survveillance Deepartment
PT.0028
MPARAÇÃ
ÃO DA CO
OBERTURA
A VACINA
AL DE CÃE
ES E GATO
OS NAS
COM
CAM
MPANHAS
S ANUAIS DE VACIN
NAÇÃO CO
ONTRA A RAIVA, NO
N
MUN
NICÍPIO DE
D SÃO PA
AULO, PER
RÍODO DE
E 2004 A 20
012.
1 1
Parannhos NT1, Bernardi
B
F1, Silva EA1, Mendes MCNC
M
- Ceentro de Co
ontrole de
Zoonnoses da Cooordenação de Vigilânccia em Saúdde - Secretarria Municipal de Saúdee de
São P
Paulo
A Caampanha dee Vacinaçãoo Contra Raiiva em Cães e Gatos noo Municípioo de São Paaulo é
uma das atividaddes do Proggrama de Co
ontrole da Raiva
R
desenvvolvido dessde 1973 peelo
m duração de
d 14 dias e foi
Centtro de Contrrole de Zoonnoses. A Caampanha tem
desceentralizada a partir de 22004, quanddo passou a ser executaada pelos Seerviços
Regionalizados de Vigilânccia à Saúde (SUVIS), sob
s a Superv
visão e Cooordenação do
m essa estratéégia um maaior número
o de postos de
d
Centtro de Contrrole de Zoonnoses. Com
vacinnação foi diisponibilizaddo. O preseente trabalhho analisou a cobertura vacinal noss 96
Distrritos Admin
nistrativos (D
DA) do Muunicípio paraa os anos 20004 a 2012,, com o objeetivo
de veerificar a ten
ndência de cobertura vacinal
v
e se a interrupçãão da campaanha em 2010 e
20111 acarretou em
e diminuição na vacinnação de 20012. Compaarando os annos de 20044 e
20099 houve um aumento dee 17,23% no
o total da coobertura vaccinal. Em 20
008 observoou-se
uma diminuição
o de 3,86% do
d número de cães vaccinados e auumento de 7,7% na
nha foi suspeensa no seuu
vacinnação de fellinos em rellação a 20077. Em 20100, a Campan
quartto dia devid
do à notificaação de evenntos adversoos acima doo esperado tendo
t
sido
vacinnados 24,755% (247.5500 animais) do
d total da m
meta estimaada. No ano de 2012 a
vacinna voltou a ser disponibbilizada e a Campanhaa foi viabilizzada no perííodo de 21 de
d
maioo a 3 de junh
ho. Os resulltados obserrvados mosttraram um decréscimo
d
de 16,6% no
n
númeero total de animais vacinados em
m relação a 2009,
2
sendo a diminuição de 21,188%
para cães e um acréscimo
a
2
2,27%
para gatos.
g
Obseervou-se aum
mento de feelinos vacinados
ndo se compparou 2012 e 2009, senndo 31 destees pertencenntes a áreass de
em 442 DAs quan
maioor exclusão (áreas homoogêneas 4 e 5). Nos DA
As Iguatemii, Pedreira, Perus e Jaççanã
a vaccinação de felinos
f
foi respectivamente 21,23%
%, 31,41%, 38,3% e 400% superiorr a
20099. Houve dim
minuição dee cobertura vacinal de gatos
g
em 53
3 Distritos de
d áreas de
maioor inclusão como
c
Tatuaapé, Santa Cecília,
C
Saúdde e Santana. Em relaçção à vacinaação
de cãães, em 2012 houve um
m decréscim
mo em geral,, sendo que em 29 DAss a vacinaçãão foi
inferrior a 70% do
d total de 2009.
2
Somen
nte os DAs Marsilac, Vila
V Curuçáá e Bela Vista
vacinnaram mais cães do quee em 2009. Observa-see uma tendêência anual de
d crescimeento
na coobertura vaccinal do muunicípio, senndo mais evvidente em felinos.
f
Com
m a suspensão da
camppanha por dois
d anos consecutivos, verificou-sse um decréscimo no tootal de anim
mais
vacinnados em 20012. Sugeree-se estudoss periódicos de estimatiiva das popu
ulações cannina e
felinaa a fim de se
s avaliar a relação
r
entrre número de
d animais vacinados
v
e população de
cães e gatos.
PT.0029
EXPLORA
ATION OF THE KNO
OWLEDGE
E, ATTITU
UDES AND
D
AN E
PER
RCEPTION
NS OF NON
N- MEDICA
ALLY TRA
AINED, AD
DULT GRE
ENADIAN
NS
ABO
OUT ZOON
NOTIC DIS
SEASES
Bidaaisee S1 - 1Stt. George's University - Departmeent of Publicc Health and
d Preventive
Mediicine
d to be awarre
Whether people own dogs, cats, birds, reptiles, rabbbits or fishh, they need
d
an
nimals can have
h
an effeect on their health by trransmitting certain diseeases.
that domestic
Anim
mal diseasess, that are zooonotic diseeases, are thhose naturallly transmittted from
verteebrate animaals to humaans. The aim
m of the reseearch was too explore th
he perceptionns of
the loocal Grenaddian public about their knowledge towards ceertain zoonootic diseasess. In
Grennada, there are
a known reservoirs
r
fo
or zoonotic diseases in the form off rabies from
m
strayy dogs and mongoose.
m
R
Rabies
is a fatal
f
viral disease
d
that infects
i
the brain
b
and sppinal
cord in mammalls includingg humans. The
T virus is transmitted
d through thee saliva of a
b
The stuudy populattion comprissed adults, non-health
n
rabidd animal, ussually by a bite.
profeessional, Grrenadian citiizens in thee parish of St.
S George. A total of 450 participaants
compprised the saample base for the colllected data. Rabies wass correctly identified ass
beingg caused byy a virus am
mong 28.4% of the sampple populatiion. Rabies as obtainedd
from
m the bite of an infectedd animal was correctly iidentified by
b 434 (96.44%) of
particcipants. Kn
nowledge off the mongo
oose as the aanimal reserrvoir of Rab
bies in Grennada
was ccorrectly reecognized byy 434 (96.4%) of the participants. Rabies prev
vention,
requiiring vaccinnation of aniimals, was identified
i
b 339 (75.33%) of the study
by
s
populaation.
For Rabies,
R
20 (4.4%)
(
of thhe populatio
on selected nnervous reaactions as th
he associatedd
sympptom.
PT.0030
BIES ANTIIGEN SPRE
EAD AMO
ONGST AP
PPARENTL
LY HEALT
THY DOGS
S IN
RAB
NIG
GERIA: A REVIEW
R
Ogunnkoya AB1, Aina OA2, Adebayo OO
O 3, Oluwaagbenga AA
A4, Tirmidhii AB5, Auduu
6
7
3
7 1
SW , Atuman YJ
Y , Babasolla OO , Garrba A - Ahhmadu Bello
o Universityy, Zaria, Niigeria
2
- Vetterinary Meedicine, Deepartment off Zoology, F
Faculty of Science,
S
Unniversity of
3
Ibadaan, Ibadan, Oyo State, Nigeria - Zoology,
Z
Deepartment of
o Veterinarry Public Heealth
and P
Preventive Medicine,
M
F
Faculty
of Veterinary
V
M
Medicine,
University
U
off Ibadan, Ibadan,
Oyo State, Nigeeria - Veterinnary Publicc Health andd Preventivee Medicine, 4Departmeent of
Virollogy, Collegge of Mediccine, Univerrsity of Ibaddan, Ibadann, Oyo Statee, Nigeria Virollogy, 5Deptt. of Animall Sciences, Faculty
F
of Agric,
A
Tarabba State Unniversity,
Jalinngo. - Animaal Sciences,, 6Dept. Vett. Surgery & Med, Fac.. Vet. Med. Ahmadu Bello
B
Univversity, Zariia-Nigeria - Veterinary Medicine, 7Diagnosticc and Extension Dept,
Natioonal Veterinnary Researrch Institutee, Vom- Niggeria - Diagnostic and Extension
E
MMARY
SUM
Rabies with a tim
meline histoory of over 2300 years BC is 100%
% preventab
ble. Yet oveer
a reported dead annuaally due to rabies.
r
Mosst of the deaaths are in thhe
55,0000 people are
developing coun
ntries, Indiaa and Africaa. In Nigeriaa, the first officially
o
documented report
o canine raabies was in
n 1925. The long historry of
of huuman rabiess was in 19112 and that of
rabiees and the ev
vidences off its endemiccity have beeen found inn all native dialects in
Nigeeria that described onlyy the violentt rabies. Ressearchers ovver the yearrs have repoorted
the ppresence of rabies
r
viruss antibodies in over 15.93 to 30.7%
% of unvacccinated dogss and
humaans respectiively, in souuth western part of Niggeria. Of reccent, researcchers in Niggerian
Univversities andd Research Institutes
I
arre experienccing and rep
porting casees of the
preseence of rabiies virus anttigen in the brains of appparently heealthy dogs and even inn the
wild life across the countryy. Some studdies of the pprevalence of
o rabies anttigens in thee
brainn and saliva of apparently healthy dogs slaughhtered for huuman consu
umption in
Nigeeria; revealeed a 28% coonsumed doggs in N/wesst, 31% - 444% from N/eeast and 24%
%
from
m N/central tested
t
positiive for rabiees antigen inn their brain
n.
Similarly, 6%-8% of the doogs had rabiies antigen in
i their salivva at the pooint of slaugghter.
n tested for rabies
r
antiggen revealedd prevalencee in mongooose to be 111%,
In wiildlife brain
jackaals 9%, squiirrels 8.3%,, hydrax linkk 17% and w
wild cats 166%.
The iintent of thiis review is to bring intto focus thee prevailing carrier statu
us of high
perceentage of Nigerian
N
doggs and wildlife and its possible
p
con
nsequences on human
beforre the rabiess interest grroups of thee internationnal communnity.
This is with a viiew to open facets for discussion
d
aand understaanding the epidemiolog
e
gy of
N
rabiees and to seeek for furtheer collaboraation on the situation off rabies in Nigeria.
It maay necessitaate drawing new protoccol for humaan prophylaaxis followinng bite from
m
appaarently healtthy dogs.
Key words: Ra
abies antigeen spread, Apparently
A
y healthy do
ogs, Nigeriia.
PT.0031
CUSSING THE RABIES SURV
VEILLANC
CE SYSTEM
M IN BRAZIL: AN
DISC
EXP
PERIENCE
E OF SURV
VEILLANC
CE TOWA
ARD BATS AFTER TH
HE HALT OF
MAS
SS VACCIN
NATION OF
O DOGS AND
A
CATS IN CAM
MPINAS, SA
AO PAULO
O
Dias RA1, De Lu
ucca T2, Roodrigues RC
CA3, Castaggna C3, Presotto D3, De Nadai DV3,
S5, Martins CM
C 5, Ferreiira F5 Fagree A4, Bragaa GB5, Guillloux AGA5, Alves AJS
1
Univversidade de
d Sao Pauloo - Departam
mento de Medicina
M
Veterinária Preventiva e
Saúdde Animal, 2Prefeitura de
d Campinaas - Vigilânccia Sanitária Norte, 3Prrefeitura de
Cam
mpinas - Cenntro de Conttrole de Zooonoses, 4Colorado Statee University
y, 5Universiidade
de Sãão Paulo - Departamen
D
nto de Mediccina Veterinnária Prevenntiva e Saúdde Animal
f
fragm
mentation thhan any otheer mammal,, and for thaat
Bats are less vullnerable to forest
ore bats are
reasoon, some species can diisperse to peri-urban orr urban areaas. Insectivo
abunndant in urbaan areas duee to the dennsity of artifficial roosts and insectss attracted by
b
city llights. Interr-species traansmission of
o the rabiess virus betw
ween bats caan occur, annd
this iis the most probable
p
mechanism of virus circuulation in baat populatio
ons. Bats caan
also ttransmit thee rabies viruus to other mammal
m
speecies, like dogs
d
and catts. With thee halt
of vaaccination campaigns
c
o dogs and cats in 20100, the imporrtance of raabies
of
surveeillance in bats
b has incrreased in Brrazil. In Cam
mpinas, Sao
o Paulo Statte, a passivee
surveeillance system for batss was impleemented in 1994 and raabies-positivve bats from
m the
famillies Molosssidae, Phylloostomidae and
a Vesperttilionidae were
w found inn a peri-urbban
area. In these areas, a vacciination blocckage in doggs and cats was
w recomm
mended afteer the
o the massiive vaccination campaiign in 2010. This contrrol strategy was able too
halt of
increease the prop
portion of vaccinated
v
a
animals
aboove a critical value whille in the resst of
the sttudy area itt did not. Thhe probability of infectiious contactt between bats
b and doggs or
cats w
was higher in the blockkage areas, evidencing the importaance of the implementaation
of coontrol measu
ures.
PT.0032
BIES IN NIIGERIA: A NEW PAR
RADIGM SHIFT?
RAB
Jahunn BM1, Ehimiyein AM
M2, Audu SW
W2, Orasetinn AT3, Adaw
wa DAY1 - 1Ahmadu Bello
B
2
Univversity Zariaa - Veterinaary Teachingg Hospital, Ahmadu Bello
B
Univerrsity Zaria Veterinary Meddicine, 3Ahm
madu Bello University
U
Z
Zaria
- Veteerinary Public Health and
a
Preveentive Meddicine
Rabies is an infecti
ious disease of the central ner
rvous
syst
tem (CNS
S) that kills
k
ov
ver 50,0000 people eacch year. The prevalencce
rate is
i between 15-20% in Nigeria,
N
how
wever, becaause of the problem
p
witth reportingg, the
true ppicture is diifficult to obbtain. Most of the casees are from the
t rural areeas and are not
n
reporrted to the relevant
r
authhorities. Reecent surveyy of dog braain from dog
g market acrross
threee states show
wed an increease in the prevalence
p
of rabies up
p to 54% froom Kaduna
Statee, 50 from Abuja
A
whilee Plateau Staate had 58%
% prevalencee. In additioon, the
emerrgence of assymptomatic rabies whhich after sevveral mousee inoculatio
ons will kill the
micee without sh
howing cliniical signs off rabies is caase of conceern. This paaper
demoonstrates thee public heaalth significcance of thee observed high
h
rabies prevalence
p
a
and
the ddangers of having
h
asym
mptomatic raabid dog in the society..
PT.0033
VA HUMA
ANA TRAN
NSMITIDA
A POR SAG
GUI-IPÚ/C
CE-BRASIL
L-2010
RAIV
Duarrte NFH1, Pinheiro KM
MA1, Rolim BN2, Targinno LMF1, Almeida
A
IL1, Duarte BH
H3 1
2
3
SES
SA-CE - Nuuvet, SESA
A-CE - LAC
CEN, UFER
RSA - FACU
ULDADE DE
D MEDIC
CINA
VET
TERINÁRIA
A
r
é umaa doença connsiderada de
d alta relevâância no âm
mbito da saúúde
Introodução: A raiva
públiica, causa grandes
g
prejuuízos econôômicos, posssui alto podder de letalidade. No Brasil,
nos aanos de 19990 a 2010, houve
h
575 caasos de raivva humana, destes 325 (56,52%) fooi na
regiãão Nordeste, sendo respponsável peelo maior núúmero de caasos, no Ceaará um totall de
44(13,53%). Deestes, 12(27,,27%) dos casos
c
a transsmissão foi por animaiis silvestres,,
x jacchus, auumentando a preocupaação ainda mais
m com essta espécie, já
j
saguiis Callithrix
que aas pessoas tem
t o hábitoo de capturaar e criar essses animais, sem ter a menor
m
noçãão do
riscoo. Desde o ano
a de 2004, houve uma mudança no perfil ep
pidemiológiico da raiva,
consiiderando um
ma redução da raiva urbbana e um aumento
a
no ciclo silvesstre. Objetiivos:
relataar um caso de raiva hum
mana em Ip
pu/CE no anno de 2010 e as ações desenvolvid
d
das.
Mateerial e Méttodos: Os daados obtidoos foram atraavés de inveestigação eppidemiológiica,
Miniistério da saaúde SVS, Unidade
U
de laboratório Animal-UN
NILAN, Sisstema de
inforrmação de agravos
a
de Notificação
N
R
: Ipu fica a 257 km de
- SINAN. Resultados
Fortaaleza, com uma
u populaação de 40.3300 habitanttes IBGE 20010. Ao passsar perto dee
uma árvore, umaa criança dee 11 anos fooi agredida no
n rosto porr um sagui, apesar da
com
mento. Apóós 60 dias, 16/11/2010,
1
graviidade da lessão, não foi iniciado neenhum tratam
inícioo dos primeeiros sintom
mas: dor de cabeça,
c
febrre, cansaço, sonolênciaa, falta de
apetiite, a criançaa foi levadaa ao hospital e tratada dde enxaquecca. No dia 17/11/2010 deud
se início a um qu
uadro convuulsivo, ficanndo internado por três dias. Os exames ante m
morte
t
RT-- PCR dos materiais:
m
saaliva, folícu
ulo piloso, liiquor foram
m
realizzados pela técnica
negaativos para raiva
r
e o tituulo de anticoorpos igual a 0,02UI/m
ml. No dia 26/11/11 o garoto
g
foi a óbito e o reesultado póss – morte fo
oi positivo para
p raiva pela técnica de IFD. Forram
v
sobrre raiva paraa alunos do
realizzadas palesttras educatiivas e apreseentação de vídeos
ensinno fundamental e médioo (1341), prrofessores e diretores, população,
p
treinamentoo
sobree raiva paraa profissionaais do PSF, busca ativaa de expostoos com idenntificação dee 13
pessooas agredidaas (mão, péé e rosto). Dose
D
por saggui e um porr cão, desses apenas doois
haviaa iniciado o tratamentoo, os demaiss iniciaram ssomente apóós orientaçãão, recolhim
mento
de noove animaiss (saguis) mantidos
m
em cativeiro coom realizaçção de coletaa de sanguee e
salivva para diagnnóstico da raiva
r
com o resultado nnegativo. Coonclusão: Apesar
A
de exxistir
todass as ferrameentas para o controle daa doença, aiinda morrem
m pessoas vítimas
v
de raaiva.
É neccessário quee haja fortallecimento das
d ações dee vigilância,, atendimennto das pessooas
agreddidas em tem
mpo oportuuno, fiscalização e apliccação de leis em relaçãão à captura e
criaçção dos anim
mais silvestrres, integraçção, sensibilização da população
p
e relação ao
em
a
riscoo que os anim
mais silvesttres oferecem e que o luugar deles é no seu hab
bitat naturall.
PT.0034
CO E MONITORAME
ENTO DA RAIVA EM
M OLINDA
A
DIAGNÓSTIC
RNAMBUC
CO – POSIT
TIVIDADE
E PARA RA
AIVA EM MORCEG
GO NÃO
PER
HEM
MATÓFAG
GO
Machhado JL1, Araújo
A
ACR
R2, Ribeiro MGB
M 3, Silvaa LAM4, Olliveira JCT5, Gomes ES
S6,
7 1
2
3
Bolleer MAA - LACEN - PE,
P LACE
EN/PE / LAN
NAGRO/PE
E, Universiidade Federral de
5
Pernaambuco, 4GEMNE,
G
U
UFPE/CAV
/ GEMNE, 6Secretaria Municipal de Saúde de São
José do Egito, 7Fundação
F
O
Oswaldo
Cruuz, Institutoo Nacional de
d Controle de Qualidaade
Saúde - Imunologia
em S
R
a Caapital
A ciddade de Olinnda, localizzada na Nordeste do Brrasil, distantte 6 km de Recife,
Pernaambucana, tem como principal
p
carracterística,, o pólo turíístico. A pop
pulação é
estim
mada em 3755. 559 habittantes. A cid
dade alta dee Olinda é formada
f
porr extensa áreea
verdee, o que prooporciona a presença e a permanênncia de quirópteros. Noo mês de Abbril
de 20012, foi enccaminhada para
p o diagn
nóstico da raaiva, um qu
uiróptero da espécie
Moloossus molosssus, fêmea,, adulta. Seg
gundo as innformações coletadas
c
noo Formuláriio
Únicco de Requissição de Exxames para Síndrome
S
N
Neurológica
, o morcegoo foi enconttrado
em uuma residência no Bairrro Amaro Branco
B
e aprresentava inncoordenaçãão e alteraçãão
compportamentall. O proprieetário da residência connfirmou quee não houve agressão ouu
contaato com o animal,
a
que veio a óbitoo. A prova de
d Imunoflu
uorescência Direta
diagnnosticou positividade para
p o vírus para raiva, o que foi coonfirmada na
n Prova
Biolóógica. Podeemos entendder que o creescimento desordenado
d
o e o desmaatamento poodem
corrooborar com o aumento de casos dee morcegos positivos paara a raiva. A vigilânciia por
sua vvez, se faz necessária
n
p que hajaa o controlee epidemiológico eliminação da dooença
para
atravvés do moniitoramento do
d vírus rábbico na regiãão através do
d envio de amostras dee
anim
mais suspeito
os, para o diiagnóstico. No Nordestte, mais preecisamente na
n Cidade de
d
Floreesta, Sertão Pernambuccano, um gaaroto de 14 aanos foi agrredido por um
u morcegoo
hemaatófago e innfectado pello vírus da raiva
r
em outtubro de 20008. O protoocolo utilizaado
foi uma adaptaçção do Protoocolo de Minwalker, e ddesta formaa foi criado o Protocoloo de
M
c
como
ficou conhecido,
c
foi o segundo no mund
do de cura da
d
Reciffe. O caso Marciano,
raivaa, porém com
m sequelas.. A partir daaí, as autoriddades sanitáárias intensiificaram a
vigilâância na tenntativa de evvitar que ouutros episóddios sucedam
m. O fato occorrido em
Olindda alerta as autoridadess sanitárias para o riscoo de contraiir a raiva, attravés de
espéccies de quirrópteros incllusive não hematófago
h
os. A educaçção ambienttal para a
popuulação em geral, se faz necessária para
p que as espécies seejam presevadas em funnção
da suua importânncia no meioo ambiente, mas, que seejam observvadas as orientações e
recom
mendações relacionadaas à presençça destes aniimais.
PT.0035
MPORTÂN
NCIA DA AVALIAÇÃ
A
ÃO SOROL
LÓGICA PARA
P
RAIV
VA E A
A IM
IMP
PLANTAÇÃ
ÃO DA TÉ
ÉCNICA NO
O ESTADO
O DE PERN
NAMBUCO
Arauujo ACR1, Machado
M
JL
LM2, Ribeiroo MGB3, Silva LAM4, Oliveira JC
CT5, Gomes ES6,
7 1
Bolleer MAA - LACEN/LA
ANAGRO--PE - DIAG
GNÓSTICO DE RAIVA
A,
2
3
4
LAC
CEN/LANA
AGRO-PE, LANAGR
RO/PE, Gruupo de Estuddos de Morccegos no
Norddeste (GEM
MNE), 5UFPE
E/CAV/GE
EMNE, 6Seccretaria Mun
nicipal de Saúde de Sãoo
7
José do Egito - Vigilância
V
S
Sanitária,
Fundação
F
O
Oswaldo
Cru
uz, Instituto Nacional de
d
Conttrole de Quaalidade em Saúde - Imuunologia
na está na Liista Nacionnal de Comppulsória Imeediata – LNCI ANEXO
O II A Raaiva Human
13. R
Raiva Humaana; PORTA
ARIA N.º 2.472, DE 311 DE AGOS
STO DE 20010 (DOU de
d
1º/099/2010 Seçãão I pág. 50)), que definne as terminoologias adootadas em leegislação
nacioonal, conforrme disposto no Regulaamento Sannitário Intern
nacional 2005 (RSI 20005),
a relaação de doeenças, agravvos e eventoos em saúdee pública de notificaçãoo compulsórria
em toodo o territóório nacionaal e estabeleecer fluxo, critérios,
c
responsabilid
dades e
atribuuições aos profissionai
p
is e serviçoss de saúde. A Organizaação Mundiaal da Saúdee
consiidera que um
m título iguual ou superrior a 0,50 UI/mL
U
repreesenta um esstado imuniitário
suficciente para proteger
p
inddivíduos exp
postos ao rssico de contaaminação pelo
p vírus
rábicco.A determ
minação dos anticorpos neutralizanntes (AcN), deve ser reaalizada em
indivvíduos previiamente imuunizados e expostos
e
aoo risco de coontrair a raivva. A profillaxia
vacinnal pré-expoosição é inddicada para pessoas
p
quee, por força de suas ativvidades esteejam
expoostos perman
nentemente ao risco dee infecção pelo vírus ráábico. Todoss os indivídduos
perteencentes aoss grupos de risco que freqüenteme
fr
ente estão em
m contato com o vírus
rábicco devem seer avaliados a cada seiss meses.O Innstituto Pasteur, no Esttado de São
Pauloo, referência Nacional para a soron
neutralizaçãão dentre ouutras técnicaas, desenvoolveu
um teeste simplifficado de inibição de foocos fluoresscentes – SF
FIMT- que vem
v sendo
utilizzado para av
valiação sorrológica parra raiva.Nesste Sentido, o Laboratóório Central LAC
CEN/PE, vem
m através do
d apoio e daa parceria ccom o Instituuto Pasteur,, realizandoo
treinaamentos com profissioonais do Estado de Pernnambuco, coom o objetivvo de implaantar
a técnnica que fav
vorecerá o monitorame
m
ento de proffissionais en
nvolvidos coom o vírus
rábicco na Regiãoo que atuam
m na área dee saúde anim
mal tais com
mo, Médicoss Veterináriios,
Médiicos Sanitarristas, agenttes de endem
mias, agentees ambientaais nos maiss variados
ambiientes de traabalho tais como:
c
diagnnóstico e caampo capturrando, vacinnando,
identtificando e classificand
c
do animais passíveis
p
dee albergar o vírus, dentrre outros
profiissionais quue estão expostos ao vírrus. No Estaado de Pernambuco, maais precisam
mente
na Capital, Reciife, de acorddo com o levvantamentoo dos Centroos de Controole de
manda para determinação dos anticcorpos neutralizantes (A
AcN), é de
Zoonnoses, a dem
4.0000 amostras/ano.A impoortância da implantação
i
o da Técnica de dosageem de anticoorpos
anti-rrábicos hum
mano no Esttado, vem no
n momentoo em que, a Vigilância e o controlee da
raivaa em Pernam
mbuco são prioridades
p
no que se trrata de Vigiilância Epiddemiologia. Com
a creescente dem
manda desta avaliação soorológica, a implantaçãão da técnicca no LACE
ENPE, ppromoverá a otimização do diagnóóstico, faciliitará maior dinâmica no que diz
respeeito ao temppo de respossta do diagnnóstico, ao custo
c
benefi
ficio, e princcipalmente a
profiilaxia da raiiva humana em nossa Região.
R
Agraadecimentoss
Ao Innstituto Passteur, Dra Neide
N
Yumiee Takaoka, Dra Ivanetee Kotait e Dra
D Maria Luuiza
Carriieri que vem
m contribuinndo de form
ma incansáveel na constrrução deste Trabalho.
PT.0036
TRIBUIÇÃ
ÃO GEOGR
RÁFICA E CARACT
TERIZAÇÃ
ÃO MOLECULAR DE
DIST
VÍRU
US DA RA
AIVA DE IS
SOLADOS
S HERBÍVO
OROS PRO
OCEDENT
TES DO
NOR
RTE DO BR
RASIL
Peixooto HC1, Gaarcia AIE2, Silva SOS2, Ramos OS
S3, Silva LP
P3, Brandão PE2,
2 1
Richhtzenhain L - FMVZ USP
U - Veterrinary Preveentive Mediicine and Animal
A
Health,
2
FMV
VZ USP - Medicina
M
Veeterinaria Preventiva
P
e Saúde Anim
mal, 3Laborratório Naciional
Agroopecuário, LANAGRO
L
O, Pará
z
com
m 100% de letalidade que
q acometee todos os mamíferos,
m
é
A Raaiva é uma zoonose
impoossível estim
mar os custoos reais de perdas
p
na peecuária, prin
ncipalmentee pelo elevaado
númeero de subn
notificações.. As ferrameentas molecculares perm
mitem identiificar as
linhaagens virais circulantess e associar essas linhaggens à origeem geográfiica. Foram
analiisadas 17 am
mostras isolladas de bovvino (11), eqquídeo (4) e bubalino (2),
( no períoodo
de 20004 a 2008, procedentees dos Estad
dos do Pará,, Tocantins e Rondôniaa. As referiddas
amosstras foram submetidass a RT-PCR
R tendo com
mo alvo um segmento
s
doo gene da
glicooproteína, reeferente a um
m fragmentto de 917pbb. Os amplifficados obtiddos foram
purifficados e seqquenciados, o dendrogrrama foi geerado utilizaando-se critéério de
otimiização de distância com
m algoritmoo Neighbor--Joining e modelo
m
evolu
utivo Kimuura 2parâm
metros, emp
pregando-see 1000 repettições de boootstrap. A análise espaacial dos
isolaados investiggados no prresente estuddo foi elaboorado, a parttir de mapas retirados do
d
IBGE
E, com auxíílio do proggrama ArcG
Gis versão 9..3. Todas ass 17 amostraas de vírus da
d
raivaa foram geneticamente agrupadas dentro
d
do clluster de Deesmodus rottundus. Denntro
dessee cluster os isolados doo presente estudo se agrruparam em
m seis subgruupos princippais:
subggrupo I, com
mposto por isolados oriuundo de Toccantins e um
m isolado prroveniente da
d
mesoorregião sud
deste do Parrá; subgrupoo II, composto de isolaado bovino procedente
p
de
Tocaantins, agrup
pou-se com
m isolados dee Desmoduss rotundus e Diphilla ecaudata
e
originários de São Miguel Tapuio
T
no Piauí;
P
subgrrupo III, form
mado por issolados
T
e um
u isolado procedente de Goiás; subgrupo
s
IV
V, compostoo por
originários de Tocantins
fo
porr amostras
isolaados de diveersas localiddades de Tocantins; subbgrupo V, formado
oriunndas de diveersas localiddades de Ro
ondônia, as quais apresentaram sim
milaridade com
c
isolaados procedeentes de Gooiânia e São
o Paulo; subgrupo VI, composto
c
poor isolados
proceedentes do Nordeste
N
Paaraense e isolados de suurtos de raivva humana ocorridos no
n
Maraanhão. A fillogenia dem
monstrou que todas as liinhagens viirais dos iso
olados do
preseente estudo estão relaciionadas à reegião geográáfica.
Supoorte Financeeiro: CAPES
S
PT.0037
RIE HISTÓ
ÓRICA DO ATENDIM
MENTO AN
NTI-RÁBIC
CO HUMA
ANO - UMA
A
SÉR
ANÁ
ÁLISE DAS
S NOTIFIC
CAÇÕES NA
N 19ª CRS
S - FREDE
ERICO WE
ESTPHALE
EN
RS.
Dornnelles M1 - 119ª Coordeenadoria Reegional de Saúde
S
- Vigiilância Ambbiental em Saúde
S
SÉRIIE HISTÓR
RICA DO ATENDIME
A
ENTO ANT
TI-RÁBICO
O HUMANO
O - UMA
ANÁ
ÁLISE DAS
S NOTIFICA
AÇÕES NA
A 19ª CRS - FREDERIICO WESTP
PHALEN RS.
R
Varggas J C A¹, Dornelles
D
M L², Bagatiini T³
¹Vigiilância Epiddemiológicaa ǀ E-mail: [email protected]
²Vigiilância Ambbiental em Saúde
S
ǀ E-m
mail: mauro--dornelles@
@saude.rs.goov.br
³Imuunizações ǀ E-mail:
E
imuunizacao19@
@saude.rs.ggov.br
INTR
RODUÇÃO
O
O Attendimento Anti-Rábicco Humano, é um dos Agravos
A
de maior notifficação nos
muniicípios da árrea de abranngência da 19ª CRS.
A 199ª CRS é connstituída poor 28 municípios, localiizada na reggião do méd
dio alto urugguai,
sendoo Frederico
o Westphaleen o municíppio sede da 19ª CRS.
O Attendimento Anti-Rábicco Humano ocorre em diversos
d
perríodos do anno, após as
pessooas sofrerem
m agressõess principalm
mente por cãães e gatos, necessitanddo a utilizaçção de
vacinna e soro an
nti-rábicos para
p a preveenção da raivva.
OBJE
ETIVOS
Conhhecer o perffil epidemioológico dos atendimenttos através da
d avaliaçãoo de alguns
aspecctos do Aten
ndimento Anti-Rábico
A
Humano naa 19ª CRS de
d 2007 a 2011.
Tom
mar medidas preventivass, buscandoo a diminuiçção do númeero de dosess de vacinass,
atravvés da corretta observação do animal agressor,, bem como da correta indicação do
d
imunnobiológico.
MÉT
TODOS
Foi rrealizado um
m estudo obbservacionall descritivo de análise quantitativa
q
a, analisandoo as
inforrmações conntidas no Siistema de Innformações de Agravoss de Notificação (SINA
AN ),
no peeríodo de 20
007 a 2011,, da ocorrên
ncia nos munnicípios perrtencentes a 19ª CRS. No
N
períoodo foram notificados
n
n SINAN 1822 casos de Atendim
no
mento Anti-R
Rábico Hum
mano.
RESU
ULTADOS
S
Dos 1822 casos de Atendim
mento Anti-Rábico Hum
mano 54,9%
% foram do sexo mascuulino,
% a espéciee agressora foi
f o cão, a
sendoo 50,8% mooradores daa zona urbanna, em 94,4%
conddição do anim
mal para finns de condu
uta do tratam
mento em 866,2% eram sadios, 94,77%
dos aanimais eram
m passíveis de observaação, sendo que 53,1% do tratamennto indicadoo foi
obserrvação + vaacina, sendoo 3019 doses de vacinas aplicadas no período.
CON
NCLUSÕES
S
A auumento no número
n
de Notificações
N
s, bem comoo a quantidaade de Vaciinas utilizaddas,
tem levado
l
a Vigilância em
m Saúde da 19ª
1 CRS a innvestir na sensibilizaçã
s
ão de
profiissionais de saúde e enttidades direeta e indiretaamente envo
olvidas, atraavés de curssos
de caapacitação em
e sala de vacinas,
v
saliientando paara que nos municípios
m
haja
profiissionais quue efetivameente realizem
m a correta observação
o do animal agressor beem
comoo a correta indicação
i
doo imunobio
ológico, dim
minuindo as doses de vaacinas utilizzadas,
consiiderando a atual
a
ausênccia de circu
ulação de vírrus rábico em
e populaçõ
ões de caninnos
doméésticos no Rio
R Grande do Sul.
BIBL
LIOGRAFIA
A
ARA
AÚJO, F.A. A Situaçãoo da Raiva no
n Brasil. Inn: Seminárioo Internacioonal da Raivva,
20000. p. 22.
ATL
LAS de Saúdde do Brasill, Brasília: Secretaria
S
d Vigilânciia em Saúdee, 2004.
de
GUIA
A de Vigilâância Epidem
miológica. Brasília:
B
Fuundação Naccional de Saaúde, 2002. 920
p.
mas de Vaciinação. Brasília: Fundaação Nacionnal de Saúdee, 2002.
Manuual de Norm
PT.0038
DUÇÃO DA
A EXPOLIAÇÃO E DO
D RISCO
O DE OCOR
RRÊNCIA DA RAIVA
A
RED
EM HERBÍVO
OROS PELO CONTR
ROLE POP
PULACION
NAL DE DE
ESMODUS
S
TUNDUS NA
N REGIÃO NOROE
ESTE DO ESTADO
E
D RIO DE
DO
E JANEIRO
O.
ROT
ROM
MIJN PC1, Bruno
B
LM de
d P2, Carneeiro I de C, Bruno AM de P2, Cattaaneo CAM,,
Rubiião EC, Kim
mura LMS3 - 1PESAGR
RO-RIO - Centro
C
Estaddual de pesqquisa em
Saniddade Animaal, 2SEAPEC, 3PESAG
GRO-RIO
A preevenção da difusão de enfermidaddes infecto-ccontagiosass pelo controole populacional
dos ttransmissorees/vetores é uma práticca secular dee comprovaada eficácia, seja aplicaada a
roedoores, carnívvoros, inverttebrados e pragas
p
de veegetais. Paraa reduzir aoo máximo a
expoosição de heerbívoros à espoliação
e
p morcegoos hematófaagos da espécie Desmoodus
por
rotunndus e conseqüentemennte, ao risco
o de infecçãão por Lyssaavirus, foram
m identificaados
abriggos com collônias de moorcegos hem
matófagos, e estes monnitorados sem
mestralmennte,
com capturas e controle
c
poppulacional eventuais,
e
e três micrrobacias do Estado do Rio
em
R
de Jaaneiro. Verificou-se que na microbbacia de Mirracema, os casos de Raaiva transmiitidos
por D
D. rotundus foram se reeduzindo a partir
p
de 19995, quandoo foi iniciadoo o
moniitoramento dos abrigoss e controle populacionnal da espéciie. No interrvalo de 36
mesees (anos de 2005
2
a 20077) em que não
n houve controle
c
poppulacional, os
o casos de
Raivva em herbívvoros tornarram a se eleevar. Verificcou-se que o repovoam
mento dos abbrigos
moniitorados lev
vava de 2 a 5 anos. Na microbacia
m
envolvendoo o municíp
pio de Aperiibé, o
moniitoramento dos abrigoss e eventual controle poopulacional por 30 messes coincidiuu
com a ausência de Raiva em
m herbívoroos ao longo de 5 anos (d
de 1996 a 2000).
2
Tambbém
c
de Raaiva em herbbívoros durrante o
não fforam notifiicados/diagnnosticados casos
moniitoramento dos abrigoss e eventual controle poopulacional,, realizado por
p 30 mesees
nos municípios
m
de Três Rioos e Sapucaiia. Conclui--se que o coontrole popuulacional dee D.
rotunndus por téccnicos experrientes reduuziu substanncialmente o contato diireto dos
morccegos com animais
a
dom
mésticos sussceptíveis nas
n microbaccias trabalhadas, e
conseeqüentemen
nte o risco de
d infecção quando da presença dee Lyssaviruss na saliva dos
d
morccegos. Enquuanto era reaalizado o coontrole popuulacional biianual de D.. rotundus nos
n
três aambientes distintos
d
e issolados, nãoo foram diaggnosticadoss casos de Raiva
R
em
herbíívoros doméésticos ao loongo dos peeríodos de monitorame
m
ento. Em não
o se realizanndo
o conntrole popullacional durrante um míínimo de 488 meses, cassos de Raivaa em herbívvoros
passaaram a acon
ntecer nessees ambientess sob estudoo. Em nenhuum dos D rootundus
exam
minados foi encontrado Lyssaviruss na saliva pela
p IFD, neem anticorp
pos específicos
contrra a virose no
n soro de 163
1 indivíduuos examinaados. É imp
prescindívell ampliar a
discuussão quantoo ao significado epizoó
ótico da detecção de annticorpos coontra antígennos
de Lyyssavirus em
m mamíferoos silvestress, e a possibbilidade desttes estarem infectados sem
apressentação de quadro clínnico (“portaadores assinntomáticos”)).
PT.0039
TORES ASS
SOCIADO
OS À OCOR
RRÊNCIA DE RAIVA
A EM REG
GIÃO RUR
RAL
FAT
DE C
CAMPINA
AS/SP/BRA
ASIL EM 2.001 e 2.0022.
Ram
mos LHM1, Donalísio
D
M 2, Lourennço RW3 - 1Prefeitura Municipal
MR
M
d Campinass de
Secreetaria Muniicipal de Saúúde, 2UNIC
CAMP - Uniiversidade Estadual
E
de
Cam
mpinas/Faculldade de Ciêências Méd
dicas - Depaartamento dee Saúde Colletiva, 3UNE
ESP
- Uniiversidade Estadual
E
Paaulista/Faculldade de Enngenharia Ambiental
A
e
virral, com altaa taxa de lettalidade e ocorrência
o
em
m
A raiiva é uma zoonose de etiologia
bovinnos e equídeos, que cauusa prejuízo
os sociais e econômicos, tendo com
mo transmisssor
o moorcego Desm
modus rotunndus. Em árrea rural do município de
d Campinaas estado dee São
Pauloo, foram reggistrados 299 casos de raaiva confirm
mados laborratorialmentte em
herbíívoros, em 2001
2
e 20022. Técnicos da Secretarria de Saúdee e da Agriccultura
realizzaram um trrabalho inteenso e conjuunto na prevvenção da raaiva animall e humana na
n
regiãão. O objetiv
vo deste esttudo é descrrever o perffil epidemioológico da ep
pizootia e
analiisar variáveiis associadaas aos óbitos na área dee ocorrênciaa. Foi realizzado estudo
transsversal com a aplicaçãoo de questio
onário a 7300 proprietáriios e morado
ores de imóóveis
da árrea, sendo que
q 5 propriedades não foram incluusas. As varriáveis colettadas foram
m:
espéccie animal acometida,
a
ó
óbitos
na prropriedade nos
n últimoss 6 meses, ex
xistência e tipo
t
de abbrigos de morcegos
m
hem
matófagos, número de pessoas exppostas ao víírus. Foram
calcuulados coefiicientes de mortalidade
m
e por raiva por
p espécie animal por 1000
indivvíduos. Foi ajustado
a
moodelo de reg
gressão logíística múltippla, tendo como variávvel
respoosta a ocorrência de óbbitos por raivva, e variávveis preditorras as acimaa referidas,
utilizzando-se os programas EPI-INFO 6.04 e, R para digitaçãão e análisess estatísticaas. O
coefiiciente de mortalidade
m
(CM) total foi de 2,3/11000 animaiis (n = 82300), em equíddeos
foi 17,9/1000 e em bovinoss 1,97/1000. Em 81(11,,1%) das prropriedades foi referidaa a
existtência de abrigos de moorcegos, senndo 27 (46,66%) do tipoo árvore, 25 (43,1%)
consttrução habittada e 6 (100,34%) consstrução abanndonada. Fooram contab
bilizadas 12278
pessooas que lidaavam com os
o animais, sendo
s
que 38
3 exerciam
m atividades nas
proprriedades com óbitos, portanto exp
postos diretaamente ao riisco de raiva humana. As
A
variááveis estatisticamente significantes
s
s associadass aos óbitos obtidas no modelo forram:
existtência de abrigos de moorcegos hem
matófagos nna propriedaade (OR = 3,2 IC 95% 1,03
– 9,991) e a ocorrrência de óbbitos em aniimais nos últimos
ú
6 meeses (OR = 32,64 IC 955%=
9,11 – 116,90). A análise de
d CM por espécie
e
anim
mal pode traazer informaações relevaantes
C em equíddeos 9 vezees maior quee o de bovin
nos na área,,
na reegião. Obserrvou-se o CM
diferrente de daddos da literatura. O estuudo desta eppizootia mosstra o poten
ncial de
ocorrrência em humanos
h
vivvendo e trab
balhando naas proximidaades dos casos em
anim
mais. A referrência de moortes de aniimais em peeríodo recen
nte (6 mesess) foi predittora
de raaiva, sugerinndo a circullação préviaa do vírus e a falta de conscientização dos
proprrietários em
m manterem os animais vacinados. A associaçção entre a raiva
r
animall e
preseença de morrcegos tem sido relatadda em outros estudos, reeforçando a necessidadde de
moniitoramento de colôniass de morcegos em área rural. As açções de vigiilância
epideemiológica,, educação em
e saúde e profilaxia animal
a
e hum
mana pressuupõem açõees
intersetoriais de relevância para a prevvenção e conntrole da raiiva humanaa e animal.
PT.0040
CO Y CARA
ACTERIZA
ACIÓN MO
OLECULA
AR DE VIR
RUS RÁBIC
CO
DIAGNÓSTIC
MUESTRA
AS EN AVA
ANZADO ESTADO
E
D DESCO
DE
OMPOSICIIÓN
EN M
Beltrrán F1, Gury
y Dohmen F1, Del Pietrro H2, Cisteerna DM3 - 1Instituto dee Zoonosis Luis
Pasteeur - Diagnóóstico, 2Servvicio Nacioonal de Saniidad y Calid
dad Agroalim
mentaria 3
Proggrama Nacioonal de Rabiia Paresiantte, INEI-AN
NLIS Dr. Carlos
C
G. Malbrán
M
Serviicio de Neuurovirosis, Buenos
B
Airees, Argentinna
Argentina ex
xiste una Reed de Laborratorios Reggionales de rabia
r
que reealizan el
En A
diagnnóstico de rutina
r
en aniimales a parrtir de muesstras de cereebro median
nte
inmuunofluoresceencia directta (IFD) y aislamiento
a
m
mediante
ennsayo biológico en ratoones
lactaantes (EBRL
L). Las conttramuestras de los posittivos y/o los aislamienttos de RAB
BV se
envíaan al Laboraatorio Nacioonal de Refferencia quee es el Serviicio Nacionaal de Sanidaad y
Caliddad Agroaliimentaria (S
SENASA) o al Institutoo de Zoonossis Luis Passteur (IZLP))
perteeneciente al Gobierno de
d la Ciudadd Autónomaa de Buenoss Aires (GC
CABA), parra
identtificar la esp
pecie reservvorio usandoo un panel de
d anticuerp
pos monoclo
onales
(caraacterización
n antigénica)). La toma, conservacióón y transpoorte adecuaddos de las
muesstras y/o aisslamientos son
s factoress decisivos para
p la correecta realizacción e
interppretación dee estos ensaayos. Las elevadas tem
mperaturas, como
c
las qu
ue se registraan en
las prrovincias deel norte de Argentina,
A
p
pueden
ocasionar el deeterioro de los cadáverees de
los annimales inv
vestigados, provocando
p
o que las muuestras de ceerebro preseenten desdee una
licueefacción leve hasta un avanzado
a
esstado de desscomposició
ón. Estas co
ondiciones
afecttan la sensibbilidad de laas pruebas diagnósticas
d
s dado que provocan
p
laa degradacióón de
la esttructura viraal y la produucción de to
oxinas bacteerianas. Asiimismo, si los
l aislamieentos
de RA
ABV no se conservan a muy bajass temperatuuras (-70°C)), pierden ráápidamentee su
viabiilidad lo quee ha provoccado la pérddida de muchhas coleccioones de RA
ABV en
laborratorios quee carecen dee la infraestrructura adeccuada. Se evvaluó una téécnica de RTR
PCR
R de un pasoo para el diagnóstico y caracterizac
c
ción molecu
ular en muesstras de tejido
cerebbral en avannzado estadoo de descom
mposición y en aislamieentos antiguuos. Se tom
mo un
grupoo de 10 cep
pas de rabia aisladas en cerebro de ratón lactannte, de las variantes
v
de
mayoor circulació
ón en nuestrro país, 3 ceerebros caninos expuesstos a descoomposición
contrrolada y 14 cepas antigguas. La caracterizaciónn antigénicaa se realizó mediante laa
técniica de inmun
nofluoresceencia indireccta usando un
u panel dee 19 anticuerrpos
monooclonales (C
CDC, USA)). La caracteerización molecular
m
dee una regiónn de 159
nucleeótidos corrrespondiente al gen de la nucleoprroteína fue analizada
a
y se confecciionó
un árrbol filogen
nético. La caaracterizació
ón antigénicca y molecu
ular se correespondió enn
todoss los aislam
mientos. En este
e estudioo pudo efecttuarse la carracterización
n molecularr de
los aislamientoss de mayor circulación
c
en Argentinna, en muesstras en avan
nzada
ue utiliza unna
descoomposiciónn y en cepas antiguas enn forma direecta, con unna técnica qu
pequueña porciónn del gen dee la nucleopproteína viraal en el 100%
% de las mu
uestras.
PT.0041
TIGENIC VARIANTS
V
S OF RABIIES VIRUS
S IN VENE
EZUELA. 2000-2012
ANT
PERE
EZ M1, HID
DALGO M1, BOYER L1 - 1Nationnal Institute of Agriculttural Researrch
(INIA
A) - Rabies Laboratoryy
ype of the geenus
Rabies is a fatal zoonotic diisease, caussed by the raabies virus, the prototy
he Rhabdoviiridae familly, with a sinngle-strandded negativee-sense RNA
A
Lyssaavirus of th
genoome, surroun
nded by a bullet shape capsid. In Venezuela
V
f many yeears rabies has
for
h
occuurred in endeemic and eppidemic form
m, constitutting a socioeconomic problem
p
thatt
affeccts human health
h
and caauses lossess in livestocck. It is distrributed throoughout the
counntry. The dettection of raabies antigeen and antiggenic characcterization of
o field straiins
allow
wed the idenntification of
o animal sppecies that serves as a reservoir ressponsible foor an
outbrreak of rabiies in a giveen area. Thee aim of thiss study was to perform the antigennic
charaacterizationn of 34 fields isolates off rabies viruus from diffferent animaal species, states
s
and yyears, to know which antigenic
a
vaariants were circulating in our counntry. The
detecction of rabiies antigen was perform
med by direect immunoffluorescence test of nerrve
tissuee imprints of
o animals with
w sympto
oms of the disease.
d
Thee viral amplification waas
perfoormed by innoculation inn suckling mice.
m
Antigenic characcterization was
w perform
med
by inndirect imm
munofluoresccence impreessions brainns of mice inoculated
i
w field sttrains
with
that had
h obviouss symptomss of the diseease. Only variants
v
1 annd 3 were fo
ound. It wass
concluded that the
t antigenic variant 1 (canine) waas located exxclusively in
i Zulia Statte,
(
w present in several states,
was
s
so thee common vampire
v
batt D.
whilee variant 3 (vampire)
rotunndus was th
he main trannsmitter of rabies
r
for livvestock in th
hat period.
Ackn
nowledgem
ment: Nationnal Institutee of Agriculltural Reseaarch (INIA),, National
Instittute of Integ
gral Agriculltural Health
h (INSAI). Funding: INIA.
I
Mattos C. OM
MS pg 30(1989). Hidaalgo M. Revv. Fac. Cs Vets
V UCV 466:33.
DeM
(20044) Hidalgo M. Rev. Faac. Cs. Vetss. UCV. 49((2):121.(20008). Meslin FX WHO 476p
(1996). Hidalgo M. Med.Vet al día. 1:19 (2011).
PT.0042
PERIMENT
TAL ANTIIVIRAL TH
HERAPY AGAINST
A
DIFFEREN
NT RABIE
ES
EXP
VIRU
US LINEA
AGES USIN
NG TRANS
SFECTION
N WITH AN
NTI-RABIIES
ANT
TIBODIES
Castiilho JG1, Baatista HBCR
R1, Rodriguues AC1, Caarnieli Jr P1, Oliveira RN
N1, Silva ACR
A 2,
2
1
1
3 1
Capoorale GMM , Carrieri ML
M , Kotaitt I , Brandãoo PE - Insstituto Pasteeur - Virologgia,
2
Instiituto Pasteuur - Imunoloogia, 3Faculldade de Meedicina Veteerinária e Zootecnia daa
Univversidade dee São Paulo
t develop a new mechhanism for antiviral
a
theerapy againsst
The aim of this study was to
w a cationnic reagent (lipofectami
(
ine
rabiees based on the introducction by traansfection with
20000) of antiboddies into neuuronal cellss infected with
w the rabiees virus. N22A cells werre
infeccted using 96-well platees and diffeerent viral cooncentrationns (0.1, 1.0,, 10 and 1000
TCID
D50) of threee lineages of
o rabies viru
us circulatinng in Brazill (dog, Desm
modus rotunndus
and E
Eptesicus fu
urinalis). Affter incubation for 24 hh, the cells were
w transfeected with antia
rabiees-virus poly
yclonal antiibodies and lipofectamine 2000. These
T
cells made
m
up thee
treatm
ment group (TG). The cells in the negative coontrol group
p (CG) weree treated wiith
only Minimum Essential
E
M
Medium.
Aftter 11 hourss, the plates were fixed with 80%
acetoone and anaalyzed by direct immunnofluorescennce using a fluoresceinn isothiocyannateconjuugated antinnucleocapsiid rabies anttibody. The effectiveneess of the traansfection and
a
subseequent neuttralization of
o the virus was
w determ
mined by calcculating thee percentagee
inhibbition of fluorescent focci. This wass done by m
measuring thhe difference in the num
mber
of fluuorescent fooci in the tw
wo groups (C
CG and TG). The resullts show thaat for lower viral
concentrations (0.1
( and 1.0 TCID50), viral
v
inhibitiion was 100
0% for all thhe lineages
gher virus concentratio
c
ons were useed (10 and 100 TCID500), inhibitionn
testedd. When hig
varieed accordingg to the viraal load and lineage
l
of raabies virus used. With an infectiouus
dose of TCID50, inhibition varied
v
from
m 82.7% to 100%
1
for th
he lineages tested.
t
Withh a
TCID50 dosse, inhibitionn was 90.7%
% for the D.. rotundus lineage,
l
90.33% for the dog
d
100 T
lineaage and 67.0
0% for the E.
E furinalis lineage. It can
c be concluded from these resultts
that, irrespectivee of the viraal load the patient
p
is exxposed to, trransfection with
w antiboddies
m
f use in anntiviral theraapy against rabies in caases where the
for
t
is an efficient mechanism
D rotundus or the dog as inhibition only varieed
transsmitter is thee hematophhagous bat D.
from
m 89.2% to 100%
1
when these lineages were ussed. Howev
ver, if the paatient has beeen
expoosed to the liineage assoociated with the insectivvorous bat E.
E furinalis,, inhibition
variees with virall load. Thesse findings show
s
that trransfection with
w antibodies is a
prom
mising mech
hanism that could be ussed to develop an antiviral therapyy against rabbies.
Furthher studies are
a requiredd to assess th
he efficienccy of transfeection with antibodies in
i
vivo.
P
Finanncial Suppoort: FAPESP
PT.0043
ASSIFICAT
TION AND
D POSITIV
VITY RATE
E OF BATS
S RECEIV
VED FOR
CLA
RAB
BIES DIAG
GNOSIS
Limaa JYO1, Schheffer KC1, Achkar SM
M1, Kotait I1, Carrieri ML
M 1 - 1Instituuto Pasteur
d
The ddiversity off bat speciess in Brazil iss great, andd there are 172 species distributed
amonng nine fam
milies. The Phyllostomid
P
dae family is the most numerous, followed byy
Vesppertilionidaee and Molosssidae. According to feeeding habitts, the majority of bats are
insecctivorous, foollowed by frugivorouss, nectarivorrous, carnivvorous and hematophag
h
gous.
As thhese animalls are considdered reserv
voirs of rabiies virus it is
i essential to
t correctlyy
identtify the speccies and knoowledge of the biologyy, behavior and
a adaptation of bats in
urbann environm
ments. The obbjective of this
t study was
w to identiify the seveeral specimeens of
bats rreceived forr rabies diaggnosis at thhe Pasteur Innstitute, meeet the positiive index off
them
m and assess their distribbution on th
he feeding habits
h
in relation to the seasons. Inn the
periood between January andd Decemberr 2011, speccimens of bats were recceived for thhe
rabiees diagnosiss and submittted to identificate the family, gennus and/or sppecies,
accorrding to ideentification keys,
k
journaals and scieentific bookss. We also recorded
r
thee
feediing habits an
nd month of
o capture off animals. The
T period of
o this studyy was divideed
accorrding to thee seasons. During
D
this period,
p
28466 specimenss of bats weere identified,
beingg 77.6%,11.4% and 11% belonginng to the Moolossidae, Vespertilioni
V
idae
and Phyllostom
midae families, respectivvely. Of thee total of baats identified
d 88.53% haad
% frugivorouus, 4.39% suucking necttar and 0.28%
habitts insectivorrous, 6.82%
hemaatophagous.. In the warrmer seasons of the yeaar (spring annd summer) it was receeived
18088 bats being 1727 insecctivorous, 488 frugivorouus, 29 suckiing nectar and
a two
hemaatophagous.. In the coldder seasons (autumn annd winter), it
i was receivved 1038 baats
beingg divided in
nto 791 inseectivorous, 145
1 frugivorous, 95 succking nectarr and 6
hemaatophagous.. In relationn to the rabiees diagnosiss, 37 specim
mens (1.3%)) were
diagnnosed as po
ositive, incluuding the faamilies Vesppertilionidaee (17), Phylllostomidaee (13)
and M
Molossidae (7). Accordding to feed
ding habits bats
b were cllassified into 30
insecctivorous annd seven fruugivorous sp
pecimens. The
T positivee rate in the winter, werre
four insectivorous and one frugivorouss, and it seeems smaller than other stations.Thee
t summerr was higherr in Vesperttilionidae thhan in Molosssidae. Thiss
posittive rate in the
resullt was differrent that waas observed by Constanntine in the USA.
U
He obbserved the
higheer rate posittive in Moloossidae. Thee differencees observed in this studdy suggestedd the
impoortance of feeeding habitts and breedding seasonn of bats, con
nsidering prrincipally thhe
supply of food and
a warm seeasons. Thee knowledgee of bats andd seasonal identificatio
i
on
mprove the epidemiolog
e
gical surveilllance progrram used inn
studiies can conttribute to im
rabiees managem
ment in deterrmining the existence of
o bats of diifferent fam
milies and/orr
feediing habits in
n different seasons.
s
PT.0044
UDY AND DISTRIBU
D
UITION OF
F RABIES VIRUS
V
IN NON NEU
URONAL
STU
ORG
GANS IN BATS
B
SENT
T TO LAB
BORATORY
Y DIAGNO
OSIS IN PA
ASTEUR
INST
TITUTE
Limaa JYO1, Achhkar SM1, Scheffer
S
KC
C1, Castilho JG1, Rodriggues AC1, Kotait
K
I1 ,
1 1
Carriieri ML - Instituto Paasteur
Bats are consideered importaant reservoiirs of rabiess virus, whicch is paramount in the
o this zoono
osis disease, through thhe use of sen
nsitive systeems.
studyy of the pathhogenesis of
The ppresence off viral antigeen in these species
s
show
ws that non
n-neuronal viral
v
spread is
efficiient in diffeerent organss that particiipate effectiively in the eliminationn of rabies virus,
v
such as, salivary
y glands andd bladder. This
T study aimed to invvestigate thee presence of
o
b submittted for laborratory diagnnosis, as weell as, to studdy
rabiees virus in saamples of bats
the ppathogenesiss of the diseease through
h the use off laboratory animals. 3,930 routinee
diagnnostic speciimens of batts were proccessed durinng the periood between January 2011
and M
May 2012 by
b direct im
mmunofluoreescence (DIIF) and virall isolation on
o murine
neurooblastoma cells
c
(VICC
C) techniquees. 58 samplles were diaagnosed rab
bies positivee
from
m 37 bats in 2011 and 21 samples were
w diagnoosis rabies positive
p
in 2012,
2
repreesenting a positivity ratte of 1.80%. We random
mly selected 28 bats frrom rabies
posittive which were
w submittted to colleect organs foor preparatiion of inocu
ulum in the
propoortion to thee ratio of 1::10 for salivvary glands and tongues, and 1:20 for bladderrs,
whicch were inocculated in volume of 0..03 mL by the
t intracereebral route in
i post-weaaning
Swisss mice (21 days old annd weighingg 11g and 144g). Clinicaal observatioon was
perfoormed durinng 30 days and
a the pressence of thee virus was verified
v
by the DIF
technnique in diseased and dead
d
animalls. 60.7%, 50% and 42..8% of anim
mals selectedd for
the sttudy were rabies
r
positiive by viral isolation inn the salivarry glands, to
ongue and
bladdder, respectively. The minimum
m
inncubation peeriod was seven days and
a maximuum
incubbation periood varied beetween 17 annd 21 days.. The presen
nt study dem
monstrated the
t
preseence of rabiies virus in non-neuron
n
nal organs (ssalivary glan
nd, bladder and tonguee) in
67.8%
% rabies po
ositive anim
mals in central nervous ssystem (CN
NS).The deteection of rabbies
viruss in non-neu
uronal organns by DIF and
a virus isoolation has been
b
observved in severral
studiies. For stud
dies of pathoogenesis off rabies in baats, these reesults demonnstrate that the
use oof mice is sttill a good alternative.
a
Due
D to lackk to use CNS
S in routine practice in bats
for reeasons of pooor preservaation of the specimen, it may be necessary
n
to use nonneuroonal organss in order to obtain the positive
p
rabbies diagnossis.
PT.0045
RAB in selecct non-targ
get wildlife species
Effeccts of ONR
Fry TL
T 1, VanDaalen K1, Dunncan C2, VeerCauteren K1 - 1USDA
A-APHIS-W
WS-Nationaal
Wilddlife Researcch Center, 2Colorado State
S
Univerrsity
RAB® is a reecombinantt rabies vacccine used too as an oral vaccine
v
in wildlife
w
speecies
ONR
such as: fox (Vu
ulpes vulpess), raccoons (Procyon lotor), and striped
s
skunnks (Mepthiss
v
vector in the ONR
RAB® vaccinne is humaan adenoviru
us type 5 (H
Had5)
mephhitis). The viral
with the gene fo
or rabies glyycoprotein in
ncorporatedd into its gennome. Had
d5 is a relatiively
u
in manyy vaccine foormulationss. Canadian
safe and well-stuudied virus,, which is used
reseaarchers (e.g.., Knowles et al. 2009) have conduucted vaccinne efficacy and safety
studiies using ON
NRAB® in 18 species of
o animals. Our researcch expands on the species
previiously evaluuated. We studied
s
the vaccine
v
as it
i relates to its safety inn wildlife
®
speciies likely too contact thee ONRAB vaccine duuring oral raabies vaccinne (ORV)
camppaigns in the United Sttates. We investigated the
t effects of
o high dosees of the
ONR
RAB® vaccine in wood rats (Neotooma spp.), eastern
e
cottoontail rabbitts (Sylvilaguus
floriddanus), Virgginia opossuums (Didelp
phis virginiiana), Easteern wild turrkey (Meleaagris
galloopavosilvesttri), and foxx squirrel (S
Sciurus nigeer), whose range overlaaps with OR
RV
targeet species inn the Unitedd States. Affter inoculattion of the animals
a
we performed
p
r
realtime PCR on feccal swabs, oral
o swabs, and tissues to detect viiral DNA. Our
O preliminnary
d
postresullts mostly cooncur with the findingss of Knowlees et al. (2009). By 7 days
vacciination, turk
keys, opossums, and coottontails haad all stoppeed sheddingg viral DNA
A.
One woodrat annd five fox squirrels
s
stilll had detecttable levels of viral DN
NA in fecal
% of fox sqquirrels weree co-infecteed
swabbs on 7 dayss post-innocculation. Hoowever, 45%
with Leptospira interroganns, which maay be a confounding faactor to the prolonged
p
f
swabs from these animals. Th
here were no
n significannt
detecction of viraal DNA in fecal
findings on grosss histologyy of liver, kidney, smalll intestine, large
l
intestine, and lungg in
W are currrently comppleting PCR
R analysis off the tissuess
any oof the speciees studied. We
listedd above as well
w as nasaal turbinatess. Initial ressults suggesst low likelihhood of
®
persiistence of ONRAB
O
inn the environnment or inn individual animals thaat contact thhe
vacciine. Our preeliminary coonclusions suggest
s
thatt non-targett species will not be
negaatively impaacted by the distributionn of ONRA
AB® as part of
o ORV proograms in thhe
Uniteed States.
PT.0046
LATION AND
A
DETE
ECTION OF
O RABIES
S VIRUS IN
N THE FEC
CAL
ISOL
CON
NTENTS OF
O NATUR
RALLY INF
FECTED FRUIT
F
BAT
TS
Scheeffer KC1, Fahl
F WO1, Iaamamoto K1, Rodriguees AC1, Carrnieli Jr P1, Carrieri
C
ML
L1 ,
1
2 1
2
Oliveeira RN , Itto FH - Insstituto Pasteeur de São Paulo,
P
Facuuldade de Medicina
M
Veterinária e Zoootecnia da Universidad
de de São P
Paulo
The sspread of thhe rabies virrus into variious tissues, organs and
d secretionss in infectedd
anim
mals occurs with
w the proogression off clinical dissease. Fecall samples may
m have low
wer
viral concentratiion and the objective of
o this study was to isollate and deteect rabies virus
in thee fecal conttents of fruitt bats through virus isoolation in ceell culture teechnique annd by
RT-P
PCR, hemi-n
nested RT-P
PCR (hnRT
T-PCR) and Real Time RT-PCR. Thirty
T
specimens
of the genus Arttibeus bats, previously identified as
a positive for
f rabies byy FAT and
m
neurooblastoma cell
c line (N22A) were seelected and the
t intestinee was
inocuulation of murine
remooved from every
e
animaal and was scraped off, in order to collect the fecal
f
contennts.
The ffecal conten
nts were weeighed, hom
mogenized annd diluted 1:10
1
(w/v) using
u
a dilueent,
consiisting of 0.8
85% saline solution, suupplementedd with 2% Bovine
B
Fetaal Serum freee of
rabiees virus-speccific antibodies and 0.11% of gentaamicin sulfaate. The susppensions were
kept at 4°C for 30
3 minutes and centrifuuged at 800xg for 30 minutes
m
at 4°°C, filtered with
a 33mm length
l
syrinnge providedd with filterr Millex® with
w porosityy of
the aaid of 5-ml and
0.45μ
μm. The susspensions were
w inoculaated into muurine neurobblastoma ceells (N2A) for
f
viral isolation. For
F molecullar techniquues, extractioon of total RNA
R
and th
he reverse
o followeed by PCR aand hnRT-P
PCR targetinng to gene N.
N
transscription weere carried out,
The R
Real Time RT-PCR
R
tecchnique was performedd on the prooduct generaated from thhe
reverrse transcripption. Of thee 30 suspen
nsions inocuulated, only one (3.33%
%) was posittive
for vvirus isolatioon. None off the samplees was posittive by RT-P
PCR, howevver, 13 sam
mples
(43.333%) were positive
p
for rabies by hnRT-PCR
h
aand Real Tiime RT-PCR
R techniquees.
The ffact that onlly one sampple was positive by viruus isolation can be exp
plained by a
varieety of interfe
ferents foundd in this sub
bstrate, suchh as the pressence of baccteria and also
a
different degrad
ded productss of food thaat can causee inhibition of the reacttions. It is also
a
belieeved that theese interfereents may inffluence the results of th
he RT-PCR
R, hnRT-PCR
R
and tthe Real Tim
me RT-PCR
R techniquess, demonstrrating that th
he fecal con
ntents are off the
mostt complex biological saamples for amplificatio
a
on techniquees used as diagnostic
d
methhods. Neverttheless, thiss study dem
monstrated thhat both thee hnRT-PCR
R and Real Time
T
RT-P
PCR techniqques were sensitive forr the detection of rabiess virus. Thuus, we concllude
that these
t
techniiques can bee used as co
omplementaary tools in laboratory diagnosis
d
annd
fecall samples may
m also be used
u
for diaagnosis of raabies.
PT.0047
urvey of thee reportingg of human
n rabies
A woorldwide su
2
Tayloor LH1, PRP
P2 - 1Globall Alliance for
f Rabies Control,
C
Parrtners for Rabies
R
Preveention
r
cases, it is imposssible to asssess the truee
Withhout accuratte reporting of human rabies
burdeen of the disease on coountries, to assess
a
how much resou
urces a goveernment shoould
alloccate to rabiees control, or to carry ou
ut cost-beneefit analyses on rabies prevention
efforrts.
Global Alliance for Raabies Controol (GARC) with the Paartners for Rabies
R
The G
Preveention is coonducting a global surv
vey of humaan rabies repporting pracctices. We want
w
to gaain a global picture of where
w
humaan rabies is a notifiablee disease and
d to assess
whetther the systtems in placce for the reeporting of cases
c
are peerceived to be
b
adeqquate. Netw
works of rabiies experts have
h
been aasked to com
mplete a sho
ort survey which
w
colleected inform
mation on whhether hum
man rabies was
w notifiablle, which auuthorities weere
respoonsible for the
t collationn and reportting of casees, whether the
t system was
w effectivve,
and ssome detailss of what daata is colleccted and how
w it is disseminated. Thhe survey iss
availlable online in a user-frriendly form
mat in Engliish, French or Spanish.
ults from 1004 respondeents in 69 coountries havve been anallyzed and shhow
Preliminary resu
h
rabiees is a notiffiable diseasse in 61 (88%) of thesee countries. However,
that human
respoondents indiicated that the
t reportinng system was
w ineffectivve for 23% of the counntries
wherre rabies waas notifiablee. A regionaal analysis suggests
s
thaat the counttries where
rabiees is not nottifiable, or where
w
the syystem is ineeffective aree almost all in Africa annd
Asia where the burden
b
of human
h
rabies is highest, and more investment in rabies coontrol
is badly needed..
PT.0048
RADERMA
AL-INTRA
AMUSCUL
LAR SWIT
TCH EFFIC
CACY IN RABIES
R
PO
OST
INTR
EXP
POSURE PR
ROPHYLA
AXIS.
Poujool P1, Simons de Fanti A1, Goeschh J1, Ribadeeau Dumas F2 - 1Pasteu
ur Institut Rabies Center, 2Pasteur Insstitut - Rabiees Center/ N
National Reeference Cennter for Rabbies
bjectives:
Introoduction-ob
Becaause of the high
h
costs of
o rabies intrramuscular (IM) post exposure
e
pro
ophylaxis (P
PEP)
protoocols, cheap
per intraderm
mal (ID) prootocols usinng reduced doses
d
were developed in
somee countries and
a validateed by WHO
O. Due to lacck of data on
o the efficaacy of a prottocol
usingg ID and IM
M route succcessively, WHO
W
recom
mmends usinng only one route of
admiinistration per
p protocoll. In France and many other
o
high resource couuntries, no
packkaging adaptted to ID rooute is availaable and onnly IM regim
mens are autthorized by the
markketing authoorization, leading doctoors not to foollow WHO recommendations for
patieents having started an ID
D protocol abroad. As there is no study availaable on ID-IM
switcch efficacy, we sought to evaluate it with twoo objectives:: to describee serologicaal
efficaacy of our daily
d
practicce and to asssess if theree is a need to
t alert our national
n
heaalth
authoorities abouut the need of
o authorizin
ng the ID rooute.
m
Mateerials and methods:
In ouur rabies cennter, PEP innitiated abro
oad with ID route are sy
ystematically switchedd to
IM and
a one seroology (ELIS
SA) is prescribed 7 to 14
1 days afterr the end off this mixedd
protoocol to contrrol its efficaacy. We reaalized a nonn interventioonal descrip
ptive prospective
studyy including all 15 patieents of our center
c
who began
b
ID PE
EP abroad between
b
07/01/2010 and 08/31/20111.
ults:
Resu
The rrabies serollogy, carriedd out in med
dian 13 dayys after the end
e of the PEP,
P
was
availlable for 12 of the 15 patients
p
whoo received an
a ID-IM prrotocol (3 others were lost
l
to follow-up). Mean
M
age off the patientss was 38.8 years
y
(6.5 too 61.5 yearss) and the
S Lanka, Thailand,
T
Madagascar and
a India. The
T antibodyy
counntry of exposure were Sri
levell was higherr than the onne recommeended (0.5 EU/ml)
E
in all
a cases. No
o patient
discllaimed any side
s effect.
Concclusion:
This study is lim
mited by a sm
mall numbeer of patientts but to ourr knowledgee it is the onnly
n this topic. We found no
n evidencee of a lack of
o efficacy due
d to the ID
D to
one aavailable on
IM rooute switch
h during rabiies PEP. Coonsequentlyy this study does
d
not proovide any
argum
ment favoriing an alert of the healtth authoritiees concernin
ng this practtice in counntries
wherre only the IM
I route is authorized. These resuults need to be
b controlleed by largerr
studiies.
The aauthors discclose no connflicts of intterest for thhis work.
PT.0049
PER
RFIL EPIDE
EMIOLÓG
GICO DO ATENDIM
A
MENTO AN
NTIRRÁBIICO HUMA
ANO
EM A
AGRESSÕ
ÕES POR CÃES
C
E GA
ATOS NA 10ª REGIÃ
ÃO DE
SAÚ
ÚDE/LIMO
OEIRO DO NORTE, NO
N PERÍO
ODO DE 20007 A 2011..
Chavves CS1, LIM
MA FMG2, RODRIGU
UES VC3 - 1SECRETA
ARIA DE SA
AÚDE DO
ESTA
ADO DO CEARÁ
C
- 100ª COORDE
ENADORIA
A REGION
NAL DE SA
AÚDE DE
LIMO
OEIRO DO
O NORTE, 2SECRETA
ARIA DE SA
AÚDE DO ESTADO DO
D CEARA
A10ª C
COORDENADORIA REGIONAL
R
L DE SAÚD
DE DE LIM
MOEIRO DO
O NORTE,
3
SEC
CRETARIA
A DE SAUD
DE DO EST
TADO DO CEARÁ
C
- 10
0ª COORDENADORIA
REG
GIONAL DE
E SAÚDE DE
D LIMOEIIRO DO NO
ORTE
A raiva é transsmitida ao homem pe
ela inoculaçção do víru
us presentte na saliva
ae
u animal infectado,, e tem leta
alidade de aproximad
damente
secrreções de um
100%
%2. Sua prrevenção é baseada no tratame
ento profilá
ático quand
do houver
susp
peita de exxposição ao
o vírus da raiva, send
do este ba
aseado nass
cara
acterísticass da lesão provocada
a pelo anim
mal agresso
or, tipo de animal e sua
s
posssibilidade de
d observa
ação¹. O esstudo tem como obje
etivo conhe
ecer o perfil
epidemiológico
o do atendimento anttirrábico hu
umano em
m agressões por cãess e
os na 10ª Região
R
de S
Saúde de Limoeiro do
d Norte, no período de 2007 a
gato
2011
1.Realizou-se a partir de dadoss do Sistem
ma de Inforrmação de
e Agravos de
d
Notifficação(SIN
NANNET,1
10ª CRES)) dos onze município
os da Regiã
ão de Saúd
de
Fora
am analisados o tipo,, local, exte
ensão e prrofundidade das lesõ
ões, a cond
duta
de p
profilaxia da
a raiva ado
otada; e também o m
motivo de in
nterrupção
o do
trata
amento. Ass agressõe
es por cães
s representtaram 62,7
7% dos aciidentes,
enqu
uanto 31,2% foram por
p gatos. Predomino
P
ou a morde
edura como
o tipo de
agre
essão com 86,9% dass notificaçõ
ões, e as lesões de ferimento
f
ú
único
com
51,8
8% dos cas
sos. Dos lo
ocais ataca
ados, mãoss e pés tive
eram 41,8%
% das
agre
essões, seg
guidos porr 34,2% da
as lesões em
e membro
os inferiore
es. Quanto
oà
profu
undidade do
d ferimento, 50,6% foi lesão superficial.
s
Um total de
d 4605
pesssoas foram
m atendidass no períod
do e o trata
amento foi instituído para 83,7%
%.
Apóss a condutta terapêuttica, 59,5%
% dos pacie
entes conccluiu o trata
amento, 1%
%
deles o interrompeu (0,4% por indicação da unidade
u
de
e saúde) e 39,5% com
inforrmação ign
norada. Co
onsiderando-se o loca
al de agres
ssão, 47,3% dos cassos
foram
m definidos como gra
aves, realizzando-se o tratamen
nto associa
ado de soro
oe
vacin
na em 8% das pesso
oas. A indiccação de vacina
v
fez parte da conduta
c
de
quasse 100% dos tratame
entos instittuídos. A conduta de observaçã
ão e vacina
a foi
apliccada em 52
2% das ag
gressões. Um
U percen
ntual de 78,8% foram
m considera
ados
sem suspeita de
d raiva no
o momento
o da agresssão e 11,8
8% clinicam
mente
peitos. Nota-se que as
a agressõ
ões por cãe
es lideram o número de acidentes.
susp
O tra
atamento profilático
p
p
pós-expos
ição institu
uído é na maioria
m
de vezes
base
eado na utilização da
a vacina, e que a con
nduta de ap
penas obsservação, que
q
deve
e ser utiliza
ada em acidentes levves não foii relevante. Todavia, se 78,8% dos
anim
mais foi con
nsiderado clinicamen
c
nte sadio no momentto da agresssão e 50,6
6%
dos acidentes foram sup
perficiais, sugere-se que
q grande
e parte dessses pacie
entes
eria ter sido dispensa
ada do trattamento. Ressalta-se
R
e que a conduta de
pode
obse
ervação foi aplicada em
e apenass 10% dass notificaçõ
ões. Torna--se necesssária
a correta aplica
ação da no
ormatizaçã
ão de profilaxia antirrrábica, com
mo também
ma
ação multid
disciplinar. Além disso, o preenchimento adequado
a
da ficha de
e
atua
notifficação é im
mportante registro do
os dados e avaliação
o dos mesm
mos,
oporrtunizando a tomada de decisões.
PT.0050
STA
ABILITY EVALUATIION OF FL
LUORESCENT ANTII-RABIES VIRUS
CON
NJUGATE PRODUCE
ED IN INS
STITUTO PASTEUR
P
R OF SÃO PAULO
P
Freittas AP1, Peixoto ZMP1, Silva ACR
R1, Schefferr KC1, Chavves LB1, Capporale GMM
M1 1
Instiituto Pasteuur
The ffluorescent anti-rabies conjugate (FARC)
(
prooduced in In
nstituto Pastteur de São
Pauloo (IP-SP) iss obtained by
b fluorescein isothiocyyanate (FITC
C) conjugattion with
speciific polyclonal antiboddies purified
d and it recoommended storage
s
temp
perature of 4°C
proteected from light
l
incidennce. Given the
t fundam
mental imporrtance of thiis reagent foor
rabiees laboratoriial diagnosiis, the aim of
o this studyy was evaluaate its stabillity in relatiion to
physical factors that can afffect the quaality and maaintenance of
o product characteristi
c
cs.
C batch IP-T
TOT1/11 subbmitted to temperatures of
To do this, diffeerent aliquotts of FARC
8
for 1hoour or exposed to ambiient light for 10, 20, 30
0, 45, 60, 900,
37, 556, 65 and 80°C
120, 150 minutees and 24 hoours were used on direcct immuneffluorescencee (DIF) test on
prints of micce infected with Challeenge Virus
slides with centrral nervous system imp
PV),
Standdard (CVS)) and in monnolayers off BHK-21 ceells infectedd with Pasteeur Virus (P
with titers previously defined of 1:160
0 and 1:80, respectively
r
y. Also, withh one aliquoot
r
ded conditioons, the sam
me tests weree performedd and exposed
retainned on the recommend
for 1, 3, 5, 8 e 10 minutes to ultravioleet (UV) radiiation on fluuorescence microscopy
m
y. The
w adoptedd to evaluating the fluoorescence in
ntensity obseerved in thee
crossses system was
reacttions, whichh was attribuuted 4+ for high fluoreescence intennsity, 3+ to
o media intensity,
2+ loow intensityy and 1+ to total
t
loss off fluorescennce. For ligh
ht exposition
n, the samples
expoosed of 10 to
o 90 minutees received 4+,
4 the sam
mples exposeed to 120 annd 150 minuutes
receiived 3+ and
d the samplees exposed for
f 24 hourss received 2+.
2 The sam
mples submittted
to tem
mperatures of 37°C andd 56°C receeived 4+ andd the samplles submitteed to 65 andd
80°C
C showed onnly traces off residual FITC on the reactions, being
b
imposssible observved
the ccharacteristics rabies virus citoplassmatic incluusions. And with the inccreasing of
expoosition time to UV it waas observedd the graduaal loss of fluuorescence intensity
i
reachhing to 1+ after
a
10 minnutes. Thesee results shoowed that FA
ARC testedd kept the saame
qualiity characteristic even exposed
e
to room light uuntil 90 min
nutes and 566°C for 1 hoour,
show
wing the sam
me florescennce intensityy of reagentt kept on reccommendedd conditionss,
althoough it doesn’t be expoosed to UV for
f long perriods duringg the readingg of reactionns on
fluorrescence microscopy, showing be a stable reaagent and as it has a higgh value, it must
m
be reevaluated whhen submittted to adverrse conditioons, avoiding
g waste.
Finanncial Suppoort: Institutoo Pasteur
PT.0051
ALUATION
N OF THE INTERFE
ERENCE OF
O ALKAL
LOIDS OBT
TAINED
EVA
FRO
OM THE SK
KIN SECR
RETION OF
F Ameeregga picta IN THE PEN
NETRATIO
ON
OF R
RABIES VIRUS IN BHK-21
B
CE
ELLS
Freittas AP1,2, Siilva ACR2, Scheffer KC
C2, Pimentaa DC1 - 1Instituto Butanntan, 2Instituuto
Pasteeur
Rabies is an infeectious diseease caused by a virus bbelonging too the familyy Lissaviruss that
mals includiing humanss. Rabies traansmission occurs
o
by virus-contai
v
ining
affeccts all mamm
salivva , through biting or liccking of an injured tisssue. The viruus replicates at the site of
inocuulation untill it reaches the nerve en
ndings. At the
t neuromu
uscular juncctions, the virus
v
bindss specificallly to the niccotinic acetyylcholine reeceptor. On the other haand, alkaloiids
have yielded intteresting ressults in term
ms of drug ddevelopmentt. Since rabies virus
infecction is, at leeast, partially associateed to the niccotinic acetyylcholine reeceptor, the
preseent study fo
ocus on the evaluation
e
of
o moleculees obtained from
f
the skin secretionn of
Ameeerega picta,, in the proccess of peneetration andd infection of
o rabies viru
us in Baby
Ham
mster Kidneyy (BKK-21)) cells. The secretion was
w collected
d though stimulation byy
manuual compresssion of the amphibian glands, andd then separrated into 14
4 fractions by
b
reverrsed-phase high
h
perform
mance liquiid chromatoography (RP
P-HPLC). The
T fractionss
weree characterizzed by masss spectromeetry and testted for their cytotoxic effect
e
on BH
HK21 ceells. Fractio
on 3 was thee only one presenting
p
cyytotoxicity,, by means of
o arising
morpphological changes
c
wheen compareed to the neggative contrrol (cells + MEM)
M
and the
posittive control (cells + 20%
% DMSO). Subsequenntly, the fracctions were screened foor
their ability to innterfere in the
t process of virus cellular penetrration. The rapid
r
test foor
3 6 and 7 were
w able to
inhibbition of fluorescent focci (RFFIT) showed thaat fractions 3,
decreease the viraal infection. Analyses by
b mass speectrometry showed
s
thatt the molecuules
preseent in the acctive fractioons ranged from
f
200 to 500 Da, whhich is in th
he alkaloid
moleecular mass range. Mollecules are undergoing
u
purification
n for properr virologicall
assayys and strucctural characcterization.
C
FAP
PESP, CNPq.
Finanncial Suppoort: Institutoo Pasteur, CAPES,
PT.0052
NSORIAME
ENTO REM
MOTO EM
M MORCEG
GOS HEM
MATÓFAGO
OS Desmoddus
SEN
rotun
ndus NO MUNICÍPIO
M
O DE POTIRETAMA
A, CEARÁ,, BRASIL, 2008 – 20009
Mouura FBP1, Silva MVS1 - 1Secretariaa da Saúde do
d Estado do
d Ceará - 1ª Coordenaddoria
Regional de Saú
úde de Fortaaleza
RODUÇÃO
O - Estudos de sensoriaamento remooto tornaram
m-se cada dia
d mais pressente
INTR
entree os trabalho
os desenvollvidos na áreea animal. Carini
C
conclluiu que a raiva
r
em bovvinos
era trransmitida pelo
p morceggo Desmodu
us rotunduss¹ e estudos relacionadoos ao controole
dessaa doença vêêm sendo direcionados paralelamente aos estu
udos de com
mportamentoo,
contrrole e dinâm
mica de populações desssa espécie. Em 2008 em Potiretam
ma-Ceará fooi
realizzado um traabalho com
m sensoriameento remotoo numa colôônia de D. rotundus com
m,
aproxximadamennte, trezentoos indivíduo
os numa furnna chamadaa Furna da Onça.
O
Apesaar da
amplla distribuiçção desta esppécie, poucco estudo se propôs a esstudar seu comportame
c
ento
de accordo com o sexo e fase lunar atraavés do senssoriamento remoto.
r
OB
BJETIVOS –
GER
RAL - Preveenir casos dee Raiva hum
mana originnários do cicclo silvestre através do
moniitoramento e controle populaciona
p
al de morceggos hematófagos. MAT
TERIAL E
MÉT
TODO – Foi utilizado sistema
s
de rádio-teleme
r
etria terrestrre com codiificação diggital,
frequuência de op
peração 4333.92 MHZ (UHF); receeptores manuuais com an
ntena direcional
YAG
GI de três elementos; reeceptores au
utônomos Data-loggers
D
s com registtros dos anim
mais
em vvários interv
valos de tem
mpo diferenttes e radio trransmissorees (Chips) pesando
p
3g.Para
análiise das perm
manências dos
d quirópteros fora, deentro e na en
ntrada do abbrigo foram
m
convvencionadoss parâmetross de tempo constituindo períodos Longos
L
≥ 50 minutos,
Médiios 30 ≥ e < 50 minutoos e Curtos < 30 minutoos. RESULT
TADOS – Os
O morcegoos
hemaatófagos D. rotundus appresentaram
m, segundo sexo, comportamentos distintos naas
diferrentes fases lunares. CO
ONCLUSÃO
O - Na lua N
Nova ambos, macho e fêmea
apressentaram atiividade ao longo
l
da nooite (antes, durante
d
e appós o apareccimento da lua),
l
perm
maneceram na
n entrada do
d abrigo quuando não havia
h
risco de
d predação
o. Na lua Chheia
houvve pouca ou nenhuma atividade
a
aliimentar do macho
m
com
m pouca ocorrrência de saída
de loonga duração antes da lua
l aparecerr e sem perm
manência na
n entrada do abrigo, teendo
em vvista a alta inntensidade do
d luar que favorecia o risco de prredação. Naa lua Minguuante
o maacho apresenntou atividaade alimentaar significannte antes daa lua apareceer e moderaada
ou neenhuma apóós o seu apaarecimento, permanecendo na entrrada do abrigo quando não
n
haviaa risco de prredação. O conhecimennto da dinâm
mica de trannsmissão dee Lyssaviruss
entree morcegos hematófagoos e o homeem, pode peermitir mudaanças na dinnâmica da
enferrmidade e a readequaçãão, em temp
po hábil, das estratégias de controlle.
Agraadecimentoss
Aos moradores do município de Potireetama, pela disponibiliddade em cedder seu temppo
A técnico
os do Núcleo de Controole de
e connhecimento para a realiização dessee trabalho. Aos
Vetoores (NUVE
ET) - Secretaaria da Saúd
de do Estaddo do Ceará (SESA), prrincipalmennte,
Mariia Mariza dee Lima e Sillva e Estevãão Curado D
Domingues,, que acomp
panharam o
prim
meiro autor em
e todas as atividades, visitas e cooletas. Aos técnicos
t
vetterinários e
demaais servidorres da 10ª Coordenadorria Regionall de Saúde de
d Limoeiroo do Norte. Aos
serviidores da Seecretaria Muunicipal de Saúde de Potiretama.
Referências Bib
bliográficas
F.; Aguiar, E.C.E.;
E
Marrtorelli, L.F.A.; Silva, M.M.S..
M
Diagnóstico
1. Allmeida, M.F
laborratorial de raiva
r
em quiirópteros reealizado em área metrop
politana na região sudeeste
do Brasil. Revissta de Saúdee Pública, p. 341-344. 1994.
1
PT.0053
BIES IN CA
ANADA – 2011
2
RAB
Know
wles MK1, Kush
K
J2, Iqbbal Z2, Ham
mill P1, Fehlnner-Gardineer C1 - 1Can
nadian Foodd
Inspeection Agen
ncy - Centree of Expertise for Rabiees, 2Canadiaan Food Insspection Aggency
- Letthbridge Labboratory
uspect rabidd animals were
w submittted for testinng to the Caanadian Foood
In 20011, 4397 su
Inspeection Agen
ncy diagnosstic laboratoories in Ottaawa, Ontarioo and Lethbbridge,
Albeerta. Of thesse, 115 (2.66%) tested positive
p
in thhe fluoresceent antibody
y test
(FAT
T). Additionally, enhannced wildliffe surveillannce sampless from Ontaario (ON,
n=1885) and Alberta (AB, n=
=88) were tested
t
and found
f
to be negative.
n
The
T majorityy of
rabiees cases werre detected in
i the proviince of Saskkatchewan (n=34),
(
follo
owed by ON
N
(n=26), Manitobba (n=21), Quebec
Q
(n=17), Britishh Columbia (n=7), Nortthwest
N cases w
were found inn AB, Yukoon Territories, or
Terriitories (n=6) and Nunavvut (n=4). No
the A
Atlantic provvinces, although with the
t exceptioon of AB, saamples subm
missions weere
very low (1 to 40 samples) from these regions. Thhe striped skunk (Mephhitis mephittis)
wed by big brown bat
was tthe species most frequeently found positive (nn=42), follow
(Epteesicus fuscuus, n=35), Arctic
A
fox (V
Vulpes lagoppus, n=11) and red foxx (V. vulpes,,
n=5). Spillover of wildlife rabies variaants into doomestic anim
mals was ob
bserved in 1
bovinne and 1 doog (Arctic foox variant), and in 4 caats, 2 horses and 1 dog (Western skkunk
variaant). The reemaining poositive casess were found in variouss bat species. Positive
casess involving human exposures origiinated from
m 33 differennt counties, whereas those
with only domesstic animal exposures or
o no exposures noted came
c
from 58 and 14
t 105 couunties repressented only 6 were com
mmon to twoo or
counnties, respecctively. Of the
moree exposure categories.
c
While 1% of
o samples with human
n exposuress tested posiitive,
10.6%
% of those with
w domesstic animal exposure,
e
annd 5.2% of those with no
n exposuree
indiccated tested positive forr rabies. In addition to the animal samples, seeven humann
suspeect cases weere tested by RT-PCR and/or FAT
T on saliva and
a nuchal skin biopsyy
sampples; all werre negative for rabies.
PT.0054
n experimeental mice rabies
r
viruss infection
Shorrt-Interferiing RNA evvaluation in
caused by varia
ant 2 and variant
v
3
M1, Allendorrf SD1, Perees MG1, Anntunes JMAP
P1, Vicente AF1, Pantoj
oja
Appoolinario CM
1
1 1
JCF , Megid J - Unesp- Faculdade
F
dee Medicina Veterináriaa e Zootecniia Depaartamento de
d Higiene Veterinaria
V
e Saude Pubblica
Objeective
n no effeective treatm
ment is availlable. Treattment
Rabies is an anccient diseasee and until now
A (siRNA) to
t inhibit raabies virus (RABV)
(
repplication shoowed
usingg short-interrfering RNA
prom
mising resultts in vitro. Our
O purposee was to evaaluate the effficacy of siiRNA in treeating
micee experimen
ntally infecteed with diffferent streett RABV straains.
M
Mateerials and Methods
C
miice, SPF, 4--6 weeks-olld were inocculated in gaastrocnemiuus
3 grooups of 20 C57/BL6
musccle with 3 different
d
RA
ABV strains.. A variant 2 isolated frrom a dog [dv2 (LD50 103.39//0.03 mL)], a variant 2 isolated froom a humann [hv2 (LD550 10-6.66/0
0.03 mL)] aand a
variaant 3 isolateed from a huuman [hv3 (LD50
(
10-66.66/0.03 mL
L)] . For eaach group, 10
1
micee remained untreated
u
annd 10 mice were
w treatedd with a mixx of 3 differrent siRNA
A
sequeences (3.3 µM
µ each) asssociated with
w lipofectaamine (Branndão et al. 2007)
2
basedd on
rabiees virus N gene as a target. Animalls received a single dosse of siRNA
A mixture, via
v
intrapperitoneal route,
r
24h post
p RABV inoculationn (p.i) and were
w observeed during 300
days. Cox Propo
ortional Hazzards modells were usedd to estimatte lethality rates
r
and Haazard
ween groupps.
Ratioos (HR) betw
Resuults
d
lethalitty was 37.55% in the innoculated grroup and 50% in the siRNA groupp (P=
For dv2,
0.71;; HR= 0.75)); For hv2, lethality waas 100% in the inoculaated group and
a 70% in tthe
treateed group (P
P= 0. 27; HR
R=0.57); Foor hv3, lethaality was 600% in the innoculated grroup
and 880% in the treated
t
grouup (P= 0. 211; HR= 1.977).
Concclusion
ms to be asssociated to the
t RABV strain
s
once the results of
The eefficacy of siRNA seem
surviival was varriable in thee groups sub
bmitted to siRNA and infected
i
witth different
RAB
BV strain . The
T siRNAss used were designed based on Passteur virus N gene
sequeence, a fixeed strain whhile in our sttudy street RABV
R
strains were useed. Even
consiidered as a conserved gene,
g
studiees showed significant genetic
g
variaability. A nearly
n
perfeect complem
mentary sequuence betw
ween siRNA molecule and
a the virall RNA targeet is
necessary for mR
RNA cleavage. Our RA
ABV N genne sequencees showed 85.7% - 95.22% of
1
to v2, confirmingg this
homoology betweeen v3 and siRNAs seqquences andd 95.2 % - 100%
naturral variability and the better
b
results obtained with
w the varriant 2. In this
t study, a nonbioloogical deliveery system was
w used annd an imporrtant point is
i the difficuulty of siRN
NAs
delivvery within CNS being this a majoor problem in
i practical therapeutic.
PT.0055
i the state of São Pau
ulo - data frrom the Paasteur Instiitute
Caniine and feliine rabies in
of Sãão Paulo, 1999-2012
Carriieri ML1, Achkar
A
SM1, Castilho JG
G1, Oliveiraa RN1, Carn
nieli Jr P1, Macedo
M
CI1,
2
2
2 1
Omooto TM , Taakaoka NY , Kotait I - Instituto Pasteur
P
- Seção de Diag
gnóstico,
2
Instiituto Pasteuur
P
cooordinated by the Pasteuur Institute of
The São Paulo State Rabiees Control Program,
Paulo, has been
b
involved in endeaavors to impprove the eppidemiologiccal surveillaance
São P
of rabbies in dogss, cats and bats,
b
particu
ularly in urbban areas. Since 1998, when
w
the laast
case of canine raabies due too antigenic variant
v
2 occcurred in thhe state, the Institute haas
mples, 17,7
796 sampless from cats aand
receiived 66,352 canine cenntral nervouss system sam
38,1333 from batt specimens. Using connventional teechniques for
f rabies diagnosis, 11
dogs, 8 cats and
d 554 bats were
w found to be positivve for this disease.
d
Wheen moleculaar
nd genetic ssequencing, were used, antigenic
technniques, suchh as antigennic typing an
variaant 3, which
h is associated with Dessmodus rotuundus, was detected in 11 dogs annd 7
cats; this variantt has also beeen identified in bat sppecies from the genus Artibeus
A
sppp.,
w
is
whicch has synannthropic habbits. Only one cat was ppositive forr variant 4, which
assocciated with Tadarida brasiliensis.
b
It should bee noted thatt the signs and
a symptom
ms of
cats aand dogs innfected by raabies variannts associateed with batss were quitee different frrom
thosee in dogs inffected with variants associated with dogs (varriants 1 andd 2). In geneeral,
sympptoms were atypical, thhe most com
mmon cliniccal picture being
b
that off paralysis and
a
hemoorrhagic gasstroenteritiss, and cases were nearlyy always isoolated. Posiitive samplees
from
m cats and doogs frequenntly came froom animalss that had beeen run overr and died.
Thesse findings show
s
the im
mportance off maintaininng active eppidemiological surveillaance
of caats and dogss and of sending samplees for rabies diagnosis,, particularly in regionss
wherre rabies is known
k
to occcur in urbaan areas.
PT.0056
EVA
ALUATION
N OF HUM
MORAL IM
MMUNE RE
ESPONSE CONFERR
RED BY
PUR
RIFIED CH
HICK EMB
BRYO CEL
LL VACCIN
NE (PCEC
CV) AND PURIFIED
VER
RO CELL VACCINE
V
(PVCV) USED
U
IN CO
OMPLETE
E AND MIX
XED PRE-EXP
POSURE SC
CHEMES AT PASTE
EUR INSTIITUTE OF
F SÃO PAU
ULO
Cunhha Neto RS1, Cunha RS
S2, Chaves LB
L 1, Schefffer KC1, Caaporale GMM
M1, Silva ACR
A 1
1
2
- Insstituto Pasteeur - Laboraatório de Soorologia, Innstituto Pastteur - Área Clínica
C
The most
m widelyy used vacccines in the Western
W
woorld for hum
man rabies prophylaxis
p
are
Puriffied Chick Embryo
E
Celll Vaccine (PCECV)
(
annd Purified Vero Cell Vaccine
V
(PVC
CV), which is used at Pasteur
P
Instiitute of São Paulo sincee 2000. Whhereas in Braazil
theree are no repo
orts of the use
u of PCEC
CV in replaccement or complement
c
tation to PV
VCV,
the aaim of this study was too evaluate humoral
h
imm
mune responnse of people vaccinateed
againnst rabies in
n complete and
a mixed pre-exposur
p
re schemes using
u
PCEC
CV and PVC
CV
with different ro
outes of appplication, inttramuscularr (IM) and intradermal
i
(ID). Accorrding
r
1115 serum sam
mples from
m different inndividuals were
w
to thee vaccinatioon scheme received,
distriibuted in seeven groups: three containing serum
m samples from
f
patiennts who receeived
compplete schem
mes with onee vaccine an
nd one routee of applicattion - 3 PVC
CV (IM), 3
PVC
CV (ID) e 3 PCECV (IM
M) -, used as reference;; and four containing seerum samplles
from
m patients whho receivedd both vaccinnes and twoo routes of application,
a
featuring a
mixeed scheme - 2 PVCV (IID) + 1 PCE
ECV (IM), 1 PVCV (ID
D) + 2 PCEC
CV (IM), 2
PCEC
D) e 1 PCE
ECV (IM) + 2 PVCV (ID). The hum
moral immuune
CV (IM) + 1 PVCV (ID
respoonse was an
nalyzed baseed on the levels of rabiies virus neuutralizing an
ntibodies
(RVN
NA) obtaineed by rapid fluorescentt focus inhibbition test (R
RFFIT); thrrough
calcuulation of itss maximum
m and minim
mum values,, median, staandard deviiation, 1st and
a
3rd qquartiles, intterquartile range
r
and Kruskal-Wal
K
llis test. Diffferences staatistically
signiificant (p-vaalue < 0.05)) were obserrved when tthe group 2 PVCV (ID) + 1 PCEC
CV
(IM) was compaared with thhe groups 3 PVCV (IM
M) and 1 PCE
ECV (IM) + 2 PVCV (ID).
(
m samples from
f
patientts who receiived 2 PVC
CV (ID) + 1
The ggroup contaaining serum
PCEC
CV (IM) is the one responsible for statistical differencess observed, because
b
it is the
groupp in which higher
h
RVN
NA titers weere detectedd, ie, the grooup with thee best humooral
immuune responsse. Despite the differennces observeed, in all serrum samplees, RVNA tiiters
obtaiined can be consideredd adequate (≥
≥0.5 UI/mL
L), regardlesss of the vacccine and rooutes
of appplication ussed. The use of PCECV
V for rabiess vaccination
n, in complete or mixed
pre-eexposure schheme, show
wed satisfacttory results in the induction of hum
moral immuune
respoonse and its use may bee adopted when
w
it is neecessary to complemen
c
nt or replacee
PVC
CV.
P, Brasil
Finanncial Suppoort: Institutoo Pasteur, Sãão Paulo/SP
PT.0057
ALYSIS OF
F DOG AN
ND CAT PO
OPULATIO
ON SUBMIITTED TO
O
ANA
VAC
CCINATIO
ON CAMPA
AIGNS AG
GAINST RA
ABIES IN A SMALL CITY FRO
OM
BRA
AZIL
Babbboni SD1,2, Fornazari
F
F1, GONZAL
LEZ GK2, V
VICTORIA
A C1, Modolo JR1 - 1UN
NESP
2
-Botuucatu - Deppartamento de
d Higiene Veterinária
V
a e Saúde Púública, Preffeitura
Munnicipal de Bootucatu - Viigilância Am
mbiental em
m Saúde
In many developping countries domestic dogs and cats are thee most impoortant
r
to meen. The annu
ual vaccinattion constituutes an important
transsmitters of rabies
preveentive activ
vity, and thee knowledgee of animal population profile can assist futurre
prophhylatic prog
grams, not just for rabiees but also ffor other infectious disseases and social
probllems involvving dogs annd cats. In most
m Braziliian cities there is no ceensus that caan
proviide these daata. One alteernative, altthough limitted, is to co
ollect inform
mation from
m the
vacciination cam
mpaign itselff, realized by
b the publicc service. The
T present study
s
aimedd to
analyyze: the num
mber of vaccinated doggs and cats in
i a small Brazilian
B
cityy; their sex and
reprooductive staatus; and evaaluate if theere was a deecrease in vaaccinated annimals afterr two
yearss without caampaign. Thhe study waas realized inn the city of Botucatu (22°53’09”
(
S
48°26’42”W), São
S Paulo Sttate, Brazil,, which com
mprises an area
a of 1,4822.87 km2, has a
on of approximately 12
27,370 hab. (last censu
us, 2010), an
nd its dog annd
humaan populatio
cat ppopulation are
a estimatedd to be 29,1197 and 7,5442 animals, respectivelly (Pasteur
Instittute estimattive,2010). Data
D from the
t campaiggns of 2009 and 2012 were
w obtaineed
from
m the recordss of the Envvironmentall Vigilance Department
D
t of the cityy. The total of
o
vacciinated anim
mals in 20099 was 26,6644 (23,755 dogs
d
and 2,9
909 cats) and in 2012 was
w
23,5884 (20,195 dogs and 3,,389 cats). The
T profile of the vacciinated dog population
p
w
was
as foollow: in 20009 – 12,2844 females annd 11,471 m
males; 3,9666 castrated and
a 19,789
intacct. In 2012 – 10,659 fem
males and 9,563
9
males;; 5,281 casttrated and 14
4,914 intactt.
The pprofile of vaaccinated caat populatio
on was as foollow: in 20009 – 1,699 females andd
1,1922 males; 1,3307 castrateed and 1,602
2 intact. In 2012
2
– 1,9007 females and
a 1482 males;
m
1,5377 castrated and
a 1,852 inntact. The number
n
of fe
females wass higher thann males for both
speciies and yearrs. The propportion of inntact animalls was also higher thann castrated ones,
o
especcially for doogs. These facts
f
shown
n that the poopulation off dogs and cats
c submitteed to
vacciination agaiinst rabies has
h a high proportion
p
o individualls able to brreed. This
of
patteern probably
y also occurr in the popuulation of unnvaccinated
d animals. Towards
T
theese
data, local autho
orities shoulld be aware of the probblematic invvolving uncoontrolled annd
i
noot just rabiess control, buut
undeesirable reprroduction off these speccies, which involves
also other aspeccts of publicc health. During the yeaars 2010 andd 2011 there was no
mpaign againnst rabies inn Botucatu. Nevertheless, only a sm
mall decreaase in
vacciination cam
the nnumber of vaccinated
v
animals was observed inn 2012. Thiis differencee is probably not
signiificant, and can be obseerved amonng previous years
y
with successive
s
campaigns.
c
Also, during 2012 there waas no reinforrcement in divulging
d
th
he campaign
n against raabies.
mpaign, thee human
Thuss, despite the two-year interruptionn of the vaccination cam
popuulation appeears to continue to conssider it as ann important activity.
PT.0058
BIES SEAS
SONALITY
Y IN BATS
S (Chiroptera, Mamm
malia) FROM
M
RAB
NOR
RTHWEST
T OF SAO PAULO
P
ST
TATE, BRA
AZIL
Pedroo WA1, Biaagi MB2, Caarvalho C3, Perri SHV3, Queiroz LH
L 3 - 1UNES
SP - Faculddade
e Meedicina Veteerinária de Araçatuba
A
- Bolsista dee Produtividdade em Pessquisa CNP
Pq 2
Depaartamento de
d Apoio, Prrodução e Saúde
S
Anim
mal, UNESP
P - Faculdadde e Medicina
Veterinária de Araçatuba
A
- Graduando do Curso de
d Medicinaa Veterinária - Bolsistaa
U
- Faaculdade e Medicina
M
V
Veterinária
d Araçatub
de
ba PIBIC/CNPq, 3UNESP
d Apoio, Prrodução e Saúde
S
Anim
mal
Depaartamento de
Rabies is an impportant zoonnosis whichh has the batts as one off the main reeservoirs off the
fi
losses to the liivestock inddustry
viral agent. In thhe XXI centtury still cauuses huge financial
A
altthough it is a disease th
hat has prev
vention, by
and ppublic healtth in Latin America,
preveentive vacciination in cattle and huuman populaations in rissk areas, and
d post
expoosition treatm
ment with serum in hum
man beings.
Conssidering the importancee of the diseease, the objjective of th
his study waas to test
statisstically the correlation
c
between itss occurrencee in bats andd seasonalitty (rain
preciipitation) inn the northw
west of São Paulo,
P
Brazzil.
To teest this hypoothesis we used
u
the reccords of resuults diagnosstic for samp
ples of batss that
weree sent for ex
xamination to
t the Rabiees Laboratorry at UNES
SP (Araçatub
ba Campus)), by
healtth or agricullture teams services off the municippalities in thhe northwesstern regionn of
São P
Paulo, Brazzil. Some samples camee from bats found insidde the housees or in the
streets by the geeneral popullation or capptured by thhe staff of th
he Laboratoory of
ESP (Araçaatuba Campu
us), betweeen the years 1999 and 2010,
2
in the same
Chirooptera, UNE
regioon. The rain
nfall data weere providedd by the stattion of INM
MET (Instituute of
Meteeorology) Votuporanga
V
a in the sam
me period. Foor statisticall analysis thhe data weree
groupped in perioods of rain (October-M
(
March) and ddry (April to
o Septemberr), year to year,
y
and w
we used thee Spearman correlation coefficient to test the correlation
c
between
b
raiinfall
and tthe occurrennce of rabiees in bats.
The aanalysis ressulted in a negative
n
andd significantt correlation
n using the average
a
rainnfall
durinng the dry and
a rainy seaasons and th
he rabies poositivity (p = 0.0185, R = - 0.477, N =
24). T
Therefore, the
t positivitty of rabies increases during
d
the drry season. The
T weak buut
signiificant negaative correlaation betweeen rainfall aand the occu
urrence of raabies suggeests
that this
t is one of
o the factorrs that affect the tempooral distributtion of this zoonosis inn bats
in thee northwesttern region of
o São Paullo, Brazil.
It is aargued heree, by hypothhesis, that duuring the drry seas
on thhe dislocatioon of bat poopulations frrom their shhelters occu
urs due to thhe water scarrcity
and rresources, and
a also duee to the form
mation of neew colonies by young bats
b that leaave
their original shelters. Thesse displacem
ments lead too dispersionn of the viruus to new arreas,
o bat
as weell as an inccrease in thee intraspeciffic and interrspecific intteractions of
popuulations, thaat will comppete for spacces in shelteers, and connsequently resulting
r
an
increeased occurrrence of rabbies in differrent speciess. In additioon to rainfall, other facttors
and pprocesses prrobably affeect or determ
mine the disstribution an
nd occurren
nce of rabies,
incluuding the avvailability off shelters annd resourcess, and the methods
m
for control of
hemaatophagous species.
Acknnowledgmen
nts: To the Institute off Meteorologgy (INMET
T) for rainfalll data. To
CNP
Pq for provid
ding scholarrships to Peedro WA annd Biagi MB
B.
PT.0059
TS AND RA
ABIES IN THE
T
NORT
THWESTE
ERN REGIION OF SÃ
ÃO PAULO
O
BAT
STA
ATE, BRAZ
ZIL.
Casaagrande DK
KA1, Favaro ABBBC2, Carvalho
C
C3, Picolo MR4, Lopes AP
A 1, Favoreetto
5
6
7
7
4
3
SR , Campos AC
CA , Hernaandez JCB , Lot MS , Albas
A
A , Pedro WA , Queiroz LH
H3 1
UNE
ESP - Faculldade e Meddicina Veterrinária de Araçatuba
A
- Mestranda do Program
ma de
Ciênncia Animal, 2UNESP - Faculdade e Medicinaa Veterinária de Araçattuba Gradduanda de Medicina
M
Veeterinária, 3UNESP
U
- Faaculdade e Medicina
M
V
Veterinária
d
de
Araççatuba - Deppartamento de Apoio, Produção
P
e Saúde Anim
mal, 4APTA
A - Agência
Pauliista de Tecnnologia Agrropecuária de
d Presidentte Prudente - Pólo Regional de
Deseenvolvimentto Tecnológgico dos Ag
gronegócios da Alta Sorocabana, 5Instituto
I
Paasteur
de Sãão Paulo e Universidad
U
de de São Paaulo - Núcleeo de Pesquuisas em Raaiva -ICB, 6USP
U
- Uniiversidade de
d São Paullo - Núcleo de Pesquisaas em Raivaa -ICB, 7UN
NIP - Araçattuba
- Graaduanda do Curso de Farmácia e Bioquímica
B
l decadess three distinct cycles of
o rabies weere identifieed in the
Durinng the two last
northhwestern reg
gion of São Paulo Statee: the urbann cycle characterized prredominantlly by
caninne rabies froom 1993 to 1997 and th
he aerial annd rural cyclles starting in
i 1998, witth
the ppredominancce of cases in bats in urrban areas aand in herbiivores. Tweenty six bat
speciies were preeliminarily reported
r
in this geograaphical regioon, includinng D. rotunddus
and D
Diaemus yooungi and frrom 1998 too 2007, a tottal of 4,035 bat samples were testeed for
rabiees resulting 50 (1.2%) positive
p
cases in nine nnon-hematopphagous speecies: three
Moloossidae, fivee Vespertiliionidae and one Phyllostomidae. The
T aim of the
t present
reseaarch was to describe thee occurrencce of rabies in non-hem
matophagouss bats and thhe
preseence of antib
bodies against rabies virus
v
in vam
mpire bats inn the northw
west of São Paulo
P
Statee, during thee period from
m January 2008
2
to Julyy 2012. Rabbies virus was
w detectedd in
22 (11.97%) out of
o 1117 nonn-hematophhagous bats and none off the 190 vaampire bats
exam
mined. 82% of the bat positive
p
sam
mples was submitted
s
too antigenic and
a genetic
charaacterizationn and the varriant of D. rotundus
r
waas identifiedd in 28% off them. Seruum
from
m 125 vampiire bats capttured in fouur different sshelters werre tested forr rabies viruus
neutrralizing antiibodies andd 28% (35/12
25) resultedd negative; 65%
6
(81/12
25) resulted
posittive with titeer from 0.100 to 0.5UI/m
ml and 7% (9/125)
(
highher than 0.55UI/ml. Moost of
vamppire bats preesenting anttibody weree female (611%) from a natural shellter located in a
tree role.
r
Althou
ugh no vamppire bat wass found possitive for rabbies, four poositive cases of
rabiees transmitteed by that sppecie were detected in the studied
d region: three bovine cattle
c
and oone cat. Thee presence of
o high perccentage of vampire
v
batss with viruss neutralizinng
antibbody indicattes a recent exposure too rabies viruus, which coonfirms thatt, although this
t
geoggraphical areea is considdered as low
w or negligibble risk for rabies
r
in herbivorous,
surveeillance meaasures shouuld be mainttained.
P (Process 2008/00976
2
6-0) and CN
NPq (Processs 578281/20008Finanncial Suppoort: FAPESP
2)
Pq Technicaal Support Fellows: Carrvalho C, Piicolo MR, Hernandez
H
J Lot MS
JC,
CNP
PT.0060
VA EM MORCEGOS
S (MAMM
MALIA: CH
HIROPTER
RA) NO ES
STADO DE
E
RAIV
PER
RNAMBUC
CO: PROBL
LEMAS E PERSPEC
CTIVAS FU
UTURAS
Silvaa LAM da1, Machado JLM
J 2, Araujjo ACR2, Oliveira
O
J doo CT de3, Silva-Filho TPda3,
3
3 1
Silvaa EMVG daa , Silva RR
R da - CAV
V/UFPE - Núcleo
N
de Biologia, 2LA
ANAGRO/P
PE 3
SECR
RETARIA ESTADUA
AL DE SAU
UDE, CAV/UFPE - GE
EMNE
n região Noordeste do Brasil
B
possu
uindo 186
O Esstado de Perrnambuco loocaliza-se na
muniicípios distrribuídos em
m cinco mesoorregiões (A
Agreste, Meetropolitana, São Franccisco,
Sertãão e Zona daa Mata), cadda uma apreesentando distintas
d
varriações sociooeconômicaas,
sanittária, climátticas e fitogeeográficas que
q podem influenciar na dinâmicca da raiva local.
A raiiva pode serr subdivididda em quatro
o ciclos o urbano,
u
o sillvestre, o ru
ural e o aéreo,
este uultimo tem como princcipal agente participantte os morceggos não hem
matófagos. O
preseente trabalho levantou o recebimennto de amosstras de morrcegos pelo LANAGRO/PE
para análise rábiica entre 19991 e 2011 identificand
i
do localidadde de encam
minhamento,,
e
da. Nesse inttervalo foraam
espéccimes positiivos e quanndo possívell a espécie encaminhad
regisstradas 31.322 amostrass para analise, destas 2,64% eram morcegos (n
( = 827) coom
uma maior repreesentação das regiões metropolitan
m
nas (n = 4433 ) e sertão (n = 291). Foi
F
m a 25 espéécies e cincoo famílias.
possíível identifiicar 322 esppécimes quee pertenciam
Moloossus molosssus foi a esppécie mais enviada (n=
=151) seguida por Glosssophaga
soriccina (n=32).. Houve um
m crescimen
nto acentuaddo no envio de morcegos ao longoo dos
anos, partindo de
d uma única amostra em
e 1991 até 240 em 2011, os maioores valores
m para os úlltimos quatrro anos. Enttretanto, essse envio não
o é bem disttribuído um
ma
foram
vez qque apenas 52
5 municípios encaminnharam amoostras, com apenas nov
ve enviando mais
de deez amostras, permaneceendo muitos municípioos com a vig
gilância desscoberta nessse
grupoo. Das 827 amostras reecebidas 299 não se enccontravam em
e condiçõees adequadaas
para análise, esttando mal coonservadas,, em elevado estágio dee putrefaçãoo ou
C
a
amostras
forram positivaas com ocorrrências da
extreemamente reessecadas. Cinquenta
regiãão metropolitana ao serrtão, 29 delaas não foram
m identificaadas, as dem
mais pertenciam
as seeis espécies (Molossus molossus,
m
D
Desmodus
rootundus, Arrtibeus plannirostris, Myyotis
sp, Glossophaga
G
a soricina e Eptesicus furinalis),
f
m
muitos
dos registros
r
ocoorreram no
ambiiente urbanoo. Os princiipais problemas detectaados devem
m-se ao baixo
o número dee
amosstras de morrcegos recebbido e ao ellevado núm
mero de munnicípios sem
m
encam
minhamento que faz coom que a sittuação do conheciment
c
to da raiva nesse
n
grupoo no
estaddo de Pernam
mbuco aindda seja incippiente. Haveendo assim a necessidaade de ter coomo
resollução futuraa, para minim
mizar esse efeito,
e
se inntensificar as ações de monitorame
m
ento
da raaiva em morrcegos bem como form
mar protocolos para o reegistro, encaaminhamennto e
identtificação daas espécies enviadas
e
parra análise, implantando
i
o equipes treinadas na
identtificação de morcegos e na resoluçção de probllemas assocciados a esse grupo noss
órgãoo responsávveis pelo moonitoramentto da raiva ffacilitando assim
a
o dessenvolvimennto
do trabalho.
ves: quiróptteros, ciclo aéreo, sinanntrópicos, vigilância
v
eppidemiológiica
Palaavras – Chav
PT.0061
DEMIOLO
OGIC, SOR
ROLOGIC AND MOL
LECULAR
R STUDIES
S OF RABIIES
EPID
VIRU
US ISOLA
ATED IN BA
AT COLO
ONIES OF Histiotus
H
veelatus, BRA
AZIL
1
Marttorelli LFA1, Kataoka APAG
A
, Cam
mpos ACA2, Sodre MM
M1, Araujo DB2, Rosa AR1,
1
3
1
Trezzza Netto J , Oliveira RN
R , Almeidda MF , Saccramento DR
RV4, Durigo
on EL2,
2,3 1
2
Favooretto SR - Centro dee Controle de
d Zoonosees-COVISA
A-PMSP, Universidadee de
Sao P
Paulo, 3Insttituto Pasteuur de Sao Paaulo, 4Genoomic Engenhharia Moleccular
Rabies was deteected in twoo bats coloniies of insecttivorous Hisstiotus velatus that useed as
o buildingss in the sam
me park in thhe North reggion of São Paulo City,,
sheltter ceilings of
Brazzil in 2001 and
a 2009. Thhis park althhough locatted in an urbban area, haas a large arrea of
rainfforest. Rabiees diagnosiss was made by FAT (Fluorescent Antibody
A
T
Test)
and MIIT
(Mouuse Inoculattion Test) teechniques. The
T numberr of bats sub
bmitted to rabies
r
diagnnosis
was fforty-three in the first colony
c
and forty in the second. Beefore it, batss received
anestthesic and blood
b
samplles were colllected by cardiac punccture in sixty
y animals
(thirtty-eight of the
t first coloony and tweenty-two off the second
d colony) an
nd the presennce
of rabbies virus neutralizing
n
antibodies was determ
mined by SFIMT (Simp
plified
Fluorrescent Inhiibition Micrrotest). The antigenic ccharacterizaation of the isolates
i
wass
madee using a paanel of monoclonal antiibodies, whhich was prooduced and provided byy
Centters for Diseease Control and Preveention (Atlannta, U.S.A), as establisshed by PannAmeerican Healtth Organizattion for chaaracterizatioon of rabies isolates in Americas.
A
F
Five
bats w
were positivve to rabiess by FAT annd MIT in each colony,, 12% in thee first colonny
and 112.5% in the second. However,
H
tw
wo bats deadd of the seco
ond colony were unsuittable
for raabies diagnoosis by tradditional tech
hniques and their brainss were subm
mitted to RT
TPCR
R with positiive results, totalizing
t
seeven positivves bats indiicating 23.33% of rabiess
viruss positivity. All blood samples
s
anaalyzed preseented neutraalizing antib
bodies titerss and
sixteen animals (40%) from
m the first coolony and tw
wo (4.6%) of
o the secon
nd presentedd
mL. There was a positivve correlatioon between the
t incubatiion period inn
titerss ≥0.5 UI/m
micee and the anttibodies titeers observedd in the batss. The sampples with thee higher
incubbation periood for MIT (29
( days) were
w from baats that show
wed the higghest
neutrralizing antiibody titer. Some bats negative
n
byy MIT and FAT
F
and app
parently
healtthy, presented high antiibodies titerrs. The antiggenic charaacterization showed onlly
one aantigenic prrofile (positiive just to MAb
M C12) observed
o
in previous sttudies with
sampples isolated
d in the sam
me species of bats in Braazil. Genettic characterrization wass
perfoormed by seequencing of
o a fragmen
nt of N proteein region and
a the rabies genetic
lineaage identifieed in these study
s
were segregated
s
w isolatees obtained from
with
f
other
Histiiotus velatuss samples issolated in otther regionss of Brazil. These resullts show thee
impoortance of thhese methoddologies forr the epidem
miological suurveillance of rabies viirus
in baats and the necessity
n
off the monitooring of bat colonies in parks and environmen
e
ntal
reserrves frequen
nted by hum
mans and whhere living other
o
wildlife species as
a preventivve
actioons of rabiess control.
PT.0062
ESTIGAÇÃ
ÃO DE CA
ASO DE RA
AIVA EM FELINO, MUNICÍPI
M
IO DE SÃO
O
INVE
PAU
ULO, 2011
Menddes MCNC1, Bernardi F1, Paranho
os NT1, Alvves GM2, Olliveira JL3 - 1Centro dee
Conttrole de Zoo
onoses da Coordenação
C
o de Vigilânncia em Saúúde - Secretaaria Municippal
2
de Saaúde de São
o Paulo, Suupervisão dee Vigilanciaa em Saúde de Lapa/Pin
nheiros Secreetaria de Saaúde do Munnicípio de São
S Paulo, 3Supervisão de Vigilancia em Saúdde de
Jabaqquara/Vila Mariana
M
- Secretaria
S
dee Saúde do Município de
d São Paullo
9 casos de
d raiva anim
mal e cinco
Em 11969, no Muunicípio de São Paulo ocorreram 989
casoss de raiva humana. Enttre 1969 e 1973 (fundaçção do Centtro do Conttrole de
Zoonnoses- CCZ/SP), o núm
mero de casoos de raiva humana
h
aum
mentou 2,2 vezes, o núúmero
de annimais vacinnados cresceu cinco veezes e obserrvou-se um decréscimo dos casos de
d
raivaa animal, ch
hegando a 566% do totall ocorrido em
m 1969. A partir
p
de 1981 não
ocorrreram mais casos humaanos e entree 1983 e 20110 não foram
m registrados casos
autócctones em cães
c
e gatos. O perfil ep
pidemiológiico da raivaa vem mudaando em toddo o
Brasiil, com restrrição da áreea de circulaação da cepa canina doo vírus. Nas regiões em
m que
a raivva foi contrrolada nos animais
a
dom
mésticos, os casos de raaiva em hum
manos
diminnuíram e oss animais sillvestres passsaram a reppresentar um
m novo desaafio. Em Sãão
Pauloo a variantee canina nãoo tem sido mais
m detectaada. Atualm
mente as variiantes
circuulantes são relacionadas
r
s a quirópteeros, ocorrenndo anualm
mente, em média,
m
dois a
quatrro casos em
m morcegos não hematóófagos. Em 01/12/2011 o CCZ/SP foi comuniicado
de diiagnóstico positivo
p
para raiva de um
u felino, coom histórico de contatoo com
quiróóptero e morte sem sinttomatologiaa. O animal foi a óbito no dia 3/10
0/2011 e
encam
minhado no
o dia 04/10//2011 para a Faculdadee de Medicinna Veterináária e
Zoottecnia/USP com suspeita de envennenamento. A liberaçãoo do resultaddo positivo
ocorrreu em 01/1
12/2011. O felino, umaa fêmea, castrada, dez anos,
a
tinha livre
l
acesso à
rua e histórico de
d vacinaçãoo anterior a 2010. No im
móvel situaado em área estritamentte
residdencial no Distrito
D
de Moema,
M
viveem cinco cãães e 23 felinos. A regiãão é bastantte
arborrizada, com
m árvores quue podem offerecer abriggo e alimen
nto para difeerentes espéécies
de m
morcegos, naas proximidades de um
m parque arbborizado, Paarque do Ibirrapuera (à 750m
7
de diistância). Frrente à conffirmação da variante Deesmodus rotundus/Artiibeus liturattus
desennvolveram-se ações dee bloqueio em área de 500m
5
de raio, a partir do
d foco. Forram
realizzadas visitaas domiciliares, levantaamento de abbrigos, avalliação e orieentação paraa
encam
minhamento médico dee moradores e freqüenttadores da casa
c
que tiveram contatto
com o animal dooente, vacinnação contraa raiva e ideentificação de
d todos os animais daa
bservação por
p 180 diass a partir do óbito do an
nimal positiivo. Todos os
o
moraadia, com ob
imóvveis da área de abrangêência foram visitados, totalizando 1.277 imóveis trabalhaados,
769 ffechados e 140 recusass. Houve disstribuição de
d material educativo,
e
e foram
vacinnados contraa raiva 102 cães e 16 gatos,
g
com hhistórico de mais de seiis meses de
vacinnação, no raaio de coberrtura de focoo. Os animaais contactaantes foram acompanhaados
pelo CCZ, no peeríodo de obbservação, mantendo-s
m
e saudáveiss. Recomendda-se o
a
de viggilância: - laaboratorial; - das agresssões; - de ru
umores e caasos
impleemento de ações
suspeeitos de anim
mais com morte
m
súbitaa ou históricco de contatto com quiró
ópteros ou
outroos animais silvestres
s
e a revisão dee estratégiass do controlle da raiva devido
d
à
mudaança da situ
uação epidem
miológica da
d doença no municípioo.
PT.0063
TIGENIC AND
A
GENE
ETIC STAB
BILITY OF RABIES
S VIRUS AF
FTER
ANT
CON
NSECUTIV
VE PASSAG
GES IN MICE AND IN
I CELLS
S
Batissta HBCR1, Oliveira RN
N2, Carnielii Jr P2, Roddrigues AC2, Santos SO
O2, Fahl WO
O2, de
2
2
2
2 1
Paulaa FC , Carriieri ML , Kotait
K
I , Casstilho JG - Instituto Pasteur
P
- Virrologia,
2
Instiituto Pasteuur
ognized stabbility of the rabies viruss (RABV), antigenic an
nd genetic
Desppite the reco
differences amo
ong strains isolated from
m different species havve been founnd. Differennt
i
in generating heterogeneity in RABV
V, includingg duration of
o
factoors may be involved
infecction, virus load
l
and hoost immune response. This
T work was
w carried out
o in orderr to
exam
mine the antigenic and genetic
g
stabbility of RA
ABVs isolateed from diffferent naturaal
reserrvoirs and to
o help the understandin
u
ng of viral pathogeny
p
a
after
consecuutive passagges in
different system
ms. In this stuudy were ussed tree RA
ABV strains, one isolateed from cannine,
m haematopphagous batt and the staandard rabiees virus straain (Challennge
one iisolated from
Virus Standard - CVS). Theese strains were
w submittted to five consecutivee passages in
i
micee and in cells. The conssecutive passsages in miice were maade by intraccerebral rouute,
for thhat, groups of six mice were subm
mitted to fivee inoculations with eacch one of thee
threee RABV straains. The innoculated mice
m were obbserved dailly and the dates
d
of deatth
weree recorded. The
T consecuutive passagges in cells were made in “Neurob
blast albino
mousse” (N2a) cell
c lineage, for that, thee strains weere inoculateed in suspen
nsion cells and
a
incubbated for 72
2 hours, subbsequently, cells
c
were frozen
f
and thawed
t
threee times. Both
micee and cell paassages werre submittedd to antigeniic and genettic characteerization. Thhe
antiggenic characcterization was
w determiined by indiirect immunnofluorescennce (IIF) wiith a
panel of eight monoclonal
m
a
antibodies
(
(Mabs)
raiseed to RABV
V antigens. For the gennetic
NA was exttracted withh Trizol and
d submitted to reverse
charaacterizationn the total RN
transscription-po
olymerase chhain reactioon (RT-PCR
R) with prim
mers targetinng the N andd the
G geenes, the am
mplicons obttained were subjected to nucleotide sequence analysis. Thhe
RAB
BV sequencees were anaalyzed usingg Bioedit paackage. Nonne significannt modificattion
was ddetected in the RABV pathogeny after five coonsecutive passages,
p
foor all RABV
Vs
analyyzed. The N gene, show
wn the samee characteristics after five
fi consecuutive passagges in
both systems an
nalyzed (micce and cells) for the treee RABV strrains. The RABV
R
isolaated
m canine and
d the CVS, shown
s
the same
s
nucleootide sequennce for the G gene afterr five
from
passaages in micee and in cellls. The RAB
BV isolatedd from haem
matophagous bat shownn one
aminno acid chan
nge in the G gene after five passagges, as in cellls as in micce. Our resuults
confi
firm the high
h stability of
o RABV an
nd indicate minor
m
genettic stability of RABV
isolaated from haaematophaggous bat thann RABV isoolated from
m canine, moore experim
ments
couldd be made to
t confirm thhese resultss.
Finanncial support: Instituto Pasteur
PT.0064
ORRÊNCIA
A DE RAIV
VA EM HU
UMANO TRANSMIT
TIDA POR ANIMAL
OCO
SILV
VESTRE, EM
E ABRIL
L 2012, NO
O MUNICIP
PIO DE TA
APURAH-M
MT.
Reis VGL1, Roccha SM2, Allmeida NN1, Barros MC1, Reis RC
C3 - 1SES-M
MT - Vigilânncia
Saúde Ambiiental, 2Secrretaria de Vigilância
V
em
m Saúde do Ministério - GT Raivaa,
em S
3
Secrretaria Mun
nicipal de Saaúde de Cuiiabá - CCZ
SUMO
RES
A raiiva é uma doença
d
infeccciosa agudaa, causada ppor um víru
us, que comppromete o
sistem
ma nervosoo central (SN
NC), levanddo a um quaadro de enceefalomielite aguda, cujaa
letaliidade é de aproximadam
a
mente 100%
% (Brasil, 20005; Brasil,, 2009 e Kootait et all,
20099). Os anim
mais doméstiicos, silvesttres terrestrees e morceg
gos são os maiores
m
transsmissores deessa doençaa aos seres humanos
h
priincipalmentte pelo conttato da salivva
contaaminada por meio de mordedura,
m
l
lambedura
o arranhaddura (KOTA
ou
AIT et all, 2009
2
e WA
ADA et all, 2011). O vírus
v
da raivva apresentaa uma alta caapacidade de
d adaptaçãoo a
diferrentes espéccies de mam
míferos, senddo esta doennça com am
mpla distribuuição mundiial,
ocasiionando graande impactto socioecon
nômico e geerando um grave
g
problema de saúdde
públiica (KOTAIIT et all, 20009; e WAD
DA et all, 20011). Diantee da problem
mática da
ocorrrência destaa doença em
m escala munndial este estudo teve como
c
objetiivo avaliar o
númeero de atend
dimentos poor agressõess causadas por
p animais silvestres, sobretudo por
p
morccegos, no esstado do Maato Grosso no
n período de
d janeiro de
d 2007 a juulho de 20122,
atravvés dos dado
os registrados no SINA
AN/DATAS
SUS/MS/SE
ES-MT. De acordo
a
com
m os
regisstros do SIN
NAN, para o período annalisado foraam notificadas 688 agrressões a
humaanos por an
nimais silvesstres. Destes casos de agressões
a
no
otificadas, 3,0%
3
(21/6888) a
espéccie agressorra foi a rapoosa, 23,5% (162/688)
(
fo
foram por prrimatas não-humanos e
73,4%
% (505/6888) foram porr morcegos.. As agressõões por morrcego corresspondem em
m
médiia 84,1 (5055/6) agressõões por ano, sendo 64,7% (327/5055) destes ateendimentos
regisstradas na zoona urbana e 35,2% (1778/505) na zona rural. Este estudoo corrobora com
os reealizados poor Reis et all, 2010; e Wada
W
et all, 2011, em que
q as agressões por
morccego segue um
u padrão onde
o
os maaiores registrros foram na
n zona urbaana, sugerinndo
uma ascendente procura ao atendimentto do serviçço público de
d saúde. Esste fato podee ser
u
como
o também, a
conseequência daa acessibiliddade ao servviço de saúdde na área urbana
consccientização da populaçção urbana em
e relação ao
a contato com
c
estes annimais. No
períoodo avaliadoo observou--se que os maiores
m
conttatos foram principalm
mente por
morccegos não hematófagoss, insetívoro
os e frugívorros, comunss em ambien
ntes urbanoos,
comoo edificaçõees e em árvoores frutíferras, utilizadaas em paisaagismo cultu
ural
regioonalizado, como
c
em poomares e praaças, facilitaando a interração entre estas
e
espéciies, e
com os caninos,, felinos e huumanos, poodendo ocassionar gravees consequêências, fatoss
n estudos de Sodré ett all, 2010 e Ribeiro et all, 2010.
estess observados também nos
Anallisando os casos
c
de raivva em morcego, no perríodo estudaado foram reegistrados em
e
20088, 02 casos em
e morcegoo não hemattófagos. Em
mbora não teenha observ
vado registroo de
posittividade em
m morcegos hematófago
h
os, no estadoo de Mato Grosso
G
tem ocorrido caasos
de raaiva em herbbívoros, trannsmitidos por
p Desmoduus rotunduss, sugerindo
o assim umaa
subnnotificação nos
n sistemass de informaação. Este ffato indica que
q o vírus rábico silveestre
– com
mpatível com a variantte 3, tem cirrculado no estado
e
repreesentando asssim um graande
riscoo para a população hum
mana. As nottificações dde agressõess a humanoss, os casos de
d
raivaa em morceggos, a ocorrrência de raiiva em herbbívoros, denntre outros fatores,
f
evideenciam o reegistro de um
m caso hum
mano no munnicípio de Tapurah,
T
diaagnosticada por
variaante compattível de anim
mal silvestree (variante 3)
3 com sugeestiva transm
missão por
cervíídeo. Estas situações
s
diiscutidas, su
ugerem que outros casoos suspeitoss de raiva
podeerão ocorrerr, sendo necessária e dee fundamenttal importânncia a impleementação de
d
ações de vigilânncia e conduutas oportunnas frente a contato porr morcegos e outros aniimais
a
a todaa sociedade sobre o riscco de transm
missão da dooença
silvestres, sinaliizando um alerta
hos desenvo
olvidos peloos
por eestas espéciees. Este estuudo teve como incentivvo os trabalh
profeessores Wilson Uieda e Ricardo Moratelli
M
Roocha, e o apooio instituciional da
Secreetaria Estad
do de Saúdee do Estado de Mato Grrosso.
Palaavras-chavee: Raiva, Viigilância, Animais Silvvestres, Quirrópteros.
PT.0065
OSPHOPRO
OTEIN P GENE
G
IS NOT
N
ASSO
OCIATED WITH
W
RAB
BV
PHO
INCU
UBATION
N PERIOD AND LETHALITY
Fahl WO1, Scheeffer KC1, Iaamamoto K1, Carnieli Jr
J P1, Castillho JG1, Oliiveira RN1,
1
1
2 1
K
I , Braandão PE - INSTITUTO PASTE
EUR - VIRO
OLOGIA,
Carriieri ML , Kotait
2
Univversidade de
d São Pauloo - FMVZ-U
USP
The pphosphoprootein P is muultifunctionnal and the lless conservved among Rabies
R
viruss
(RAB
BV) proteinns, associateed with the ribonucleop
r
protein (RN
NP), with a role
r in the
bindiing of the polymerase L in the RN
NP. Interactiions involviing P are rellated to RABV
tropism, the celll-to-cell viruus spread annd inhibition of innate immune ressponse that
o stops the replication of viruses. P is also inv
volved in thhe trans-axoonal
interferes with or
BV and inteerferon (IFN
N) inhibitionn, which coould modulaate the
transsport of RAB
pathoogenesis virral. There arre hypothesses that the RABV
R
protteins polym
morphisms caan
lead to variation
ns in the repplication effi
ficiency of different
d
strrains of RABV in diffeerent
hostss, culminatinng in variabble viral titeers, modulatting viral paathogenesis and leadingg to
different efficien
ncies of intrra-axonal trransport andd function as a co-factoor in the
T
synthhesis of viraal RNA. Hoowever theree is still muuch to be claarified on thhis subject. To
this eend, 29 RAB
BV strains from
f
bats of the genus Artibeus spp. isolated in
n mouse braain
weree selected with
w variablee lethality (L
LET) and inncubation peeriods (IP) in
i days afterr
inocuulation in th
he central neervous systeem of mice.. The samplles were sub
bjected to
titrattion in N2a cell cultures and compplete sequenncing of the P gene. Data analysis
incluuding LET, IP and titer in associatiion with thee pattern of segregationn of the viraal
strainns studied inn the phyloggenetic treee showed noo associationn between these
t
variabbles
and ppolymorphisms of the P Gene. It can
c be specuulated that other
o
proteinns, such as the
polym
merase L, currently
c
under consideeration by thhe authors, may
m have an
n importantt role
in thee modulatio
on of incubaation and letthality, wheen consideriing viral strains that aree
very closely relaated regardiing the P geene. This infformation iss applicablee for discusssions
oirs, the dynnamic of thee virus and the
t differentt
abouut the importtance of rabbies reservo
form
ms of host-viirus co-evollution, contrributing to further
f
studies on RAB
BV pathogennesis.
S
Finanncial Suppoort: Institutoo Pasteur dee São Paulo and CAPES
PT.0066
RVEY OF THE
T
SPAT
TIAL DISTR
RIBUTION
N OF BATS ACCOR
RDING TO
SUR
TYP
PE AND PO
OSITIVE FOR
F
RABIE
ES IN BOT
TUCATU-S
SP THE PE
ERIOD 20006
TO 22011.
Langgoni H, Men
nozzi BD, Vitória
V
C
Bats are consideered to be thhe man transmitters andd disseminaators of rabiies virus in
c
particularly in regions
r
wheere canine rabies has beeen controllled.
Brazzil’s urban centers,
This brings up a great fear for
f the publlic health auuthorities inn the country
y about thiss
w
be the possible reintroductio
r
on of rabiess in domestiic animals due
d to
diseaase, which would
easieer interaction with the bats,
b
and alsso from conntact with hu
umans. Cleaarly increasing
the nnumber of caases of various speciess of bats thaat were diagnnosed as poositive for raabies
throuughout the country
c
espeecially in thhe state of S
São Paulo. The
T municippality of
Botuucatu – SP iss located in the midwesstern regionn and is locaated at 22 53 09 south
latituude and 48 26
2 42 west longitude,
l
8 meters above
804
a
sea leevel, and makes
m
an
impoortant surveiillance workk, since 20003, where annimals are collected
c
on
n public roaads or
residdences and then
t
are subbmitted for diagnosis
d
att Faculty off Veterinaryy Medicine and
a
Anim
mal Science of UNESP, Botucatu, where is innstalled the Zoonosis
Z
D
Diagnostic
Labooratory. Am
mong theirs 2006
2
and 20
011 where received
r
9844 bats for diiagnosis, beeing
in 20006, 278 batts, 182 in 20007, 115 in 2008, 116 in
i 2009, 166 in 2010 annd 127 in 2011.
Onlyy in 2008 an
nd 2011 did not find poositive for raabies bats. In others yeaars there weere
11 poositive diaggnoses, charracterized att least three genres: Arttibeus, Myo
otis and
Nyctinomops an
nd two speciies, Artibeuss lituratus aand Myotiss nigricans. The positivve
bats (all non-hem
matophagouus) are distrributes in thhe central paart of the citty, in areas whit
w
he city. In thhe spatial diistribution of
o species inn the
largee trees, two creeks that surround th
counntry, we find
d the homoggeneous disttribution off at least 40 genus/species of bats,
beingg the most frequent
f
55%
% (557) bellonging to thhe genus Molossus,
M
inccluding speccies
Moloossus molosssus and Moolossus rufus, distributeed in equal uniformity
u
throughout
t
the
muniicipality. Reevealing thaat although this distribuution, and th
he large num
mber of
genera/species involved, thhe virus is reestricted to only
o
five geenera/speciees, also
nicipality. Also
A important to highllight the nonnrestriicted to the central areaa of the mun
posittivity of the bats of the genus Molo
ossus, despiite being thee greater quuantity and better
b
distriibuted by co
ounty.
1
UN
NESP - São Paulo
P
State University – Veterinarry Medicinee and Anim
mal Productioon
Colleege – Publicc Health andd Veterinaryy Hygiene Department
D
t – Botucatuu – SP, Brazzil.
PT.0067
V
RÁ
ÁBICO EM
M INDIVÍDU
UOS E CO
OLÔNIAS DE
D
ESTUDO DO VÍRUS
OS NO RIO
O GRANDE
E DO SUL BRASIL
QUIRÓPTERO
R
JCA2, Ferreira
F
JC2, Batista HB
BCR3, Rigo
oletti R2, Rooehe PM2,4 Pachheco SM1, Rosa
1
2
3
4
Instiituto Sauver, FEPAGR
RO - IPVDF
F, Institutoo Pasteur, UFRGS
U
O núúmero de esppécies de quuirópteros infectadas pelo
p vírus daa raiva (RABV) tem siddo
cresccente. Até o momento, 42 espéciess de três fam
mílias foram
m identificad
das no Brasil. Os
dadoos sobre a prrevalência do
d vírus nesstas espéciess e a importtância destaas na
manuutenção doss diferentes ciclos da raaiva, ainda são
s escassoss. O Institutto de Pesquisas
Veterinárias Desidério Finaamor (IPVD
DF/FEPAGR
RO) é refereencia para o diagnósticco de
d Brasil. Deesde agostoo de 2007 o
raivaa no Rio Graande do sul (RS), sul do
IPVD
DF/FEPAGRO faz a iddentificação morfológicca dos quiróópteros enviiados ao
diagnnóstico de raiva,
r
cujos espécimes são proveniientes de divversos munnicípios do
Estaddo. A identiificação dass espécies dee quirópteroos é fundam
mental tanto para avaliaar a
circuulação do RA
ABV, comoo para identtificar possívveis impacttos do estressse nas colôônias
de quuirópteros das
d diferentees espécies. De 2007 a 2011 houvee 47 morceggos positivoos
para o RABV vaariando de 07
0 a 13 indiivíduos/anoo. Cerca de 300
3 morceggos/ano foraam
boratório dee Virologia do
d IPVDF/F
FEPAGRO para o diaggnóstico de
enviaados ao Lab
raivaa, provenien
ntes de 115 municípios
m
. Das 38 esppécies que ocorrem
o
no RS, nove foram
fo
identtificadas inffectadas com
m o RABV.. As espéciees são Artibeeus lituratuus e Desmoddus
rotunndus pertenccentes à fam
mília Phyllo
ostomidae; E
Eptesicus fuurinalis, Hisstiotus velattus,
Lasiuurus ega, Myotis
M
levis e Myotis nig
gricans da família
f
Vesppertilionidaae, e
repreesentantes da
d família Molossidae,
M
Molossus molossus
m
e Tadarida
T
brrasiliensis. As
A
espéccies A. liturratus, E. furrinalis e Lassiurus ega foram
f
relataadas como agressoras.
a
Porém
m, tal agresssão ocorreuu no momen
nto da captuura. As outras espécies identificadaas
posittivas para o RABV nãoo apresentarram relatos dde agressão
o, apesar de terem sido
identtificadas mu
uitas fraturaas ósseas e cranianas
c
(ààs vezes, quase sem cérrebro), lesõees
internnas (órgãoss com hemattomas) ou carcaças
c
seccas. As espéécies Molosssus molossuus e
Tadaarida brasiliensis são as
a espécies mais
m frequenntes com diiagnóstico positivo
p
para
raivaa e, também
m as mais comuns no Riio Grande do
d Sul. Nos últimos doiis anos (2011020111) o IPVDF//FEPAGRO
O vem receb
bendo indivííduos proveenientes de colônias
c
e,
entãoo, é realizadda a análise a fim de veerificar se ass colônias possuem
p
circculação do vvírus
rábicco. Os resulttados mostrram que collônias até 700 indivíduoss não possu
uem morceggos
posittivos; no enttanto, em allguns casos, morcegos encontradoos mortos e provenientees de
colônnias com mais de 500 ou
o 1000 moorcegos, posssuem indivvíduos positiivos, e nessse
caso,, a espécie é T. brasilieensis. Os daados demonsstraram quee nem semprre os relatoss de
agresssões por morcegos
m
esttão relacionnados com a infecção peelo RABV. Portanto,
estuddos devem ser
s realizadoos para avalliar se o imppacto na rem
moção dos morcegos
m
d
de
seus abrigos diuurnos, sem aguardar
a
suaa dispersão natural e saazonal, acarrreta o aumeento
c
e consequente
c
e queda na imunidade,
i
que pode in
nfluenciar no
n
do esstresse nas colônias,
aumeento do núm
mero de indiivíduos infeectados peloo RABV.
PT.0068
BEH
HAVIOR FOR SIX YE
EARS OF ATTACKS
A
S A HUMA
AN BY VAM
MPIRE BA
ATS
IN N
NORTHER
RN JALISC
CO, MEXIC
CO, AFTER
R THE IMPLEMENT
TATION OF
O A
INTE
ENSIVE CAMPAIGN
C
N TO CON
NTROL TH
HIS POPUL
LATION AND
A
ITS
REL
LATIONSH
HIP WITH THE EXIS
STENCE OF
O REFUG
GES AND LOCAL
L
LIVE
ESTOCK NEARBY.
N
Moraales Rodrígu
uez JA1, Goonzález Lóp
pez AA1, Roodríguez Coobián FJ2 - 1Secretaría de
d
Saludd Jalisco - Región
R
Saniitaria 01 No
orte Colotlánn, 2Secretarría de Agriccultura,
Ganaadería, Desaarrollo Ruraal, Pesca y Alimentació
A
ón de Jaliscoo - Distrito de Desarrolllo
Ruraal 08
f six yearss of attacks on humanss by
Absttract. Objective. To deetermine thee behavior for
vamppire bats in northern Jaalisco, Mexiico, after thee implemen
ntation of ann intensive
camppaign to conntrol their population by
b using an ointment
o
made
m
of warffarin suspennded
in vaaseline ( Vam
mpirinip II)), and the reelationship of
o such attacks in the presence
p
of
refugges and nearrby livestocck. Materiaal and Meth
hods. From June 2006 to April 20008
was made
m
an inttensive searrch campaiggn bats refugges in northhern Jalisco,, recording your
locattion on GPS
S and applyiing ointmen
nt of anticoaagulant only
y those classified as
vamppire bats by
y their externnal morphological featuures. In add
dition, a seaarch was
condducted and timely moniitoring of atttacks on huumans by vaampire bats during the years
20066 to 2011, an
nd the amouunt of livesttock in the area
a in the same
s
periodd to find anyy
relatiionship betw
ween these variables. Results.
R
Duuring the six
x years studiied there weere
864 aattacks on humans
h
by vampire
v
batts. The ten municipaliti
m
ies of northeern Jalisco
reporrted attacks, although in
i different amounts. Frrom 2008 thhere was a reduction
r
inn the
numbber of attackks, with thee overall redduction at thhe end of 20011, 50%. We
W identified
551 rrefuges with
h bats, but only
o
13.25%
% of them had
h the preseence of vam
mpire bats. We
W
obtaiined a positive correlattion betweenn aggressionn to humans and the prresence of
refugges in the prresence of vampire
v
bats (r2 = 0.699, p = 0.00). There was no correlation
betw
ween aggresssion and thee presence of
o livestock in the regioon. The mosst affected
muniicipalities were
w Bolañoos Chimaltittán and Sann Martin de Bolaños.
B
Co
onclusions.. The
ointm
ment of antiicoagulant Vampirinip
V
II has proveen useful in
n controllingg vampire bats.
b
The nnearby refu
uges identifyy the presennce of these bats, can seensitize the population to
carryy out strateg
gies to reducce the attackks on humann and on local livestock.
Ackn
nowledgem
ments. The authorities
a
o the Healthh Region 1 of the Heallth Departm
of
ment
of Jaalisco, the Rural
R
Develoopment Disttrict 08 of thhe Ministry
y of Agriculture, Livesttock,
Ruraal Developm
ment, Fisherries and Foood of Jaliscoo, and H. Municipalitie
M
es of northerrn
Jaliscco-Bolaños, Colotlán, Chimaltitán
C
n, Huejucar,, Huejuquillla el Alto, Mezquitic,
M
S
San
Marttin de Bolañños, Santa Maria
M
de loss Angeles, Totatiche
T
annd Villa Gueerrero- durinng
the yyears 2006 to
t 2008, for his technical, methodoological and
d / or financcially unquaalified
support to perfo
orm this worrk.
PT.0069
TECTION OF
O RABIE
ES VIRUS IN
I INSECT
TIVOROU
US BATS FR
ROM THE
E
DET
URB
BAN AREA
A IN MONT
TERREY NUEVO
N
LE
EON, MEX
XICO
Reynna EJ1, Péreez JZV1, Gaalindo EIG1, Rosales PC
C2 - 1Laboraatorio Estataal de Salud
Públiica - Secretaría de Saluud del Estaddo de Nuevoo León, 2Ceentro de Investigación
Biom
médica del Noreste
N
- Instituto Mexxicano del Seguro
S
Sociaal
Mexico theree are 154 speecies of bats, of these, 37 are distrributed in Nuevo
N
Leon,,
In M
whicch can be innsectivorouss, frugivoroous, or hemaatophagous.. The most common
c
baat in
Nuevvo Leon is Tadarida
T
braasiliensis mexicana,
m
it is and insecctivorous, an
nd it is alsoo the
smallest free-taiiled bats. Thhere are othher species oof insectivorrous bats, suuch as
ous palliduss,
Nyctticeius humeeralis, Myotis sp, Lasiuurus ega, Euumops perootis, Antrozo
Coryynorhinus toownsendii, Lasiurus
L
cinnereus, and Leptonycteeris nivalis, which are uuseful
in maaintaining ecological
e
b
balance,
conntributes to the
t spread of
o seeds, and
d eat insects that
harm
m agriculturee. However, in 1984 th
he rabies virrus was repoorted in Taddarida
brasiiliensis mex
xicana , and in 1998 thee first humaan death cauused by rabiies transmittted
by baats insectivoorous was reported
r
in Latin
L
Ameriica. Betweeen 2009 andd 2010, 7 caases
of the variant V--9 were diaggnosed andd they corressponded to the
t same doomestic speccies,
c
In rural areas of N
Nuevo Leon
n, as well ass in the urbaan
Canis familiaris and Felis catus.
s
of bats.
b
Monterrrey is the
area of Monterreey there aree habitats foor different species
t
most populous
p
citty in Mexicco, there, thee highest
capittal of the staate and the third
numbber of casess have been detected. Therefore,
T
thhe objectivee of this stud
dy was to detect
casess of rabies in the urbann area of Moonterrey beccause of pottential risk for
f humans and
domeestic animalls. During 2011
2
and 20012, 21 bat samples
s
weere analyzedd by using
immuunofluoresccence, antiggenic characcterization and
a genotypping techniqques,.
Identtification off species shoowed 14 Taadarida brassiliensis, 3 Lasiurus
L
sp, 1 Nycticeiuus
humeeralis, 1 Lep
ptonycteris nivalis, 1 Lasiurus
L
egaa, and 1 Lassiurus cinereeus. From thhese,
8 batts were posiitive for rabbies virus, annd an antiggenic characcterization was
w
perfoormed using 8 monocllonal antiboodies, with this study, the variantt 9 (V-9) waas the
mostt predominaate. Genotyyping of Lassiurus ega annd Lasiuruss cinereussee samples
show
wed 99% of similitude with
w Lasiuruus intermeddius. This sttudy demonnstrates the high
h
prevaalence of raabies in inseectivorous bats,
b
which threatens
t
bo
oth humans and domestic
anim
mals, and sugggests the im
mplementattion of sustaainable prog
grams to preevent cases of
rabiees in these species.
Acknnowledgements: We arre grateful too Miguel Angel
A
Zunigaa, Isabel Agguilar Tavitaas
and A
Alma Lilian
na Lizarán Meneses
M
forr their suppoort in the diiagnosis of rabies
r
viruss.
Finanncial support: This worrk was supp
ported by thhe Rabies Prrogram of thhe Ministry of
Heallth of Nuevo
o Leon and Health Servvices of Nuevo Leon.
PT.0070
a historic review
r
Publlic producttion of antirrrabies vacccines in Arrgentina: an
Perezz OR1, De Filippo
F
J1, Jurado
J
R1, Micucci
M
M1, Rudak L1, Fernandez G1, Zanetti F2,
Calam
mante G2 - 1ANLIS "D
Dr. Carlos G.
G Malbrán" - Servicio Vacuna
V
Anttirrábica - IN
NPB,
2
Instiituto Nacion
nal de Tecnnología Agropecuaria (IINTA) - Insstituto de Biotecnología
In 19976, the most importannt outbreak of
o rabies occurred in Argentina.
A
Thhe governm
ment
adoppted differen
nt strategiess to find an answer
a
to thhis sanitary problem. The
T main
impoortant was a vaccinationn campaignn using first generation vaccines Fu
uenzalidaPalaccios type (F
FP), that havve been prod
duced in ratt sucking brrain. At natiional level, this
produuction has been
b
done inn the INPB-ANLIS “D
Dr. Carlos G.
G Malbrán” and resulteed in
an efffective conntrol of the outbreak.
o
Unntil 2009, 100000 hum
man doses (oonly for natiional
use) and 50000 veterinary doses
d
(only for outbreaak control) have
h
been produced
p
peer
wing OPS-O
OMS´ recom
mmendationns, the goveernment is
year.. Since 20099, and follow
leadiing for a rep
placement of
o FP vaccinnes for impoorted second
d generationn ones for
humaan use and stressed
s
vacccination annimal campaaigns using similar vacccines produuced
by prrivate vacciine manufacctures. Althhough endem
mic areas sttay in the no
orth of the
counntry, modificcations in bats´ migratiion patterns have to be consider, due
d to climaate
channges.
This scene entaiils an urgent need of veeterinary vaaccines to make
m
a real sanitary
s
imppact.
his challenge, we have been
b
develooping cell-cculture platfforms to
Lookking after th
approoach more technologic
t
cal vaccines.
The ddevelopmennt of veterinnary vaccinees using BH
HK cells as substrate haas been donne on
microocarriers ceell culture. We
W have woorked with cell
c densitiees between 5 and 8 x 1006
cells//ml generatting, at leastt, four harveests with titlles of 106 LD
L 50/ml or higher.
h
Threee
consiistent vaccine batches have
h
been produced
p
with a potenccy of 1 IU/ddose or highher.
For new
n nationaal vaccines, we have deeveloped a rrecombinan
nt canarypoxx virus in
collaaboration wiith the INTA
A. This viruus has the gllycoprotein
n rabies genee among its
DNA
A, extracted of CVS-Malbrán rabiees strain, deeveloped in our laboratoory. The virrus is
grow
wn over prim
mary culturee of chickenn embryo fibbroblasts. Inn non avian hosts, thesee
virusses produce a suicidal infection:
i
thhey can’t coomplete its infection
i
cyycle, but therre’s
expreession of viiral proteinss that are proocessed by host cells, generating
g
a humoral and
a
celluular immunee response. This
T potential vaccine has shown impressive antigenic
valuees, higher thhan 3.5 IU/m
ml and the measuremen
m
nt of neutraalizing antibbodies doesnn’t
decreease significcantly after a year. An advantage of
o these viru
uses is the resistance
r
too
liofillization proccess using lactose
l
10 mg/ml
m
withoout losing itts immunogenicity. In this
t
way,, we have reealized potency assays that show similar
s
valuees in compaarison with not
nother
liofillizated virusses. By thesse days, we are workingg hard in deeveloping an
recom
mbinant can
narypox viruus containinng only the rabies nucleoprotein gene.
g
The finnal
objecctive would
d be a canaryypox antirabbies vaccinee containing
g both the raabies
nucleeoprotein annd glycoprootein genes.
PT.0071
PLEO DE LA
L TÉCNIICA DE PC
CR EN MU
UESTRAS CLÍNICAS
C
S PARA
EMP
DIAGNÓSTIC
CO DE RAB
BIA - DESC
CRIPCIÓN
N DE CASO
O
M 1, Ram
mírez-Torress J1, Venegaas-Cureño E1, Ortiz-Ricco A1,
Roblles-Pesina MAG
1
Delggadillo-Álvaarez JB , Moontaño-Hiroose JA1 - 11Centro Naccional de Seervicios de
Diaggnóstico en Salud
S
Anim
mal (CENAS
SA / SAGA
ARPA / SEN
NASICA) - Carretera
C
Fedeeral México - Pachuca Km
K 37.5, Tecámac de Villanueva,
V
, Estado de México, México
M
c
de laa polimerasaa (PCR) se aplica exitoosamente en
n el diagnósstico
La reeacción en cadena
confi
firmatorio y la vigilanciia epidemioológica de laa rabia. A peesar de no ser
s una técnnica
presccrita por la OIE
O ni por la
l OMS, cu
uando se reaaliza por perrsonal entrenado y bajoo
estricctos controlles de calidaad resulta seer de gran utilidad
u
cuanndo el diagnnóstico por
técniicas tradicioonales no ess muy claro.. Adicionalm
mente, una vez amplifiicado, el gennoma
se puuede secuennciar para coonfirmar el origen del virus
v
y provveer informaación muy
valioosa para estu
udios epidem
miológicos y de evoluución. A conntinuación se
s describe un
u
ejem
mplo: Se reciibió un encééfalo de cannideo. La historia sólo refería
r
un annimal vacunnado,
con uun cuadro de
d dos semannas de deprresión, pérdiida de peso, tristeza y falta
f
de apetito
pocoo antes de laa muerte. Noo hubo expo
osición de personas.
p
Laa muestra fuue procesadda en
menoos de 24 horras por inm
munofloureseencia segúnn la técnica descrita
d
porr la OIE (Maanual
of Diiagnostic Teests and Vaaccines for Terrestrial
T
A
Animals,
6thh Edition. Paaris, Francee,
20088). Los resulltados obtennidos fueronn discordanttes entre do
os técnicos: para uno fuue
negaativa, mientrras que otroo observó unn foco fluorescente en una
u de las im
mpresiones. La
pruebba biológicaa para aislam
miento viraal en ratoness CF1 resulttó negativa aun después de
un paase ciego y la muestra se dio comoo negativa al
a no haber virus
v
viablee. Para
compplementar el
e diagnósticco, la muesttra fue proceesada en el laboratorio de biologíaa
moleecular del Centro, segúnn la técnicaa descrita poor Heaton P.R. (Journall of Clinicall
Micrrobiology. Nov.
N
1997: pag
p 2762- 2766).
2
Para la validacióón del proceeso de
extraacción y PC
CR fue proceesada a la paar una muesstra de viruss de rabia vacunal.
v
Adiccionalmentee, como testiigo interno se empleó un
u juego de primers paara detecciónn
de A
ARN 18S 3224 pb con laa finalidad de
d descartarr la posibiliddad de tenerr un falso
negaativo debido
o a posibles factores de inhibición de la muesttra. Los productos
obtennidos fueron
n visualizaddos en un geel de agarossa al 2%. Enn la muestraa problema y en
el tesstigo positivvo de extraccción se obsservó un am
mplificado de 606 pb. Asimismo,
A
see
obserrvó una ban
nda de 495 bp
b en la muuestra probleema y testig
go de extraccción. Es bieen
sabiddo que la sensibilidad y especificad de las técnnicas de PC
CR son más elevadas quue las
técniicas como detección
d
dee anticuerpo
os fluorescenntes, inmunnoperoxidassa o ELISA..
Adiccionalmentee, no se requuiere que el virus permaanezca viabble para su identificacióón y
clasifficación. Ell eventual esstado de descomposición del tejiddo tampoco presenta
p
unn
probllema para el
e diagnósticco. Al tenerr estos primeers con la capacidad dee identificarr seis
diferrentes genottipos de rabiia, se reduce la cantidaad de reactiv
vos necesariios para su
identtificación. Actualmente
A
e la muestraa está en prooceso de seccuenciaciónn. Este trabaajo
fue realizado con recursos del
d gobiernoo federal dee México, a través de laa Secretariaa de
ural, Pesca y Alimentacción.
Agriccultura, Gannadería, Deesarrollo Ru
PT.0072
ESTRATÉGIA
A DE AÇÃO
O PARA DIMINUIR
D
O NÚMER
RO DE TR
RATAMENTOS
TIRRÁBIC
COS HUMA
ANOS PÓS
S-EXPOSIÇ
ÇÃO
ANT
Friass DFR1, Nunnes JOR2, CARVALH
C
O AAB2 - 1EMBRAPA
A CAMPO GRANDE,
2
UNE
ESP - FACU
ULDADE DE
D CIÊNCIIAS AGRÁ
ÁRIAS E VE
ETERINÁR
RIAS DE
JABO
OTICABAL
L - MEDIC
CINA VETE
ERINÁRIA PREVENT
TIVA
A avvaliação dos casos de aggressão por animais, seem critérios bem definiidos, pode leevar
à inddicação equiivocada de profilaxia
p
a
antirrábica.
O número de
d tratamenttos pósexpoosição, em muitos
m
muniicípios em todo
t
o Brasiil, tem preocupado as autoridades
a
sanittárias. Assim
m, o presentte trabalho objetivou
o
avvaliar os reggistros e a conduta
c
da
profiilaxia antirráábica humaana no Muniicípio de Jabboticabal, no
n período de
d 2007 a 20010.
Para tanto, foram
m analisadaas as inform
mações das fichas
fi
de ateendimento e dos resultaados
a
Os dados fooram tabulaados
de viisitas domicciliares e obbservação doos animais agressores.
em E
Excel e analisados com
m softwares EpiInfo
E
e Action.
A
Tamb
bém calculoou-se o custto
diretoo das vacinas antirrábicas humanaas aplicadass. Foram elaaboradas a nova
n
ficha de
d
atenddimento anttirrábico hum
mano a ser proposta, a ficha referêência de ateendimento a
agresssão animall e a ficha de visita zoo
ossanitária, todas
t
com seus
s
respecttivos manuaais de
instruução para preenchimennto. Além disso, foi reaalizada a cappacitação doos responsááveis
pela aplicação das
d mesmas e as visitas zoosanitáriias. Em 13666 fichas annalisadas,
m agravo paara cada 2122 habitantes; das pessoaas agredidass, 74,3%
compputou-se um
recebberam profilaxia pós-exxposição, nu
um total de 1.251 dosees de vacinaa. A análise
detallhada dos daados, aliadaa ao fato de Jaboticaball ser consideerada regiãoo controladaa
para a raiva, perrmitem quesstionar que 894 pessoas podem terr recebido vacina
v
sem
necessidade, totalizando 10094 doses a um custo approximado de R$20.1334,70. Nas
a
agressores, 78,,1% eram
visitaas zoosanitáárias, constaatou-se que,, dentre os animais
domiiciliados; 844,4% estavaam sadios nos dias zeroo e após perríodo de obsservação; 555%
dos aataques foraam provocaddos e 73% das
d agressõees ocorreram
m nas próprrias residênccias.
Vale ressaltar quue esta pesqquisa compllementou um
ma anterior,, onde o índdice de indiccação
ós-exposiçãão, no períoddo de 2000 a 2006, foi 81,6%%. A redução desse
d
de traatamento pó
índicce nos anos seguintes demonstrou
d
que o conheecimento doos dados peelos profissionais
envoolvidos no attendimento antirrábicoo humano leevou a uma atuação maais criteriosaa com
conseequente dim
minuição daa taxa de ind
dicação de vacinas
v
e dee erros no prreenchimennto
das ffichas. Concclui-se, entãão, que umaa maior atennção deve ser dispensaada ao
atenddimento a agravos por animais, coom adequaçõões na fichaa de registro
o, aplicação das
fichaas referênciaa de atendim
mento a agreessão animaal e de visita zoosanitárria. É essenncial a
interaação dos seerviços médiicos e médiicos veterináários e a avaliação de todos
t
os
aspecctos envolvidos em cadda caso; isso
o dará subsíídios e seguurança para se instituirr o
tratam
mento antirrrábico pós-exposição com
c
critérioo e segurançça. Por outro
o lado,
progrramas educativos sobree comportam
mento anim
mal, cuidadoos e posse reesponsável
devem ser intensificados coom a expecttativa de dim
minuir o núm
mero de cassos de agresssão
mal.
anim
PT.0073
ANT
TI-RABIES
S ANTIBOD
DIES SEROLOGICA
AL INVEST
TIGATION
N FROM
DOG
GS OF A RURAL
R
AR
REA IN SÃO
O PAULO STATE - BRAZIL
B
Peress MG1, Baccchiega TS1, Martorelli LFA2, Kataaoka APAG
G2, Vicente AF1,
Apolllinario CM
M1, Allendorf SD1, Antuunes JMAP1, Megid J1 - 1UNESP - School of
Veterinary Meddicine and Animal
A
Sciennce - Deparrtament of Veterinary
V
Hygiene
H
andd
2
B
P, Brasil, Zoonosis
Z
Coontrol Centeer - Rabies Laboratory
L
Publiic Health- Botucatu-SP
- São
Pauloo-SP, Brasil
Rabies is one off the most im
mportant zooonoses, witth fatal proggnosis in alm
most 100% of
mated that evvery 10 to 15
1 minutes one
o person dies from a rabies viruus
casess. It is estim
infecction and 10000 people receive posst-exposure treatment by
b hour in thhe world. D
Dogs
are thhe main tran
nsmitters too man by bitte, so it is esstimated thaat approxim
mately 55,0000
casess of human rabies in thhe world aree caused by dog viral vaariant. The main
m prevenntive
meassure of hum
man cases is the rabies vaccination
v
campaign of
o dogs and cats, whichh was
respoonsible by thhe decreasee of dog andd consequenntly human rabies
r
numbber . The last
humaan rabies caase caused by
b canine raabies virus in the state of São Paullo occurredd in
19977 and in 199
98 the last caase canine rabies.
r
In thhis context the
t present work
w
aimedd to
evaluuate the effeectiveness of
o protectivee vaccinatioon of dogs from
f
rural faarms in the
counnty of Anhem
mbi - SP, affter two yeaars without being
b
immuunized againnst rabies with
w a
vacciine campaiggn. During 2011
2
an epiidemiologiccal question
nnaire was applied
a
to 222
farm
ms to collect data about the health of animals and
a hygieniic handling of propertyy.
One hundred peercent of dogg’s owners declared thaat their dog
gs only receiived anti-rabbies
p
camppaing that was
w stopped two years before.
b
Fiftyy one dogs
vacciine during public
bloodd samples were
w collectted by jugullar vein punncture. The blood
b
collecction tube was
w
centrrifuged and the serum stored
s
at -200 ° C. Sera were
w testedd for neutralization in ceell
cultuure for the determinatio
d
on of neutralizing antibbodies to rabbies virus byy rapid methhod
focuss inhibition of fluorescence (RIFF
FT). Of the 51
5 analyzedd dogs, 68%
% presented antirabiees antibodies titers aboove 0.5 UI/m
ml, indicatinng that they are still pro
otected but on
the oother hand 32%
3
of analyyzed populaation was detected unpprotected annd therefore
susceeptible to in
nfection. Ouur results aggree with liteerature dataa that indicaates that rabies
vacciine induce immunity
i
loonger than one
o two yeaars and conssider the revvaccination
everyy three yearrs but at the same time demonstratte that a percentage of the
t animals don't
manttain protectiive titers aft
fter two yearrs indicatingg that the reevaccinationn protocol m
must
be caarefully evaaluated conssidering the epidemioloogical situattion.
PT.0074
ÓRUM DE QUALIDA
Q
ADE E BIOSSEGURA
ANÇA NO DIAGNÓS
D
STICO DA
l FÓ
RAIV
VA EM RE
ECIFE: PE
ERSPECTIIVAS E MU
UDANÇAS
S DE PARA
ADIGMA
Arauujo ACR1, Machado
M
JL
LM2, Ribeiroo MGB3, Silva LAM4, Oliveira JC
CT5, Gomes ES6,
Bolleer MAA7 - 1LACEN/LA
ANAGRO--PE - DIAG
GNÓSTICO DE RAIVA
A, 2LACEN
N/PE/
3
LAN
NAGRO/PE, Universiddade Federaal de Pernam
mbuco. Cen
ntro Acadêm
mico de Vitóória,
4
Gruupo de Estuddos de Morccegos no No
ordeste (GE
EMNE), 5UF
FPE/CAV/G
GEMNE,
6
Secrretaria Mun
nicipal de Saaúde de São
o José do Eggito, 7Fundaação Oswalddo Cruz
uação atual do rebimennto e das conndições de
Diannte da necessidade de ellencar a situ
amosstras para raaiva no Laboratório Naacional Agroopecuário em Pernamb
buco
LAN
NAGRO/PE, amostras estas,
e
potencialmente innfectantes que
q vem senndo expostaas de
modoo indiscrimiinado em allguns casos, se fez neceessária a reaalização do l Fórum dee
Quallidade e Bio
ossegurançaa no Diagnóstico da Raiva, entre os dias 15 e 16 de Noveembro
de 20011 em Reccife-PE. Este Fórum tevve como objjetivo prom
mover a discussão e o deebate
sobree a Qualidadde e a Biosssegurança no
n diagnóstiico da Raivaa, destacanddo de modo
incluusivo a coletta, acondiciionamento e envio do m
material, vissando aindaa a
impleementação do método para a padronização e conformidaades das am
mostras,
garanntindo assim
m a Biosseggurança paraa os profissiionais envollvidos diretta e
indirretamente no
o monitoram
mento e conntrole da Raaiva no Estaado. Durantee o evento, que
teve como públiico alvo Méédicos Veterrinários, Méédicos Sanittaristas, Epiidemiologisstas,
n controle da Raiva, foram
f
Gestoores das Reegionais de Saúde e Proofissionais eenvolvidos no
levanntados temaas relevantess sobre a realidade e poor vezes, o obstáculo
o
d envio de
do
amosstras de alguuns Municíppios de Pern
nambuco. Na
N ocasião, pudemos coonceber quee, o
contrrole e o mon
nitoramentoo da raiva, é uma realiddade ainda pouco
p
difunndida e
priorritária em allguns Municcípios mais distantes da Capital. A distância geográfica
g
ea
ausênncia de treinnamento paara profissioonais envolvvidos, tambéém foi destaacado comoo
sendoo um dos im
mpedimentoos para o ennvio das amoostras. Através das paleestras e daddos
comppartilhados com o publlico, foi elabborada umaa pactuação entre os Geestores
Munnicipais e a Secretaria
S
d Saúde do Estado juntto ao LACE
de
EN/PE, as estratégias
e
d
de
melhhorias para a consolidaçção da qualiidade e biosssegurança na Vigilânccia
epideemiológica da raiva. Fooi acordadoo o comprom
misso do Esttado: Acom
mpanhar e
orienntar os trabaalhos de cappacitação veerificando o cronogram
ma de execuçção das
atividdades; Forn
necer normaas e instruçõões para exeecução das atividades;
a
L
Liberar
os
recurrsos financeeiros previsttos para a exxecução dass capacitaçõões e treinam
mentos;
Proporcionar coondições físiicas e financeiras para a execução das capacittações e
R
de Saúde parra o envio das
d amostrass dos
superrvisões; Esttruturar as Regionais
Munnicípios da sua Regionaal. As amosttras, deverão chegar ao
o laboratórioo em condiçções
adeqquadas de seerem trabalhhadas e paraa que isso occorra, se fazz necessário
o que as Normas
de Q
Qualidade e de
d Biosseguurança, sejaam obedeciddas e os proffissionais reesponsáveiss pela
coletta, acondicioonamento e envio destaa(s) amostraa(s) tenham
m o conhecim
mento básicco
sobree a zoonosee e a técnicaa de coleta padronizada
p
a, através daa efetiva reaalização destta
pactuuação. Podeemos avaliarr que, aparttir deste eveentos, e no cumpriment
c
to dos acorddos
entree as partes envolvidas,
e
ocorreu um
ma melhora significativa
s
a. Sabemos que o ideall
aindaa está em coonstrução m
mas, o passoo inicial se deu,
d no mom
mento em qu
ue a discusssão
sobree a importanncia da vigiilância epideemiológica da Raiva, foi
f democraatizada e toddos se
fazem
m protagoniistas nessa luta,
l
que é a erradicaçãão da raiva no
n Estado de
d Pernambuuco.
PT.0075
ÁLISE ESP
PACIAL DA
A EPIZOO
OTIA DE RAIVA
R
OCORRIDA NO
N
ANÁ
MUN
NICÍPIO DE
D CAMPIINAS/SP/BRASIL NO
OS ANOS DE
D 2.000 A 2.002.
Ram
mos LHM1, Donalísio
D
M 2, Lourennço RW3 - 1Prefeitura Municipal
MR
M
d Campinass de
Secreetaria Muniicipal de Saúúde, 2UNIC
CAMP - Uniiversidade Estadual
E
de
Cam
mpinas/Faculldade de Ciêências Méd
dicas - Depaartamento dee Saúde Colletiva, 3UNE
ESP
- Uniiversidade Estadual
E
Paaulista/Faculldade de Enngenharia Ambiental
A
ovinos e equuinos (tambbém chamadda de raiva dos
A raiiva é uma zoonose quee acomete bo
herbíívoros ou doos Animais Domésticoos de Interessse Econôm
mico - ADIE
E), de grandee
relevvância para a saúde púbblica e anim
mal. É caractterizada porr encefalite viral e apreesenta
elevaado coeficieente de letallidade. Nesstes animaiss tem como transmissorr o morcegoo
Desm
modus rotun
ndus. A criaação e manu
utenção de herbívoros
h
o
ocorre
em áreas
á
rurais, com
cenárrios diferen
ntes, isto é, pastagens,
p
m
matas
nativaas, refloresttamentos, vaales e rios.
Ferraamentas do geoprocesssamento poddem ser úteiis para o connhecimentoo da
epizoootiologia da
d doença em
m áreas exteensas ou redduzidas. No
os anos 2.0000 a 2.002 foi
f
regisstrada epizootia de raivva em herbívvoros em região rural do
d município
o de Campiinas,
todoss confirmaddos laboratoorialmente. O objetivo deste
d
traballho é caracteerizar a
distriibuição espaacial dos óbbitos de herbbívoros e doos abrigos de
d morcegos hematófaggos.
A loccalização e informaçõees dos óbitos animais e dos abrigoss de desmoddus foram
colettadas pela Secretaria
S
M
Municipal
dee Saúde e peela Secretaria de Agricu
ultura ambaas
utilizzando-se GP
PS (Global Position Syystem). Padrronizou o lo
ocal de óbito
o a porta de
entraada da proprriedade em que o anim
mal estava alojado. Foraam utilizadoos os prograamas
compputacionais Microsoft Excel®,
E
ArrcGIS® verssão 9.2, para transferên
ncia, e análiise
dos ddados. As variáveis estuudadas foraam: ano e mês
m da ocorrrência do óbbito, espéciee
anim
mal, localização espaciaal dos óbitoss e dos abriggos de Desm
modus e tipoos de abrigoos
destees últimos. Foram
F
delim
mitadas áreaas de influenncia (bufferrs) de 3 km dos abrigoss de
morccegos. Como resultadoss, foram geoorreferenciaados 66 (98,5%) registrros de mortes de
anim
mais em 52 focos,
f
sendoo acometidoos 40 (59,7%
%) bovinos e 25 (37,3%
%) equinos. Não
foi evvidenciada sazonalidadde da doençça. Houve deeslocamento dos casoss na direção
sudeste-noroestee, contornanndo a zona urbana,
u
suggerindo que esta sirva como
c
obstácculo
morcegos heematófagos. A maioriaa dos abrigos localizavaam-se próxiima a corpos
aos m
d’águua. Foram identificados 10 abrigos de Desmoodus, todos do
d tipo artifficial. O núm
mero
médiio de morceegos foi de 11
1 por abrig
go. Os buffeers dos abriggos 3, 5 e 6 concentram
ma
maioor número de
d óbitos reggistrados. Os
O resultadoss mostraram
m que 25 (37
7,9%) óbitoos
estavvam contidoos em áreas de influênccia de 3 km de três abrigos próxim
mos (abrigos 3
e/ou 5 e/ou 6). Não
N foram considerado
c
os os abrigos de Desmoodus localizaados nos
muniicípios vizinnhos. Verifiica-se o carááter rural daa doença, poorém há várrios bairros
residdenciais circcundados peela área ruraal, o que eviidencia o rissco de infeccção humanna.
Este trabalho refforça que a incorporaçãão das ferraamentas e téécnicas de
m na compreeensão da coomposição da paisagem
m e padrõess
geopprocessamennto auxiliam
ambiientais, da ocupação
o
doo solo sendoo de grande utilidade naa vigilânciaa epidemiolóógica
da raaiva como de
d outras zooonoses. Verrifica-se a escassez de trabalhos
t
uttilizando
ferraamentas do geoprocessa
g
amento e dee análise esppacial na occorrência daa raiva.
PT.0076
O RABIES
S AND EASTERN AN
ND WESTE
ERN EQUIINE VIRA
AL
DIAGNOSIS OF
CEPHALIT
TIDES IN EQUIDS
E
BY MULTIP
PLEX HEM
MI-NESTE
ED RT-PCR
R
ENC
TEC
CHNIQUE
Iamaamoto K1, Durymanova
D
a EA2, Carriieri ML1, Oliveira
O
RN1, Carnieli Jr
J P1, Fahl WO
W 1,
1
3
1
2 1
Kotaait I , Silva MLCR
M
, Caastilho JG , Ito FH - Instituto
I
Passteur - Diaggnóstico,
2
Univversidade de
d São Pauloo - FMVZ-U
USP, 3UFCG
G-PB - CST
TR-UFCG/P
PB
Seveeral viral zooonoses affect the equidds causing nneurologicall diseases, in
ncluding raabies
and E
Eastern andd Western eqquine encepphalitides (E
EEE and WE
EE). Clinicaal diagnosiss is
oftenn not conclu
usive, in a way
w that labooratory diaggnosis is esssential. Dataa from the
Labooratory of Rabies
R
Diagnnosis at the Pasteur Insstitute of São Paulo, bettween 20000 and
20100, demonstraate that approximately 75% of subbmitted equiid samples, wich animaals
preseented neurological sym
mptoms, werre negative for rabies, emphasizing
e
g the imporrtance
of acchieving a differential
d
d
diagnosis
foor equine enncephalitis caused
c
by allphaviruses. The
aim of
o this studyy was to tesst the suitabiility of usinng multiplex
x hemi-nesteed RT-PCR
R for
the ddiagnosis off rabies, EEE
E and WEE
E in equids central
c
nerv
vous system samples. W
We
used the primerss 21G, 304 and 504 dirrected to thee N gene off rabies viruss, and the
mers cM3W, M2W, nEE
EE and nWE
EE directedd the NSP1 gene
g
of WE
EE and EEE
E
prim
virusses. A prelim
minary studdy of the primers was caarried out, as
a well as thheir use in a
hemii-nested RT
T-PCR, evaluuating the optimal
o
annealing temp
perature, thee analytical
sensiitivity and specificity
s
a the reprooducibility of the techn
and
nique in possitive field
sampples for rabiies and EEE
E. Based on the establisshed protoco
ol for the heemi-nested RTPCR
R, variations in reagentss concentrattions for thee multiplex hemi-nested
h
d RT-PCR
protoocol were peerfomed. After establisshing the prrotocol for th
his reaction
n, the tests too
veriffy the analyttical sensitivvity and speecificity andd reproducib
bility were performed and
the reesults were compared to
t those obttained by heemi-nested RT-PCR.
R
Inn the detectiion
thresshold test, thhe analyticaal sensitivityy was similaar for both techniques,
t
resulting inn 101.7
foor the three standard
s
virrus CVS, EE
EEV and WEEV.
W
In th
he detection threshold teest
usingg a sample with
w the three viruses, a high specificity of the primers was
w verified and
the multiplex
m
heemi-nested RT-PCR
R
waas able to deetect the thrree viruses simultaneou
s
usly.
Therre was no diifference in the proporttions of sam
mples detected as positiv
ve for rabiees
obtaiined by both
h techniquees, according
g to the Fishher exact test (P = 1.00000). Howevver,
for E
EEE positivee field sampples, the prooportion of samples dettected as po
ositive by thhe
hemii-nested RT
T-PCR a diffference in th
he proportioon obtained
d by multipleex hemi-nessted
RT-P
PCR (P <0.00001) was observed.
o
A
Although
it was
w not possible to use WEE posittive
field samples in this study, the results suggest thatt its detectio
on would bee possible bby
m
heemi-nested RTmultiiplex hemi--nested RT-PCR. Thus,, data suggeest that the multiplex
PCR
R technique could
c
be appplied to dettect rabies and
a WEE, but with som
me limitationns for
the E
EEE detectioon, in a wayy that new studies
s
will be carried out.
o
I
of São
S Paulo
Finanncial support: CNPq annd Pasteur Institute
PT.0077
PLEMENTA
ATION OF
F THE FLU
UORESCE
ENT ANTIB
BODY TEC
CHNIQUE
E
IMP
NEU
UTRALIZA
ATION VIR
RUS TEST
T IN RABIE
ES LABOR
RATORY DIAGNOSI
D
IS
OF P
PASTEUR INSTITUT
TE OF SÃO
O PAULO / BRAZIL
Ferreeira JS1, Tro
otti ACP1, Scheffer
S
KC
C1, Caporalee GMM1, Siilva ACR1, Chaves LB1 1
Instiituto Pasteuur - Laboratóório de Diagnóstico dee Raiva
Fluorescentt Antibody Virus
V
Neutrralization Test (FAVN)) is used rou
utinely in many
m
The F
laborratories for the referencce measurem
ment of rabies virus neeutralizing antibodies
a
(VNA
A) in serum
m of animalss to confirm
m the efficaccy of vaccinnation against rabies, which
w
is reqquired to au
uthorize the transit of thhese animalls in countriies free of raabies. The
Worlld Organizaation for Annimal Health
h (OIE) recoommends a VNA title ≥ 0,5 IU/mL
L to
ensurre that the animal
a
has immunity
i
aggainst the raabies virus. The rabies laboratory
diagnnosis of Passteur Instituute of São Paaulo/Brazil (IP/SP) is a national reeference of the
Miniistry of Heaalth for rabiees diagnosiss and perforrms the meaasurement of
o VNA in
humaans and anim
mals serum
m samples foor proof of im
mmunizatio
on against raabies. The
objecctive of thiss study was to implemeent the FAV
VN in the laaboratory rooutine of rabbies
diagnnosis at IP/S
SP using as reference th
he Rapid Flluorescent Focus
F
Inhibition Test
(RFF
FIT). Initiallly, the Challlenge Viruss Standard (CVS)
(
was titrated by the
t FAVN and
a
RFFIIT methods, performinng serial diluutions from 10-1 to 10-112 and determ
mining the
dilutiion of 100 TCID50
T
(500% infectiouus dose in tissue culturre) or 100 FFD50 (50%
% of
the ddose formingg focus) forr FAVN and
d RFFIT, reespectively. A total of 97
9 serum
sampples from annimals vaccinated againnst rabies with
w differen
nt ranges off VNA
previiously titratted by RFFIIT, and 15 samples
s
from
m unvaccinated animalls were seleected.
Statistical analysis of agreeement was performed
p
c
considering
the results in
i a qualitattive
-6
a ³ 0,5) ussing the Kapppa test. Thhe CVS titlee was 10 inn TCID50 fo
for
analyysis (< 0,5 and
FAV
VN and 10-5 in FFD50 for
f RFFIT. The
T FAVN showed higgh specificitty with titerrs
<0,099 IU/mL an
nd LogD50 <0,83 in sam
mples from
m unvaccinatted animals. Qualitative
analyysis of the results
r
show
wed a substaantial agreem
ment betweeen the two methods
m
(kaappa
= 0,666, p <0,0011). The titles of the seraa from vacccinated anim
mals were 0,,12 IU/ml too
5,92 IU/ml (GM
M = 0.92 IU//mL) for FA
AVN and 0,13 IU/mL to
t 9,55 IU/m
mL (GM = 11,34
mL) for RFF
FIT. The diluution factorr in LogD500 values varried from 0,74 to 2,27 (GM
(
IU/m
= 1,555) for FAV
VN and 1,088 to 2,48 (GM = 1,84) for
f the RFFIT. The obtained resullts
show
wed concorddance withinn the param
meters of speecificity and
d sensitivityy between FA
AVN
and R
RFFIT meth
hods. In thiss context, thhe implemenntation of FAVN
F
in thee rabies
diagnnosis laboraatory IP/SP has a great importancee for rabies epidemiological
invesstigation by
y the VNA evaluation
e
inn animal serum samplees and may complemennt the
technniques alreaady in use inn the IP/SP.
PT.0078
OFILAXIA PRE EXPOSIÇÂO DA
D RAIVA
A HUMANA
A ATEND
DIDOS NO
O
PRO
AMB
BULATOR
RIO MÉDIC
CO DO IN
NSTITUTO PASTEUR EM 20111
Cunhha RS1 - 1innstituto pasteeur
p necessiddades
O Innstituto Pastteur atende em seu ambbulatório médico as pesssoas, que por
profiissional ou de
d lazer proocuram proffilaxia da raaiva humanaa, realizada através de
esqueema de pré exposição de
d acordo com a normaa técnica esppecifica. O atendimentto
médiico é dispon
nível diariam
mente, inclu
usive aos finnais de semana e feriaddos, facilitanndo o
acessso à vacina contra raivaa. O esquem
ma de pré exxposição consiste na ad
dministraçãoo de
três ddoses de vaccina de céluulas VERO((VERORAB
B®), via inntradermica,, nos dias 0,, 7 e
28. A avaliação do titulo dee anticorposs neutralizanntes pelo método simpllificado de
soronneutrlizaçãoo é realizadaa dez dias após
a
a adminnistração daa ultima dosse da vacinaa
contrra raiva hum
mana. Nestee trabalho annalisamos oos dados obttidos dos prrontuários das
d
pessooas atendidaas que iniciaram o esquuema de vaccinação durrante o ano de
d 2011. Fooram
atenddidas 1221 pessoas,
p
senndo 69,50%
% do sexo feeminino e 30
0,50% sexo
o masculino,, nas
seguiintes faixas etárias: 16,,30% menorres de vintee anos, 79,922% entre 200 a 49 anos e
3,77%
% 50 anos e mais. Quaanto às atividades inform
madas a disstribuição fooi de 54,50%
% de
profiissionais e estudantes
e
d biologia e medicina veterinária;; 16,90% dee agentes dee
de
zoonnoses, 11,200% de funcioonários de pet
p shops/faaculdades/addestradores; 4,60% de
viajaantes; 5,60%
% de estudanntes de cursso técnico de auxiliar de veterináriia; 5,20% dee
volunntários de centro
c
de controle de zooonoses/zooológico e 2,000% a inforrmação não
dispoonível. Do total
t
de pesssoas atendid
das, 1086 peessoas iniciaram esqueema de pre
expoosição em 20011, e dentrre estas 885 (82,87%) completaram
c
m o esquem
ma proposto de
três ddoses da vaccina contra raiva humaana. A avaliação soroló
ógica foi reaalizada por 610
6
pessooas (60,93%
%) e o título satisfatórioo foi obtido por 96,56%
%.
PT.0079
A DO ESTA
ADO DE SÃ
ÃO PAULO
O PARA A VACINAÇ
ÇÃO CON
NTRA
ESTRATÉGIA
AIVA EM CÃES E GATOS
G
EM
M SITUAÇÃ
ÃO DE AB
BASTECIM
MENTO
A RA
PAR
RCIAL DE VACINA
Takaaoka NY1, Reichmann
R
M 1, Omo
MLB
oto TM1, Kotait
K
I1 - 1In
nstituto Pastteur - CCD SES//SP
a
de vaccina contra a raiva em cães
c
e gatoss para o Estaado de São
A neecessidade anual
Pauloo (ESP) é de
d 8 milhõess de doses, para
p todas as
a atividadess (campanh
ha, repasse,
rotinna, bloqueio de foco, essterilização e adoção officiais etc.). Como em 2010
2
a
Cam
mpanha de Vacinação
V
fooi suspensa, frente aos eventos advversos pela vacina
v
utilizzada,
e em
m 2011 não havia
h
disponnibilidade de
d vacina dee boa qualid
dade, o ESP, assim com
mo
outroos estados em
e que a raiiva pela variiante caninaa encontra-sse controladda, durante dois
d
anos não recebeeu outra vacina. Foi neccessário, enttão, o estabelecimento de estratégias
u melhor vigilância epidemiológ
e
gica da raivva por parte dos municíípios,
para estimular uma
mais suspeitos, b) que
com o envio de material paara diagnóstiico laboratoorial: a) anim
or serem aggressores, c)) com causaa de morte
morrreram no peeríodo de obbservação po
descoonhecida, d)
d atropeladoos e e) perceentual dos apreendidos
a
s, não regataados ou
adotaados. No caaso de recebbimento de parcela
p
da quantidade
q
d vacina neecessária, fooram
de
priorrizados os loocais com circulação
c
viral, em quaalquer espéccie, estabeleecendo-se metas
m
de vaacinação maais baixas, baseadas
b
no
os números de cães e gaatos vacinaddos nos anoos de
20077, 2008 e 20009. Foram, também, esstabelecidoss contatos com
c
o Centrro Panameriicano
de Feebre Aftosaa e Zoonosess (PANAFT
TOSA/OPA
AS/OMS), Faculdade dee Medicina da
Univversidade dee São Paulo (FM/USP) e Faculdadde de Mediccina Veterin
nária e Zooteecnia
da U
Universidadee de São Pauulo (FMVZ
Z-USP), paraa viabilizar estudos sobbre análises dos
fatorres de risco da raiva em
m animais dee estimação, tanto por variante
v
de morcego,
quannto pela reinntrodução daa variante canina. A etaapa inicial dos
d estudos deve ser a
deterrminação daa proporçãoo de amostraas a serem encaminhad
e
das ao laboraatório e o
aprim
moramento das estimativas das pop
pulações caaninas e, priincipalmentte, felinas. Em
E
inícioo de 2012 o Ministérioo da Saúde disponibilizo
d
ou 3 milhõees de doses de vacina
impoortada de bo
oa qualidadee, para o ES
SP. Nesta opportunidadee, foi efetuad
da a
distriibuição, dan
ndo prioridaade aos munnicípios quee demonstraaram circulaação viral (22011
e 20112), seguido
os dos que vacinaram
v
em
e 2010 maais de 20% dos
d cães, co
om uma
estim
mativa da po
opulação cannina de pelo
o menos de 6 mil. Isto, pelo receioo de que se as
vacinnas que viesssem a comppletar o quaantitativo neecessário tivvessem exceesso de
album
mina bovinaa, propicianndo maior possibilidadee de choquee anafilático
o aos animaiis
pela hipersensib
bilidade à vaacina anterio
or. Pela vinnda de mais 1.090.000 doses,
d
até
o, foram atendidos 163 municípioss, incorporaando aqueless com
meaddos de julho
diagnnóstico labo
oratorial possitivo para raiva,
r
de fevvereiro a meeados de jullho deste anno,
além
m daqueles com
c
circulaçção viral em
m 2009 e 2010. Essa esttratégia fez com que oss
muniicípios ficasssem mais atentos
a
aos sintomas
s
dee raiva nos animais,
a
meelhorando a
vigilâância epidemiológica, com mais amostras,
a
dee todas as esspécies, envviadas para
diagnnóstico labo
oratorial e, consequente
c
emente, maior quantidaade de casos positivos para
raivaa (2011 – 1116 casos e 2012
2
– 174 casos
c
até finnal de julhoo).
PT.0080
ENTOS AD
DVERSOS PELA
P
VAC
CINA CON
NTRA A RA
AIVA EM CÃES E
EVE
GAT
TOS NO ES
STADO DE
E SÃO PAU
ULO - 20100
Takaaoka NY1, Reichmann
R
M 1, Omo
MLB
oto TM1, Kotait
K
I1, Gomes LH2, Aranda
A
CM3 1
2
Instiituto Pasteuur - CCD - SES/SP,
S
Ceentro de Coontrole de Zoonoses do Município de
3
São P
Paulo, Cooordenadoria de Controle de Doenças - SES/SP
P
o Paulo (ES
SP) coordena a Campannha de Vacinação contrra a raiva em
m
O Esstado de São
cães e gatos desde 1975. Noo inicio da década
d
de 80,
8 atingiu-sse 80% de cobertura
c
vaacinal
p
com
m a particippação de toddos os muniicípios. A vacina
v
utilizada
(1 cãão para 10 pessoas),
no peeríodo 1975
5 a 2009 foii tipo Fuenzzalida & Pallácios. O im
munobiológico utilizadoo nos
estaddos é forneccido pelo Ministério da Saúde (MS
S), que decid
diu em 2010
0 substituir a
vacinna por uma produzida em
e culturas de células BHK,
B
que não
n deveria causar reaçção
adversa neurológica e propiiciaria uma melhor respposta imunoológica. Noo Brasil todoo
v
d
deve
ser aprrovado peloo Ministérioo da Agriculltura, Pecuáária e
produuto de uso veterinário
Abasstecimento (MAPA)
(
e a vacina adqquirida peloo MS produuzida por um
m Laboratórrio
privaado nacionaal tinha regisstro no MA
APA e os lotes foram approvados seggundo
parâm
metros de potência, inoocuidade, essterilidade e vírus residdual, conforme
recom
mendações da Oficina Internacionnal de Epizoootias (OIE)). É importaante destacaar que
no ESP a campaanha de vaciinação é reaalizada pelaa maioria doos município
os no mês de
d
agostto, antes doos demais esstados. Já noo final de juulho, algunss municípioss passaram a
referrir ocorrênciia de eventoos adversos após a apliccação da vaacina; em innício de agosto o
muniicípio de Gu
uarulhos, seegundo maioor do estadoo, relatou muitas
m
reações à vacina,,
suspeendendo a campanha
c
e em meadoss desse mêss na capital do ESP, ao iniciar a
vacinnação, ocorrreram inúm
meras notificcações dessees eventos. Diante
D
desssa situação, a
camppanha de vaacinação conntra raiva em
m cães e gaatos foi susp
pensa em 20
0/08/2010. O
muniicípio de Sãão Paulo, poor intermédiio do Centroo de Contro
ole de Zoonooses, já conntava
com uma forma de a populaação notificcar problem
mas com seuss animais ap
pós vacinaçção,
d
mais consistentes
c
s para essa ddecisão de suspensão
s
d campanhaa
da
forneecendo os dados
(altísssima incidêência de eveento adversoo de quase 3 casos por 1.000 cães vacinados e
cercaa de 30 por 1.000 gatoss vacinados)). Do total de
d animais a serem vaccinados em todo
t
ESP (cerca de 6.500.000), aproximada
a
amente 10%
% receberam
m a vacina. Foram
F
tomaadas
proviidências, deentre as quaais se destaccam reuniõees com especialistas, alggumas
necroopsias dos animais
a
morrtos (em Faculdades dee Medicina Veterináriaa), testes em
m
anim
mais de laborratório com
m diferentes vias de adm
ministração, testes de
citotooxicidade, assim comoo dosagem da
d concentrração de sap
ponina e pro
oteínas, por
métoodos bioquím
micos. Os estudos
e
realiizados indiccavam que a vacina prooduzia evenntos
adversos e especcialmente a alta concenntração de albumina
a
boovina, sendoo que esta última
usa do probblema. Essass providênccias adotadaas pelo ESP,, fizeram coom
podeeria ser a cau
que sse intensificcasse os estuudos e fosseem realizadoos mais testtes com a vaacina, antess de
sua liberação, beem como hoouvesse melhoria no eddital de aquisição pelo MS.
a
Atuaalmente estáá em mudannça o processso de produução da vacina de uso animal,
produuzida em cu
ultivo celulaar por laborratório nacioonal.
PT.0081
S 5 CLAVE
ES PARA LA
L PREVENCIÓN DE
E MORDE
EDURAS DE PERROS
LAS
Gebaara RR1, Gaamboa M2, Mora
M
R3, Vigilato MA4, Tamayo H5 - 1WSPA
A Brasil
(Sociiedade Mun
ndial de Prooteção Anim
mal), Brasil - Gerente de Programaas Veterinários WSP
PA Brasil, 222 WSPA M
Mesoamérica (Sociedadd Mundial para
p la Proteección Anim
mal),
3
Costaa Rica - Gerente de Proogramas Veeterinários - WSPA Meesomérica, 3- WSPA
Suram
mérica (Socciedad Munndial para laa Protecciónn Animal), Colombia
C
- Gerente de
Proggramas Veteerinários - WSPA
W
Suram
mérica, 4PA
ANAFTOSA
A/OPS (Orgganización
Panaamericana de
d la Salud) - Especialissta em Saúdde Pública Veterinária
V
5
PAN
NAFTOSA/O
OPS, PANAFTOSA/O
OPS (Organnización Pan
namericana de la Saludd) Coorrdenador dee Zoonoses - PANAFTO
OSA/OPS
La innformación es esencial para el éxitto de prograamas de preevención de Rabia
transsmitida por mordeduras
m
s caninas. Estudios
E
cieentíficos dem
muestran quue la mayorría de
las m
mordeduras de perros see dan por unn desconociimiento sobre la manerra correcta de
d
interaactuar con ellos.
e
Las leesiones caussadas por mordeduras
m
son
s un probblema globaal,
ponieendo a personas en riessgo de contrraer enferm
medades com
mo la Rabiaa.
Este proyecto bu
usca identifficar los mennsajes más importantess y relevantes para infoormar
e manejo dee los perros usando meensajes simpples. Ello buusca
y eduucar a persoonas sobre el
tambbién preveniir situacionees que puedden desencaddenar morddeduras caniinas,
conseecuentemennte reducienndo el numeero de casoss de rabia humana.
h
Los pprincipales mensajes iddentificadoss como " Laas 5 claves" fueron diseeñados a parrtir
de unna investigaación biblioggráfica teniendo en cueenta las prin
ncipales cau
usas de los
acciddentes por mordeduras
m
y la interprretación prinncipal de los errores deel idioma caanino.
LOS MOLE
ESTES: No molestes o acaricies peerros cuand
do estén com
miendo,
NO L
durm
miendo o enffermos, o cuuando se enncuentren coon sus cachoorros o jugu
uetes; No tee
acerqques a perro
os que esténn dentro de un
u automóvvil, detrás dee una reja, amarrados,
a
o que
sean desconociddos; No haggas movimientos o sonnidos brusco
os cerca de perros.
p
TIENDE SU
U LENGUAJE: No te accerques a peerros que deemuestren miedo.
m
Estoos se
ENT
echann para atráss, se lamen la
l cara, bajaan la cola, eechan las oreejas para atrrás y muesttran
los ddientes; No te
t acerques a perros ag
gresivos. Esstos fruncenn los labios, enseñan los
dienttes, gruñen, se les erizaa el pelo y mantienen
m
laa cola erguida y erizadaa.
QUÉ
ÉDATE TOT
TALMENT
TE INMÓV
VIL SI SE ACERCA UN
N PERRO SUELTO:
S
N
No
corraas ni grites. Baja la cabbeza y no mires
m
directaamente a loss ojos del peerro; si estás de
pie, qquédate quieto con los brazos pegaados al cuerrpo como un
n tronco; si caes al pisoo,
proteege tu cara y vientre y permanece
p
inmóvil com
mo una rocaa; cuando ell perro hayaa
perdiido interés, aléjate lentamente y enn silencio.
EVIT
TA ACCIDENTES: Loos niños siem
mpre debenn estar acom
mpañados po
or adultos
cuanndo estén cerrca de un peerro, aún si este es connocido; Siem
mpre pide peermiso al duueño
antess de acariciaar a un perroo; Antes de acariciar a un perro, permite que huela tu maano y
acaríícialo en la espalda
e
o laados del peccho; Manténn tu cara aleejada de la cabeza
c
del perro;
p
Si el perro se rettira, no tratees de reteneerlo.
CASO DE SER
S
MORD
DIDO, SIGU
UE ESTAS RECOMEN
NDACIONE
ES: Lava laa
EN C
heridda inmediataamente por 15 minutoss con abunddante agua y jabón; Reecolecta
inforrmación quee identifiquee al perro (iincluyendo su estado dee vacunació
ón) y a su duueño;
Acudde al centro de salud más
m cercano lo antes possible; Inform
ma al centro
o de salud y a la
autorridad sanitaaria competeente sobre el
e incidente de mordeduura y el lugaar en dondee se
encuuentra el perrro. No lo molestes,
m
maaltrates o mates;
m
La auutoridad maantendrá al perro
p
en unn lugar segu
uro con agua y comida para observvación.
LIOGRAFIA
A:
BIBL
1. A communityy approach to
t dog bite prevention.
p
V
Medical
M
American Veterinary
Canine Interractions .Veet
Assoociation. Tassk Force onn Canine Agggression annd Human-C
Med Today: Cannine Aggresssion Task Force
F
JAVM
MA, Vol 21
18, No. 11, June
J
1, 20001. At
ma.org/public_health/ddogbite/doggbite.pdf
: httpp://www.avm
2. Prreventing do
og bites in children:
c
ranndomised coontrolled triial of an eduucational
intervvention , Siimon Chapm
man,John Cornwall,Joaanne Righettti,Lynne Suung BMJ
20000;320:1512-1513 doi:100. bmj.320.77248.1512 ((Published 3 June 20000) At:
http:///www.bmj.com/contennt/320/7248
8/1512.1.fulll.pdf
3. Asspectos epiddemiológicoos de las mo
ordeduras caaninas. Jorgge Palacio / Marta Leóóna /
Sylviia García-B
Belenguer Departament
D
to de Medicina y Ciruggía Animal. Facultad dee
Cienncias Experimentales y de la Saludd. Universiddad Cardenaal Herrera-C
CEU. Moncaada.
Valencia. Españ
ña.Departam
mento de Pattología Aniimal. Faculttad de Veterrinaria.
Z
Z
Zaragoza.
E
España.
Univversidad de Zaragoza.
4. Brritish Kenneel Club. In Safe
S and souund websitee: http://ww
ww2.thekennnelclub.org.u
uk/safe_andd_sound.htm
ml
5. C
Centers of Disease Conttrol and Prevention; US
SA Gov . Dog Bite Preevention .
At w
website: httpp://www.cdc.gov/HomeandRecreaationalSafetty/DogBitess/bitepreven
ntion.html
6. Thhe blue dog project – The
T developm
ment of a dog
d bite prevvention proggramme aim
med
at yooung childreen T De Keeuster, K Meeints, R Buttcher . Journnal of Injurries
Preveention 10;116:A169, 20010
7. Doog bite prev
vention: whaat children know
k
.C A Dixon, E M Mahabee--Gittens, C J
Linddsell At : Jouurnal of Inuuries Prevenntion 2010;; 16:A169-A
A170
8. W
World report on child injjury preventtion , M. M.
M Peden,K. Oyebite, World
W
Healthh
Orgaanization, 20
008
9.httpps://ebusineess.avma.orrg/EBusinesss50/files/prroductdownnloads/dog_bbite_brochuure.p
df
Doggone Saafe Program
m. Dog bite prevention
p
t
through
eduucation/Dog
g Safety for kids
10. D
at : http://www
h
.be-a-tree.com and ww
ww.doggoneesafe.com
11. hhttp://www.w
worldrabiessday.org/asssets/files/CooloringBookk_DogBiteP
Prevention.ppdf
12. H
Humane Dog Populatioon Managem
ment Guidannce. Internaational Com
mpanion Aniimal
Manaagement Cooalition. ICA
AM , 2007. At : www.wspa-intern
national.orgg
13. F
Feldman ED
D, Nelson RW.
R Caninee and feline endocrinoloogy and repproduction. 3 ed.
St Loouis: Sauderrs; 2004. p. 1088-90.
14. ““Criando um
m amigo“ víídeo. Realizzação Centroo Controle de
d Zoonosees-SP (Centrro
Colaaborador da OPAS/OM
MS) -2004. Produção
P
Insstituto Ninaa Rosa e WS
SPA –Socieedade
Munndial de Protteção Anim
mal.
15. ““Be safe aroound dogs” Wake Coun
nty, North Carolina
C
Hu
uman Servicces pamphleet.
16. ““Manual Báásico de Tennencia Respponsable” A
Asociación Nacional Protectora dee
Anim
males de Coosta Rica.
17. ““Lenguaje Canino”
C
Sociedad Munndial para laa protección
n Animal, WSPA.
W
18.
vos/pdf/ano10_n02_sit__epidemiol__zoonoses__br.p
http:///portal.saudde.gov.br/portal/arquiv
df
PAL
LABRAS - CLAVE:
C
preevención, mordeduras
m
caninas, rabbia
1.
2.
Sociedad
d Mundial para la Proteección Anim
mal – WSPA
A
Organizaación Panam
mericana de la Salud OPS
O
PT.0082
ALUATION
N OF CLIN
NICAL ASP
PECTS, FL
LUORESC
CENT ANTIBODY TE
EST
EVA
AND
D N GENE
E QUANTIF
FICATION
N OF RABV
V VARIAN
NTS ISOLA
ATED FRO
OM
DIFF
FERENT ANIMAL
A
S
SPECIES
Allenndorf SD1, Harary
H
CMA2, Vicentee AF2, Antuunes JMAP2, Peres MG
G2, Fonseca CR2,
2 1
2
Megiid J - FMV
VZ-UNESP
P - DHVSPM
MI, FMVZ
Z/UNESP - DHVSPMI
m different mammals
m
seeem to havee unique
The rrabies viruss (RABV) issolated from
charaacteristics th
hat influencces the outco
ome after annimals get infected.
i
Thhe RABV
circuulates in natu
ure and is maintained
m
b reservoirrs that are reesponsible for
by
f persuadiing
the ddisease for almost
a
4,0000 years. As different cllinical aspeccts are obserrved in anim
mals
naturrally and exxperimentallly infected the
t aim of this
t study was
w to evaluaate clinical
aspeccts, quantifyy the viral replication
r
in brain tissuue of experiimentally innfected mouuse
assocciated with FAT resultss of RABV isolated froom 5 animall species. The selected
viruss were isolatted from soome of the main
m reservooirs in Brazil being relaated to a batt
(Myootis spp.), to
o a fox (Cerrdocyon thous), to a dog (Canis fam
miliaris), too a bovine
(relatted to Desm
modus rotunndus) and too a monkey (Challithrixx jacchus). The
T mice were
w
40, fe
female, speccific pathogen free (SPF
F), swiss albbino mice, 45
4 days oldd that were
dividded into 5 grroups of 6 animals.
a
Thhe intracerebbral mouse inoculation
i
med
was perform
with 0,03mL of the inoculuums (standarrd dilution) for clinicall observatioon and sampple
ough all virrus resulted in an infecttion with 1000% of lethaality, it is
colleection. Altho
remaarkably notaated that thee time of evolution of some
s
samplees is much longer
l
than
otherrs variants. When the animals
a
get sick after beeing inoculaated with saamples
originated from fox and moonkey it takes more tim
me to death to
t occur whhen compariing to
d samples. The
T monkeyy and fox saamples had similar evoolution periood,
the oother studied
for m
monkey sam
mple the sym
mptoms initiiated after 10
1 days and the evolutioon to death of
all innoculated annimals weree of 4 days, in the fox sample
s
the symptoms
s
innitiated afteer 9
days and the evoolution in thhis case wass of 8 days. Otherwise the sampless isolated frrom
d bovine inittiated symptoms after 8 days and had
h an evolu
ution periodd of
the bbat, dog and
threee days. The presence off the RABV
V was investtigated by FAT.
F
The viirus was
identtified by thiis techniquee in all sampples. Some differences
d
could be ob
bserved. Thhe
patteern of positiv
vity in some infected tiissues is sm
moother wheen comparinng samples
relateed to dog, fox,
f
and vam
mpire bats. The
T virus issolated from
m Challithrixx jacchus annd
Myottis spp. pressented a discrete fluoreescence and some pointts of fluoresscence in the
tissuee, while thee samples isolated from
m the bovinee, a variant probably
p
related to
Desm
modus rotun
ndus (V3), the
t corpusclles were bigg and with strong
s
fluoreescence, succh as
thosee observed in
i canine reelated samplles. In orderr to quantifyy the viral replication inn the
brainn of the infeected animalls the Real Time
T
was pperformed, the
t cDNA was
w made ussing
oligoo dt and Supper Script III (Invitrogenn), and the reaction
r
waas performedd using GoT
Taq
qPCR
R Mster Miix (Promegaa). The dog,, fox and boovine samplle presentedd a higher raate of
repliccation, whicch could bee observed by
b the detecction of viral antigen inn brain of
infeccted animalss. The time to death waas not relateed to the viral replicatioon once the virus
isolaated from Ceerdocyon thhous presentted replicatiion similar to
t the dog and
a Desmoddus
rotunndus and waas the sampple with the longest incuubation andd evolution time.
t
On the
otherr hand viruss isolated froom monkey
y had the low
west replicaation compaaring to the other
isolaated virus foollowed by the
t virus isoolated from the bat (Myyotis spp.). This
T resultss
demoonstrated th
hat virus isollated from different
d
species have unique
u
charracteristics that
t
can rreflect on th
he viral repliication but not
n necessaary on the evvolution perriod and
lethaality of the infection.
PT.0083
DEMIOLO
OGICAL ANALYSIS
A
OF RABIE
ES IN HER
RBIVORES
S IN THE
EPID
REG
GION OF ARAÇATU
A
UBA – SÃO
O PAULO, BRASIL:
B
PRELIMIN
P
NARY
RES
SULTS
Alvees AJS1, Carrvalho C2, Casagrande
C
DKA3, Souuza Filho VN
N4, Pedro WA
W 5, Queirooz
2 1
LH - USP - Faaculdade de Medicina Veterinária
V
e Zootecniaa - Doutoran
nda do
2
d Medicinaa Veterináriaa Preventivaa e Saúde Animal,
A
UN
NESP Depaartamento de
Facuuldade e Meedicina Veteerinária de Araçatuba
A
- Departameento de Apo
oio, Produçãão e
Saúdde Animal, 3UNESP - Faculdade
F
e Medicina Veterinária
V
de Araçatub
ba - Mestraanda
do Prrograma de Ciência Annimal - Deppartamento de
d Apoio, Produção
P
e Saúde
S
Anim
mal,
4
Cooordenadoria de Defesa Agropecuár
A
ria - SAA/S
SP - Coordennador do Prrograma
Estaddual de Raivva dos Herbbívoros - ED
DA de Baurru, 5UNESP
P - Faculdad
de e Medicinna
Veterinária de Araçatuba
A
- Bolsista de Produtividade em Pesquisa CNPqq d Apoio, Prrodução e Saúde
S
Anim
mal
Depaartamento de
or zoonosis of great imppact on bothh economicc and publicc health and
Rabies is a majo
ores is distriibuted in alm
most all Lattin Americaan
veterrinary mediccine. Rabies in herbivo
counntries and itss control constitutes a major
m
challeenge for Livvestock Deffense. This study
s
aims to make ann epidemioloogical analyysis of Rabiies in herbivvores in ruraal areas of most
m
t Office of
o Agricultuure and Liveestock Defennse Araçatuuba
muniicipalities inncluded in the
(EDA
A). To this end,
e an epiddemiologicaal questionnnaire was elaborated annd applied too
obtaiin informatiion related to
t rabies in herbivores. The follow
wing variablles were
analyyzed: presennce, frequenncy and period of occuurrence of baat bites marrks on the
anim
mals, the presence and tyypes of shelters for batts; recent occcurrence off death of
anim
mals with neu
urological signs
s
on thee property, use
u of vacciine against rabies,
r
use of
methhods for batss control annd environm
mental channge in the arrea of the prroperty. Thee
resullts show thaat out of 1111 properties sampled, 223.4% of theem observedd marks of bat
b
bitingg in the aniimals, and of
o these, 23% observedd the marks of bites daaily, 4% weeekly,
8% m
monthly andd 65% rarelyy. The obseervation of bbite marks occurs
o
moree during thee
rainyy season (73
3.1%). 11.7%
% of ownerrs surveyed report the presence
p
of shelters of bats
b
and tthe main typpes of shelteers are aban
ndoned housses (38%), hollow
h
trees (38%) andd
otherrs (15%). Only
O
70.9% of
o those surrveyed vacccinate the annimals againnst rabies annd
76.322% of thesee vaccinate the
t entire heerd. In 4.5%
% of the prooperties it was
w reportedd the
use oof some metthod for thee control of bats. Approoximately 60% of properties had some
kind of environm
mental channge, particullarly with reespect to the advance of
o sugarcanee in
s
werre the number of animaals in the
the reegion. The quantitativee variables studied
propeerties sampled, accordiing to the kiind, with ann average off 243 animaals per farm
m (237
cattlee and six hoorses) and median
m
93 (990 and 3), reespectively. We also performed thhe
assocciation betw
ween properrties where it
i was foundd seropositiive bats with
h the variabbles
of the questionnnaires, by thhe method of chi-squaree test or Fishher. The vaariables that
wed a strongg associationn with seroppositivity off bats were the presencce of shelterr and
show
vacciination. Alll statistical analyzes
a
weere performed using SP
PSS 20.0. Inn each of thee
propeerties were obtained geeographic co
oordinates using
u
a GPS
S, allowing a spatial
charaacterizationn by plottingg the points with the heelp of the program ARC
CGIS 10.0.
PT.0084
EVIEW OF
F THE CLA
ASSIFICA
ATION OF RABIES VIRUS
V
LIN
NEAGES
A RE
MAIINTAINED
D BY INSE
ECTIVORO
OUS BATS IN BRAZIIL
Oliveeira RN1, Castilho
C
JG1, Batista HB
BCR1, de Paaula FC1, Carnieli Jr P1, Lima JYO
O1 ,
1
2
1
1
3
3
Maceedo CI , Meenozzi BD , Carrieri ML
M , Kotait I , Mirandaa CCP , Branndão PE 1
2
Instiituto Pasteuur, Universidade Estaddual Paulistaa - Faculdad
de de Mediccina de
Botuucatu, 3Univversidade dee São Paulo - Faculdadee de Mediciina Veterináária e Zooteecnia
wn about thee importancee of nonhem
matophagouus bats in thee epidemiollogy
Littlee was know
of rabbies in Brazzil and mostt of Latin America
A
untiil the 1980ss. From thatt decade on,, as
caninne rabies caame under control in maany municippalities andd molecular and antigennic
typinng was incorrporated in surveillance programs, the importtance of nonnhematophaagous
bats in the epideemiology off the diseasee began to be
b appreciatted in these countries. In
I
s
baseed on gene N have show
wn that diffe
ferent lineag
ges are
Brazzil, genetic studies
circuulating in insectivorouss bats from the
t species Tadarida brasiliensis, Nyctinomopps
laticaaudatus and
d genus Myootis, Eptesiccus, Molosssus, Histiotuus and Lasiuurus. In most
studiies, the charracterizationn of these lineages is baased on only 264 nt of the
carbooxyterminall region of the
t viral nuccleoprotein, when the ideal
i
wouldd be to use thhe
compplete N gene. The aimss of the pressent study w
was to review
w the genettic classificaation
of the RABV isoolated from
m insectivoro
ous bats from
m Brazil baased on currrent literaturre,
p
DNA
A sequencingg of the nuccleoprotein comparing the
Genbbank dataset and new partial
phyloogenetic annalysis of N gene based
d on 1218 nuucleotides (nnt 203 to ntt 1420) withh that
basedd on 264 nuucleotides (nnt 1157 to nt
n 1420), corrresponding
g respectiveely to aminoo
acidss 45 to 450 and 363 to 450
4 of the viral
v
nucleooprotein. Phyylogenetic analysis
a
demoonstrated th
he existence of at least eleven
e
lineaages of RAB
BV associatted with
different generaa and species of insectivvorous bats.. Nine of these lineagess have alreaady
beenn described in
i literaturee while two of them weere herein ch
haracterizedd for the firsst
time and associaated to the genus
g
Myotiis and Lasiuurus. There were no diffferences inn the
s
by coomparing thhe two align
nments usedd, but changges
classsification off Brazilian strains
weree observed in
n phylogenetic relationnships between the clussters, with bootstrap
b
vaalues
alwaays greater regarding
r
thhe 1218 nt trree. Two sequences of RABV from
m the genuss
Myottis from Uru
uguay and Chile
C
did noot keep the same
s
classiffication afteer the analyses
with the two alig
gnment lenggths. These findings shhould be tak
ken into acco
ount in
s
of innfections might
m
be deteermined in a
moleecular epideemiology off rabies, as sources
moree accurate way
w and alsoo in the corrrect use of fragments
f
of the N Genne for the
classsification off lineages off RABV.
PT.0085
SITU EVAL
LUATION OF CYTO
OTOXIC IM
MMUNE RESPONSE
R
E IN CENT
TRAL
IN S
NER
RVOUS SY
YSTEM OF
F HUMAN RABIES
R
T
TRANSMIT
TTED BY DOG
D
AND
D
VAM
MPIRE BA
AT
Gueddes F1, Fern
nandes ER1, Martins JF
F2, Pagliari C3, Duarte MIS
M 3 - 1Insttituto Pasteuur de
2
São P
Paulo - Imu
unologia, Innstituto de Infectologia
I
a Emílio Ribbas, 3Faculd
dade de
Mediicina da Unniversidade de São Paullo
C (+) T cells
c
and nattural killer (NK)
(
cells are
a immunee effectors thhat,
Introoduction: CD8
by cyytokine prodduction or cytotoxicity
c
y, help to coontain a viraal infection.
Objeective: To quantify
q
andd compare thhe T CD8 lyymphocytess, natural kiiller cells annd B
granzzyme expreession in cenntral nervouus system leesions of human rabies transmittedd by
dog aand vampiree bat.
Meth
hods: Five fragments of
o central neervous systeem (CNS) were
w selecteed (cortex,
hippoocampus, basal gangliaa, cerebellum
m and meduulla oblongaata) from eaach specimeen of
the fo
four human rabies casess transmitteed by dog annd four casees by vampiire bat
(Desmodus rotuundus). The fragments were
w subjeccted to imm
munohistocheemistry withh
C
CD57 and B granzzyme. For each
e
specim
men, cells weere quantified
antibbodies for CD8,
by coounting the number of immunolabbelled cells in
i thirty fiellds considerring the
parennchyma. Foor normalizing, a x10 ocular lens w
was used wiith a square grid in a x440
objecctive markin
ng an area field
f
of 0.06
625 mm2. Sttatistical an
nalysis was performed
p
b
by
Grapph Pad Prism
m version 5.0 for Winddows (Graphh Pad softw
ware, San Diiego, Ca,
U.S.A
A.) using thhe nonparam
metric Mannn-Whitney test.
t
Samplees were connsidering
different at the 95%
9
(p≤0.05) level of significance
s
e.
ults: The nu
umber of CD
D8+ T lympphocytes in human rabiies transmittted by dog w
was
Resu
loweer (p<0.00011) than in thhose with huuman rabiess transmitted by vampire bat. No
signiificant difference in thee number off CD57+ naatural killer cells (p>0.005) and the
numbber of B graanzyme-exppressing cellls (p>0.05) was observ
ved betweenn samples
evaluuated of thee human rabbies transmittted by dog and vampirre bat.
Discu
ussion and Conclusions: In the present
p
studyy, we comppared lesions in CNS off
humaan rabies traansmitted by dogs and vampire baats by quanttitative exam
mination of the
“in situ” cytotoxxic immunee response. Rare
R NK ceells and B grranzyme-exxpressing ceells in
o
b there weere no signifficant statisttical differeence
but
cerebbral parenchhyma were observed,
betw
ween the hum
man rabies transmitted
t
by dog andd vampire baat. This couuld reflect ann
immuune evasion
n mechanism
m triggered by rabies virus,
v
prevennting these cells arrive at
the site of injury
y, or that theeir cytotoxicc function would
w
be alttered. CD8+
+ T lymphoocytes
pire bat, whhich appeareed
weree more abunndant in the human rabiies transmittted by vamp
relateed to the virral variant type
t
involveed in infectiion, howeveer eventuallyy the functiion
thesee cells may be impairedd. So, we caan speculatee if this fact also could be due to loonger
surviival of thesee patients coompared to those bittenn by dog.
PT.0086
ular growth
h in differen
nt bioreacttors to rabies virus production
cellu
Lantiieri VS1, Medeiros
M
FM
M2, Frazatti-G
Gallina NM
M2 - 1Intituto
o Butantan - Seção de
2
Raivva, Instituto
o Butantan - Seção de Raiva
R
o virus production proocess involvves differentt challengess, mainly whhen
The sscaling up of
is useed an animaal cells origgin with a suubstrate. The growing of
o the animaal cells in hiigh
densiities depend
ds on the beeads and theese cells preesent high suusceptibilityy to the sheaar
stress that occurrs in the proocess realizeed in bioreactors. The objective
o
off this study was
w
g
of Vero
V
cells in
i the scalinng up processs of rabies virus
to evvaluate the growing
produuction in biioreactor. Tw
wo bioreacttors were ussed in this study,
s
one of
o 30 L (Bio
Flow
w 4500, NBS
S) and otherr of 150 L (Bio
(
Flo PR
RO Industriaal, NBS). Th
hese bioreacctors
have different aggitation systems: whilee the 30 L hhas a “Cell Lift
L Impeller”, the
S
has pittched blade impellers. This
T differeence was im
mportant to select
s
indusstrial, one STR,
the vvelocity of agitation
a
neccessary to maintain
m
thee beads in suuspension and
a to minim
mize
the shhear stress and
a bead coollisions. Veero cells addded to solidd microcarriiers, Cytodeex 1
(2g/L
L), infected with PV raabies virus (MOI 0,02) were cultiv
vated in seruum-free meddium
VP S
SFM AGT in the two bioreactors. Were
W realizzed seven cyycles in eachh bioreactorr
type and the inittial cellular concentratiion was 13-14 cell/microcarrier. Supernatantss of
w harvested on days 2 and 3 afteer start the cycle
c
of prooduction.
thesee cultures were
Sampples of these cultures were
w taken every
e
day duuring the prroduction cyycle to
deterrmine the ceellular concentration. Itt was studieed too the ceellular loss in the first day
d
after the cell inooculation to analyze thee cell difficuulty for spreead on the microcarrier
m
rs.
The aaverages off the values of cell speccific grow raate found beefore the haarvest beginnning
weree 0.025 h-1 and
a 0.023 h-1
in the induustrial and 30
3 L bioreactors respecctively. Thee
perceentage averaages of celllular loss in the first dayy after cell inoculation
n were 37% (±
16%)) in the induustrial bioreeactor and 52%
5
(± 21%
%) in the biooreactor of 30
3 L. The
analyyze of the data
d found too cell speciffic grow ratte and cellullar loss in th
he rabies virrus
produuction cyclees showed that
t the spreead and grow
wth cellularr were not affected
a
by the
t
bladees of the im
mpeller of industrial bioreactor. In conclusion,
c
the expansion of Veroo cell
grow
wth for rabiees virus prodduction in bioreactor
b
of 150L was satisfactoryy because inn this
systeem the valuees of cellulaar loss and cell
c specificc growth ratte were simiilar or betteer
than the values found
f
in thee 30 L bioreeactor.
Finanncial support: Fundaçãão Butantan
PT.0087
v
anttirrábicas veterinária
v
s, nacionais e importa
adas realizaadas
Avalliação das vacinas
pelo Ministério
o da Agricu
ultura Pecu
uária e Abaastecimento
o do brasil no
n período de
20099 a 2011
Cam
mpos APG1, Gonçalves G1, Zaroni MMH1, Sirrineu JC1, Buzatto
B
DF1, Faria AM1 1
Minnistério da Agricultura
A
P
Pecuária
e Abastecime
A
ento - Lanaggro-SP - Co
ontrole de
Vacinas Antirráábicas
Introodução e ob
bjetivo: Noo Brasil as vacinas
v
a serrem usadas na profilax
xia da raiva
anim
mal são subm
metidas a coontrole por órgão
ó
goverrnamental. Em
E cada lotte de vacinaa
nacioonal e importada são reealizadas am
mostragens destinadas
d
à análise fisscal. Dentre os
testes aplicados estão o de esterilidade
e
e, inocuidadde, inativaçãão viral, pH e o teste dee
c
de
NIH.. Para o testte de NIH faaz-se a deteerminação dda dose efetiiva (DE50) capaz
proteeger 50% doos camundoongos vacinaados com a vacina a seer testada e uma
u vacinaa
referrência. A paartir da compparação estaatística entrre a DE50 daas duas vaciinas, calculaa-se a
potênncia relativaa da vacina teste. A anáálise fiscal das
d vacinas, realizada pelo
p MAPA
A no
Lanaagro-SP, é de
d extrema importância
i
a, uma vez que,
q promovve os interessses nos
objettivos da saúúde pública, economia e para a inddústria produ
utora indicaa pontos crííticos
que ssão mereced
dores de meelhorias. Deeste modo, foram
f
realizzadas estatíssticas descriitivas
dos rresultados obtidos
o
dos lotes
l
de vaccinas amostrradas entre 2009 a 2011. O objetivvo
destee trabalho fooi avaliar o desempenhho das mesm
mas, além dee comparar as vacinas
nacioonais e impoortadas. Resultados: Durante
D
estee período fooram realizaados 227 tesstes,
nos qquais foram
m avaliadas amostras
a
dee lotes de 144 tipos de vaacinas impo
ortadas e 18
nacioonais (com seus respectivos lotes). As vacinas nacionais totalizaram
m 592 partiddas e
as im
mportadas 1554. O teste de inocuidaade, que avaalia a toxicid
dade da vaccina antirrábbica,
inativvação viral e a mensuraação do pH, não apreseentaram rep
provação. Poor sua vez, o
teste de esterilid
dade apresenntou em méédia 0,5% dee reprovaçãão nas vacinnas nacionaiis. No
e média 9%
9 de retestte interno (R
RI). Em 36%
% dos
teste de NIH as vacinas aprresentaram em
os limites estabelecidos e em 16% a DL50. A
testes a DE50 daa VRN estavvam fora do
N mostrouu que 22% das vacinass nacionais não
n apresenntaram nenhhuma
avaliiação pelo NIH
reproovação, nas importadass 86% apressentaram-se satisfatóriaas. Para difeerenciar o
desem
mpenho dass vacinas, quanto à potêência relativva, estas forram separaddas nos gruppos:
“vaciinas com 10
00% de aproovação” e “vacinas
“
com
m alguma reeprovação”.. As potênciias
médiias de cada vacina, por grupo, foraam comparaadas por um
m teste estatíístico de
compparações mú
últiplas de médias.
m
O 1º.
1 Grupo fooi constituíddo por 11 tippos de vacinnas
impoortadas e cinnco nacionaais, com desstaque a um tipo de vaccina importaada que é
distinnto dos outrros 15, deviido a sua vaariabilidade da potênciaa (superior) relativa e suua
médiia. No 2º. Grupo
G
foi composto porr 13 tipos dee vacinas naacionais e trrês importaddas,
diferrenciando-see em oito tippos de compportamentoo com relaçãão a potênciia relativa.
Concclusão: Os produtos avvaliados atenderam aoss critérios dee aceitabilid
dade de acoordo
com a Portaria Ministerial
M
n
n°288,
conttudo consideerando os diferentes
d
prrodutos e
espéccies a que se destinam,, as vacinas importadass demonstraaram o maioor índice de
aprovvação. Resssalta-se a neecessidade de
d uniformidade e conssistência nos produtos a fim
de see obter vacin
nas de elevaada capaciddade imunoggênica, assim
m como, deesenvolvimeento
de m
métodos paraa avaliar a qualidade
q
do
o antígeno e concentraçção proteicaa nas vacinaas
avaliiadas. Agra
adecimento: MAPA.
PT.0088
GS AND CA
ATS RESC
CUED IN A SMALL BRAZILIA
B
AN CITY: POTENTIA
P
AL
DOG
RAB
BIES TRAN
NSMITTER
RS
Babbboni SD1, Fo
ornazari F2, Gonzalez GK
G 1, Modoolo JR3 - 1Ennvironmental Health
Survveillance Deepartment, Municipal
M
P
Prefecture
off Botucatu, São Paulo State,
S
Braziil.,
2
Schoool of Veterrinary Mediicine and Animal
A
Sciennce - UNES
SP - Botucattu - Veterinnary
Hygiiene and Pu
ublic Health, 3School off Veterinaryy Medicine and Animalls Science UNE
ESP - Botuccatu - Veteriinary Hygieene and Pubblic Health
Domestic dogs aand cats aree the most iimportant rrabies transmitters to m
men in Brazzil. des the annual vaccination realizeed by the pu
ublic service
e, many anim
mals remain
n Besid
unprrotected and
d exposed tto rabies. Sttray animalss are even m
more vulnerable to rab
bies due tto sanitary conditions w
which they are exposeed, such as ccontact with
h bats, bite injuries caused by other strray dogs and cats potentially infeccted, and ab
bsence of wards thesee facts, the population of domestic dogs and cats is an vacciination. Tow
impo
ortant aspecct in rabies prevention. The present study aim
med to analyze the num
mber of do
ogs and catss rescued in
n a small Braazilian city. The study w
was realized
d in the cityy of Botucatu (22°53
3’09”S 48°26’42”W), Sãão Paulo Staate, Brazil, w
which comp
prises an area of 2.87 km2, and has a hu
uman population of approximatelyy 127,370 h
hab.. The citty 1,482
realizzes annual vvaccination
n campaign against rabies since 19
968, and the
e last positivve diagn
nosis for rab
bies in a dog was in 1985. Althouggh canine/fe
eline rabiess remain controlled since
e this date, tthe infectio
on is presentt in bats, bo
ovines and h
horses, as confiirmed by seeveral positiive diagnosis performeed annually.. The animaal rescue is perfo
ormed by th
he Environm
mental Heallth Surveillaance Departtment of the Municipal prefeecture, and includes stray or unwaanted animals. The lastt group con
nsists mostlyy of dogss and cats ab
bandoned b
by their own
ners, usually declaring a sever disease, lack o
of physical space, eexcessive agggression, aamong otheers. Data weere recordeed from the mber of rescued animaals was as fo
ollow: in 20
006 – 2,298 dogs depaartment’s filles. The num
and 5
560 cats; in 2007 – 2,066 dogs and
d 533 cats; in 2008 – 1,271 dogs aand 421 cats; in 2009
9 – 688 dogss and 149 cats; in 2010
0 – 655 dogss and 204 cats; and in 2011 – 641 dogs and 1
199 cats. Th
hese resultss show a greeat numberr of animals rescued byy the public service in Botuccatu. A decrease in the number of animals waas observed
d along the 9. This fact iis a result of a municipal law (num
mber 12.916
6) yearss, especiallyy after 2009
estab
blished in 2008, which defined new criterionss for animal euthanasia, and consequently reeduced the amount of this proced
dure. As a co
onsequencee, the Health Survveillance Department aadopted a new policy, aand started to Envirronmental H
rescu
ue only straay animals ccausing public disturbance or visib
bly sick. Unw
wanted anim
mals, which were freq
quently ordered to be eeuthanatizeed (by their owners) be
efore 2008, nimals led to
o a weree also not alllowed to be rescued. TThus, the reeduction of rescued an
decreease in euth
hanasia pro
ocedures, w
which is in acccordance w
with the new
w law. The resullts of this ne
ew measuree, as well ass the law, w
were not evaaluated, and
d one must consider the possible increase of strayy animal population, an
nd also the risk of rabiees esponsible o
ownership is an importtant aspect that must be evaluateed infecction. The re
afterr the implem
ment of thiss new policyy, once thatt unwanted animals aree not being allow
wed to be ab
bandoned aand sent to the Environ
nmental Health Surveilllance Depaartment.
PT.0089
NHECIMENTO SOBR
RE RAIVA
A AVALIA
ADO A PAR
RTIR DE ALUNOS
A
D
DO 2º
CON
ANO
O FUNDAM
MENTAL DE
D UMA ESCOLA
E
M
MUNICIPA
AL DE
JABOTICABA
AL/SP
Moraaes FC1, Piccinato MAC
C1, Grisólio APR1, Nunnes JOR1, Frrias DFR1, Cruz
C
CA,
2
3
1
1
Meirrelles-Bartoli RB , Roddrigues TO , Bürger KP
P , Carvalho
o AAB 1
FCA
AV/UNESP
P/Jaboticabaal/SP, 2Univversidade Feederal de Gooiás,
3
DAP
PSA/FMVA
A/UNESP/A
Araçatuba/S
SP
00 habitantees, a qual
Jabotticabal posssui uma poppulação urbaana estimadda em 70.00
convvive com um
ma grande cooncentração
o de cães e gatos,
g
sugerrindo a neceessidade de
conhhecimento soobre controle da reproddução dessees animais, posse
p
respo
onsável e
preveenção de im
mportantes zoonoses.
z
N
Neste
cenárioo, teve origeem o projeto “Formaçãão de
multiiplicadores em controlee de zoonosses urbanas e em possee responsáveel de animaiis de
estim
mação no Município
M
dee Jaboticabaal/SP”, que se
s encontra em execuçãão como proojeto
pilotoo em uma escola
e
do muunicípio, poossibilitandoo a aprendizzagem de ed
ducandos, por
p
meioo de um cursso semi--preesencial de 180 horas, com posterior interaçãão e
disseeminação deesses conhecimentos aoos alunos, toornando-os transmissorres de
inforrmação paraa a sociedadde. Por consstituir a raivva urbana um
ma importannte zoonosee e
anuaalmente ser responsável
r
l por milharres de mortees de seres humanos
h
noos países nãão
indusstrializados, ela se tornna um gravee problema dde saúde pú
ública em innúmeras regiões
do m
mundo, tendo
o o cão com
mo um dos principais
p
reeservatórioss e fonte de infecção.
Assim
m, parte desste projeto teve
t
como objetivo
o
avaaliar o conhecimento prrévio dos
escollares do 2º ano
a fundam
mental de um
ma escola municipal
m
daa cidade de Jaboticabal/
J
/SP
sobree a raiva, paartindo do princípio
p
de que a escolla é responssável pela fo
ormação de
cidaddãos conscieentes, críticcos e ativos na sociedadde. Visto quue o contatoo direto com
m cães
e outtros possíveeis reservatóórios é cada vez mais frrequente, fooi desenvolvvido e aplicaado
um qquestionário
o a 43 alunoos do 2º ano fundamenttal, questionnando se posssuem anim
mais
doméésticos em sua
s residênccia, históricco de vacinaação contra raiva e se existe o
conhhecimento por parte dessses alunos sobre essa enfermidad
e
e. Os resulttados obtidoos
demoonstraram que
q 95,34% dos entreviistados posssuem animaais de estimaação e que,
apesaar de 35% desses
d
anim
mais ter livree acesso a ruua, 13,95% dos alunos afirmaram que
seus cães e gatos não são vaacinados coontra raiva e 27,90% diisseram nunnca ter ouviddo
falar sobre a doeença. Os entrevistados também ressponderam sobre a ocoorrência de
miliares, obseervando um
ma porcentaggem
acideentes (morddeduras) entre seus fam
signiificativa de 18,60% de acidentes reelatados oriiundos de attaques sofridos pelo próprio
anim
mal de estimação e 16,27% causadoos por anim
mais desconhhecidos. Essses dados sãão
preoccupantes, um
ma vez que o contato entre
e
essas ccrianças e seeus animaiss é muito
próxiimo, e sem um controle efetivo coom a vacinaação o risco de infecção
o persiste
necessitando aboordagens esscolares sobbre esta e ouutras importtantes zoonooses, as quaais os
alunoos estão sussceptíveis.
PT.0090
ETROSPECTIVE ST
TUDY OF ANIMAL
A
R
RABIES
DIIAGNOSIS
S IN THE
A RE
CEN
NTRAL RE
EGION OF SÃO PAU
ULO STATE
E, BRAZIL
L, DURING
G A 10-YEAR
PER
RIOD
Langgoni H1, Forrnazari F1, Marson
M
PM
M1, Guimarãees FF1, Men
nozzi BD1, Da
D Silva DB
B1,
1
1
1 1
Paiz LM , Wandderley GG , Cavalcantee RV - School of Vetterinary Medicine and
mal Science - UNESP - Botucatu - Veterinaryy Hygiene annd Public Health
H
Anim
a
cann transmit raabies to men
n. The
Seveeral species of domesticc and wild animals
epideemiology prresent differrent patternns accordingg to each reggion, and thhe identificaation
of infected anim
mals has greaat importan
nce to assist prophylaticc measures. The presennt
mal rabies ddiagnosis reealized by th
he Zoonosees
studyy consists inn a retrospecctive of anim
Diaggnosis Service (ZDS), from
f
the Veeterinary Hyygiene and Public
P
Health Departm
ment,
locatted at the Scchool of Veeterinary Meedicine and Animals Sccience (FMV
VZ), São Paaulo
Statee University
y (UNESP), Botucatu city
c (22°53’009”S 48°26
6’42”W), Brrazil. The ZDS
Z
is linnked to Pastteur Institutee of São Pauulo State, coontributing to rabies coontrol. Mostt of
the samples subm
mitted to raabies diagno
osis (over 955%) come from
f
cities located
l
in thhe
B
(arounnd
centrral region off São Pauloo State, whicch is the moost populouss State of Brazil
41,2552,160 habiitants), has a territory of
o 248,209.4426 km2, an
nd its animaal trade has great
g
impoortance in th
he national economy.
e
T immunoofluorescentt antibody test (IFAT) and
The
the bbioassay in mice
m were the
t laboratorial tests em
mployed forr rabies diaggnosis. Dataa
weree recorded from
fr
2002 too 2011. A tootal of 51466 samples were
w analyzeed (2,409 baats,
1,6699 dogs, 512 cats, 329 bovines,
b
1000 terrestrial wild mamm
mals, 95 horrses, 38 sheeep,
19 gooats and 6 pigs).
p
The sppecies that presented
p
positive diag
gnosis were bats (n=24),
bovinnes (34), hoorses (n=6) and pigs (n=
=1), totaliziing 65 posittive animalss. The
propoortion of po
ositivity amoong each sp
pecies were: 1.0% for bats
b (24/240
09), 10.3% for
f
bovinnes (34/329
9), 6.3% for horses (6/995), 16.6% for
f pigs (1/66) and 0.0%
% for dogs, cats,
c
sheepp, goats and
d terrestrial wild mamm
mals. The reesults indicaate that boviines, horsess and
bats are infectedd by rabies virus
v
in thiss region. Altthough the proportion
p
o positive pigs
of
p
%), only 6 annimals weree sampled, aand a study
y including a representaative
was hhigh (16.6%
numbber of pig saamples couuld provide a more reliaable data, noot only for pigs
p but alsoo for
sheepp, goats and
d terrestrial wild mamm
mals. Bats presented
p
a lower
l
propoortion of positive
anim
mals when coompared to bovines and horses. Thhis fact is a result of thhe high num
mber
of baats submitteed to rabies diagnosis without
w
cliniical suspicioousness of the
t disease,
sincee any bat fouund in urbaan areas by the
t Environnment Healthh Departmeent of the ciity is
sent tto rabies diaagnosis. Thhe absent off positive doogs and cats indicates th
he success of
o the
preveentive meassures adopteed for these species in tthe last yearrs, especiallly the municipal
vacciination cam
mpaigns, whhose focus arre these aniimals.
PT.0091
TIGENIC CHARACT
C
TERIZATION OF RA
ABID ANIM
MAL ISOL
LATES,
ANT
NOR
RTHERN BRAZIL,
B
2008/2012.
Casseb LMN1, Coelho
C
TFS
SB1, Pereiraa AS1, Ferroo MNP1, Lop
pes NRS2, Begot
B
AL3,
4
1
1 1
FC , Travasssos da Rosaa ES - Insstituto Evandro Chagas Limaa RJS , Vasconcelos PF
Seçãão de Arbov
virologia e Febres
F
Hemorrágicas, 2Universidadde Federal Rural
R
da
Amaazônia, 3Seccretaria de Estado
E
de Saaúde Pública - Núcleo de
d Doençass Endêmicass de
Estaddo do Pará, 4Núcleo dee Doenças Endêmicas
E
dde Estado doo Pará
T identificcation of thee antigenic variants of rabies viruss furnish
Introoduction: The
impoortant inform
mation on thhe origin off the circulatting rabies virus
v
in a deetermined
regioon and, in coonsequencee, the existin
ng cycles annd the involv
ved speciess on them.
Aimiing to exten
nd the know
wledge of thee epidemiollogy of the rabies
r
in no
orthern Brazzil,
we inncorporated
d the techniqque of antiggenic characcterization for
f the laborratorial
diagnnosis of the rabies. Objjective: To determine
d
thhe epidemioologic profiile of the annimal
rabiees in northerrn Brazil, duuring the peeriod of Jannuary 2008 through
t
Julyy 2012.
Meth
hod: A total of 32 rabiees virus isollates from wild
w and dom
mestic anim
mals were ussed
for anntigenic chaaracterizatioon as follow
ws: 21 dogs,, a cat, five bovine, threee equine annd
two bbats all of th
hem from Pará
P State. The
T Indirectt Imunofluorescence Assay (IIFA)) was
perfoormed using
g a panel off monoclonaal antibodiess against thee rabies viruus nucleoprrotein
(prodduced by CD
DC/Atlanta, USA), Onne dog rabidd sample waas genetic chharacterizatted
perfoormed using
g the assay of
o RT-PCR in two stepps seconds Barbosa
B
et al
a (2007).
Resu
ults: The 200 dog isolatees and one cat
c sample w
were identiffied as Variaant 2, strainn
comm
monly founnd among doogs; all boviine, equine and bat rabbid samples (nonhemaatophagous)) were identtified as Vaariant 3, whoose reservoiir is the hem
matophagouus bat
Desm
modus rotun
ndus, a sampple of dog, not compattible with thhe panel, waas characteriized
genetically as VAg3.
V
Concclusions: Thhese results suggest thaat in the nortthern regionn of
p
in
n dogs; the other
o
strainss
Brazzil the antigeenic variantt 2 continues to be the prevalent
weree antigenicallly characteerized as bellong to the variant
v
3 (fr
from vampirre bats). Thee
genetic characteerizations off all studiedd strains are on going annd should complement
c
t this
studyy.
MCT and IEC/SVS/MS
Finanncial support: CNPq/M
PT.0092
TIGENIC AND
A
GENO
OTYPIC CHARACT
C
ERIZATIO
ON OF RA
ABIES VIR
RUS
ANT
ISOL
LATES FR
ROM BATS
S FROM BOTUCATU
B
U CITY AN
ND REGIO
ON
Menoozzi BD1, Carrieri
C
ML, Oliveira RN,
R Pereira VBR, Langgoni H - 1Unnesp - DHV
VSP
s
time ago,
a bats aree receiving an
a increasedd importancce in Publicc Health beccause
For some
they are consideered the maiin reservoirs of rabies virus
v
aroundd the world, including
h bats remain the rabies epidemiollogical cyclles for centuuries, only in the
Brazzil. Although
last ddecades the rabies in baats had its recognition, throughoutt researchess focused onn the
role oof these aniimals in the epidemioloogical cyclee of the disease and alsoo on their
variaants and their implicatioons in the development
d
t of new resservoirs for rabies viruss,
mainnly in regionns where thee disease inn dogs was controlled.
c
T state off São Paulo,,
The
coorddinated by the
t Institutee Pasteur, peerforms epiddemiological surveillannce for rabiies
throuugh registerred laboratorries for its diagnosis.
d
T
The labs of Zoonosis
Z
D
Diagnostic
Serviice, School of Veterinaary Medicinne and Anim
mal Science - UNESP - Botucatu - SP
is reggistered for receiving material
m
for diagnosis of
o some munnicipalities of the regioon.
Betw
ween 2003 and
a 2010, 199 samples of
o non-hemaatophagous bats were diagnosed
d
ass
posittive by IFD and Biologgical Evidennce. These isolates were antigenicaally
charaacterized by
y the panel oof monoclonnal antibodies donated
d by PAHO--CDC (Centters
for D
Disease Conntrol and Prrevention, Atlanta,
A
GA, USA Pan-A
American Health),
H
reveaaling seven of these isoolates belon
nged to variaant 3, which
h has the vaampire bat
Desm
modus rotun
ndus as reseervoir, as weell as an inssectivorous bat of the genus
g
Myotiss
preseenting variaant 4, characcterisitc of the
t other inssectivorous bat Tadaridda brasiliennsis,
and oother three profiles
p
are not compattible, NC-1,, NC-2 and NC-3. All isolates
i
werre
assayyed by RT-P
PCR and their productss had the nuucleoproteinn gene (N) viral
v
partiallly
sequeenced, geneerating a phylogenetic tree
t that groouped thesee isolates intto four clustters,
desiggnated as lin
neages Nycttinomops, Myotis,
M
Desm
modus rotunndus, and a new lineagge of
rabiees virus not previously characterized, which aapparently has
h the bats of the genuus
Myottis as reservvoir. This linneage show
wed an averaage intraspecific identitty of 99.8%
%,
rangiing from 99
9.6 to 100% for nucleottides and 1000% amino acids.
PT.0093
EXP
PRESSÃO DA
D GLICO
OPROTEÍN
NA DO VÍR
RUS DA RAIVA
R
POR
R CÉLULA
AS
DE D
DROSÓFIL
LA E ANÁ
ÁLISE DA PRODUÇÃ
P
ÃO DE ANTICORPO
OS EM
CAM
MUNDONG
GOS IMUN
NIZADOS
Venttini-Monteirro DC2,1, Leemos MAN
N2, Jorge SA
AC2, Tonso A1, Pereira CA2, Astrayy
2 1
RM - USP - Escola Politeecnica/Labooratório de Células
C
Animais, 2Instituto Butanttan Labooratório de Imunologia
I
Viral
m sendo
Introodução: céllulas de Droosophila meelanogasterr Schneider 2 (S2) vêm
utilizzadas para a expressão de proteínaas heteróloggas, por atinggirem altas densidadess,
serem
m de fácil cuultivo e exppressarem estavelmentee o gene de interesse. A glicoproteeína
do víírus da raivaa (RVGP) é um produtto de interessse biotecnoológico por ser responssável
pela indução da resposta im
mune protetoora contra a infecção pelo vírus daa raiva.
GP através de
d cultivos de
d células S2
S transfectaadas e analiisar a
Objeetivos: prodduzir a RVG
respoosta imunológica de annimais imunnizados com
m essa glicop
proteína. Métodos:
M
céllulas
S2 fooram transfeectadas com
m um plasmídeo contenndo o gene da
d RVGP co
ontrolado por
um ppromotor dee expressão indutível e selecionadaas primeiram
mente por reesistência à
higroomicina. A nova
n
linhaggem, S2MtR
RVGP, foi cultivada
c
em
m biorreatorr Bioflo, vollume
de traabalho 1L, com inóculoo inicial de 5x105 cel/m
mL. Após in
ndução com
m CuSO4 (2-6
5x100 cel/mL) as
a células fooram cultivaadas até iníccio da fase estacionária
e
a de crescim
mento
celullar (28°C, 90 rpm, OD 10% de satturação de ar).
a A RVGP
P foi dosadaa por ELISA
A. Os
camuundongos fo
oram imunizzados (dias 0, 7 e 14) com
c
vacina comercial (controle
(
posittivo), RVGP
P recombinaante e salinaa (controle negativo). Após
A
as trêss doses do
agennte imunizan
nte os camuundongos recceberam a cepa
c
CVS inntracerebrall para ensaio
desaffio. Os anticcorpos foram
m dosados em amostraas de soro po
or ELISA (P
Platelia).
Resu
ultados: O Xmax
X
obtiddo nos ensaiios em biorrreator foi 3,,1x107 cel/m
mL e a
RVG
GPmax 3,9 mg/L.
m
Apóss esquema vacinal
v
de trrês doses a vacina
v
comeercial produuziu
4,17 UI/mL de anticorpos
a
a
anti-RVGP
(apresentanndo resultaddos superiorres a 0,5 UI//mL,
mo exigido pela OMS,, após a prim
meira dose vacinal).
v
A RVGP recoombinante, após
mínim
três ddoses, apressentou 3,71 UI/mL (sup
perior a 0,5 UI/mL apóós segunda dose).
d
O gruupo
salinna não demo
onstrou proddução de an
nticorpos. No ensaio deesafio, os annimais
imunnizados com
m vacina com
mercial tiveerem 100% de sobrevivvência, gruppo RVGP
recom
mbinante 800% e zero no
n grupo sallina. Discusssão: frente aos resultaddos obtidos de
expreessão de pro
oteínas, proodução de annticorpos e alta porcenttagem de soobrevivência no
teste desafio, as células S2 demonstram
m ser um pootencial sisttema de exppressão paraa a
GP. A menor produção de anticorppos nos anim
mais imunizzados com RVGP
R
RVG
recom
mbinante, em
e comparaação ao grup
po vacina coomercial, poode estar rellacionada ao
fato dde nossas preparações ainda não conterem
c
puureza adequada e não estarem
assocciadas a adjuvantes.
SP (2009/099327-7) e CN
NPq (1427229/2010-8).
Supoorte financeiiro: FAPES
PT.0094
AVA
ALIAÇÃO DO
D PROG
GRAMA DE
E ATENDIM
MENTO ANTIRRÁBICO A
PES
SSOAS EN
NVOLVIDA
AS EM AGR
RAVOS CO
OM ANIMA
AIS NO MUNICÍPIO DE
DES
SCALVADO
O, SÃO PA
AULO, BR
RASIL
B
KP1, Souza RR
R2, Nunes JO
OR1, Picinatto MAC1, Moraes
M
FC1,
Grisóólio APR1, Bürger
1
1
1
1 1
Friass DFR , Maachado MA , Bichuette MA , Carvvalho AAB - Faculdadde de Ciênccias
Agráárias e Veterrinárias, FC
CAV/Unesp, Câmpus Jaaboticabal/S
SP - Departamento de
Mediicina Veteriinária Preveentiva, 2Graaduanda do curso de Medicina Vetterinária da
UNIC
CASTELO,, Descalvaddo/SP
No Brasil,
B
a raiva urbana
a encontra--se controlada na ma
aioria dos estados,
e
poré
ém o núme
ero de trata
amentos prrofiláticos pós-expos
p
ição efetua
ados em
deco
orrência do
o envolvimento de pe
essoas em acidentess com cães
s ou gatos
ainda é elevad
do. Este tra
abalho obje
etivou avalliar os registros de profilaxia
mana pós-e
exposição no Municíípio de Desscalvado/S
SP, no perííodo
antirrrábica hum
de 2
2005 a 2009. Para tan
nto, foi rea
alizado, em
m parceria com
c
a Seccretaria de
Saúd
de do Mun
nicípio, um levantame
ento de casos registrrados nas fichas
f
de
invesstigação de atendime
ento huma
ano pós-exxposição, com
c
inform
mações
provvenientes do
d SINAN (Sistema
(
d Informaçção de Agravos e No
de
otificações).
®
Esse
es dados fo
oram tabulados no software Microsoft Exxcel 2007 e analisad
dos
em ttabelas e gráficos.
g
No
os resultad
dos de 752
2 casos no
otificados destacou-se
d
e no
ano de 2005 o elevado n
número de pessoas que
q procurraram atendimento
dico. Os ma
aiores agra
avos (75% dos casoss) foram re
egistrados na zona
méd
urba
ana, o que indica que
e a maioria
a dos agresssores eram domicilia
ados. A faixa
etáriia com mais acidente
es foi de 0--15 anos, com
c
245 agravos. Fa
ato explicado
pela intensa attividade da
as criançass. Para o sexo
s
mascu
ulino, 53% dos
entes, na faixa
f
etária
a de 31-45 anos. Com
m relação ao animal,, 85% dos
acide
agra
avos foram causadoss por cães e 9,4%, po
or gatos. Ainda,
A
46,8
8% dos cãe
es e
33,8
8% dos gattos envolvidos nos ac
cidentes fo
oram decla
arados vacinados no
mom
mento da agressão. No
N entanto
o, há falhass de preenchimento do
d campo
relaccionado a vacinas
v
na
as fichas de notificaçção. Para os
o cães, 39
9,15% dela
as
estavam incom
mpletas e, para os ga
atos 46,47%
%. Em 70,6% dos ca
asos, o animal
s
no momento
m
do
o agravo e 85,3% ma
antiveram-se sadios
foi declarado sadio
ante o perío
odo de obsservação; no entanto
o, 29,4% das fichas estavam
e
dura
incompletas co
om relação
o a esta infformação. Por fim, 55
53 (73,54%
%) pessoass
olvidas noss acidentess foram submetidas à profilaxia
a com uso de vacinass, e
envo
os ca
asos conduzidos ape
enas com a observaçção do animal, ou se
eja, sem
instittuição de vacina,
v
fora
am 199 (26
6,5%). Com
m relação ao soro an
ntirrábico, foi
f
indiccado para 70
7 pessoa
as (9,3%). Os
O resultad
dos indicam que a in
nstituição d
de
profiilaxia pós-e
exposição,, na maioriia dos caso
os, foi feita
a com base
e apenas na
n
cara
acterização
o dos acide
entes, não se levando
o em conssideração a condição
o
epidemiológica
a da área e nem a co
ondição do
o animal no
o momento
o do agravo
o. É
ssidade da
a atuação de
d forma in
ntegrada entre os serrviços méd
dico
visívvel a neces
e mé
édico veterrinário na conduta
c
do
o atendime
ento e da capacitação
c
o dos dem
mais
profiissionais envolvidos no tema, o que favorrece a corrreta aplicação do
trata
amento anttirrábico pó
ós-exposiç
ção.
PT.0095
N-SPECIFIIC STIMUL
LATION INDUCED BY CLOST
TRIDIOSIIS VACCIN
NE
NON
IN A
ANTI-RABIIES ANTIB
BODIES TITERS
T
IN VACCINA
ATED MIC
CE
Appoolinario CM
M1, Allendorrf SD1, Perees MG1, Maartorelli LFA
A2, Kataokaa APAG2,
1
1
1 1
Vicente AF , Anntunes JMA
AP , Megid J - Unesp--Faculdade de Medicin
na Veterinárria e
2
d Higiene Veterinariaa e Saude Pu
ublica, Cenntro de
Zoottecnia - Deppartamento de
Conttrolede Zoon
noses de Sãão Paulo - Laboratório
L
de Zoonosees e Doençaas Transmitiidas
por V
Vetores
In prrevious studdies an unexxpected incrrease in antii-rabies titerrs in vaccinaated calves and
also in not vacciinated calvees that weree born from previously vaccinated dams was
i
occcurred one month
m
after clostridiosiis vaccinatioon and perssisted
obserrved. This increase
until the 12th month
m
of agee, suggestingg that this vaccine
v
had stimulated the
t specificc
W
the expperiment waas performeed with not vaccinated
v
immuunity against rabies. When
dams
and ccalves, no inncrease in anti
a rabies antibodies
a
titers was observed sugg
gesting that
closttridiosis vacccine was abble to increaase the levels of anti-raabies antibodies only inn
anim
mals that werre previouslly primed with
w the rabiies vaccine or that weree actively
vacciinated. In th
his context, the aim of the
t present study was to
t confirm, in another
anim
mal model, th
he effect off clostridiosiis vaccine as
a a non speecific stimu
ulus for anti
rabiees antibodies titers. Forrty eight swiiss mice, female, 60 daays-old divided into sixx
groupps of eight animals
a
eacch were used
d. The grouup A was noot vaccinated, group B and
C waas vaccinateed with anti rabies and clostridiosis vaccine, respectively
r
y, on day 0 aand
sacrificed at day
y 15; group D and E waas vaccinateed with antii rabies and clostridiosiis
d 30 and group
g
F wass vaccinatedd
vacciine, respecttively, on daay 0 and saccrificed at day
with clostridiosiis vaccine on
o day 0 folllowed by anntirabies vaccine on daay 15 and
y 30. It wass used a com
mmercial ceell culture annti-rabies vaaccine and a
sacrificed on day
i a dose off 0.04 mL byy intramusccular route.
polyvvalent closttridiosis vacccine, both in
Anim
mals were saacrificed according ethhical commiittee guideliines and blo
ood sampless
weree collected. Serum sampples were obtained, stoored at -20oC and anti-rrabies
antibbodies titers were evaluuated using RIFFT moddified test. Animals
A
nott vaccinatedd or
vacciinated againnst clostridiiosis didn´t present protective leveels of anti raabies antibodies.
Micee vaccinated
d against rabbies presentted protectivve levels off antirabies antibodies
a
t
titers
at daay 15 that inncreased on day 30. Titer of antirabbies antibod
dies 15 dayss after antiraabies
vacciine was signnificantly higher
h
in aniimals that reeceived closstridiosis vaaccine beforre,
comppared to thee group onlyy vaccinatedd against rabbies and evaaluated 15 days
d
after
vacciination. Theese results agree
a
with previous
p
repports and reiinforce a noon specific
stimuulation induuced by closstridiosis vaaccine. More studies arre needed cooncerning thhe
closttridiosis vacccine compoonents respoonsible for the
t non specific stimulation and thhe
interference of this
t in seroloogical tests in practice..
PT.0096
quência de Quirópteroos Positivoss para Raivva Examinaados no Esttado de Gooiás
Freq
de 20007 a 2011..
Chaggas IN1, Roccha CGN1, Aguiar MS1, Resende RM
R 1, Vieiraa RC1, Costta YL1, Sanntos
1 1
MFC
C - AGRO
ODEFESA
Os m
morcegos peertencem à Ordem
O
Chirroptera, e podem ser veetores ou reeservatórios de
váriaas doenças importantes
i
em saúde pública,
p
denntre elas a raaiva. Existem dados dee
morttes, por raiva, de conquuistadores esspanhóis e seus
s
animaiis, sugerindoo que os
morccegos já tran
nsmitiam a raiva antes da colonizaação das Am
méricas. Moorcegos, de
diferrentes hábitoos alimentarres, foram descritos
d
como transmiissores da raaiva, já tenddo
sido estabeleciddo que os moorcegos hem
matófagos sejam
s
responnsáveis pelaa maioria doos
ntre os morrcegos hemaatófagos, a espécie
e
relatoos de casos de raiva noo Brasil. Den
Desm
modus rotun
ndus é a prinncipal transsmissora da raiva para os
o herbívoroos, acarretanndo
granddes prejuízoos econômiccos na área rural. O Esttado de Goiiás possui uma
u pecuária
impoortante e exppressiva, sendo um graande forneceedor de carnne, tanto naccional comoo
internnacionalmeente. Portantto, é fundam
mental o connhecimentoo da frequênncia de
ocorrrência de raaiva em quirrópteros. Fooram analisaados os exam
mes realizaados pelo
laborratório do seerviço oficial de Defessa Sanitária Animal do Estado de Goiás
G
entre os
mesees de janeiro
o de 2007 a dezembro de
d 2011. Neeste períodoo foi realizaado o diagnóóstico
de raaiva em 8100 morcegos no
n Laboratóório de Anáálise e Diagnnóstico Veterinário LAB
BVET, da Agência Goiaana de Defeesa Agropeccuária - AG
GRODEFESA. Estes
diagnnósticos forram realizaddos através das técnicass de Imunoffluorescência Direta – IFD
e da Inoculação Intracerebrral em Camuundongos – IICC. Dos morcegos recebidos,
r
8
806
m negativoss e quatro fooram positivvos para raivva, o que reepresenta 0,449% do totaal das
foram
amosstras. Dos 810
8 morcegoos analisadoos no LABV
VET, 748 (992,35%) eraam oriundoss do
Estaddo de Goiáss e 62 (7,65%
%) eram dee outros Estaados da Fedderação; 5199 (64,07%) eram
hemaatófagos e 291
2 (35,93%
%) eram morcegos não hematófagoos. Todos os morcegos
posittivos eram hematófago
h
Desmodus rotundus,
r
e provenientees do
s, machos, da espécie D
Estaddo de Goiáss. Houve um
ma amostra positiva
p
no ano de 20007, provenieente do
muniicípio de Niiquelândia e três amosttras no ano de 2009, sendo uma doo municípioo de
Mam
mbaí e duas do municíppio de São Domingos.
D
A
Apesar
de teer sido encoontrado som
mente
0,49%
% de morceegos positivvos, em todo
os os examinnados, os riiscos à saúdde pública e para
a poppulação anim
mal são reais, sendo neecessária a manutenção
m
o da vigilânccia
epideemiológica,, com ênfasee no controlle de morceegos, diminu
uindo assim
m a circulaçãão
viral e reduzindoo as chancees de ocorrên
ncia de raivva em seres humanos e animais.
PT.0097
RIE HISTÓ
ÓRICA DOS
S CASOS DE
D RAIVA
A HUMANA
A NO ESTADO CEA
ARÁ
SÉR
Nayllê FHD1, Pin
nheiro KMA
A1, Damascceno KA1, Barros
B
AS1, Oliveira FB
B1, Oliveiraa RJ1,
Oliveeira RJ1, Du
uarte BH2 - 1SESA-CE - NUVET, 2UFERSA - FACULD
DADE DE
MED
DICINA VE
ETERINÁR
RIA
r
é umaa zoonose de notificaçãão compulsó
ória, que aprresenta quaase
Introodução: A raiva
100%
% de letalidaade. No Muundo, segun
ndo a Organnização Munndial de Saúúde, ocorre entre
e
50.0000 a 70.0000 mortes porr Raiva Hum
mana ao anoo, principalm
mente nos continentes
c
Asiáttico, Africaano e na Am
mérica Latinna. No Brasiil, de 2000 a 2011, foraam confirmaados
168 ccasos de raiiva humana e 167 (99,44%) óbitos. A região Nordeste
N
representa 55%
% dos
casoss registrado
os entre 1980 e 2008; a região Nortte, no mesm
mo período, 19%; regiãão
Sudeeste, 16%; reegião Centrro-Oeste, 100%; e a regiião Sul, mennos de 1%. Entre 1980 e
20088, cães e gattos foram reesponsáveis por transm
mitir 79% doos casos; moorcegos, 11%
%;
outroos animais, 10%. No anno de 2010, o Brasil reggistrou 3 (trrês) casos humanos
h
da
doennça, 2 (dois)) no Ceará e um no Rioo Grande doo Norte. No ano de 2011, foram
regisstrados 2 (do
ois) casos no
n Estado do
o Maranhãoo. Objetivoss: Realizar o estudo
epideemiológico da Raiva noo Ceara no período de 2000 a 2011, visando realizar
r
açõões
educativas e conntrolar os faatores de risco relacionaados à doen
nça, preveniindo a
a
descritiva
ocorrrência de caasos humanos. Materiaal e Métodoos: Foi realiizada uma analise
das 114 fichas dee Investigaçãão Epidemiiológica de Raiva
R
do SIINAN (Sistema de
Inforrmação de Agravos
A
de Notificaçãoo) e dos daddos provenieentes do settor de estatísstica
do N
Núcleo de Co
ontrole de Vetores
V
da Secretaria
S
dda Saúde do Ceara. Ressultados: Fooram
confi
firmados, noo período dee 2000 a 2011, 15 óbitoos humanos por raiva, sendo
s
12 (80%)
ocasiionados porr agressões de cães e 033 (20%) porr sagui. Os casos
c
foram
m diagnosticcados
em 10 municípioos do Estaddo: Fortalezaa, Caucaia, Maracanaú
ú, Camocim,, Pindoretam
ma,
d Curu, Ipuu e Chaval. O maior nú
úmero de caasos foi
Umirrim, Tururuu, São Luís do
confi
firmado em Fortaleza
F
(qquatro casoss; 27%), segguido de Caaucaia (três casos; 20%
%). Os
outroos municípioos registraraam um casoo (6,66%) caada. O maioor número de
d casos porr ano
foi coonfirmado em
e 2003, seendo sete (446,6%), todoos transmitiidos por cãees. Conclusão:
O cão, dentro daa série históórica analisaada, ainda reepresenta o principal trransmissor de
d
m, principalm
mente até o ano de 20033. A partir de
d 2003, hoouve
raivaa ao homem
intennsificação daas ações de vigilância e controle da
d raiva urbana e uma mudança
m
noo
perfil epidemiollógico, na quual o sagüi passou a seer o principaal transmisssor, visto quue
h
dee 2005 a 20011, 3 (75%) foram proovenientes dde
dos qquatro (quattro) óbitos humanos,
agresssões provoocadas pelo mesmo. En
ntretanto, em
m 2010 foi registrado
r
um
m óbito hum
mano
proveeniente da agressão
a
por cão, evideenciando a necessidade
n
e de analisarr ações
desennvolvidas no
n ciclo urbaano para evitar a ocorrêência de nov
vos casos.
E-maail: [email protected]
PT.0098
DEMIOLO
OGICAL ASPECT
A
OF
F RABIES IN THE REGION
R
OF VALE DO
D
EPID
PAR
RAIBA DUR
RING THE
E PERIOD
D OF 1992 T
TO 2011.
Souzza MCAM1, Torres Junnior MD2, Lombardi
L
Feerreira PA3, Nassar AFC4, Villadobbos
5
6
7
7 1
EMC
C , Ferrari JJ
J , Lara MC
CCSH , Cunnha EMS - Polo APT
TA Vale do Paraiba 2
Labooratorio de Sanidade
S
Annimal, Seccretaria da Agricultura
A
e Abastecim
mento Escriitorio de Deefesa Agroppecuaria, 3Cooperativa
C
de Lacticin
nio Médio Vale
V Paraibaa,
4
5
Instiituto Biolog
gico - Laborratorio de Bacterioses,
B
Instituto Biologico
B
- Laboratorio
L
o de
6
Raivva e Encefalites virais, Secretaria Municipal
M
d Saude dee Jacarei - SP,
de
S 7Institutoo
Biolóógico - Laboratorio de Raiva e Enncefalites Viirais
Vale do Parraiba, situatted in the eaast of Sao Paaulo State, is
i a region where
w
the
The V
anim
mal rabies occcurs, in its endemic foorm, for deccades. The control
c
meassurements
adoppted, mainlyy the vaccinees in animaals and the systematic population
p
c
control
of thhe
hemaatophagous bats from the
t species Desmodus
D
r
rotudus,
havve not achieeved the
expected successs to impedee and limit the
t infectionn cases of thhe virus. Thhis can be
explaained by thee fact that thhe region haas favorablee environmeental condittions for
perpeetuation of shelters andd abundant food
f
for thee maintenan
nce of hemaatophagous bat.
b
The oobjective off this study is to report the occurreence of rabies during th
he period off
19922 to 2011 in suspected samples
s
from 41 cities from metroopolitan reg
gion of Valee do
Paraiiba - SP. 54432 sampless were analy
yzed by the Direct Imm
munofluoresscence (DIF)
technnique and intracerebraal inoculatioon of mice. In
I the samp
ples analyzed, 567 in 54432
(10.44%) gave po
ositives resuults being 21.7% (527/22421) from herbivores, 2.4% (23/9972)
from
m chiropterass, 3.7% (6/1162) from other
o
speciess (swine, caaprine, ovinee, primates and
rodennts) and 0.6
6% (11/18777) from dog
gs and cats. The obtaineed results in
ndicate that the
sampples of herbivores show
wed the greaater rate of ppositive resuults when compared
c
too the
sampples of the other
o
animall’s species analyzed,
a
w
which
indicaates the endeemicity of the
t
diseaase in these species. Baased in this observationn, it becomees importantt and essenttial to
the reegion the coontinuous adoption andd the improvvement of epidemiolog
e
gical supervvision
systeems, such ass: the encouurage of the education programs
p
annd populatio
on awareness,
mainnly in the ruural areas, thhe realizatioon of hematoophagous bat control, and
a the
vacciination of suusceptible animals.
a
Forr an effectivve performaance and succcess, thesee
actioons must be conducted with the parrticipation of
o the State institutionss of
envirronment, heealthy and agriculture.
a
Bibliiographical citation
TISTA, H. B.
B de C. R. Raiva:
R
uma breve revissão. Acta. Sci.
S Vet. v.335, n.2, p.1225,
BAT
20077.
DEL
LPIETRO, H.
H A. Aspecctos econôm
micos y sanittários del prroblema de la rabia
parallítica y de laa agresión del
d vampiro. In: SEM. IINTER. DA
A RAIVA, 2000,
2
São Paulo,
P
SP. P
Programas e Resumos: São Paulo: De Paula Print
P
Artes Gráficas,
G
20
000. v.1.
DEA
AN, D. J. Thhe fluoresceent antibodyy test. In: WORLD
W
HEA
ALTH ORG
GANIZATION.
Laboor. Techn. in
n Rabies. 4 ed. Genevaa: World Heealth Organiization,19966. p.88
GOD
DWASSER,, R. A. Fluoorescent antiibody stainiing of streett and fixed rabies
r
víruss
antgeens. Proc. of the Soc. Exp.
E Biol. Med.,
M
v.98, p.219,
p
19588.
KOP
PROWSKI, H. The mouuse inoculattion test. Inn: MESLIN, F. X.; KAP
PLAN, M. M.
M H.
Laboor. Techn. in
n Rabies. 4 ed. Genevaa: World Heealth Organiization, 19996. p.80.
KOT
TAIT, I. Co
ontrole da raaiva dos herrbívoros. Man.l Técn. do
d Inst. Pastteur, n.9, p.8,
20100.
RUP
PPRECHT, C.
C E. Rabiees re-examinned. Lanc. Infec.
I
Dise... v.2, n.6, p.327,
p
2002.
SOU
UZA, M. C. A. M. Epiddemiology of
o rabies: bioological andd serologicaal aspects off
rabiees in vampirre bats Desm
modus rotun
ndus(E.Geooffroy) captuured in Valee do Paraibaa,
Southheastern reggion of Brazzil. Arq. do Inst. Biol., São Paulo, v.64, n.2, p.91,
p
1997.
SUG
GAY, W. Isoolamento doo vírus da raaiva de morrcegos hemaatófagos do Estado de São
S
Pauloo, Brasil. Bol. de la Offic. San. Pannam. v.50, n.4,
n p.310, 1966.
1
.
PT.0099
BIA: DIAG
GNOSTICO
O y CARAC
CTERIZAC
CION ANT
TIGENICA
A
RAB
Russso SE1, Novvaro LP2, Buurkhard IR2, Centuriónn G2, Luque AA2 - 1SEN
NASA - Rabbia,
DLA
A -DILAB, 2SENASA - Rabia, DL
LA-DILAB
dades de Peqqueños Aniimales perteenece a la
El Departamentoo de Rabia y Enfermed
ntrol Técnico (DILAB), del Serviccio Nacionall de
Direccción Gral. de Laboratoorios y Con
Saniddad y Calid
dad Agroalim
mentaria (SE
ENASA) quue depende del Ministeerio de
Agriccultura, Gannadería y Peesca de la República
R
A
Argentina.
Desdde el año 20000 integram
mos el Labooratorio Naccional de Reeferencia paara el
Diaggnóstico de Rabia
R
juntoo con el Servvicio de Neuurovirosis del
d INEI-AN
NLIS.
Posteeriormente junto
j
con ell Departameento de Zooonosis y Con
ntrol de Vecctores del
Miniisterio de Saalud de la Nación
N
se coonformó el Centro
C
Naciional de Referencia y
Coorrdinación (C
CNRC), donnde se incorrporó como Laboratorioo Coordinaddor al
Depaartamento de
d Diagnóstiico y Produucción del Innstituto de Zoonosis
Z
Luuis Pasteur
(IZL
LP) que depeende del Miinisterio de Salud del Gobierno
G
dee la Ciudad Autónoma
A
d
de
G.C.A.B.A), y la Red Nacional
N
de Laboratorioos de Diagn
nóstico de Rabia
R
Buennos Aires (G
integgrada por los laboratoriios provinciales y regioonales.
Com
mo Laboratorrio Nacionaal de Refereencia:
Reallizamos y confirmamos el diagnósstico de rabiia por la téccnica estánddar de
inmuunofluoresceencia directta (IFD), el aislamientoo del virus por
p los métoodos clásicoos
(inocculación en ratón o en cultivos
c
celu
ulares) y la caracterizaación antigén
nica mediannte el
panel de 8 anticuuerpos monnoclonales (AcMc),
(
proovistos por el
e C.D.C., Atlanta,
A
Geoorgia,
A.
EUA
Prodducimos reacctivos para uso interno y de los labboratorios de
d la red de diagnósticoo:
Sueroo Antirrábicco de origenn equino co
onjugado conn isotiociannato de fluorresceína, virus
CVS
S (Virus Stanndard de Deesafío) y CR
RN (cerebroo de ratón normal).
n
La mayor
m
parte de las muesstras recibiddas para diagnóstico poor nuestro Laboratorio
L
s
son
de boovinos y de otros animaales de prodducción. Se han diagno
osticado y caracterizadoo la
mayooría de ellass como variante 3 (Vam
mpiro), peroo cada vez más
m frecuenntemente, y
proveenientes de distintos luugares dentrro de la zonaa donde habbitan los vam
mpiros
(Desmodus rotuundus), se haan caracteriizado varianntes donde el
e AcMc 15 ha dado
posittivo.
Hem
mos aislado en
e más de 40
4 bovinos esta
e variantee así como también
t
en caprinos y
equinnos.
Ello ha dado lug
gar a la derivación de laas muestrass para un poosterior anállisis molecuular y
A
integ
grante tambbién del Labboratorio Naacional de
filoggenético porr parte del ANLIS,
DC.
Referencia, con la colaboraación del CD
PT.1100
VA EM HE
ERBÍVOR
ROS NO MU
UNICÍPIO
O DE GRAN
NJA - CEA
ARÁ
RAIV
VAS
SCONCELO
OS DC1, DU
UARTE NFH2, OLIVE
EIRA FB2, MAIA
M
JUNIIOR JE3,
MOR
RAIS NB3 - 1ADAGRII - ADAGRI, 2SECRET
TARIA EST
TADUAL DE
D SAUDE
E DO
CEA
ARÁ - NUV
VET, 3ADAG
GRI
a
nose caracteerizada por uma encefaalite aguda e quase sem
mpre
A Raaiva é uma antropozoon
fatal,, causada poor uma popuulação de vírus da fam
mília Rhabdooviridae, gên
nero Lyssavvirus
(ACH
HA; SZYFR
RES, 2003). A principaal forma de transmissão
o desta zoon
nose ocorree nas
agresssões aos annimais susceeptíveis, atrravés da inooculação do agente etioológico presente
na saaliva dos annimais infecttados (VER
RONESI & FOCACCIA
F
A, 1997). No
N municípioo de
Grannja, localizado na regiãoo norte do estado
e
do Ceará distantte a 353,2 Km
K de Fortaaleza,
ocorrreu a notificcação em noovembro dee 2011 sobree mortes de animais co
om sinais
clíniccos de enceefalite. Equippe da Secreetaria Estaduual de Saúdde e Agênciaa de Defesaa
Agroopecuária. Nas
N proprieddades foram
m identificaddos animaiss expostos como: equíddeos,
bovinnos, ovinos, caprinos e suínos. Os casos ocorrreram em 03
0 (três) propriedades nno
distriito de Parazzinho, nas loocalidades de
d Lagoa Reedonda, proopriedade 01 (morte de 06
bovinnos), propriiedade 02 Lagoa
L
redonda (morte dde 01 bovino
o com confiirmação
laborratorial), proopriedade 03
0 Lagoa Coomprida ou “Carro Queebrado” (moorte de 05
bovinnos e 01 asiinino). Em uma
u cacimbba foi identiificado colô
ônia de morccegos
hemaatófagos da espécie Deesmodus rottundus, posssível responnsável da traansmissão da
d
doennça para os animais.
a
Foi realizada captura
c
na referida
r
caccimba próxim
ma na
Locaalidade de “Terra Dura da Sucupira ou Carro Quebrado”.. A colônia possuía
aproxximadamennte 100 morccegos, foram
m capturadoos 11, a pasta vampiriccida foi apliccada
em 005 morcegoss e os outros 06 foram enviados paara diagnósttico laboratorial da Raiiva,
dos qquais 01 foi confirmadoo positivo para
p raiva attravés de Im
munofluoresscência Direeta e
Provva Biológicaa. Foram adootados os seeguintes enccaminhamentos: educaação em saúúde
atravvés de palesttras com as equipes de saúde da faamília e peqquenos criaddores de
bovinnos. Os criaadores foram
m orientadoos para adoçção de práticcas profilátiicas na
imunnização do rebanho,
r
bem
m como, o uso
u de pastaas vampiriccida nos anim
mais agrediidos,
e nottificação im
mediata dos animais
a
susspeitos nos escritórios
e
da
d Adagri, além
a
de
encam
minhamento a Unidadee de Saúde de pessoas expostas aoo risco de accometimentto da
Raivva por contatto com anim
mais suspeittos. Os casoos de raiva ocorreram
o
n proprieddades
nas
em qque os produ
utores não realizavam
r
v
vacinação
ccontra a raivva, foi de funndamental
impoortância as ações
a
integrradas entre a Saúde e Agricultura.
A
PT.1101
GILÂNCIA AMBIENT
TAL DIAN
NTE DO SU
URTO DE RAIVA
R
SIL
LVESTRE NO
VIG
MUN
NICÍPIO DE
D SÃO BE
ENEDITO NO ESTAD
DO DO CE
EARÁ EM 2010
Silvaa MVS1, Mooura FBP1 - 1Secretariaa da Saúde do
d Estado do
d Ceará - 1ª Coordenaddoria
Regional de Saú
úde de Fortaaleza
RODUÇÃO
O – Casos reelatados de raiva
r
em raaposas e trannsmissão em
m humano é
INTR
relataada nos estaados do Ceaará¹, Paraíbaa², Pernambbuco, Bahia e Minas Geerais3. Na Serra
da Ibbiapaba nos meses de maio
m e junhoo de 2010 no
n município
o de São Beenedito no
Estaddo do Cearáá ocorreram
m seis casos de raiva em
m raposa guaaxinim nas localidades
l
de
Muriicituba, Corrquinho, Sannta Luzia e em Barroqquinha. OBJJETIVO – Diagnosticar
D
ra
situaação populaccional dos animais
a
silvvestres em áreas
á
com diiferentes graaus de alterração
(morradias, planttações) onde ocorrem agressões,
a
aanalisando fatores
f
de risco da raivaa.
MAT
TERIAL E MÉTODO
M
– Foi utilizaado GPS naa marcação dos locais onde
o
ocorrerram
ataquues para anaalisar as áreeas de vida de
d cada espéécie animall agressora. Vacinação de
bloquueio em cãees e gatos e trabalho edducativo foram realizaddos. Armadiilhas para
captuurar raposass foram insttaladas paraa pesquisa de circulaçãoo do vírus ráábico dentroo dos
limittes marcado
os por GPS. RESULTA
ADOS - A siituação epiddemiológicaa da raiva em
m
São B
Benedito fooi grave, devvido o grand
de número de
d ataques em
e seres huumanos caussados
por rraposas e guuaxinins. Diiagnósticos laboratoriaiis positivos de diferenttes animais
silvestres de com
mprovaram a circulação
o viral na reegião. A utiilização antrrópica na Seerra
m alterado o meio ambieente para ateender os obj
bjetivos com
m fins
da Ibbiapaba tem
econnômicos, o que
q comprovva a adaptaçção dos cannídeos em árreas alterad
das pelo hom
mem,
crianndo legítimo
os corredorees ecológicoos, os quais possibilitam
m às espéciees silvestress
transsitarem livreemente paraa as áreas haabitadas porr pessoas. Os
O animais silvestres
s
esstão
com o potenciall de dispersãão e colonizzação limitaadas devido à fragmenttação de habbitat
mentação de habitat as doenças
d
emeergentes e re-emergent
r
tes
na zoona rural. Com a fragm
que jjá não possu
uíam importtância epideemiológica, aparecem em
e surtos, epidemias
e
o
ou
epizoootias numaa populaçãoo e região co
omo em Sãoo Benedito. CONCLUS
SÃO - A raiiva
aindaa é um prob
blema imbattível em mu
uitos lugaress e um consstante desafiio para os
serviiços de saúd
de pública, porém,
p
um fato permannece óbvio, o de que a erradicaçãoo da
raivaa humana deepende, fundamentalmeente, do conntrole da raiiva animal, inclusive coom a
utilizzação da viggilância ambbiental.
Agraadecimentoss
Aos moradores do município de São Benedito,
B
peela disponibilidade em ceder seu teempo
A técnico
os do Núcleo de Controole de
e connhecimento para a realiização dessee trabalho. Aos
Vetoores - Secrettaria da Saúúde do Estaddo do Cearáá, que acomp
panharam o primeiro autor
a
em toodas as ativ
vidades. Aoss técnicos e demais serrvidores da 13ª Coordennadoria
Regional de Saú
úde de Tiangguá. Aos seervidores daa Secretaria Municipal de Saúde dee São
Beneedito.
Referências Bib
bliográficas
F
C.E
E.A.A.; Soussa, F.S. (19889). Raiva em
e animais silvestres no
n
1. Baarros, J.S.; Freitas,
Estaddo do Cearáá particularm
mente na rapposa (Dusiccyon vetulu
us). Zoon. Rev.
R Int., 1, 99
13.
B. (2004). Epidemiolog
E
gia da raivaa: caracterizzação de víruus isolados de
2. Goomes, A.A.B
anim
mais doméstiicos e silvesstres do sem
miárido paraaibano da reegião de Pattos, Nordestte do
Brasiil. São Paullo, Brasil, 107p. (Thesiis, Faculdadde de Mediccina Veterinnária e
Zoottecnia, Univversidade dee São Paulo).
3. Arraújo, F.A.A
A. (2002). Raiva
R
humanna no Brasil: 1992-20001. Belo Ho
orizonte, Braasil,
90p. (M.Sc. Dissertation, Escola
E
de Veeterinária, Universidad
U
de Federal de Minas
Geraais).
PT.1102
SER
ROPREVAL
LENCE OF
F RABIES VIRUS IN
N WILD CA
ARNIVORE
ES
1
1
2
2
Peress MG , Baccchiega TS , Martorelli LFA , Kataaoka APAG
G , Vicente AF1,
1
1
1
1 1
Apolllinario CM
M , Allendorf SD , Antuunes JMAP , Megid J - UNESP - School of
Veterinary Meddicine and Animal
A
Sciennce - Deparrtament of Veterinary
V
Hygiene
H
andd
B
P, Brasil, 2Zoonosis
Z
Coontrol Centeer - Rabies Laboratory
L
Publiic Health- Botucatu-SP
- São
Pauloo-SP, Brasil
All m
mammals arre susceptible to rabies virus, especially thosee of the ordeer Carnivoraa and
Chirooptera. In th
he wild cyccle animals that
t act as rreservoirs vaary accordinng to regionn. In
Latinn America, the
t bats act as main resservoirs of tthe virus in the wild cyycle, howeveer the
rabiees virus has been isolateed from oth
her wildlife species such Cerdocyoon thous (Crrabeatinng Fox), Proocyon cancrrivorus (Craab-eating Raaccoon), annd Callithrixx jacchus
jacchhus (Comm
mon marmose). In this context, this work aimed to analyzee the
seropprevalence for
f rabies viirus in 9 carrnivores capptured in areas of largee and small
foresst fragmentss in the ruraal area in cenntral west reegion of Sãão Paulo statte. Were
captuured 4 Nasuua nasua (C
Coati), 4 Cerrdocyon thoous (Crab-eaating Fox) and
a a Leopaardus
parddalis (Ocelott), with the use of Tom
mahawk trapps containing chicken leeg as bait. They
T
weree anesthetizeed with Zoleetil® (Tiletamine and Zolazepam)
Z
) and had bllood samplees
colleected by jug
gular vein puuncture. The blood wass placed in collecting
c
tuube withoutt
EDT
TA, and seruum in microotube. Sera samples
s
were stored at -20 ° C andd than were
testedd for neutraalization in cell
c culture for the deteermination of
o neutralizing antiboddies to
rabiees virus by rapid
r
methood focus inhhibition of flluorescencee (RIFFT). In
I Cerdocyoon
thouss, that is connsidered as an importannt reservoirr for a particcular rabies virus variannt in
Brazzil, 75% shoowed anti-raabies antibodies titers above
a
0,5 UI/mL. Thesee data suggests
that rabies
r
virus is presenteed in the anaalyzed regioon and someehow these wildlife speecies
get inn contact with rabies virus. It’s im
mportant to note
n that theese animals were captuured
in a rregion of close contact with other humans andd domestic animals, reppresenting a
possiible epidem
miological risk.
PT.1103
LORACIÓN
N DE UN TEST
T
RÁP
PIDO DE IN
NMUNOCR
ROMATOGRAFÍA EN
E
VAL
PLA
ACA PARA
A LA DETE
ECCIÓN DE
D RABIA EN
E MUESTRAS FRE
ESCAS Y EN
E
AVA
ANZADA DESCOMP
D
POSICIÓN DE ARGE
ENTINA
Guryy Dohmen F1, Barcos O2, Cisterna D3, Cicuttiin G1, Menaa Segura C1, Beltrán F4 1
Instiituto de Zoo
onosis Luis Pasteur - Diagnóstico,
D
, 2Laboratorrio Colón, 3INEI-ANLIIS
Dr. Carlos
C
Malbbrán - Serviccio de Neurrovirosis, 4Innstituto de Zoonosis
Z
Lu
uis Pasteur,,
Buennos Aires, Argentina
A
En ell presente estudio se evvaluó un tesst de inmunoocromatogrrafía (RIDT)) para rabiaa
(Rabbies Ag Testt Kit, Bionoote Inc., Korrea) de las principales
p
c
cepas
viralees circulantees
en laa República Argentina, en muestraas frescas quue incluyero
on a los reseervorios máás
frecuuentes, como así tambiéén en cerebrros en avannzado estadoo de descom
mposición baajo
conddiciones de laboratorio.
l
Para ello see realizó el diagnósticoo sobre un lo
ote de 50 dee
muesstras clínicaas frescas poor las técniccas de IFD, RIDT y EB
BRL entre loos años 2011 y
20122. Para evaluuar las cepaas de Lasiurrus sp (V6) y terrestre silvestre (V
V2) se utilizaaron
aislam
mientos en cerebro de ratón, con la
l intenciónn de incluir en
e el ensayoo las varianttes
de m
mayor circulaación del paaís. Posterioormente se ddescongelarron 5 cerebrros guardaddos
a -700°C de un brrote de rabia canina (V
V1) 2002-20008. La senssibilidad dell test se valooró
con uuna cepa dee virus fijo utilizado
u
en la producciión de vacu
una CRL, prreviamente
titulaado en ratonnes de 21 días
d y RT-PC
CR en form
ma paralela. La
L concordaancia del RIIDT
con lla IFD y EB
BRL fue del 100% y puudo detectarrse hasta la dilución
d
10-4
del virus fijo,
f
que ccorrespondiió a 100 DL
L50 en 0,03m
ml. Existen regiones dee explotacióón ganaderaa en
el noorte del país con rabia paresiante
p
y otras con aantecedentess de rabia de
d ciclo terreestre
de diifícil acceso
o cerca de laa frontera coon Bolivia een donde el veterinario del municiipio
podríía realizar un
u primer diiagnóstico diferencial
d
a fin de dar parte a las autoridades
a
s
sanittarias y así aplicar
a
rápiddamente lass acciones de
d profilaxiss corresponddientes a loss
morddidos y lueg
go remitir laa muestra a los laboratoorios de refeerencia paraa confirmación y
caraccterización de
d la cepa responsable
r
del brote. En
E nuestra conclusión
c
el
e RIDT es de
uso muy
m simple y podría seer consideraado de sumaa utilidad enn muestras post-mortem
p
m
bien conservadaas o descom
mpuestas quee hayan com
mpletado el período de estado de laa
enferrmedad.
PT.1104
E OF PROP
PIDIUM IO
ODIDE LIK
KE A CELU
ULAR CON
NTRAST STAINING
S
G IN
USE
THE
E DIRECT FLUORES
SCENT AN
NTIBODY (DFA) TES
ST FOR TH
HE RABIE
ES
VIRU
USES DIA
AGNOSTIC
C.
Igualla-Vidales M1 - 1InDRE
E - Virologia
RODUCTIION.
INTR
m importtant zoonosis worldwid
de and repreesents a seriious
Rabies remains one of the most
y countries.. Into the diiagnostic toools the direcct fluoresceent antibodyy
probllem in many
(DFA
A) test is a fast
f and sennsitive method to diagnnosis rabies infection inn animals annd in
humaans (1, 3). The
T test is based
b
on miccroscopic exxamination, under ultraaviolet lightt,
imprressions, sm
mear sampless of tissue of
o the hippoocampus (flaagpole of Ammon),
A
thee
cerebbellum and the medullaa or tissue sections; anttibodies (IgG
G) used in the
t conjugaate
monooclonal allo
ow specific and uniform
m coloring without
w
inteerference fro
om the fundd.
In ceell biology, propidium
p
i
iodide
is used as a dye contrast thaat differentiiates the nuccleus
from
m the cytoplaasm (intercaalates in DN
NA) (4), theere is evidennces of its use in stains for
identtification off Herpes Sim
mplex Typee 1 (5).
AL:
GOA
Impllement the use
u of propidium iodidee in the diaggnosis of the rabies viru
us by directt
fluorrescent antibbody (DFA)) test
TERIALS AND
A
MET
THODS
MAT
70 Saamples werre selected: 50 brains: (25
( positive samples off varying deegrees of
posittivity (1 + too 4 +) and 225 samples negative)
n
annd 20 isolattes of the rab
abies virus inn
mousse neuroblaastoma cell (15
( positivee samples off varying deegrees of poositivity (1 + to 4
+) annd 5 negativve samples) of the rabiees laboratorry samples bank.
b
The direct
d
fluorescent
antibbody (DFA)) test was appplied as inddicated by the
t supplier Anti-Rabiees Monoclonnal
Globbulin (IDF, FUJIREBIO
F
O.) (DIAGN
NOSTICS, IInc.); to the end of the test,
t
added 20
µL oof the propiddium iodidee solution too a final conncentration of
o 0.3 µg/m
mL (in each
imprrint of brain or well celll culture) annd it was incubated at room
r
tempeerature for 5
minuutes, it was eliminated
e
b rinsing with
by
w PBS pH
H 7.4, let air dry and was
w added a drop
of buuffered glyccerin pH 8.44; reading was
w conducteed on an maagnificationn lens
fluorrescence microscope 100 X and 40 X. The readding was evvaluated for 4 people; 2
experts in rabiess diagnosis and 2 peoplle in the leaarning proceess.
SULTS AND
D CONCLUSION
RES
Of thhe 50 samplles brains were
w obtained the follow
wing results; 25 positiv
ve samples of
o
varyiing degrees of positivitty (1 + to 4 +) and 25 negative
n
sam
mples and all 20 isolatees of
the raabies virus of in mousee neuroblasttoma cell (115 positive samples
s
of varying
v
deggrees
of poositivity (1 + to 4 +) annd 5 negative samples; all samples coincided with
w the
previiously reporrted results.. All personnnel involveed in readingg coincided in the ease of
identtify the nuclleus of the cell
c in the brain
b
imprinnts as well as in cell cullture slides.
Becaause of the propidium
p
iodide is useed to staininng the DNA
A, were watcched the ceells
nucleeus of red-o
orange colorr; likewise facilitated
f
thhe identificaation of infeection in thee
cytopplasm of thee cell (in poositive casess) by the fluuorescent appple green contrast
c
of thhe
fluorrescein isoth
hiocyanate (FITC)
(
fluoorochrome, tthat is conju
ugated to thhe monoclonnal
antibbodies targeting the prootein of rabiies virus. Thherefore, it is
i concluded
d that the usse of
propiidium iodid
de does not interfere
i
in the DFA teest it is conccluded that the
t use of
propiidium iodid
de does not interfere
i
in the DFA teest, since alll results werre identical to
the reeported in the
t samples tested befoore; the use oof propidium
m iodide is helpful mainly
for teechnical staaff who do not
n have expperience in identifying cells in impprints and cell
c
cultuure; the use of propidium iodide allows a conttrast which facilitates thhe identification
of the fluorescennce of the raabies virus.
PT.1105
ALUATION
N OF BAIT
T FLAVOR
RS FOR PO
OTENTIAL
L USE IN ORAL
O
RAB
BIES
EVA
VAC
CCINE DEL
LIVERY TO
T FERAL
L DOGS (C
Canis familiiaris)
Bendder SC1, Bennder P2, Haausig K3, Beerentsen A4, Bergman DL
D 5, VerCaauteren K4 1
2
Navvajo Nation - Navajo Nation
N
Veterrinary Progrram, USDA
A - APHIS-W
Wildlife
3
Serviices, Phoen
nix, Arizonaa, USA, US
SDA - APHIS-Wildlifee Services, Phoenix,
P
Arizoona, 4USDA
A - APHIS- Wildlife Seervices- Naational Wilddlife Researcch Center, Fort
F
Colliins, Coloraddo, 5USDA - APHIS Wildlife
W
Servvices, Phoennix, Arizonaa
d
doogs on triball lands in thhe United Sttates
: It iss estimated that less thaan 20% of domestic
are vvaccinated against
a
rabiees. One pottential methhod to increaase vaccinattion rates m
may
be thhe distributioon of oral raabies vaccinnes (ORV). ONRAB® is the prim
mary ORV baait
used in Canada to vaccinatee striped sku
unks and raaccoons. Reesearch has suggested the
t
n
annimals that may ingest these baits are feral doomestic
mostt common non-target
®
dogs. To furtheer investigatte the potenttial use of ONRAB
O
ORV
O
baits to
o vaccinate feral
a
rabiees on tribal lands, we performed
p
a flavor prefference studdy to
domeestic dogs against
increease ORV acceptance. Seven bait flavors (bacon, cheesee, dog food, hazelnut,
marsshmallow, peanut
p
butteer and sardinne) were offfered in pairrs to 13 dom
mestic
dogs. Each dog
g was offered all possibble combinaations of baiit pairs overr a period off ten
mes each baitt was consuumed
days, with each bait offeredd six times. The proportion of tim
ong dogs weere
first bby individuual dogs wass calculatedd and compaarisons amo
condducted. Dogg food was selected
s
firsst 56% of thhe time, andd more frequuently than all
otherr bait types (F = 13.09, P = 0.00055) although bacon was close seconnd at
54%. Marshmaallow was seelected first during 14%
% of offerinngs and exhiibited the leeast
m
extensive evaluatiion is
prefeerence amonng all bait tyypes (F = 222.46, P < 0..0001). A more
plannned, prelimiinarily; dogg food or baccon flavored ORV baitts appear to be good
choicces for optim
mizing bait ingestion by
b feral dom
mestic dogs.
PT.1106
OFILAXIA DA RAIVA HUMAN
NA: COMP
PARAÇÃO
O DOS ATE
ENDIMENTOS
PRO
MÉD
DICOS NO
OTIFICADO
OS EM 200
01 E 2011 NO
N ESTAD
DO DE SÃO
O PAULO,,
BRA
ASIL
Omooto TM1, Taakaoka NY1 - 1Instituto Pasteur
p
A raiiva, doença infecciosa quase semppre fatal, pode ser preveenida com profilaxia
especcífica. A pro
ofilaxia da raiva
r
human
na pode serr realizada pré
p ou pós-eexposição aoo
víruss da raiva, utilizando-se
u
e vacina conntra raiva huumana e sorro antirrábicco de acorddo
com recomendaações especííficas. Os im
munobiólogiicos (vacinaa de cultivo celular e sooro
mente nos serviços de saúde
s
da red
de pública do
d
antirrrábico) estãão disponíveeis gratuitam
Brasiil. Os acidentes com annimais mam
míferos conssiderados dee risco para a transmisssão
da raaiva são monnitorados peelo Sistemaa de Vigilânncia Epidem
miológica, attravés de
notifficações doss atendimenntos realizaddos nas uniddades de saúúde no Sisteema de
Inforrmação de Agravos
A
de Notificaçãoo (SINAN). Nos últimoos onze anos a média annual
de attendimentoss realizados pelas unidaades de saúdde do Estad
do de São Paaulo foi de
1074491 notificaçções, com a taxa de 27 atendimenttos por 10000 habitantes; sendo quue
41% receberam imunoprofiilaxia contraa raiva e a taxa
t
de aban
ndono de 133,8%. No anno de
20011 foi notificaado o últimoo caso de raaiva humanaa no estado de São Pauulo, por variiante
de m
morcego, cujo animal aggressor foi um
u gato. Em
m 2001 foraam notificaddos 117599
atenddimentos médicos,
m
com
m 99,9% de casos de póós-exposição, de 95,8%
% dos 645
muniicípios, taxaa de profilaxxia humanaa de 35,1% e taxa de ab
bandono de 12%. Quannto às
espéccies animais envolvidaas nos atenddimentos nottificados, oss cães e gato
os foram oss
princcipais agressores responnsáveis pelaa procura doo atendimennto médico,, respondenddo
por 886% do totaal. No ano de
d 2011, daddos preliminnares revelaam que de 117486
atenddimentos nootificados, 98%
9
foram de
d pós-expoosição, em 98,3%
9
dos municípios,
m
, com
taxass de profilax
xia humana de 49,3% e 14,2% de abandono. Os
O cães e gaatos
correesponderam
m a 96% do total
t
de espéécies. Apóss o período de
d 11 anos, apesar da
discrreta diminuiição no núm
mero de aten
ndimentos notificados,
n
chama atennção o aumeento
dos ccasos em quue o morceggo foi o respponsável pella profilaxiaa em 2011; e também a
diminnuição dos registros seem especificcar a espéciee animal ennvolvida. Peersistem aindda
inconnsistências nos
n registroos dos bancoos de dadoss, gerados elletronicameente, dificulttando
uma análise rápiida e aprofuundada dos mesmos.
PT.1108
ON MOLE
ECULES ON
O RABIES
S
EFFECTS OF AMPHIBIIAN SKIN SECRETIO
US INFEC
CTION IN MAMMAL
M
LIAN CELL
LS
VIRU
1
VIGE
ERELLI H1,2
, Sciani JM
M1, Jared C3, Antoniazzzi MM3, Caaporale GM
MM2, Silva
2
1 1
ACR
R , Pimenta DC - Instituto Butanttan - Laboraatório de Biioquímica e Biofísica,
2
Instiituto Pasteuur - Laboratóório de Soroologia, 3Instituto Butan
ntan - Laborratório de
Bioloogia Celularr
Rabies is an acu
ute infectiouus disease caaused by a virus
v
that afffects the ceentral nervoous
m
o infectionn is associateed to the ceell penetratio
of
on via the
systeem, which mechanism
nicottinic acetylccholine receeptor. The genus
g
Bufo, recently split into Bufoo in the Oldd
Worlld and Rhinnella in the New
N Worldd, contain a large
l
number of alkalo
oids and sterroids
in theeir skin secrretion. The aim of this study was to
t assay moolecules extrracted from
m the
skin of amphibiaans as possiible interferring agents iin the proceess of infecttion of the rabies
r
B
(Rhineella jimi) skkin secretion
ns were colllected throuugh
viruss in mammaalian cells. Bufo
mechhanical stim
mulation. A liquid-liqui
l
d partition (H
( 2O-CH2Cl
C 2) was perrformed andd the
two rresulting so
olutions werre purified by
b RP-HPLC
C, in a C18 column. Sttructural
charaacterizationn was perforrmed by maass spectrom
metry. Cytottoxic tests of
o the isolateed
comppounds werre performedd over BHK
K-21 cells. Briefly,
B
96-w
well microttiter plates
contaaining the cells
c
were inncubated forr 24h in the media conttaining diffeerent dilutioons
of the purified molecules.
m
F the virollogic test, fixed
For
fi
strain PV
P (Pasteurr Virus) was
w both
used on fluoresccence inhibiition test annd fluorescennt foci inhibbition test, with
nd time courrse treatmen
nt of the cellls with the virus and thhe fractions.
simuultaneous an
Sixteeen fractionns were obtaained by RP
P-HPLC. Thhe cytotoxic tests revealled that 9
fractiions were tooxic to BHK
K-21cells. On
O the virollogic test, frraction 2 shoowed a lastiing
effecct, independdent from thhe simultaneeous and tim
me course trreatments in
n both tests.
Masss spectromeetric analysees showed th
hat this fracction containns a steroid
d named
helleebrigenin. Frraction 14 was
w able to reduce rabiies virus inffection in booth tests,
appaarently show
wing compettition effectts. Mass speectrometric analyses sh
howed that this
fractiion containss two indolee alkaloids, N`,N`-dim
methyl 5-hyd
droxytryptam
mione
(bufootenine) andd N`,N`,N`--trimethyl 5-hydroxytryyptamine (5
5-HTQ), whhich are currrently
undeergoing puriification. Thhe two indivvidual compponents willl be retestedd for biological
activvity in orderr to evaluatee which retaains the biollogical effecct so that more
m
in depthh
assayys can be peerformed.
ESP, CNPqq
Finanncial Suppoort: Institutoo Butantan, Instituto Paasteur, FAPE
PT.1109
O DE YODU
URO DE PROPIDIO
P
COMO CO
OLORANT
TE DE CO
ONTRASTE
E
USO
CEL
LULAR EN
N LA TECN
NICA DE INMUNOFLUORESC
CENCIA DIRECTA
D
PAR
RA EL DIA
AGNOSTIC
CO DEL VIIRUS DE LA
L RABIA
Igualla-Vidales M1 - 1InDRE
E - Virologia
ndo, y representa
La raabia continúúa siendo unna de las zoonosis más importantes en el mun
un prroblema serrio en muchhos países. Dentro
D
de laas herramienntas de diag
gnóstico la
técniica de inmun
nofluoresceencia directaa con anticuuerpos fluorrescentes (IF
FD) constituuye
un m
método rápiddo y sensible para diagnnosticar la infección
i
ráábica en anim
males y en seres
humaanos (1, 3). La prueba se basa en el
e examen microscópic
m
co, bajo luz ultravioletaa, de
imprresiones, frootis de muesstras de tejiddo del hipoccampo (astaa de Ammonn), del cerebbelo
y la medula
m
o seecciones de tejido; los anticuerpos
a
monoclonaales (IgG) utilizados
u
enn el
conjuugado perm
miten la colooración espeecífica y uniiforme sin interferenciaa del fondo..
En biología celu
ular el yoduro de propiddio se utilizza como colorante de coontraste quee
mite diferencciar el núcleeo celular deel citoplasm
ma (se interccala en el DNA) (4), exxisten
perm
evideencias de su
u utilizaciónn en tincionees para idenntificación de
d Herpes Simple
S
tipo 1(5).
OBJJETIVO:
Impllementar el uso
u de Yodduro de Proppidio dentroo del diagnóóstico del virrus de la rabbia
por IInmunofluorrescencia diirecta.
MAT
TERIALES
S Y METO
ODOS
n 70 muestrras: 50 encééfalos: (25 muestras
m
positivas de diferentes
d
grrados
Se seeleccionaron
de poositividad (1
1+ a 4+) y 25
2 muestrass negativas) y 20 aislam
mientos del virus de la rabia
r
en céélula de mieeloblastomaa de ratón (115 muestras positivas de
d diferentess grados de
posittividad (1+ a 4+) y 5 muestras
m
neg
gativas) del banco de muestras
m
del laboratorioo de
rabiaa. Se aplico la técnica de
d inmunoflluorescenciaa directa (IF
FD) según lo
l indica el
proveeedor Anti--Rabies Monnoclonal Gllobulin (FD
DI, FUJIREB
BIO. DIAGN
NÓSTICS,
Inc.), al terminoo de la técnica, se adiciiono 20 µL de la soluciión de yoduuro de propiidio a
mpronta de encéfalo
e
o pozo
p
con céllulas
una cconcentracióón final de 0.3 µg/mL (en cada im
de cuultivo )y se incubo a tem
mperatura ambiente
a
duurante 5 min
nutos, se eliimino lavanndo
con P
PBS pH 7.44 , se dejo seecar al aire y se agregoo una gota de glicerina tamponada
t
pH
8.4 ; se realizo la lectura enn un microsccopio de epifluorescencia con el objetivo
o
10X
Xy
gnóstico de rabia
r
y2
40X.. Se evaluó la lectura para 4 personnas; 2 experrtos en diag
persoonas en procceso de apreendizaje.
SULTADOS
S Y CONC
CLUSION
RES
De laas 50 muesttras de encééfalo se obtu
uvieron los siguientes resultados;
r
2 muestrass
25
posittivas de difeerentes graddos de positiividad (1+ a 4+) y 25 muestras
m
negativas y dee los
20 aiislamientos del virus dee la rabia enn célula de m
mieloblastooma de ratón
n (15 muesttras
posittivas de difeerentes graddos de positiividad (1+ a 4+) y 5 muuestras negativas; todaas las
muesstras coincid
dieron con los resultaddos anteriorrmente reportados. Toddo el personnal
invollucrado en la
l lectura cooincidió en la facilidadd de identificcar los núcleos celularees,
tantoo en impronttas de encéffalo así com
mo en las lam
minillas de cultivo celuular. Debidoo a
que eel yoduro dee propidio se
s intercala en el DNA,, se observaaron los núccleos de las
célullas de color rojo-naranjja; así mism
mo se facilitto la identifficación de la
l infecciónn en
el cittoplasma ceelular (en los casos posiitivos) por eel contraste de color veerde manzanna
fluorrescente deb
bido al fluorrocromo isootiocianato de
d fluoresceeína (FITC)) que esta
marccando a los anticuerpos
a
s monoclonaales dirigidoos a la proteeína del viru
us de la rabiia.
Por lo
l anterior se
s concluye que el uso del yoduro de propidioo no interfieere en la técnnica
de IF
FD ya que se obtienen resultados
r
idénticos a llos que se reeportaron en
n las muestrras
probaadas; el usoo del yoduroo de propidiio es de ayuuda principaalmente paraa personal
técniico que no tiene
t
la expeeriencia en identificar llas células en
e improntaas y en cultivvos
celullares; el usoo del yoduroo de propidio permite un
u contraste que facilitaa la
identtificación dee la fluoresccencia propia del virus de la rabia..
RADECIM
MIENTOS:
AGR
A toddo el person
nal del laborratorio de raabia del Insstituto de Diiagnóstico y Referenciaa
Epiddemiológicos (InDRE), al Dr. Luis Leucona reepresentantee de USDA/APHIS/IS
Méxiico, por el apoyo
a
recibiido.
PT.1110
BIES PROP
PHYLAXIS
S : EXPER
RIENCE RE
EPORT ON
N INFECT
TIOUS
RAB
DISE
EASES RE
EFERENCE HOSPIT
TAL IN NA
ATAL, RIO
O GRANDE
E DO NOR
RTE
Silvaa JDPS1, Sillva DS2 - 1State
S
Healthh Secretary, 2Municipall Health Seccretary
Hospital Giiselda Triguueiro is a refference on Rio
R Grande do Norte in
n infectious
The H
diseaases for 25 years
y
and deeployed thee service forr rabies propphylaxis in collaboratioon
with the professional veteriinarian. Oveer the years the service demand is increasing,
beingg treated moonthly betw
ween 260-3000 patients victims
v
of animals attacck, which
justiffied the servvice need foor run underr a 24-hour duty care. The
T treatmeent is develooped
throuugh various entry ports since requeests for perssonal inform
mation or by
y phone, as well
as sccientific tech
hnical suppoort for the multidiscipl
m
inary hospittal team and
d municipallities.
Guiddances are given
g
to servved patientss on the emeergency dep
partment and
d, after folloow
the eestablished treatment
t
prrotocol, theyy are forwaarded to the veterinariann, wich guiddes
abouut the pathog
genesis and symptomattology of thhe disease inn the animall, follows thhe
casess and condu
ucts detailedd research; being
b
evaluated the posssible risks of contagioon
and sstarting the appropriatee mechanism
ms to promoote the healtth of the poppulation wiith
the ccorrect appliication of thhe rules on rabies
r
preveention, workking seamleessly with health
h
serviices in the control
c
and prevention
p
of
o diseases in humans and animalss.
PT.1111
ME AND ST
TORAGE TEMPERA
T
ATURE INF
FLUENCE
E IN TITER
R OF ANTII
TIM
RAB
BIES VIRU
US FLOUR
RESCENT CONJUGA
C
ATE PROD
DUCED IN PASTEUR
R
INST
TITUTE – SÃO PAULO
Trotii ACP1, Silv
va ACR1, Peeixoto ZMP
P1, Ferreira KCS1, Chav
ves LB1, Caaporale GM
MM 1
Instiituto Pasteuur
Pasteeur Institutee, SP, Brazill
g which a product
p
or reeactant has, within
Stabiility is definned as the period during
speciified limits, the same properties
p
an
nd characterristics posseessed at the time of its
manuufacture. Thhe determinnation of thee validity ennsures that th
he reagent maintains
m
thhe
charaacteristics of
o strength, purity
p
and quality
q
durinng the proposed periodd when storeed
undeer appropriaate conditionns. It is recoommended for
f maintennance of connjugated
polycclonal or an
ntiribonucleoproteins raabies virus, at 4°C and protection of light
inciddence. The objective
o
off this study was
w to evaluuate the stabbility of thee title of lotss of
rabiees virus fluoorescent connjugate prodduced at thee Pasteur Insstitute of Sãão Paulo (IP
P/SP),
adeqquately storeed for differrent periods and temperratures. Thee titles of th
he conjugatees
used in this stud
dy, so produuced and maaintained unnder appropriate condittions were
d
immuunefluoresceence (DIF) test
t on slidees with decaals of the ceentral
deterrmined by direct
nervoous system (CNS) of mice
m positive for rabies, and we used three battches, the which
w
weree named antiribonucleooproteins: Feebruary/20006 (L.RNP: 02/2006), rabies
r
virus:
Januaary/2010 (L
L.TOT: 01/22010) and raabies virus: April/20111 (L.TOT: 04/2011),
0
wiith
titerss 1:140, 1:1000 and 1:800, respectiveely. Aliquotts of these conjugates
c
was
w also stoored
at - 220°C, after production.
p
Aliquots keept at 4°C to - 20°C were
w evaluated by the IF
FD,
in Juuly 2012, using decals CNS
C
of bovvine positivee for rabies,, as the titers obtained
aliquuot (L.RNP:: 02 / 2006), (L.TOT: 01/2010)
0
and (L.TOT: 04/2011),
0
after periodss of
storaage at 4°C, 1:70,
1
1:80 and
a 1:80, annd aliquots of
o the preserrved -20°C,, bonds 1:1440,
1:1000 and 1:1000, respectiveely. The resuults of this study show
wed a small decrease
d
of the
title aaliquots of conjugates
c
p
produced
inn the IP/SP, when propperly stored for long perriods
at 4°C, this mayy be due to changes
c
in temperature
t
e through su
uccessive op
penings of thhe
o
b
better
preserrvation of thhe titles of this
t reagent when aliquuots
refriggerator, by observing
are kkept frozen, for single use
u at - 20°C
C without undergoing
u
r
repeated
thaawing.
PT.1112
AIVA EM SÃO JOSÉ
É DO EGIT
TO, SERTÃ
ÃO PERNA
AMBUCO:: UMA
A RA
ANÁ
ÁLISE DAS
S AMOSTR
RAS ENVIIADAS
Machhado JL1, Araújo
A
ACR
R2, Ribeiro MGB
M 3, Silvaa LAM4, Olliveira JCT5, Gomes ES
S6,
7 1
2
3
Bolleer MAA - LACEN - PE,
P LACE
EN/PE / LAN
NAGRO/PE
E, Universiidade Federral de
4
5
Pernaambuco. Ceentro Acadêêmico de Viitória, GEM
MNE, UFP
PE/CAV / GEMNE,
G
6
Secrretaria Mun
nicipal de Saaúde de São
o José do Eggito, 7Fundaação Oswalddo Cruz,
Instittuto Nacion
nal de Contrrole de Quallidade em Saúde
S
A raiiva é uma an
ntropozoonnose altamennte letal quee acomete a todos os mamíferos,
m
a sua
transsmissão se dá
d principalm
mente pela mordedura de um anim
mal infectaddo e envolveem
nos sseus ciclos diferentes
d
g
grupos
de mamíferos
m
enntre eles aniimais domésticos e
silvestres. Apesaar de ser um
ma doença grave
g
é de fáácil profilaxxia e possível de ser
ma das propoostas de moonitoramentoo é o encam
minhamento de amostraas
moniitorada. Um
para análise rábiica de difereentes espécies de mam
míferos a fim
m de se estuddar a circulaação
do víírus nos difeerentes cicloos da doençça (urbano, silvestre, aééreo e rural)). O municíppio,
em eestudo, situaa-se no sertãão pernambu
ucano e reppresenta o mais
m efetivo no
encam
minhamento de amostrra da regiãoo, dista da caapital pernaambucana ceerca de 400
quilôômetros, loccalizando-see a uma latittude 07º28'444" sul e a uma
u longitu
ude 37º16'288"
oestee, estando a uma altitudde de 585 metros.
m
No ultimo
u
censo
o sua populaação foi
estim
mada em 35..792 habitanntes ocupanndo uma áreea de 792,00
0 km². As innformações das
amosstras foram coletadas nos
n livros dee registro doo LANAGR
RO/PE e no banco de daados
da V
Vigilância Saanitária do município.
m
O primeiro registro de amostra encaminhada pelo
muniicípio de Sãão José do Egito
E
para annálise rábicca refere-se a um bovinno enviado em
e
19799, e até dezeembro de 20011, o Muniicípio envioou 414 amosstras agrupaadas em cincco
categgorias animaais (Criaçãoo = 56, Dom
méstico = 855, Humana = 1, Quirópptero = 251 e
Silveestre = 21), do total enccaminhado 42
4 foram poositivas. Annalisando seeparadamente
cada categoria teemos que enntre os dom
mésticos destacaram-se os cães com
m uma maioor
p
a
com en
ncaminhameento de
freqüüência (ocorrrendo em praticament
e todos os anos
amosstras) e um maior númeero de amosstras (n=80)) encaminhaadas. Para os
o animais de
d
criaçção, os bovinnos, represeentaram o maior
m
númerro de encam
minhamentos (n=40) beem
comoo a maior frreqüência annual (nove anos).
a
O maaior númeroo de amostraas para os
morccegos foi dee não-hemattófagos (n=145). Os sillvestres foraam represenntados apenaas
pelass raposas (n
n=21) encam
minhadas em
m oito anos, e amostra de
d humano foi
encam
minhada um
ma única vez. Raposa e bovinos reegistraram o maior núm
mero de casoos
posittivos, ambos com 12 reegistros, segguidos peloss morcegos (n=11). Os cães
regisstraram apennas três amoostras positiivas, o ultim
mo em 20077, e o único caso positivvo
humaano ocorreuu em 1992. Apesar
A
da aparente
a
erradicação daa raiva urban
na (canina) o
víruss se mostra circulante
c
a
ainda
nesse ambiente, ppelos registrros de morccegos positivvos
nos úúltimos anos, destes cinnco ocorreraam em 20111. E, pela diiversidade de
d amostrass
encam
minhadas pelo
p municíppio, pode-see verificar que,
q o mesm
mo está efetiivando o
moniitoramento da raiva.
PT.1113
PROGRAM
MA DE EM
MPLEO TE
EMPORAL
L (PET): CO
OMO UNA
A
EL P
HER
RREMIENT
TA PARA LA VIGIL
LANCIA EP
PIDEMIOL
LÓGICA DE
D
COM
MUNIDAD
DES CON FOCO
F
RAB
BICO EN EL
E ESTAD
DO DE GUE
ERRERO,
MÉX
XICO
Doraantes SL1, Carrillo
C
PS2 - 1Secretariia de Salud--Guerrero-M
Méxcico - Zoonosis,
2
Coleegio Superior Agropeccuario del Estado de Guuerrero
C
Chica,, del
Dadaa la situacióón Epidemioológica de laa Jurisdicción Sanitariaa 06 en la Costa
estaddo de Guerrero, Méxicoo, en donde se ha identtificado áreaas de alto rieesgo, debiddo a
la preesencia y ciirculación de
d virus rábiico, lo cual se manifiessta en el altoo índice de rabia
r
parallitica bovinaa registradoo en la regióón, así comoo el incremeento de agreesiones por
especcies silvestrres, 7 por murciélagos
m
y 5 por tejóón; se establleció el siguuiente programa
de inntervención emergente para interruumpir la caddena de tran
nsmisión de la rabia conn el
apoyyo de la Secrretaria de Desarrollo
D
Social del Goobierno del estado de Guerrero
G
(SED
DESOL).