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INTRODUCTION: Rabies is an important zoonosis that causes thousands of deaths worldwide each year. Although the terrestrial cycle, mainly transmitted by dogs, is controlled in Brazil, the aerial cycle remains a serious public health issue, besides the economic problem. In the aerial cycle, the haematophagous bat Desmodus rotundus is the main source of infection, where several different species of non-haematophagous bats can be infected and can transmit the virus. METHODS: The aim of this work was to study the epidemiological pattern of rabies using antigenic characterization with monoclonal antibodies and genetic characterization by reverse-transcriptase polymerase chain reaction followed by sequencing and phylogenetic analysis of non-haematophagous bats' and herbivorous animals' central nervous system samples from the western region of the State of São Paulo, Brazil. RESULTS: From 27 samples, 3 antigenic variants were identified: AgV-3, AgV-4, and AgV-6; and from 29 samples, 5 different clusters were identified, all belonging to the rabies virus species. CONCLUSIONS: Although only non-haematophagous bats were evaluated in the studied region, the majority of samples were from antigenic and genetic variants related to haematophagous bats Desmodus rotundus. Samples from the same antigenic variant were segregated in more than one genetic cluster. This study demonstrated the diversity of rabies virus genetic lineages presented and circulating in non-haematophagous bats in the studied region.

Made available in DSpace on 2013-09-27T14:54:15Z (GMT). No. of bitstreams: 1 WOS000303308000001.pdf: 489226 bytes, checksum: a19b099ce32617e5d8603d2470bc1a6c (MD5) Previous issue date: 2012-05-01 Made available in DSpace on 2013-09-30T18:26:43Z (GMT). No. of bitstreams: 1 WOS000303308000001.pdf: 489226 bytes, checksum: a19b099ce32617e5d8603d2470bc1a6c (MD5) Previous issue date: 2012-05-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:40:07Z No. of bitstreams: 1 WOS000303308000001.pdf: 489226 bytes, checksum: a19b099ce32617e5d8603d2470bc1a6c (MD5) Made available in DSpace on 2014-05-20T13:40:07Z (GMT). No. of bitstreams: 1 WOS000303308000001.pdf: 489226 bytes, checksum: a19b099ce32617e5d8603d2470bc1a6c (MD5) Previous issue date: 2012-05-01 Bats are main reservoirs for Lyssavirus worldwide, which is an important public health issue because it constitutes one of the big challenges in rabies control. Yet, little is known about how the virus is maintained among bats, and the epidemiological relationships remain poorly understood. The aim of the present study was to investigate the distribution of the rabies virus (RABV) in bat tissues and organs and to genetically characterize virus isolates from naturally infected non-hematophagous bats. The heminested reverse transcriptase polymerase chain reaction (hnRT-PCR) and sequencing using primers to the nucleoprotein coding gene were performed. The results showed a dissemination of the RABV in different tissues and organs, particularly in the salivary glands, tongue, lungs, kidneys, bladder, intestine and feces, suggesting other possible forms of RABV elimination and the possibility of transmission among these animals. The phylogenetic analysis confirmed that different variants of RABV are maintained by non-hematophagous bats in nature and have similar tissue distribution irrespective of bat species and phylogenetic characterization. (C) 2012 Elsevier B.V. All rights reserved. UNESP, Dept Higiene Vet & Saúde Publ, Fac Med Vet & Zootecnia, BR-18618970 Botucatu, SP, Brazil Univ Santo Amaro, Escola Vet Santo Amaro, São Paulo, Brazil Universidade Federal de Minas Gerais (UFMG), Escola Vet, Dept Vet Prevent Med, Belo Horizonte, MG, Brazil Ctr Controle Zoonoses São Paulo, São Paulo, Brazil UNESP, Dept Higiene Vet & Saúde Publ, Fac Med Vet & Zootecnia, BR-18618970 Botucatu, SP, Brazil

INTRODUCTION: This paper presents the first report of rabies in three bat species, Molossus molossus, Molossops neglectus and Myotis riparius in the city of São Paulo, Brazil. METHODS: Bats were diagnosed as positive for rabies using the fluorescent antibody test and mouse inoculation test. The isolates were characterized antigenically using a panel of eight monoclonal antibodies. The samples were also genetically analyzed by partial sequencing of the portion of nucleoprotein gene between positions 1157 and 1445nt. RESULTS: Analysis of the results verified that the sample isolated from the species M. molossus presented antigenic variant 6, while the other two samples showed a different profile from that established in the panel, one not previously reported in the literature. The results of genetic analysis revealed that the M. molossus sample segregated with Lasiurus sp. isolates, M. neglectus segregated with a subgroup of Eptesicus furinalis isolates and the Myotis riparius sample segregated with Myotis sp. isolates. CONCLUSIONS: The cases reported in this paper emphasize the need for clarification of the circumstances in which cases of rabies in wildlife occur, principally in urban areas. INTRODUÇÃO: Esse trabalho apresenta o primeiro registro de raiva em três espécies de morcegos: Molossus molossus, Molossops neglectus e Myotis riparius na Cidade de São Paulo, Brasil. MÉTODOS: Os morcegos foram diagnosticados como positivos para raiva usando as técnicas padrão de imunofluorescência direta e o teste de inoculação em camundongo. Os isolados foram caracterizados antigenicamente usando um painel de oito anticorpos monoclonais (CDC/Atlanta/USA). As amostras também foram analisadas geneticamente por sequenciamento parcial do gene da nucleoproteína entre as posições 1157 e 1445nt. RESULTADOS: O resultado das análises mostrou que as amostras isoladas da espécie M. molossus apresentou variante antigênica 6, enquanto as outras duas amostras mostraram um perfil diferente daquele estabelecido no painel e ainda não registrado em literatura. Os resultados da analise genética revelaram que a amostra de M. molossus segrega com isolados de Lasiurus sp., M. neglectus segrega com o isolado do subgrupo de Eptesicus furinalis e uma amostra de M. riparius segrega com isolados de Myotis sp. CONCLUSÕES: Os casos relatados neste estudo enfatizam a necessidade do esclarecimento da ocorrência de casos de raiva em morcegos, principalmente em áreas urbanas.

INTRODUCTION: Bats are one of the most important reservoirs and vectors of the rabies virus in the world. METHODS: From 1988 to 2003, the Zoonosis Control Center in São Paulo City performed rabies diagnosis on 5,670 bats by direct immunofluorescent test and mouse inoculation test. Blood samples were collected from 1,618 bats and the sera were analyzed using the rapid fluorescent focus inhibition test to confirm rabies antibodies. RESULTS: Forty-four (0.8%) bats were positive for rabies. The prevalence of rabies antibodies was 5.9% using 0.5IU/ml as a cutoff. Insectivorous bats (69.8%) and bats of the species Molossus molossus (51.8%) constituted the majority of the sample; however, the highest prevalence of antibodies were observed in Glossophaga soricina (14/133), Histiotus velatus (16/60), Desmodus rotundus (8/66), Artibeus lituratus (5/54), Nyctinomops macrotis (3/23), Tadarida brasiliensis (3/48), Carollia perspicillata (3/9), Eumops auripendulus (2/30), Nyctinomops laticaudatus (2/16), Sturnira lilium (2/17) and Eumops perotis (1/13). The prevalence of rabies antibodies was analyzed by species, food preference and sex. CONCLUSIONS: The expressive levels of antibodies associated with the low virus positivity verified in these bats indicate that rabies virus circulates actively among them. INTRODUÇÃO: Morcegos são um dos mais importantes reservatórios e vetores do vírus da raiva no mundo. MÉTODOS: No período entre 1998 e 2003, o Centro de Controle de Zoonoses da Cidade de São Paulo realizou o diagnóstico de raiva em 5.670 morcegos utilizando as técnicas de imunofluorescência direta e inoculação intracerebral em camundongos. Sangue foi coletado de 1.618 espécimes para pesquisa de anticorpos pela técnica de inibição de foco de fluorescência rápida. RESULTADOS: Quarenta e quatro (0,8%) morcegos foram positivos para raiva. A prevalência de anticorpos foi de 5,9% usando 0,5UI/ml como ponto de corte. Os morcegos de hábito alimentar insetívoro (69,8%) e os morcegos da espécie Molossus molossus (51,8%) representaram a maioria da amostra. Entretanto, as mais altas prevalências de anticorpos foram observadas nos morcegos Glossophaga soricina (14/133), Histiotus velatus (16/60), Desmodus rotundus (8/66), Artibeus lituratus (5/54), Nyctinomops macrotis (3/23), Tadarida brasiliensis (3/48), Carollia perspicillata (3/9), Eumops auripendulus (2/30), Nyctinomops laticaudatus (2/16), Sturnira lilium (2/17) e Eumops perotis (1/13). A prevalência de anticorpos foi analisada por espécie, hábito alimentar e sexo. CONCLUSÕES: O expressivo nível de anticorpos associado à baixa positividade para o vírus da raiva entre os morcegos estudados indica que o vírus circula ativamente entre morcegos.

The Polo da Alta Sorocabana Laboratory in Presidente Prudente, SP, in partnership with other research institutions, conducted studies related to bats from the western region of the State of Sao Paulo, Brazil. Thus, certain situations were investigated, including: a) isolation of the rabies virus from 2006 to 2008; b) identification of respective antigenic variants; and c) characterization of daytime shelters of Desmodus rotundus vampire bats.Samples for examination originated from nonhematophagous bats forwarded to the laboratory and subjected to direct fluorescent antibody test and mouse inoculation test. Positive samples were characterized by the monoclonal antibody test. Regarding the bats, they were identified and classified and mapping of their shelters was also performed.The laboratory received 1,113 nonhematophagous bats for rabies diagnosis, 11 (1%) of which were positives, and among the positive samples, 5 (45.5%) presented antigenic variant 3 (from the bat Desmodus rotundus) and 4 (36.5%) were compatible with samples derived from Brazilian insectivorous bats. Sixteen vampire bat shelters were investigated and observation confirmed the presence of another 3 species of nonhematophagous bats coexisting with them.The experiments showed that at least 3 antigenic variants of rabies virus are circulating in the region and that the cohabitation of vampire bats with nonhematophagous bats could be related to the dissemination of the rabies virus.

Some bat species have adapted to the expanding human population by acquiring the ability to roost in urban buildings, increasing the exposure risk for people and domestic animals, and consequently, the likelihood of transmitting rabies. Three dead bats were found in the yard of a house in an urban area of Jundiaí city in the state of São Paulo in southeast Brazil. Two of the three bats tested positive for rabies, using Fluorescent Antibody and Mouse Inoculation techniques. A large colony of Eptesicus furinalis was found in the house's attic, and of the 119 bats captured, four more tested positive for rabies. The objectives of this study were to report the rabies diagnosis, characterize the isolated virus antigenically and genetically, and study the epidemiology of the colony. Algumas espécies de morcegos têm se adaptado ao uso de abrigos em construções urbanas, aumentando a possibilidade de contato desses morcegos com pessoas e animais domésticos e conseqüentemente, o potencial risco de transmissão de raiva. Três morcegos foram encontrados no jardim de uma casa na área urbana da cidade de Jundiaí, Estado de São Paulo, Sudeste do Brasil, dois deles foram positivos para raiva pelas técnicas de imunofluorescência e inoculação em camundongos. Uma grande colônia de E. furinalis foi identificada, vivendo no sótão da casa e 119 morcegos foram encaminhados para diagnóstico de raiva, com mais quatro morcegos positivos. O objetivo desse estudo é apresentar a caracterização genética e antigênica do vírus da raiva isolado desses morcegos e o estudo epidemiológico da colônia.

Esse artigo apresenta uma lista atualizada de espécies positivas para raiva no Brasil e foi desenvolvida a partir da base de dados na internet da literatura nacional, internacional e dos anais das mais importantes reuniões técnicas e científicas, envolvendo raiva e morcegos no Brasil durante o período de 1996 a 2009. A nova lista de morcegos positivos para raiva consiste de 41 espécies, pertencentes a 25 gêneros e três famílias: Phyllostomidae 43.9%, Vespertilionidae 29.3% e Molossidae 26.8%. Também foram discutidas questões como a falta de dados sobre sexo, faixa etária e circunstâncias de captura dos animais e identificação incompleta ou desatualizada das espécies. Resultados dos estudos genéticos e antigênicos realizados em amostras de morcegos brasileiros positivos para raiva foram apresentados. This paper presents an updated list of bat species positive for rabies in Brazil. It was developed based on database research via the internet, of international and national literature and annals of the most important technical and scientific meetings related to rabies and chiroptera in Brazil from 1996 to 2009. The new list of rabies positive bats consists of 41 species, belonging to 25 genera and three families: Phyllostomidae 43.9%, Vespertilionidae 29.3% and Molossidae 26.8%. In addition, questions were raised regarding the lack of data, including sex, age, circumstances and location of bat capture and incomplete and outdated species identification. Results of genetic and antigenic studies performed on Brazilian rabies positive bats were shown.

O Laboratório de Virologia Clínica e Molecular do Instituto de Ciências Biomédicas da Universidade de São Paulo, utilizando-se da técnica de anticorpos monoclonais, tipificou 18 amostras de vírus rábico provenientes de morcegos não hematófagos de várias espécies provenientes da Região de Presidente Prudente, SP, Brasil. Destas amostras, 15 (82,3%) foram definidas como variante 3 (compatível com amostras isoladas de morcegos Desmodus rotundus) e 3 (16,7%) como variante 4 (compatível com amostras isoladas de morcegos Tadarida brasiliensis). Using the monoclonal antibody technique, the Clinical and Molecular Virology Laboratory of the Institute of Biomedical Sciences of the University of São Paulo typed 18 rabies virus samples from non-hematophagous bats of several species from the region of Presidente Prudente, SP, Brazil. Among these samples, 15 (82.3%) were defined as variant 3 (compatible with samples isolated from Desmodus rotundus bats) and three (16.7%) as variant 4 (compatible with samples isolated from Tadarida brasiliensis bats).

Thomas' Mastiff bat, Eumops maurus (Thomas, 1901) is known from northern South America in Ecuador, Venezuela and Guiana and so it probably occurs in Brazil, yet it has not been reported. Here, we present the first record for E. maurus in Brazil. Captures were from the Brazilian states of Tocantins and Goiás (northern and midwestern Cerrado region of the country), and São Paulo (southeastern Brazil, Atlantic Forest). In the first two locations, all bats were roosting in palm leaves while in the later location, a single animal was found alive in a 15th-storey apartment in the city of São Paulo. This bat inhabits savannas with swamps dominated by the palm Mauritia flexuosa (buriti) and gallery forests. This record is the first of E. maurus in an urban area. Brazilian specimens are apparently larger than those from northern South America. Eumops maurus (Thomas, 1901) tem sua ocorrência no norte da América do Sul, no Equador, na Venezuela e na Guiana. O presente estudo descreve a primeira ocorrência de E. maurus para o Brasil, com capturas recentes de espécimes nos estados de Tocantins e Goiás (regiões Norte e Centro-Oeste do país), domínio de Cerrado, e no estado de São Paulo, região sudeste, inserido no domínio da Mata Atlântica. Nos dois primeiros casos, os espécimes estavam no interior de folhas de palmeiras, e último, o indivíduo foi encontrado vivo dentro do quarto de um apartamento na cidade de São Paulo. Dados bibliográficos revelam que esta espécie habita regiões abertas, associada às áreas periodicamente alagadas (no cerrado, veredas) dominadas por palmeiras da espécie Mauritia flexuosa (buriti) e matas de galeria. Este trabalho ressalta o primeiro registro desse morcego em local urbanizado. Os espécimes brasileiros apresentam dimensões corpóreas maiores que os espécimes provenientes na região norte da América do Sul.

Using the monoclonal antibody technique, the Clinical and Molecular Virology Laboratory of the Institute of Biomedical Sciences of the University of São Paulo typed 18 rabies virus samples from non-hematophagous bats of several species from the region of Presidente Prudente, SP, Brazil. Among these samples, 15 (82.3%) were defined as variant 3 (compatible with samples isolated from Desmodus rotundus bats) and three (16.7%) as variant 4 (compatible with samples isolated from Tadarida brasiliensis bats).

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T19:04:51Z No. of bitstreams: 1 S0036-46652002000200007.pdf: 56466 bytes, checksum: d504c9abe5e28aaa48da34e9ccf026f5 (MD5) Made available in DSpace on 2013-08-22T19:04:51Z (GMT). No. of bitstreams: 1 S0036-46652002000200007.pdf: 56466 bytes, checksum: d504c9abe5e28aaa48da34e9ccf026f5 (MD5) Previous issue date: 2002-04-01 Made available in DSpace on 2013-09-30T20:08:16Z (GMT). No. of bitstreams: 2 S0036-46652002000200007.pdf: 56466 bytes, checksum: d504c9abe5e28aaa48da34e9ccf026f5 (MD5) S0036-46652002000200007.pdf.txt: 25319 bytes, checksum: deab854ff7ac45ca2e7bee2c7ad69da2 (MD5) Previous issue date: 2002-04-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:17:26Z No. of bitstreams: 2 S0036-46652002000200007.pdf: 56466 bytes, checksum: d504c9abe5e28aaa48da34e9ccf026f5 (MD5) S0036-46652002000200007.pdf.txt: 25319 bytes, checksum: deab854ff7ac45ca2e7bee2c7ad69da2 (MD5) Made available in DSpace on 2014-05-20T15:17:26Z (GMT). No. of bitstreams: 2 S0036-46652002000200007.pdf: 56466 bytes, checksum: d504c9abe5e28aaa48da34e9ccf026f5 (MD5) S0036-46652002000200007.pdf.txt: 25319 bytes, checksum: deab854ff7ac45ca2e7bee2c7ad69da2 (MD5) Previous issue date: 2002-04-01 Amostras de vírus rábico isoladas de animais e humanos no período de 1989 a 2000 foram tipificadas antigenicamente com a utilização de um painel de anticorpos monoclonais contra a nucleoproteína viral, pré-estabelecido para o estudo da epidemiologia molecular do vírus rábico isolado nas Américas. As amostras testadas foram isoladas no laboratório de diagnóstico do Instituto Pasteur e outros centros de diagnóstico de raiva no Brasil. Além das cepas de vírus rábico fixo CVS-31/96-IP, mantida em cérebro de camundongos e a PV-BHK/97, mantida em cultura de células, cepas de vírus rábico isoladas de cães, gatos, bovinos, eqüinos, morcegos, ovinos, caprino, suínos, raposa, sagüí, coatí, guaxinim e humanos, totalizaram 330 amostras. Seis variantes antigênicas foram definidas, compatíveis com perfís observados no painel de anticorpos monoclonais pré-estabelecido utilizado, as de número 2 (cão), 3 (Desmodus rotundus), 4 (Tadarida brasiliensis), 5 (Vampiro da Venezuela), 6 (Lasiurus cinereus) e Lab (reagente a todos os anticorpos utilizados), além de outros seis perfís desconhecidos, não compatíveis com aqueles observados no painel utilizado. A maior variabilidade foi observada entre as amostras isoladas de morcegos insetívoros e a variante mais comum isolada entre as espécies foi a variante 3 (Desmodus rotundus). Estes fatos podem representar a existência de múltiplos ciclos de transmissão independentes, envolvendo diferentes espécies de morcegos. Animal and human rabies samples isolated between 1989 and 2000 were typified by means of a monoclonal antibody panel against the viral nucleoprotein. The panel had been previously established to study the molecular epidemiology of rabies virus in the Americas. Samples were isolated in the Diagnostic Laboratory of the Pasteur Institute and in other rabies diagnostic centers in Brazil. In addition to the fixed virus samples CVS-31/96-IP, preserved in mouse brain, and PV-BHK/97, preserved in cell culture, a total of 330 rabies virus samples were isolated from dogs, cats, cattle, horses, bats, sheep, goat, swine, foxes, marmosets, coati and humans. Six antigenic variants that were compatible with the pre-established monoclonal antibodies panel were defined: numbers 2 (dog), 3 (Desmodus rotundus), 4 (Tadarida brasiliensis), 5 (vampire bat from Venezuela), 6 (Lasiurus cinereus) and Lab (reacted to all used antibodies). Six unknown profiles, not compatible with the panel, were also found. Samples isolated from insectivore bats showed the greatest variability and the most commonly isolated variant was variant-3 (Desmodus rotundus). These findings may be related to the existence of multiple independent transmission cycles, involving different bat species. Instituto Pasteur Instituto Biológico Centro de Controle de Zoonoses UNESP UNESP

Rabies virus was isolated from a frugivorous bat, Artibeus lituratus, captured in downtown Rio Claro, SP, Brazil, in 1997. There was not any animal rabies since 1986, and this is the first isolation of rabies virus in a frugivorous bat in Rio Claro. Implications of Public Health were discussed. O vírus rábico foi isolado de morcego frugívoro Artibeus lituratus, capturado no município de Rio Claro, SP, em bairro residencial, em 1997. Neste município, o último caso de raiva animal ocorreu em 1986, sendo este o primeiro relato do isolamento em morcego frugívoro. As implicações em Saúde Pública foram discutidas.

Descrevem-se, pela primeira vez, o isolamento e a identificação do vírus da raiva em morcego frugívoro Artibeus fimbriatus no município de São José do Rio Preto, Estado de São Paulo. O vírus foi isolado de exemplar encontrado em área urbana, caído sob uma árvore e ainda vivo. O diagnóstico foi realizado pelas técnicas de imunofluorescência direta e inoculação intracerebral em camundongos. This is the first report of the isolation and identification of the rabies virus in the frugivorous bat Artibeus fimbriatus in the city of Sao José do Rio Preto, Sao Paulo State, Brazil. The virus was isolated from an animal found in an urban area. The animal was found on the ground under a tree, still alive. Diagnosis was made by direct immunofluorescence and intracerebral inoculation of mice.