In situ histopathological and immunohistochemical characterization of rabies in the brains of naturally infected equines and bovines / Caracterização histopatológica e imuno-histoquímica In situ do vírus da raiva em cérebros de equinos e bovinos naturalmente infectados

The present study sought to characterize the phenomena involved in the histopathology of rabies and to assess the presence and amount of viral antigen in situ in different brain regions of naturally infected equines and bovines. The histopathological examination showed several changes due to inflammation, being most often infected cells neurons. The neuronal degeneration involved 100% of cases, in addition to a diffuse lymphocytic Infiltration and gliosis, characterized by vasculitis and perivasculitis. The presence of Negri bodies was in most cases in discreet, and the fragments with higher concentrations of antigen by both techniques employed were the cerebellum and the brain stem. Immunohistochemistry test (IHC) demonstrated greater sensitivity when applied to samples of bovines. Our results showed that in 37.5% of the total number of fragments analyzed, viral inclusions were not observed, however, was the presence of inflammatory process. In relation to the species, the fragments from bovine's animals showed a slight increase when examined under this feature. These findings highlight the importance of submitting samples from suspected animals for laboratory diagnostic, even when there are no apparent abnormal histological findings. (AU)

O presente estudo buscou caracterizar os fenômenos envolvidos na histopatologia da raiva e avaliar a presença e quantidade de antígeno viral in situ nas diferentes regiões cerebrais de equinos e bovinos naturalmente infectados. O exame histopatológico demonstrou várias mudanças devido à inflamação, sendo mais frequentemente infectadas as células neuronais. A degeneração neuronal foi observada em 100% dos casos, além de uma infiltração linfocitária difusa e gliose, caracterizada por vasculite e perivasculite. A presença de corpúsculos de Negri foi observada na maioria dos casos de maneira discreta, e os fragmentos com maior concentração de antígeno, por ambos os testes empregadas foram o cerebelo e o tronco encefálico. O teste de Imuno-histoquímica (IHC) demonstrou maior sensibilidade quando aplicada em amostras de bovinos. Nossos resultados demostraram que em 37,5% do número total de fragmentos analisados, inclusões virais não foram observadas, no entanto, havia processo inflamatório. Em relação à espécie, os fragmentos de bovinos demonstraram um ligeiro aumento quando examinado sob este aspecto. Esses achados destacam a importância de submeter amostras de animais suspeitos para diagnóstico laboratorial, mesmo quando não houver nenhum achado histopatológico anormal.Palavras-chave: raiva, equinos, bovinos, imuno-histoquímica, IFD, alterações histopatológicas. (AU)

Analysis of rabies diagnosis in dogs and cats in the state of São Paulo, Brazil

The genetic lineage of rabies virus (RABV) associated with dogs has not been found in the state of São Paulo since 1998, and all cases of rabies in domestic animals reported since then have involved the RABV lineage that circulates in bats. As there has been a change in the rabies transmission cycle in cats and dogs, we decided to analyze the tests used to diagnose rabies in these animals in the 15-year period from 2002 to 2016 in the state of São Paulo. During this period, 85,508 central nervous system (CNS) samples from dogs and cats were submitted to the Rabies Diagnosis Section at the Pasteur Institute of São Paulo for testing. All of the samples were tested by the fluorescent antibody test (FAT) and at least one of the following three tests: mouse inoculation test (MIT), rabies tissue culture infection test (RTCIT) and reverse transcription polymerase chain reaction (RT-PCR). Of all the samples tested, twenty were positive in at least one of these assays. Four other positive samples were identified at other institutions in the state of São Paulo. Of the twenty samples that tested positive at the Pasteur Institute of São Paulo, nine were tested by FAT, and the results were subsequently confirmed by other techniques; five gave inconclusive results, and therefore, other techniques had to be used as soon as possible in case the samples were positive; and six were negative by FAT and positive by one or more of the following tests:...

Analysis of rabies diagnosis in dogs and cats in the state of São Paulo, Brazil.

The genetic lineage of rabies virus (RABV) associated with dogs has not been found in the state of São Paulo since 1998, and all cases of rabies in domestic animals reported since then have involved the RABV lineage that circulates in bats. As there has been a change in the rabies transmission cycle in cats and dogs, we decided to analyze the tests used to diagnose rabies in these animals in the 15-year period from 2002 to 2016 in the state of São Paulo. During this period, 85,508 central nervous system (CNS) samples from dogs and cats were submitted to the Rabies Diagnosis Section at the Pasteur Institute of São Paulo for testing. All of the samples were tested by the fluorescent antibody test (FAT) and at least one of the following three tests: mouse inoculation test (MIT), rabies tissue culture infection test (RTCIT) and reverse transcription polymerase chain reaction (RT-PCR). Of all the samples tested, twenty were positive in at least one of these assays. Four other positive samples were identified at other institutions in the state of São Paulo. Of the twenty samples that tested positive at the Pasteur Institute of São Paulo, nine were tested by FAT, and the results were subsequently confirmed by other techniques; five gave inconclusive results, and therefore, other techniques had to be used as soon as possible in case the samples were positive; and six were negative by FAT and positive by one or more of the following tests: RTCIT, MIT and RT-PCR. Genetic typing of isolates from eighteen samples identified them as the lineage circulating in bats. In light of this finding, which indicates that genetic lineages associated with bats are circulating in domestic animals in the state of São Paulo, when the results of FAT carried out with samples from aggressive cats and dogs are inconclusive, complementary tests should be used. Decomposing samples and samples for which FAT was inconclusive should be tested using molecular techniques so that a definitive result can be obtained quickly and timely post-exposure prophylaxis can be administered to exposed individuals.

A rabies virus vampire bat variant shows increased neuroinvasiveness in mice when compared to a carnivore variant

Rabies is one of the most important zoonotic diseases and is caused by several rabies virus (RABV) variants. These variants can exhibit differences in neurovirulence, and few studies have attempted to evaluate the neuroinvasiveness of variants derived from vampire bats and wild carnivores. The aim of this study was to evaluate the neuropathogenesis of infection with two Brazilian RABV street variants (variant 3 and crab-eating fox) in mice. BALB/c mice were inoculated with RABV through the footpad, with the 50% mouse lethal dose (LD50) determined by intracranial inoculation. The morbidity of rabies in mice infected with variant 3 and the crab-eating fox strain was 100% and 50%, respectively, with an incubation period of 7 and 6 days post-inoculation (dpi), respectively. The clinical disease in mice was similar with both strains, and it was characterized initially by weight loss, ruffled fur, hunched posture, and hind limb paralysis progressing to quadriplegia and recumbency at 9 to 12 dpi. Histological lesions within the central nervous system (CNS) characterized by nonsuppurative encephalomyelitis with neuronal degeneration and necrosis were observed in mice infected with variant 3 and those infected with the crab-eating fox variant. However, lesions and the presence of RABV antigen, were more widespread within the CNS of variant-3-infected mice, whereas in crab-eating fox-variant-infected mice, RABV antigens were more restricted to caudal areas of the CNS, such as the spinal cord and brainstem. In conclusion, the results shown here demonstrate that the RABV vampire bat strain (variant 3) has a higher potential for neuroinvasiveness than the carnivore variant. (AU) i

The epidemiological importance of bats in the transmission of rabies to dogs and cats in the State of São Paulo, Brazil, between 2005 and 2014

In Brazil, rabies control in dogs and cats was pioneered by the state of São Paulo with the adoption of the Pan American Health Organization recommendations for prophylaxis and control, which led to a reduction in rabies cases from 1994 onwards. As a result of these measures, the rabies virus (RABV) genetic lineage associated with dogs has not been found in the state since 1998, and all the cases in domestic animals reported since then have been caused by bat­associated lineages of RABV. In the light of this, this study sought to investigate rabies cases in dogs and cats in the state of São Paulo between 2005 and 2014 and identify the associated transmission cycles by characterizing the RABV lineages responsible for these cases. Nine samples from dogs (n = 5) and from cats (n = 4) were collected between 2005 and 2014. The tenth animal, a rabid cat, was analysed by a different laboratory. The N gene nucleotide sequences obtained were analysed with the neighbor­joining algorithm and Kimura 2­parameter model using the MEGA 6 program. Phylogenetic analysis revealed that the genetic lineages identified in all the samples were those circulating in Brazilian bats. The findings of this study demonstrate that bats play an important role in the transmission of rabies to domestic animals in São Paulo state and that emphasis should be placed on the implementation of public policies to support surveillance of chiropterans for rabies.(AU) i

A rabies virus vampire bat variant shows increased neuroinvasiveness in mice when compared to a carnivore variant.

Rabies is one of the most important zoonotic diseases and is caused by several rabies virus (RABV) variants. These variants can exhibit differences in neurovirulence, and few studies have attempted to evaluate the neuroinvasiveness of variants derived from vampire bats and wild carnivores. The aim of this study was to evaluate the neuropathogenesis of infection with two Brazilian RABV street variants (variant 3 and crab-eating fox) in mice. BALB/c mice were inoculated with RABV through the footpad, with the 50% mouse lethal dose (LD50) determined by intracranial inoculation. The morbidity of rabies in mice infected with variant 3 and the crab-eating fox strain was 100% and 50%, respectively, with an incubation period of 7 and 6 days post-inoculation (dpi), respectively. The clinical disease in mice was similar with both strains, and it was characterized initially by weight loss, ruffled fur, hunched posture, and hind limb paralysis progressing to quadriplegia and recumbency at 9 to 12 dpi. Histological lesions within the central nervous system (CNS) characterized by nonsuppurative encephalomyelitis with neuronal degeneration and necrosis were observed in mice infected with variant 3 and those infected with the crab-eating fox variant. However, lesions and the presence of RABV antigen, were more widespread within the CNS of variant-3-infected mice, whereas in crab-eating fox-variant-infected mice, RABV antigens were more restricted to caudal areas of the CNS, such as the spinal cord and brainstem. In conclusion, the results shown here demonstrate that the RABV vampire bat strain (variant 3) has a higher potential for neuroinvasiveness than the carnivore variant.

Phylogenetic analysis of rabies virus isolated from canids in North and Northeast Brazil.

Cases of canine rabies continue to occur in North and Northeast Brazil, and the number of notifications of rabies cases in wild canids has increased as a result of the expansion of urban areas at the expense of areas with native vegetation. In light of this, we performed molecular characterization of rabies virus isolates from dogs and Cerdocyon thous from various states in North and Northeast Brazil. In all, 102 samples from dogs (n = 56) and Cerdocyon thous (n = 46) collected between 2006 and 2012 were used. The nucleotide sequences obtained for the N gene of rabies virus were analyzed, and phylogenetic analysis revealed the presence of two distinct genetic lineages, one associated with canids and one with bats, and, within the canid cluster, two distinct sublineages circulating among dogs and Cerdocyon thous. In addition, phylogenetic groups associated with geographic region and fourteen cases of interspecific infection were observed among the isolates from canids. Our findings show that analysis of rabies virus lineages isolated from reservoirs such as canids must be constantly evaluated because the mutation rate is high.

Diagnóstico de cinomose canina por RT-PCR em amostras de cães do Estado de São Paulo enviadas para a vigilância laboratorial da raiva

A cinomose canina (CD) é uma das doenças infecciosas mais importantes dos cães domésticos. No Brasil é ainda a principal causa de mortalidade de cães em algumas populações urbanas. Embora sequências de diferentes genes do vírus sejam utilizadas como alvo para detecção do vírus da cinomose canina (CDV), o gene N parece ser o melhor para a amplificação de todas as suas linhagens. Utilizando-se a técnica de RT-PCR direcionada ao gene N do CDV, foram analisadas 190 amostras de sistema nervoso central (SNC) de cães do estado de São Paulo com quadros sugestivos de encefalite e que foram encaminhadas ao Instituto Pasteur para o diagnóstico da raiva, durante o ano de 2014. A positividade foi superior a 50% indicando que a cinomose continua a ser uma importante causa de mortalidade canina.

Monoclonal antibodies for characterization of rabies virus isolated from non-hematophagous bats in Brazil.

INTRODUCTION: In Brazil, various isolates of rabies virus (RABV) show antigenic profiles distinct from those established by the reduced panel of eight monoclonal antibodies (MAbs) determined by the Centers for Disease Control and Prevention (CDC), utilized for the antigenic characterization of RABV in the Americas. The objective of this study was to produce MAbs from RABV isolates from insectivorous bats with an antigenic profile incompatible with the pre-established one. METHODOLOGY: An isolate of RABV from the species Eptesicus furinalis that showed an antigenic profile incompatible with the panel utilized was selected. Hybridomas were produced utilizing the popliteal lymph nodes of mice immunized with ribonucleoproteins purified from the isolate. RESULTS: Two MAbs-producing clones were obtained, BR/IP1-3A7 and BR/IP2-4E10. Fifty-seven isolates of RABV from different species of animals and different regions of Brazil were analyzed utilizing the MAbs obtained. In the analysis of 23 RABV isolates from non-hematophagous bats, the MAbs cross-reacted with ten isolates, of which four were of the species Nyctinomops laticaudatus, one of the species Eptesicus furinalis, and five of the genus Artibeus. Of the nine isolates of non-hematophagous isolates that displayed an incompatible profile analyzed, characteristic of insectivorous bats, BR/IP1-3A7 reacted with five (55.55%) and BR/IP2-4E10 with four (44.44%). CONCLUSIONS: The MAbs obtained were able to recognize epitopes common between the three genera, Artibeus, Eptesicus, and Nyctinomops, thereby allowing the antigenic characterization of RABV isolates in Brazil.

Rabia transmitdaa por murciélagos enBBrasil / Rabies Transmitted by Bats in Brazil

Frente al desafío que la rabia representa para la industria pecuaria y la salud pública en América Latina, el presente artículo tiene como objetivo hacer una revisión de literatura amplia y crítica sobre la epidemiología de la rabia transmitida por murciélagos en Brasil. El tema es abordado inicialmente desde una perspectiva histórica hasta la caracterización molecular de aislamientos del virus, para finalmente contrastar con la situación de otros países de las Américas. La información referente a Brasil es presentada de manera separada debido a la gran abundancia de especies de murciélagos de diversos hábitos alimenticios, implicadas en la transmisión del virus de la rabia y las complejas relaciones entre los ciclos epidemiológicos revelados por estudios de tipificación antigénica y análisis filogenético, lo cual ha permitido reconocer con más nitidez, la importancia de los quirópteros como reservorios y transmisores de esta enfermedad. Este nuevo escenario epidemiológico exige reexaminar las medidas de control aplicadas hasta el momento, desde un abordaje multidisciplinar, así como cooperación intersectorial y participación por parte de la comunidad.

Considering that rabies represents a challenge for the livestock industry and public health in Latin America, this article is intended to do a comprehensive and critical literature review on the epidemiology of rabies transmitted by bats in Brazil. The subject is addressed from a historical perspective to molecular characterization of rabies virus isolates and finally making a contrast with other countries of the Americas. Information concerning Brazil is presented separately because of the abundance of bats species with different feeding habits, involved in the transmission of rabies virus and the complex relationships between epidemiological cycles in this country, which have been disclosed by antigenic typing and phylogenetic analysis. This has allowed to recognize more clearly the importance of bats as reservoirs and transmitters of this disease. This new epidemiological scenario requires reappraising current control measures, using a multidisciplinary approach, intersectoral cooperation and community participation.

Diphylla ecaudata y Diaemus youngi, Biología y comportamiento / Diphylla ecaudata and Diaemus youngi, Biology and behavior

En la presente revisión se presentan algunos aspectos de la biología y comportamiento de las especies de quirópteros hematófagos Diphylla ecaudata y Diaemus youngi. Y se analizan las principales características anatómicas que permiten hacer la diferenciación de estas especies con Desmodus rotundus. Es notable la falta de información sobre las dos primeras especies y la necesidad de capacitación para la correcta identificación de las mismas, lo que redundará en un direccionamiento más específico para controlar las poblaciones de Desmodus rotundus.

This review presents some aspects about the biology and behavior of the hematophagous bats Diphylla ecaudata and Diaemus youngi. It is notable the lack of information regarding both species, and the need of training in order to identify them properly from Desmodus rotundus. We analyze the main anatomical features that allow differentiation of these species. This will result in a more specific control of Desmodus rotundus populations.

Eastern equine encephalitis cases among horses in Brazil between 2005 and 2009.

Eastern equine encephalitis is a viral zoonosis that exhibits complex distribution and epidemiology, and greater importance should be given to this disease by the public-health authorities. In Brazil, although eastern equine encephalitis virus (EEEV) has been identified in vectors and antibodies are sometimes detected in horses and humans, there have been no records of equine encephalitis in horses caused by this virus during the last 24 years. This study describes eighteen cases of eastern equine encephalomyelitis that occurred in six Brazilian states between 2005 and 2009. Viral RNA was identified using semi-nested RT-PCR to detect members of the genus Alphavirus, and by genetic sequencing. The gene encoding NSP1 was partially amplified, and after genetic sequencing, eighteen sequences were generated. All eighteen strains were classified as belonging to lineage III of American EEEV. These findings could be an indication of the importance of this virus in animal and human public health.

Método de diagnóstico antemortem da raiva humana por meio de técnicas de biologia molecular, utilizando saliva e biópsia de pele da região da nuca / An antemortem method for diagnosis of human rabies by molecular techniques using saliva and neck-skin biopsy

Por meio das técnicas de RT-PCR com primers direcionados para o gene da

glicoproteína e RT-PCR e hemi-nested RT-PCR com primers direcionados para o gene da nucleoproteína, o RNA do vírus da raiva foi identificado em 95,2% de 21 amostras, 18 de saliva e três de biópsia de pele da região da nuca, coletadas entre a hospitalização e a morte de um paciente com sinais clínicos da raiva. O tratamento administrado ao paciente incluiu a indução de coma e terapia antiviral. Cada técnica, isoladamente, detectou RNA viral em 90,5%; 57,1% e 85,7% das amostras, respectivamente. Nossos resultados sugerem que a amplificação em paralelo de diferentes regiões do genoma do vírus da raiva pode fornecer maior confiabilidade ao diagnóstico antemortem da doença, auxiliando a decisão médica quanto à aplicação do protocolo de tratamento com antivirais.

Rabies virus in insectivorous bats: implications of the diversity of the nucleoprotein and glycoprotein genes for molecular epidemiology.

Insectivorous bats are the main reservoirs of rabies virus (RABV) in various regions of the world. The aims of this study were to (a) establish genealogies for RABV strains from different species of Brazilian insectivorous bats based on the nucleoprotein (N) and glycoprotein (G) genes, (b) investigate specific RABV lineages associated with certain genera of bats and (c) identify molecular markers that can distinguish between these lineages. The genealogic analysis of N and G from 57 RABV strains revealed seven genus-specific clusters related to the insectivorous bats Myotis, Eptesicus, Nyctinomops, Molossus, Tadarida, Histiotus and Lasiurus. Molecular markers in the amino acid sequences were identified which were specific to the seven clusters. These results, which constitute a novel finding for this pathogen, show that there are at least seven independent epidemiological rabies cycles maintained by seven genera of insectivorous bats in Brazil.

Genetic characterization of rabies virus isolated from bovines and equines between 2007 and 2008, in the States of São Paulo and Minas Gerais.

INTRODUCTION: Rabies is an acute disease of the central nervous system and is responsible for the deaths of thousands of humans, wild animals and livestock, particularly cattle, as well as causing major economic losses. This study describes the genetic characterization of rabies virus variants that circulate in Desmodus rotundus populations and are transmitted to herbivores. METHODS: Fifty rabies virus isolates from bovines and equines in the States of São Paulo and Minas Gerais, Brazil, were genetically characterized and compared with sequences retrieved from GenBank. RESULTS: Two clusters (I and II) with mean nucleotide identities of 99.1 and 97.6% were found. The first of these contained nearly all the samples analyzed. Lineages from other Brazilian states grouped in cluster II. CONCLUSIONS: Analysis of the amino acid sequences of the N proteins revealed the existence of genetic markers that may indicate possible variations between geographic regions, although the biologically active regions are conserved within the species over space and time.

Genetic characterization of rabies virus isolated from bovines and equines between 2007 and 2008, in the States of São Paulo and Minas Gerais / Caracterização genética de vírus da raiva isolado de bovinos e equinos entre 2007 e 2008, nos Estados de São Paulo e Minas Gerais

INTRODUCTION: Rabies is an acute disease of the central nervous system and is responsible for the deaths of thousands of humans, wild animals and livestock, particularly cattle, as well as causing major economic losses. This study describes the genetic characterization of rabies virus variants that circulate in Desmodus rotundus populations and are transmitted to herbivores. METHODS: Fifty rabies virus isolates from bovines and equines in the States of São Paulo and Minas Gerais, Brazil, were genetically characterized and compared with sequences retrieved from GenBank. RESULTS: Two clusters (I and II) with mean nucleotide identities of 99.1 and 97.6 percent were found. The first of these contained nearly all the samples analyzed. Lineages from other Brazilian states grouped in cluster II. CONCLUSIONS: Analysis of the amino acid sequences of the N proteins revealed the existence of genetic markers that may indicate possible variations between geographic regions, although the biologically active regions are conserved within the species over space and time.

INTRODUÇÃO: A raiva é uma doença aguda do sistema nervoso central e é responsável por mortes de milhares de humanos, animais silvestres e animais de criação - especialmente bovinos - além de causar elevadas perdas econômicas. Este trabalho descreve a caracterização genética das variantes do vírus da raiva que circulam em populações de Desmodus rotundus e são transmitidas aos herbívoros. MÉTODOS: Cinquenta isolados de vírus da raiva de bovinos e equinos provenientes dos Estados de São Paulo e Minas Gerais, Brasil, foram caracterizadas geneticamente e comparadas com sequências recuperadas do GenBank. RESULTADOS: Dois clusters, I e II, apresentando identidades médias de nucleotídeos de 99,1 e 97,6 por cento, foram obtidos, sendo o primeiro composto de quase a totalidade das amostras analisadas. Linhagens de outros estados do Brasil "clustered" no II. CONCLUSÕES: A análise das sequências de aminoácidos da proteína N revelou que existem marcadores genéticos que podem determinar uma possível regionalidade embora as regiões biologicamente ativas apresentem-se conservadas dentro das espécies ao longo do tempo e espaço.

Antigenic and genetic characterization of the first rabies virus isolated from the bat Eumops perotis in Brazil.

Although the main transmitters of rabies in Brazil are dogs and vampire bats, the role of other species such as insectivorous and frugivorous bats deserves special attention, as the rabies virus has been isolated from 36 bat species. This study describes the first isolation of the rabies virus from the insectivorous bat Eumops perotis. The infected animal was found in the city of Ribeirão Preto, São Paulo. The virus was identified by immunofluorescence antibody test (FAT) in central nervous system (CNS) samples, and the isolation was carried out in N2A cell culture and adult mice. The sample was submitted to antigenic typing using a panel of monoclonal antibodies (CDC/Atlanta/USA). The DNA sequence of the nucleoprotein gene located between nucleotides 102 and 1385 was aligned with homologous sequences from GenBank using the CLUSTAL/W method, and the alignment was used to build a neighbor-joining distance-based phylogenetic tree with the K-2-P model. CNS was negative by FAT, and only one mouse died after inoculation with a suspension from the bat's CNS. Antigenic typing gave a result that was not compatible with the patterns defined by the panel. Phylogenetic analysis showed that the virus isolated segregated into the same cluster related to other viruses isolated from insectivorous bats belonging to genus Nyctinomops ssp. (98.8% nucleotide identity with each other).

Antigenic and genetic characterization of the first rabies virus isolated from the bat Eumops perotis in Brazil

Although the main transmitters of rabies in Brazil are dogs and vampire bats, the role of other species such as insectivorous and frugivorous bats deserves special attention, as the rabies virus has been isolated from 36 bat species. This study describes the first isolation of the rabies virus from the insectivorous bat Eumops perotis. The infected animal was found in the city of Ribeirão Preto, São Paulo. The virus was identified by immunofluorescence antibody test (FAT) in central nervous system (CNS) samples, and the isolation was carried out in N2A cell culture and adult mice. The sample was submitted to antigenic typing using a panel of monoclonal antibodies (CDC/Atlanta/USA). The DNA sequence of the nucleoprotein gene located between nucleotides 102 and 1385 was aligned with homologous sequences from GenBank using the CLUSTAL/W method, and the alignment was used to build a neighbor-joining distance-based phylogenetic tree with the K-2-P model. CNS was negative by FAT, and only one mouse died after inoculation with a suspension from the bat's CNS. Antigenic typing gave a result that was not compatible with the patterns defined by the panel. Phylogenetic analysis showed that the virus isolated segregated into the same cluster related to other viruses isolated from insectivorous bats belonging to genus Nyctinomops ssp. (98.8 percent nucleotide identity with each other).

No Brasil, embora os principais transmissores da raiva sejam cães e morcegos hematófagos, o papel de outras espécies, tais como morcegos insetívoros e frugívoros, merece atenção especial, uma vez que o vírus da raiva já foi isolado em 36 espécies de morcegos. Este estudo descreve o primeiro isolamento do vírus da raiva em um morcego insetívoro Eumops perotis. O animal infectado foi encontrado na cidade de Ribeirão Preto, São Paulo. O vírus foi identificado pelo teste de imunofluorescência direta (IFD) em amostras de sistema nervoso central (SNC), e o isolamento foi realizado em cultura de células N2A e em camundongos adultos. A amostra foi submetida à tipificação antigênica, utilizando um painel de oito anticorpos monoclonais (CDC/Atlanta/USA). A seqüência de DNA do gene da nucleoproteína, localizada entre os nucleotídeos 102 a 1385, foi alinhada com seqüências homólogas presentes no GenBank, usando o método CLUSTAL/W e o alinhamento foi utilizado para a construção da árvore filogenética de distância "neighbor-joining" com o modelo K-2-P. O SNC testado foi negativo por IFD, e somente um camundongo morreu após inoculação com a suspensão do SNC do morcego. A tipificação antigênica apresentou resultado não-compatível com os padrões definidos pelo painel. A análise filogenética mostrou que o vírus isolado segregou no mesmo grupo relacionado com outros vírus isolados de morcegos insetívoros, gênero Nyctinomops ssp. (98,8 por cento de identidade de nucleotídeos entre elas).

First report of rabies in vampire bats (Desmodus rotundus) in an urban area, Ubatuba, São Paulo state, Brazil.

The purpose of this report is to record the first case of a hematophagous bat (Desmodus rotundus) infected with rabies virus in an urban area in Brazil. To the authors' knowledge, this is the first such case in Latin America. After discovering a bat in his garden at 10 o'clock in the morning, a resident of Ubatuba municipality asked the Zoonosis Control Center team to visit his home. The animal was caught alive on the same day and sent to the Pasteur Institute laboratory, where it was identified as a Desmodus rotundus specimen. Standard tests for rabies diagnosis were carried out (direct immunofluorescence and viral isolation), and the results were positive. The presence of different species of (primarily insectivorous) bats in urban areas represents a serious public health problem. This case, however, is indicative of a much greater risk because the species in question has hematophagous habits, what means this animals has a low energy reserves and, therefore, its need to feed daily.

First report of rabies in vampire bats (Desmodus rotundus) in an urban area, Ubatuba, São Paulo state, Brazil

The purpose of this report is to record the first case of a hematophagous bat (Desmodus rotundus) infected with rabies virus in an urban area in Brazil. To the authors' knowledge, this is the first such case in Latin America. After discovering a bat in his garden at 10 o'clock in the morning, a resident of Ubatuba municipality asked the Zoonosis Control Center team to visit his home. The animal was caught alive on the same day and sent to the Pasteur Institute laboratory, where it was identified as a Desmodus rotundus specimen. Standard tests for rabies diagnosis were carried out (direct immunofluorescence and viral isolation), and the results were positive. The presence of different species of (primarily insectivorous) bats in urban areas represents a serious public health problem. This case, however, is indicative of a much greater risk because the species in question has hematophagous habits, what means this animals has a low energy reserves and, therefore, its need to feed daily.

Este relato tem por objetivo fazer o primeiro registro de morcego hematófago (Desmodus rotundus) infectado com o vírus da raiva, encontrado em área urbana de um município do Brasil e, até onde os autores têm conhecimento, na América Latina. Um munícipe de Ubatuba, São Paulo, solicitou a visita da equipe do Centro de Controle de Zoonoses em sua residência, após ter encontrado um morcego em seu quintal, às 10:00 horas da manhã. No mesmo dia o animal foi recolhido, ainda vivo, para ser encaminhado ao Laboratório do Instituto Pasteur. No Laboratório foi feita a identificação do espécime, Desmodus rotundus, e realizadas as provas clássicas para diagnóstico da raiva (Imunofluorescência Direta e Isolamento Viral), que resultaram positivas. A identificação de diferentes espécies de morcegos em áreas urbanas, predominantemente espécies insetívoras, representa um sério problema para a saúde pública. Este caso, no entanto, por tratar-se de espécie com hábitos hematofágicos, indica um risco ainda maior, tendo em vista a baixa reserva energética destes animais e a necessidade que têm de se alimentarem diariamente.