Transmisión vertical del virus de la rabia cría-madre, fenómeno que podría mantener al virus en especies reservorios de vida silvestre / Vertical transmission of the rabies virus from calf to mother, a phenomenon that could maintain the virus in reservoir species of wildlife

Resumen Introducción: La prueba biológica establecida por la Organización Mundial de la Salud para aislar y amplificar el virus de la rabia consiste en inocular vía intracraneal ratones lactantes para detectar signos de rabia en un periodo de 21 días. Objetivo: Constatar el contagio viral en las madres de ratones lactantes inoculados con virus de la rabia. Método: Veintisiete aislados mexicanos de virus de la rabia se inocularon vía intracraneal en ratones lactantes, los cuales fueron observados por 21 días y sus madres, por 60 días. El diagnóstico se llevó a cabo mediante inmunofluorescencia en cerebro. El virus se caracterizó por secuenciación y anticuerpos monoclonales. Resultados: Todas las camadas presentaron rabia entre siete y 15 días posinoculación (p. i.); tres de las 27 hembras (11 %), a los días 33, 37 y 39 p. i. de sus crías. La caracterización viral mostró que las madres se infectaron con la misma variante de sus crías, dos procedían de murciélago hematófago y una de perro. Las camadas que trasmitieron rabia a sus madres fueron nueve individuos. Conclusiones: En la naturaleza, el virus de la rabia podría preservarse mediante la transmisión de los neonatos (más susceptibles de contraer y amplificar el virus) a sus madres.

Abstract Introduction: The biological test established by the World Health Organization to isolate and amplify the rabies virus consists in inoculating lactating mice by intracranial route and detecting rabies signs for 21 days. Objective: To verify viral transmission in mothers of rabies virus-inoculated lactating mice. Method: Twenty-seven Mexican rabies virus isolates were inoculated by intracranial route in lactating mice, which were observed for 21 days. The mothers were observed for 60 days. The diagnosis was established by immunofluorescence in brain tissue. The virus was characterized by sequencing and with monoclonal antibodies. Results: All litters showed rabies at between 7 and 15 days post-inoculation (p. i.). Three of the 27 females (11 %) had developed rabies at days 33, 37 and 39 p. i. of their litters. Viral characterization showed that the mothers were infected with the same variant of their offspring, two of them stemming from hematophagous bat and one from dog. The liters that transmitted rabies to their mothers were nine individuals. Conclusions: In nature, the rabies virus could be preserved by transmission from neonates (more susceptible to contracting and amplifying the rabies virus) to their mothers.

Rabies virus monitoring in bat populations in Rondônia state, Brazil

Abstract INTRODUCTION: The Jirau hydroelectric power plant built in Rondônia state has environmental impacts that could be relevant to rabies outbreaks. METHODS: Bat populations were monitored for rabies by fluorescent antibody testing and simplified fluorescent inhibition microtesting between 2010 and 2015. RESULTS: All 1,183 bats tested negative for rabies. The prevalence of rabies antibodies was 17.5% in 1,049 bats. CONCLUSIONS: The rabies antibody dosage was not reactive in samples collected before the environmental changes, and there was a progressive increase in subsequent collections that could indicate an increase in rabies virus circulation among bats and risk of a rabies outbreak.

Comparative analysis of Mouse Inoculation Test and Virus Isolation in Cell Culture for rabies diagnosis in animals of Parana, Brazil

Abstract INTRODUCTION: Rabies is an acute zoonotic disease, caused by a rhabdovirus that can affect all mammals, and is commonly transmitted by the bite of a rabid animal. The definitive diagnosis is laboratorial, by the Fluorescent Antibody Test (FAT) as a quick test and Mouse Inoculation Test (MIT) as a confirmatory test (gold standard). Studies conducted over the past three decades indicate that MIT and Virus Isolation in Cell Culture (VICC) can provide the same effectiveness, the latter being considered superior in bioethics and animal welfare. The aim of this study was to compare VICC with MIT, in terms of accuracy, biosafety and occupational health, supply and equipment costs, bioethics and animal welfare, in a Brazilian public health lab. METHODS: We utilized 400 samples of animal neurological tissue to compare the performance of VICC against MIT. The variables analyzed were accuracy, biosafety and occupational health, time spent in performing the tests, supply and equipment costs, bioethics and animal welfare evaluation. RESULTS: Both VICC and MIT had almost the same accuracy (99.8%), although VICC presented fewer risks regarding biosafety and mental health of the technicians, and reduced time between inoculation and obtaining the results (approximately 22 days less). In addition, VICC presented lower supply costs (86.5% less), equipment costs (32.6% less), and the advantage of not using animals. CONCLUSIONS: These results confirm that VICC can replace MIT, offering the same accuracy and better features regarding cost, results, biosafety and occupational health, and bioethics and animal welfare.

VIROLOGICAL AND SEROLOGICAL DIAGNOSIS OF RABIES IN BATS FROM AN URBAN AREA IN THE BRAZILIAN AMAZON / Diagnóstico virológico e sorológico de raiva em morcegos de uma área urbana na Amazônia Brasileira

The outbreaks of rabies in humans transmitted by Desmodus rotundus in 2004 and 2005, in the northeast of the Brazilian State of Para, eastern Amazon basin, made this a priority area for studies on this zoonosis. Given this, the present study provides data on this phenomenon in an urban context, in order to assess the possible circulation of the classic rabies virus (RABV) among bat species in Capanema, a town in the Amazon basin. Bats were collected, in 2011, with mist nets during the wet and dry seasons. Samples of brain tissue and blood were collected for virological and serological survey, respectively. None of the 153 brain tissue samples analyzed tested positive for RABV infection, but 50.34% (95% CI: 45.67-55.01%) of the serum samples analyzed were seropositive. Artibeus planirostris was the most common species, with a high percentage of seropositive individuals (52.46%, 95% CI: 52.31 52.60%). Statistically, equal proportions of seropositive results were obtained in the rainy and dry seasons (c2 = 0.057, d.f. = 1, p = 0.88). Significantly higher proportions of males (55.96%, 95% CI: 48.96-62.96%) and adults (52.37%, 95% CI: 47.35-57.39%) were seropositive. While none of the brain tissue samples tested positive for infection, the high proportion of seropositive specimens indicates that RABV may be widespread in this urban area.

Os surtos de raiva em humanos transmitida por Desmodus rotundus em 2004 e 2005 no nordeste do estado do Pará, Brasil, Amazônia Oriental, fizeram desta uma área prioritária para estudos sobre essa zoonose. Diante disso, o presente estudo fornece dados sobre esse fenômeno em contexto urbano, afim de avaliar uma possível circulação do vírus clássico da raiva (RABV) entre espécies de morcegos em Capanema, cidade localizada na bacia Amazônica. Os morcegos foram coletados em 2011, com auxílio de redes de espera durante as estações seca e chuvosa. Amostras de encéfalo e de sangue foram coletadas para o diagnóstico virológico e sorológico, respectivamente. Das 153 amostras de encéfalo analisadas, nenhuma encontrou-se positiva para infecção pelo RABV, porém, 50,34% (95% CI: 45,67-55,01) das amostras de soro analisadas estavam soropositivas. Artibeus planirostris foi a espécie mais comum, e seu percentual de indivíduos soropositivos foi bem elevado (52.46%, 95% CI: 52,31-52,60). Porções estatisticamente iguais de soropositivos foram registrados nas estações (c2 = 0,057, d.f. = 1, p = 0,88). Uma porção significativamente elevada de machos (55,96%, 95% CI: 48,96%-62,96%), e adultos (52,37%, 95% CI: 47,35%-57,39%) foram soropositivos. Apesar de nenhuma das amostras de encéfalo terem sido positivas para raiva, a alta proporção de espécimes soropositivos indica uma possível circulação do RABV nessa área urbana.

Fluorescent antibody test, quantitative polymerase chain reaction pattern and clinical aspects of rabies virus strains isolated from main reservoirs in Brazil

ABSTRACTRabies virus (RABV) isolated from different mammals seems to have unique characteristics that influence the outcome of infection. RABV circulates in nature and is maintained by reservoirs that are responsible for the persistence of the disease for almost 4000 years. Considering the different pattern of pathogenicity of RABV strains in naturally and experimentally infected animals, the aim of this study was to analyze the characteristics of RABV variants isolated from the main Brazilian reservoirs, being related to a dog (variant 2),Desmodus rotundus (variant 3), crab eating fox, marmoset, and Myotis spp. Viral replication in brain tissue of experimentally infected mouse was evaluated by two laboratory techniques and the results were compared to clinical evolution from five RABV variants. The presence of the RABV was investigated in brain samples by fluorescent antibody test (FAT) and real time polymerase chain reaction (qRT-PCR) for quantification of rabies virus nucleoprotein gene (N gene). Virus replication is not correlated with clinical signs and evolution. The pattern of FAT is associated with RABV replication levels. Virus isolates from crab eating fox and marmoset had a longer evolution period and higher survival rate suggesting that the evolution period may contribute to the outcome. RABV virus variants had independent characteristics that determine the clinical evolution and survival of the infected mice.

Bat-borne rabis in latin america / Rabia transmitida por murciélagos en Latino América

The situation of rabies in America is complex: rabies in dogs has decreased dramatically, but bats are increasingly recognized as natural reservoirs of other rabies variants. Here, bat species known to be rabies-positive with different antigenic variants, are summarized in relation to bat conservation status across Latin America. Rabies virus is widespread in Latin American bat species, 22.5%75 of bat species have been confirmed as rabies-positive. Most bat species found rabies positive are classified by the International Union for Conservation of Nature as “Least Concern”. According to diet type, insectivorous bats had the most species known as rabies reservoirs, while in proportion hematophagous bats were the most important. Research at coarse spatial scales must strive to understand rabies ecology; basic information on distribution and population dynamics of many Latin American and Caribbean bat species is needed; and detailed information on effects of landscape change in driving bat-borne rabies outbreaks remains unassessed. Finally, integrated approaches including public health, ecology, and conservation biology are needed to understand and prevent emergent diseases in bats.

La situación de rabia en América es compleja: la rabia en perros ha disminuido drásticamente pero los murciélagos están siendo reconocidos cada vez más como reservorios naturales de otras variantes de rabia. Aquí compilamos las especies de murciélagos reconocidas como positivas a rabia con diferentes variantes antigénicas, así como su relación con el estado de conservación de los murciélagos a lo largo de América Latina. El virus de rabia está ampliamente distribuido en las especies de murciélagos de América Latina, 22.5% (75) de las especies de murciélagos conocidas han sido confirmadas como especies positivas a rabia. La mayoría de las especies de murciélagos reportadas como positivas a rabia son clasificadas por la Unión Internacional para la Conservación de la Naturaleza como “Preocupación Menor”. De acuerdo al tipo de dieta, los murciélagos insectívoros tuvieron la mayor cantidad de especies reconocidas como reservorio del virus rabia, mientras en proporción los hematófagos fueron los más importantes. Investigaciones a escala gruesa deben buscar entender aspectos de ecología de la rabia; es necesaria la información básica sobre la distribución y dinámica poblacional para muchas especies de murciélagos de América Latina y el Caribe; y el efecto del cambio del paisaje en la generación de brotes de rabia transmitida por murciélagos permanece sin ser evaluado. Por último, para entender y prevenir enfermedades emergentes a partir de los murciélagos es necesario un enfoque integral incluyendo salud pública, ecología y biología de la conservación.

Comparison of a modified shell vial culture procedure with conventional mouse inoculation for rabies virus isolation

Rabies is a neurotropic disease that is often lethal. The early diagnosis of rabies infection is important and requires methods that allow for the isolation of the virus from animals and humans. The present study compared a modified shell vial (MSV) procedure using 24-well tissue culture plates with the mouse inoculation test (MIT), which is considered the gold standard for rabies virus isolation. Thirty brain samples (25 positive and 5 negative by the fluorescent antibody test) obtained from different animal species at the National Institute of Hygiene Rafael Rangel in Caracas, Venezuela, were studied by the MIT and MSV assays. Nine samples (36%) were positive at 24 h, 10 (40%) were positive at 48 h and six (24%) were positive at 72 h by the MSV assay. With the MIT assay, 76% were positive at six days post inoculation and 12% were positive at 12 and 18 days post inoculation. One sample that was negative according to the MSV assay was positive with MIT on the 12th day. The MSV procedure exhibited a sensitivity of 96.2%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value 80%. This procedure allowed for rapid rabies virus detection. MIT can be employed as an alternative method in laboratories without tissue culture facilities.

Circulation of the rabies virus in non-hematophagous bats in the city of Rio de Janeiro, Brazil, during 2001-2010 / Circulação do vírus da raiva em morcegos não-hematófagos na cidade do Rio de Janeiro no período entre 2001-2010

INTRODUCTION: Rabies is one of the most known lethal zoonosis, responsible for 55,000 human deaths per year. It is transmitted to humans mainly by the bite of domestic or wild animals infected with the virus. This paper shows the circulation of this virus in non-hematophagous bats in the City of Rio de Janeiro, Brazil. METHODS: A survey was performed on the number of bats that had been sent for diagnosis by the Seção de Virologia of the Instituto Municipal de Medicina Veterinária Jorge Vaitsman and were positive for rabies. The positive animals were identified, and the isolated viruses were sent for antigenic typification with indirect immunofluorescence. The results were compared with the antigenic panel of the Centers for Disease Control and Prevention. RESULTS: During 2001-2010, the laboratory received 555 non-hematophagous bats for rabies diagnosis, with 198 (35.7%) from Rio de Janeiro City. A total of 11 (5.5%) animals were positive for this disease. Antigenic typification revealed the predominance of variant 3 in 9 (81.8%) of the isolated viruses; 1 virus was classified as variant 4 and 1 variant was identified that segregated with the viruses in insectivorous bats. CONCLUSIONS: The data obtained in this study showed the presence of the rabies virus in synanthropic populations of non-hematophagous bats in the City of Rio de Janeiro. The circulation of this agent in these animals represents a serious risk to human and animal health and requires attention and control measures by the authorities.

INTRODUÇÃO: A raiva é uma das mais letais zoonoses conhecidas, responsável pelo óbito de 55 mil pessoas anualmente. É transmitida ao homem principalmente pela mordida de animais, domésticos ou silvestres, infectados pelo vírus. O presente estudo mostra a circulação deste agente em morcegos não-hematófagos no município do Rio de Janeiro, Brasil. MÉTODOS: Na Seção de Virologia do Instituto Municipal de Medicina Veterinária Jorge Vaitsman, foi realizado um levantamento do número de morcegos recebidos positivos para o diagnóstico. Os animais positivos foram identificados, e o vírus isolado foi submetido à tipificação de variantes antigênicas pela técnica de imunofluorescência indireta, com os resultados comparados com o painel antigênico utilizado pelo Centro de Controle de Doenças (Center of Disease Control -CDC). RESULTADOS: Entre 2001-2010, o laboratório recebeu 555 morcegos nãohematófagos para o diagnóstico da raiva, sendo 135 (35,5%) do município do Rio de Janeiro. Um total de 11 (5,5%) animais foram positivos para a doença. A tipificação antigênica revelou a predominância da variante 3, com nove (81,9%) vírus isolados, um pertencente a variante 4 e outro de uma variante que segrega com a presente em morcegos insetívoros. CONCLUSÕES: Os dados observados no presente estudo demonstraram a presença do vírus da raiva em populações sinantrópicas de morcegos não-hematófagos no município do Rio de Janeiro. A circulação deste agente nesses animais representa um grave risco a saúde humana e animal, e requer atenção e medidas de controle e prevenção por parte das autoridades competentes.

Molecular characterization of rabies virus isolated from non-haematophagous bats in Brazil / Caracterização molecular do vírus da raiva isolado de morcegos não-hematófagos no Brasil

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.

INTRODUÇÃO: A raiva é uma importante zoonose responsável por milhares de mortes anualmente em todo o mundo. Embora o ciclo silvestre, onde os cães são os principais transmissores esteja controlado no Brasil, o ciclo aéreo, onde o morcego hematófago Desmodus rotundus é o principal transmissor e diversas espécies de morcegos não hematófagos podem se infectar e transmitir o vírus, permanence como um importante problema econômico e de saúde pública. MÉTODOS: O objetivo deste trabalho foi a caracterização antigênica por meio da utilização de anticorpos monoclonais e a caracterização genética por meio da reação em cadeia pela polimerase pela transcriptase reversa seguida de análise filogenética em morcegos não hematófagos e animais domésticos herbívoros provenientes da região oeste do Estado de São Paulo. RESULTADOS: A análise antigênica de 27 amostras determinou três variantes distintas: Agv-3, AgV-4 e AgV-6; a análise genética de 29 amostras identificou 5 diferentes grupos, todos pertencentes a espécie Rabies virus. CONCLUSÕES: Ainda que apenas amostras de morcegos não hematófagos tenham sido analisadas, a maioria das variantes antigênicas e genéticas identificadas na região estava relacionada com a variante mantida pelos morcegos hematófagos Desmodus rotundus. Amostras de uma mesma variante antigênica segregaram em mais de um clado genético. Este estudo demonstrou a diversidade de linhagens genéticas do vírus da raiva presentes e circulantes em morcegos não hematófagos na região estudada.

Rabies virus in Molossus molossus (Chiroptera: Molossidae) in the State of Pernambuco, Northeastern Brazil / Vírus rábico em Molossus molossus (Chiroptera: Molossidae), no Estado de Pernambuco, nordeste do Brasil

Isolamento do vírus rábico em morcegos (Molossus molossus) em área urbana, no município de Recife, Pernambuco, Brasil. Quatro exemplares foram encontrados durante o dia em locais visíveis e não habituais caídos no chão ainda com vida, não havendo contato com pessoas ou animais. Destes apenas dois foram identificados; porém, não foi possível fazer a identificação em dois espécimes, pois os mesmos foram incinerados antes da identificação. O diagnóstico foi positivo para as provas de imunofluorescência direta e inoculação intracerebral em camundongos. O presente estudo apresenta o isolamento de vírus rábico pela primeira vez no Estado de Pernambuco em morcegos insetívoros.

Rabies virus was detected in bats (Molossus molossus) from an urban area in the City of Recife, State of Pernambuco, Brazil. Four individuals were found during the day in visible, non-habitual places, lying on the ground, but still alive. No contact occurred with people or animals. Of these, only two were identified; it was not possible to identify two specimens, since they were incinerated prior to identification. Diagnosis was positive by direct immunofluorescence and intracerebral inoculation in mice. This study presents the first instance in which the virus was detected in insectivorous bats in the State of Pernambuco.

Descripción epidemiológica del reservorio de rabia en murciélagos de la Región Metropolitana: Chile. 2000-2009 / Epidemiological description of rabies reservoir in bats in the Metropolitan Region: Chile. 2000-2009

In Chile, in 1985 rabies was detected in insectivorous bats. Since then the epidemiological surveillance activities was extended to these species and the epidemiological pattern was characterized as endemic in bats. In this study we analyzed positive rabies cases in the Metropolitan Region between 2000 and 2009. We identified 325 cases of rabies in 11.472 analyzed samples. We determined an increase of positivity cases in bats between the years of the study. Places with the highest number of positives cases were Las Condes, Santiago, Providencia, Puente Alto and Ñuñoa. In 147 cases we identify the circumstances in which the bat was found: 89 inside, 45 outside the building and 13 in public spaces. In 39 cases there was contact with humans or animals. These results reinforce the relevance of educating the population against the contact with bats and allow health authorities to take early surveillance and control measures.

En Chile, en 1985, se detectó por primera vez rabia en murciélagos insectívoros. Desde entonces, las acciones de vigilancia epidemiológica se ampliaron hacia estas especies, caracterizándose el patrón epidemiológico como una endemia en quirópteros. En este estudio se analizaron epidemiológicamente los casos positivos a rabia en la Región Metropolitana, entre los años 2000 y 2009. Se identificaron 325 casos de rabia en murciélagos de un total de 11.472 analizados. Se determinó un aumento de positividad en los murciélagos entre los años analizados. Las comunas con mayor número de positivos fueron Las Condes, Santiago, Providencia, Puente Alto y Ñuñoa. En 147 casos se señalan las circunstancias en que se encontró el murciélago: 89 al interior, 45 al exterior del inmueble y 13 en la vía pública. En 39 casos hubo contacto con personas o animales. Estos resultados reafirman la importancia de la educación de la población frente al contacto con murciélagos y permiten a las autoridades sanitarias tomar las medidas de vigilancia y control en forma oportuna.

Risco de transmissão do vírus da raiva oriundo de sagui (Callithrix jacchus), domiciliado e semidomiciliado, para o homem na região metropolitana de Fortaleza, estado do Ceará / Risks of transmitting rabies virus from captive domiciliary common marmoset (Callithrix jacchus) to human beings, in the metropolitan region of Fortaleza, state of Ceará, Brazil

INTRODUÇÃO: Uma variante do vírus da raivafoi identificadaem associação a casos de raiva humanos, no Estado do Ceará, transmitidos por saguis (Callithrix jacchus), primatas frequentemente criados como animais de estimação. Essa variante não apresenta proximidade antigênica ou relação genética com as variantes do vírus encontradas em morcegos e mamíferos terrestres das Américas. O objetivo do estudo foi avaliar os fatores de risco de transmissão do vírus da raiva oriundo de sagui (C. jacchus), criado como animal de estimação, para o homem na região metropolitana de Fortaleza, Ceará. MÉTODOS: Foi aplicado um questionário estruturado aos criadores de saguis, residentes nos municípios de Aquiraz e Maranguape, Ceará, enfocando o manejo e a interação desses primatas com humanos. Para avaliação da ocorrência de antígenos rábicos, através do teste de imunofluorescência direta (IFD), foram coletadas amostras de saliva dos saguis domiciliados e semidomiciliados. Com base nos resultados obtidos desses espécimes, foram analisadas amostras de sistema nervoso central (SNC). RESULTADOS: Na análise dos questionários, observou-se a proximidade dos criadores de saguis durante o manejo desses animais nos domicílios, bem como, seus conhecimentos limitados sobre a raiva, demonstrando haver risco quanto à transmissão do vírus. De 29 amostras de saliva de saguis reavaliadas, uma (3,4 por cento) apresentou reação de IFD positiva. De 11 amostras de SNC, três (27,3 por cento) apresentaram positividade. CONCLUSÕES: Os dados laboratoriais estão de acordo com os achados dos questionários, confirmando haver risco da transmissão do vírus da raiva devido à convivência de humanos com saguis (C. jacchus).

INTRODUCTION: In the State of Ceará, a new variant of the rabies virus was identified associated with cases of human rabies transmitted by common marmosets (Callithrix jacchus), which are frequently kept as pets. This new variant does not present antigenic proximity or genetic relationship to variants of the virus isolated from bats and terrestrial mammals from the American continent. The present study aimed to evaluate the risk factors of rabies virus transmission from common marmosets (C. jacchus) maintained as pets in the metropolitan region of Fortaleza, State of Ceará, Brazil, to human beings. METHODS: A questionnaire focusing on animal management and interaction between humans and primates was applied to individuals who had marmosets in the municipalities of Aquiraz and Maranguape. In order to evaluate the presence of rabies antigens by direct immunofluorescence test (DIF), samples of saliva were collected from domiciliary captive marmosets. Based on the detection of rabies antigens, biopsy samples of central nervous system (CNS) were analyzed. RESULTS: Analysis of questionnaire data verified that a close relation exists between humans and their pet marmosets, especially during management practices. Additionally, these people showed minimal knowledge regarding rabies, which represents a greater risk of infection. Of the 29 saliva samples evaluated, one (3.4 percent) was positive for DIF reaction and of the 11 CNS samples, three (27.3 percent) were positive. CONCLUSIONS: Laboratory data are in agreement with the questionnaire findings, which confirm an increased risk of rabies virus transmission due to the close relation between humans and marmosets.

Os morcegos e a raiva na região oeste do Estado de São Paulo / The bats and rabies in the Western region of the State of São Paulo, Brazil

INTRODUÇÃO: O laboratório do Pólo da Alta Sorocabana de Presidente Prudente, SP, em parceria com outras instituições de pesquisa, realizou estudos pertinentes aos morcegos da região oeste do Estado de São Paulo, Brasil. Para tal, foram pesquisadas algumas situações, tais como: a) isolamento do vírus rábico, no período 2006 a 2008; b) as respectivas variantes antigênicas; c) abrigos diurnos do morcego hematófago Desmodus rotundus. MÉTODOS: As amostras para exame foram provenientes de morcegos não hematófagos encaminhadas ao laboratório sendo submetidas aos testes de imunofluorescência direta e prova biológica. As amostras positivas foram caracterizadas antigenicamente por meio do teste de anticorpos monoclonais. Quanto aos morcegos, foram identificados e classificados, e também foi realizado mapeamento de abrigos dos mesmos. RESULTADOS: O laboratório recebeu 1.113 morcegos não hematófagos para diagnóstico laboratorial, sendo 11 (1 por cento) deles positivos, e dentre as amostras positivas, 5 (45,5 por cento) delas tiveram variante antigênica 3 associada ao morcego D. rotundus e 4 (36,5 por cento) foram compatíveis com amostras de morcegos insetívoros. Foram pesquisados 16 abrigos de morcegos hematófagos e observou-se a presença de outras 3 espécies de morcegos não hematófagos convivendo com eles. CONCLUSÕES: Os experimentos mostraram que o vírus rábico continua circulando na região com pelo menos 3 variantes antigênicas, e que, a coabitação de morcegos hematófagos com não hematófagos pode ter alguma relação com a disseminação do vírus rábico.

INTRODUCTION: 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. METHODS: 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. RESULTS: The laboratory received 1,113 nonhematophagous bats for rabies diagnosis, 11 (1 percent) of which were positives, and among the positive samples, 5 (45.5 percent) presented antigenic variant 3 (from the bat Desmodus rotundus) and 4 (36.5 percent) 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. CONCLUSIONS: 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.

Rabies diagnosis and serology in bats from the State of São Paulo, Brazil / Diagnóstico e sorologia de raiva em morcegos do Estado de São Paulo, Brasil

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 percent) bats were positive for rabies. The prevalence of rabies antibodies was 5.9 percent using 0.5IU/ml as a cutoff. Insectivorous bats (69.8 percent) and bats of the species Molossus molossus (51.8 percent) 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 por cento) morcegos foram positivos para raiva. A prevalência de anticorpos foi de 5,9 por cento usando 0,5UI/ml como ponto de corte. Os morcegos de hábito alimentar insetívoro (69,8 por cento) e os morcegos da espécie Molossus molossus (51,8 por cento) 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.

Characterization of rabies virus isolated from a colony of Eptesicus furinalis bats in Brazil / Caracterização do vírus da raiva isolado de uma colônia de morcegos Eptesicus furinalis, do Brasil

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.

Caracterização molecular do vírus da raiva isolado de Desmodus rotundus capturados no Estado do Rio de Janeiro / Molecular characterization of rabies virus isolated from Desmodus rotundus captured in Rio de Janeiro State

Caracterizou-se filogeneticamente o vírus da raiva, isolado de morcegos hematógafos (Demodus rotundus). Cento e noventa e nove D. rotundus foram capturados em cinco abrigos, no Norte e Noroeste do Estado do Rio de Janeiro e sul do Espírito Santo. Sete deles foram positivos para a raiva. Amostras desses vírus foram sequenciadas e comparadas com sequências provenientes de diversos estados brasileiros. As sequências de vírus da raiva isoladas, na região norte do Estado do Rio de Janeiro, mostraram características que as distinguem de amostras de vírus isoladas em outras regiões do país, no entanto foram idênticas às isoladas de bovinos no noroeste do Rio de Janeiro.

Rabies samples isolated from vampire bats captured in the Rio de Janeiro State were phylogenetically analyzed. One hundred and ninety nine vampire bats were captured from five shelters from North and Northwest of Rio de Janeiro and South of Espírito Santo States. Seven of them were positive for rabies. Theses samples were sequenced and compared with rabies virus sequences from several Brazilian states. The sequences of rabies virus, isolated in the present work, from North of Rio de Janeiro State, showed characteristics that differ of the sequences isolated from bats from other Brazilian regions. However, they were identical to samples isolated from cattle in Northwest of Rio de Janeiro state.

Updated list of bat species positive for rabies in Brazil / Lista atualizada das espécies de morcegos positivas para raiva no Brasil

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 percent, Vespertilionidae 29.3 percent and Molossidae 26.8 percent. 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.

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 por cento, Vespertilionidae 29.3 por cento e Molossidae 26.8 por cento. 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.

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.

Cobertura e uso do solo e sua influência na ocorrência de raiva nos municípios de Jacareí e Santa Branca, Vale do Paraíba, Estado de São Paulo, entre 2002 e 2009 / Influence of landscape and land use on the occurrence of rabies in the municipalities of Jacareí and Santa Branca, Vale do Paraíba, State of São Paulo, between 2002 and 2009

Objetivo: Realizou-se um estudo, no período de 2002 a 2009, com a finalidade de verificar se as mudanças nas classes de cobertura da terra e no uso do solo podem exercer influência na ocorrência da raiva, nos municípios de Jacareí e de Santa Branca, situados na Região do Vale do Paraíba, no Estado de São Paulo, Sudeste do Brasil. Métodos: A ferramenta utilizada para avaliar estas alterações foi o sensoriamento remoto, através de imagens de satélite Land-sat. Para pesquisar a presença do vírus da raiva (RABV) foram coletados animais silvestres atropelados nas rodovias, morcegos encontrados na área urbana em atitude suspeita, morcegos hematófagos da espécie Desmodus rotundus e animais de interesse econômico (ADIE) que vieram a óbito por enfermidade com sintomatologia nervosa. O material coletado, sistema nervoso central (SNC), desses animais foi encaminhado para o laboratório de referência nacional, o Instituto Pasteur de São Paulo (IP-SP) e para o Laboratório de Sanidade Animal da Agência Paulista de Tecnologia dos Agronegócios (APTA)- Pólo Regional do Vale Paraíba (PRDTA/VP) - Pindamonhangaba-SP. A determinação da presença do antígeno viral foi feita através da prova de imunofluorescência direta (IFD) e o isolamento do vírus através da inoculação intracerebral em camundongos (IC). Os inóculos das amostras foram submetidos à prova de reação em cadeia pela polimerase transcriptase reversa (RT-PCR) para verificar a presença do vírus da raiva. Resultados: No período de estudo tivemos a ocorrência de dez casos da doença, sendo três em morcegos insetívoros na zona urbana de Jacareí, e sete em ADIE, sendo dois, em Jacareí e cinco, em Santa Branca. Todos os animais silvestres terrestres examinados foram negativos para a presença do vírus da raiva em todas as provas realizadas (IFD, IC, RT-PCR)...

Objective: This study was carried out during the period of 2002-2009, with the purpose to verify whether the changes in types of land cover and in land use can influence the occurrence of rabies in Jacareí and Santa Branca, situated in the Vale do Paraíba, State of São Paulo, southeastern brazil. Methods: The tool used to evaluate these alterations was the remote sensing through images of satellite land-sat. In order to search for the presence of rabies virus, brain materials were collected from wild animals roadkilled in highways, bats found in the urban area in suspicious attitude, desmodus rotundus hematophagous bats captured in rural areas and farm animals of economic importance (aei) dead with suspect of rabies. For rabies diagnosis, specimens of central nervous system (cns) of these animals were sent to the Pasteur Institute of São Paulo (IP-SP) - national reference laboratory and to the laboratory of animal Paulista Agency for Agribusiness Technology (APTA) - Polo Valley Regional of Paraíba (PRDTA / VP), in Pindamonhangaba, state of SP. The presence of viral antigen was determined by the direct fluorescent antibody technique (D-FAT) and the isolation of the virus by means of intracerebal mouse inoculation test (MIT). The brain suspensions were submitted to reverse transcriptase. Polymerase chain reaction to check the presence of rabies virus. Results: In the period of study, 10 positive cases were detected, being 3 in insectivorous bats in the urbane zone of jacareí, and 7 in aei, being 2 in jacareí and 5 in santa branca. All the terrestrial wild animals examined were negative for the presence of rabies virus in all tests performed (d-fat, mit, and rt-pcr)...

Diagnóstico de un caso de rabia humana en Jujuy, 2008 / Diagnosis of a human rabies case in Jujuy, 2008

El 22 de Julio de 2008, un niño de 8 años de edad, residente en la provincia de Jujuy, Argentina, falleció por una encefalitis producida por el virus de la rabia. El diagnóstico se realizó mediante la detección de anticuerpos en suero y se confirmó por inmunofluorescencia en el cerebro. La tipificación antigénica correspondió a la variante 1 trasmitida por perros. El análisis molecular estableció que el virus detectado es de la misma variante genética que circula en Jujuy desde 2003. Este trabajo resume la evolución clínica del paciente y la posterior investigación epidemiológica que reveló el antecedente de mordedura por un perro 60 días antes de la iniciación de la enfermedad y la ausencia de un tratamiento antirrábico post-exposición.

On July 22, 2008, a previously healthy 8 years old boy from Jujuy, Argentina, died of encephalitis later confirmed as rabies. Diagnosis was made on the basis rabies-specific antibodies presence in a serum sample and it was confirmed by detection of the viral antigens in brain necropsy using the inmunofluorescent test. Antigenic characterization identified dog as source of infection. Molecular analysis recognized the same genetic variant circulating in Jujuy since 2003. This report presents the patient's clinical course and the epidemiologic investigation that revealed a dog bite 60 days before the illness onset and the lack of rabies treatment.