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Among 713 equids sampled in northeastern Brazil during 2013-2018, West Nile virus seroprevalence was 4.5% (95% CI 3.1%-6.3%). Mathematical modeling substantiated higher seroprevalence adjacent to an avian migratory route and in areas characterized by forest loss, implying increased risk for zoonotic infections in disturbed areas.

Coronaviruses belong to a well-known family of enveloped RNA viruses and are the causative agent of the common cold. Although the seasonal coronaviruses do not pose a threat to human life, three members of this family, i.e., SARS-CoV, MERS-CoV and recently, SARS-CoV2, may cause severe acute respiratory syndrome and lead to death. Unfortunately, COVID-19 has already caused more than 4.4 million deaths worldwide. Although much is better understood about the immunopathogenesis of the lung disease, important information about systemic disease is still missing, mainly concerning neurological parameters. In this context, we sought to evaluate immunometabolic changes using in vitro and in vivo models of hamsters infected with SARS-CoV-2. Here we show that, besides infecting hamster’s astrocytes, SARS-CoV-2 induces changes in protein expression and metabolic pathways involved in carbon metabolism, glycolysis, mitochondrial respiration, and synaptic transmission. Interestingly, many of the differentially expressed proteins are concurrent with proteins that correlate with neurological diseases, such as Parkinsons’s disease, multiple sclerosis, amyotrophic lateral sclerosis, and Huntington’s disease. Metabolic analysis by high resolution real-time respirometry evidenced hyperactivation of glycolysis and mitochondrial respiration. Further metabolomics analysis confirmed the consumption of many metabolites, including glucose, pyruvate, glutamine, and alpha ketoglutarate. Interestingly, we observed that glutamine was significantly reduced in infected cultures, and the blockade of mitochondrial glutaminolysis significantly reduced viral replication and pro-inflammatory response. SARS-CoV-2 was confirmed in vivo as hippocampus, cortex, and olfactory bulb of intranasally infected hamsters were positive for viral genome several days post-infection. Altogether, our data reveals important changes in overall protein expression, mostly of those related to carbon metabolism and energy generation, causing an imbalance in important metabolic molecules and neurotransmitters. This may suggest that some of the neurological features observed during COVID-19, as memory and cognitive impairment, may rely on altered energetic profile of brain cells, as well as an unbalanced glutamine/glutamate levels, whose importance for adequate brain function is unquestionable.

Screening of 533 bats for influenza A viruses showed subtype HL18NL11 in intestines of 2 great fruit-eating bats (Artibeus lituratus). High concentrations suggested fecal shedding. Genomic characterizations revealed conservation of viral genes across different host species, countries, and sampling years, suggesting a conserved cellular receptor and wide-ranging occurrence of bat influenza A viruses.

Bats are notorious reservoirs of genetically-diverse and high-profile pathogens, and are playing crucial roles in the emergence and re-emergence of viruses, both in human and in animals. In this report, we identified and characterized previously unknown and diverse genetic clusters of bat coronaviruses in the Atlantic Forest Biome, Brazil. These results highlight the virus richness of bats and their possible roles in the public health., Highlights • We describe the circulation of CoV lineages, 13 α-CoV and two β-CoV in AFB bats. • Coronaviruses were detected in 15 bat samples from eight bat species (50% positivity). • A cluster of α-CoVs were detected in Sturnira bat that grouped with α-CoVs lineage-1. • We identified two distinct β-CoV that clustered within MERS-CoV containing lineage C. • Results highlight the virus richness of bats and their possible roles in the public health.

The World Health Organization (WHO), reports that rabies causes tens of thousands of deaths every year killing humans, non-human primates and other animals. Rabies continues to be a public health issue, despite the existence of effective vaccines. The dogs remain the primary reservoir and transmitter of rabies to humans globally. In the Americas, bats are regarded as the second most common source of rabies virus to humans. The vampire bat Desmodus rotundus has been identified as a natural reservoir of rabies virus (RABV) in this region. The complete genome of the RABV variant maintained by populations of vampire bats D. rotundus has rarely been reported. In this study, we sequenced and analyzed the genome of a RABV variant detected in D. rotundus. The sample, collected from an endemic area in São Paulo State, was phylogenetically compared with the genome of the standard sample for species Rabies virus as well as other samples belonging to terrestrial and bat-associated cycles of rabies transmission, available in GenBank. Distinct patterns linked to the genetic lineage were identified. These data can aid in the understanding of the molecular epidemiology of this virus and the epidemiological importance of this species in the transmission of the RABV.

Human respiratory syncytial virus (HRSV) is an important respiratory pathogens among children between zero-five years old. Host immunity and viral genetic variability are important factors that can make vaccine production difficult. In this work, differences between biological clones of HRSV were detected in clinical samples in the absence and presence of serum collected from children in the convalescent phase of the illness and from their biological mothers. Viral clones were selected by plaque assay in the absence and presence of serum and nucleotide sequences of the G2 and F2 genes of HRSV biological clones were compared. One non-synonymous mutation was found in the F gene (Ile5Asn) in one clone of an HRSV-B sample and one non-synonymous mutation was found in the G gene (Ser291Pro) in four clones of the same HRSV-B sample. Only one of these clones was obtained after treatment with the child's serum. In addition, some synonymous mutations were determined in two clones of the HRSV-A samples. In conclusion, it is possible that minor sequences could be selected by host antibodies contributing to the HRSV evolutionary process, hampering the development of an effective vaccine, since we verify the same codon alteration in absence and presence of human sera in individual clones of BR-85 sample.

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.

This epidemiological study was conducted using antigenic and genetic characterisation of rabies virus isolates obtained from different animal species in the southeast of Brazil from 1993 to 2007. An alteration in the epidemiological profile was observed. One hundred two samples were tested using a panel of eight monoclonal antibodies, and 94 were genetically characterised by sequencing the nucleoprotein gene. From 1993 to 1997, antigenic variant 2 (AgV-2), related to a rabies virus maintained in dog populations, was responsible for rabies cases in dogs, cats, cattle and horses. Antigenic variant 3 (AgV-3), associated with Desmodus rotundus, was detected in a few cattle samples from rural areas. From 1998 to 2007, rabies virus was detected in bats and urban pets, and four distinct variants were identified. A nucleotide similarity analysis resulted in two primary groups comprising the dog and bat antigenic variants and showing the distinct endemic cycles maintained in the different animal species in this region.

The Human Respiratory Syncytial Virus (HRSV), isolated in 1955, is the main cause of hospitalization of babies and infants with respiratory illness. Several studies have been conducted worldwide aiming the development of a safe and effective vaccine against HRSV. The G2 region of glycoprotein G is used as genotyping default. In the present study, we performed a phylogenetic analysis of G protein and a comparative study between G2 region and ectodomain of attachment glycoprotein. Fifty-three nasal swab samples from children less than 5 years old and presenting symptoms of acute respiratory illness, assisted at the University Hospital (UH) of University of Sao Paulo (USP) in 2004, were submitted to sequencing by PCR and compared with GenBank sequences. We concluded that the G2 region is adequate for HRSV genotyping.O vírus respiratório sincicial humano (HRSV), isolado em 1955, é a principal causa da hospitalização de bebês e crianças pequenas com sintomas de doença respiratória. No mundo inteiro, vários estudos para o desenvolvimento de uma vacina segura e eficiente contra o HRSV têm tido alta prioridade. A região G2 da glicoproteína G é usada como padrão para genotipagem do HRSV. Neste estudo, foi realizada a análise filogenética da glicoproteína G e o estudo comparativo entre a região G2 e o ectodomínio dessa glicoproteína. Cinquenta e três amostras de swab nasal de crianças com menos de cinco anos de idade, apresentando doença respiratória aguda, atendidas no Hospital Universitário (HU) da Universidade de São Paulo durante o ano de 2004, foram submetidas a sequenciamento por PCR e comparadas com seqüências do GenBank. A região G2 mostrou ser adequada para a genotipagem do HRSV.

Dados da Organização Mundial da Saúde (OMS) mostram que a raiva é um problema de saúde pública podendo acarretar sérios prejuízos ambientais e econômicos, a despeito da existência de vacinas eficazes de uso humano e veterinário. Segundo seu último informe, estima-se que no mundo em torno de 55.000 pessoas por ano morrem de raiva. O cão permanece como principal transmissor da raiva para o homem e também como principal vítima da doença. Nos países que conseguiram controlar a raiva em animais domésticos, o vírus se mantém circulante na natureza por meio dos animais silvestres, sendo os morcegos apontados como a segunda espécie transmissora da raiva a humanos. Os Lyssavirus têm sido detectados em morcegos, em diversos continentes, sendo identificados como transmissor em dez das onze espécies de Lyssavirus. Fósseis de morcego mostram sua presença há 50 milhões de anos. Mas somente em 1911, Carini relacionou pela primeira vez a raiva aos morcegos, levantando a hipótese destes serem os transmissores da doença a outros animais. Há registros de que o vírus da raiva foi isolado em pelo menos 41 das 167 espécies de morcegos brasileiras, sendo que a maioria dessas espécies está relacionada a atividades humanas com a presença destes animais próximos ao local de trabalho e moradia das pessoas. Os morcegos hematófagos Desmodus rotundus são encontrados do norte do México até a costa norte do Chile, região central da Argentina e costa do Uruguai e com exceção do Chile. Esta espécie de morcego tem sido apontada como reservatório natural do vírus da raiva nesta região. Alguns pesquisadores observaram que a raiva em morcegos não hematófagos precede a raiva bovina e em animais de estimação, sugerindo que os morcegos não hematófagos podem ser o elo entre a raiva silvestre e a raiva urbana e o fato de se detectar a variante mantida por morcegos hematófagos Desmodus rotundus em cães e gatos mostra que o papel deste morcego no ciclo da raiva não está limitado à raiva silvestre. As características dos Lyssavirus adaptados a morcegos têm mostrado diferenças quando comparadas à raiva relacionada aos carnívoros, confirmando a necessidade do desenvolvimento de metodologias que permitam estudos complementares mais precisos a respeito da biologia e epidemiologia da raiva em quirópteros. A escassez de dados na literatura, até o momento, a respeito do genoma completo da variante do vírus da raiva mantida por populações de morcegos hematófagos Desmodus rotundus, deixa uma lacuna no entendimento da epidemiologia molecular deste vírus. A importância epidemiológica desta espécie na transmissão da raiva é inquestionável. Neste estudo foi sequenciado e analisado, o genoma da variante do vírus da raiva mantido por populações de morcego hematófago Desmodus rotundus isolado de um morcego hematófago Desmodus rotundus. A amostra, procedente de área endêmica no Estado de São Paulo, foi filogeneticamente comparada com o genoma da amostra padrão para a espécie viral 1 - Rabies virus e outras amostras pertencentes ao ciclo aéreo ou terrestre de transmissão, disponíveis no GenBank, identificando possíveis padrões de diferenciação, próprios do ciclo aéreo, e em alguns casos relacionados somente à variante estudada. Data from the World Health Organization (WHO) show that rabies is a public health problem which can cause serious environmental and economic damage, despite the existence of effective vaccines for human and veterinary use. According to WHO latest report, estimated that worldwide around 55,000 people per year died of rabies. The dog remains the main transmitter of rabies to humans as well as the main victim of the disease. In countries that were successful in controlling rabies in domestic animals, the virus is still circulating in nature by wild animals and the bats are seen as the second species transmitting rabies to humans. The Lyssavirus have been detected in bats in several continents and is identified as a transmitter in ten of eleven species of Lyssavirus. Bat fossils show their presence for 50 million years. But only in 1911, in the first time Carini related to rabies at bats, raising the possibility of these being the transmitters of the disease to other animals. Reports show that the Rabies virus was isolated in at least 41 of the 167 species of bats in Brazil, with the majority of these species is related to human activities with the animals living near the local job and houses of people. The vampire bat Desmodus rotundus is found from northern Mexico to northern Chile coast, central coast of Argentina and Uruguay and with the exception of Chile. This bat species has been identified as a natural reservoir of the Rabies virus in this region. Some researchers observed that rabies into non-hematophagous bats precedes the bovine rabies and in pets, suggesting that the non-hematophagous bats may be the link between wildlife rabies and urban rabies and the fact that detect the variant maintained by vampire bats Desmodus rotundus in dogs and cats shows that the role of bat rabies in the cycle is not limited to wildlife rabies. The characteristics of Lyssavirus bat adapted have been shown differences when compared to rabies related to the carnivores, confirming the need to develop methods that enable more accurate follow-up studies about the biology and epidemiology of rabies in bats. The paucity of data in the literature to date about the complete genome of the Rabies virus variant maintained by populations of vampire bats Desmodus rotundus leaves a gap in understanding the molecular epidemiology of this virus and the epidemiological importance of this species in the transmission of Rabies virus is unquestionable. In this study we sequenced and analyzed the genome of the Rabies virus variant maintained by populations of bat Desmodus rotundus isolated from a bat Desmodus rotundus. The sample, coming from an endemic area in São Paulo, was phylogenetically compared with the genome of the standard sample for spcies 1 - Rabies virus and other samples belonging to the Terrestrial and Aerial cycles of transmission, available in GenBank, to identify possible patterns of differentiating themselves Aerial cycle and in some cases linked only to variant studied.

To the Editor: Bats are now recognized as natural reservoirs for many families of viruses that can cross species barriers and cause emerging diseases of humans and animals. Protecting humans against emerging diseases relies on identifying natural reservoirs for such viruses and surveillance for host-jumping events. The emergence of the Middle East respiratory syndrome coronavirus (MERS-CoV) on the Arabian Peninsula (1) further justifies increased surveillance for coronaviruses (CoVs) in bats. MERS-CoV most likely is a zoonotic virus from a bat reservoir and is associated with high case-fatality rates among humans. The existence of a diverse array of alphacoronaviruses in bats in the United States, Canada, and Trinidad has been reported (2–6). Recently, a possible new alphacoronavirus was detected in an urban roost of bats in southern Brazil (7), and a survey of bats in southern Mexico reported 8 novel alphacoronaviruses and 4 novel betacoronaviruses, 1 with 96% similarity to MERS-CoV (8). These findings expand the diversity and range of known bat coronaviruses and increase the known reservoir for potential emerging zoonotic CoVs. Expanding on our previous work (2,3), we analyzed samples from 97 bats from Brazil and 75 bats from Mexico (Technical Appendix). During 2007–2010, intestinal samples were collected from bats of 10 species in northwest Sao Paulo state in southeastern Brazil. These bats had been submitted to the University Estadual Paulista for rabies testing as a result of epidemiologic surveillance or, in some cases, because of possible or known contact with humans. During 2011–2012, as part of an ongoing rabies surveillance project, intestinal samples were collected from bats of 12 species in their usual habitats in Jalisco state in midwestern Mexico. Bats from a variety of species, including insectivorous, nectarivorous, frugivorous, and hematophagous bats, were included in this study for the purpose of obtaining a diverse array of potential exposures. Intestines were collected and stored, and RNA was purified as described (2). CoV RNA was detected by using a pancoronavirus PCR selective for the RNA-dependent RNA polymerase gene, and amplicons were sequenced as described (3). Virus isolation was not attempted as part of this study. From 1 of 17 Molossus rufus bats and 1 of 8 Molossus molossus bats, an identical novel alphacoronavirus was detected (BatCoV-M.rufus28/Brazil/2010, GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"KC886321","term_id":"529367586"}}KC886321). Both specimens were collected in Brazil during 2010 from adult male bats that had been found in urban areas on residential property. The 412-nt sequence of this virus was most closely related to alphacoronaviruses detected in Eptesicus fuscus bats in North America (82% nt identity), Myotis australis bats in Australia (77% nt identity), Miniopterus bats in Kenya (77% nt identity), and Rhinolophus bats in Hong Kong (77% nt identity) (Figure). Bats of the genus Molossus are insectivorous; their geographic range is restricted to the New World, from northern Mexico to northern Argentina. Figure Phylogenetic tree showing relationships based on 412-nt and 439-nt sequences of a conserved region of gene 1b of BatCoV/Molossus rufus28/Brazil/2010 (alphacoronavirus) and BatCoV/Pteronotus davyi49/Mexico/2012 (betacoronavirus) to other known coronaviruses. ... A novel betacoronavirus (presumably group C) was detected in a specimen from 1 of 4 Pteronotus davyi bats (BatCoV-P.davyi49/Mexico/2012, GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"KC886322","term_id":"529367588"}}KC886322). This specimen was collected in 2012 from an adult male bat roosting in a cave in La Huerta, Mexico. The 439-nt sequence of this virus has 71% nt identity to the novel human group C betacoronavirus MERS-CoV and 72% nt identity to various group D betacoronaviruses detected in Rousettus, Pipistrellus, and Tylonycteris bats in the Philippines, China, and Kenya (Figure). Bats of the species P. davyi (Davy’s naked-backed bat) are insectivorous and are found from southern Mexico to the northern parts of South America. They prefer to roost in caves and man-made structures, such as mines. In summary, we found a novel alphacoronavirus in bats from Brazil and a novel betacoronavirus in a bat from Mexico. Both viruses were detected in bats with known or potential contact with humans. Because the bats we sampled were mostly adult males, the prevalence of CoVs that we identified is probably an underestimation of the true incidence of CoVs in these bat populations. For bats of other species, incidence of CoVs among juvenile and female bats is higher (2,9). Furthermore, we used a non-nested, broadly conserved CoV PCR, which might have limited the sensitivity of CoV RNA detection. The finding of a novel betacoronavirus in insectivorous bats in the New World is noteworthy. Three human CoVs (229E, SARS-CoV, and MERS-CoV) all have animal reservoirs of closely related viruses in Old World insectivorous bats (10) from which they most likely emerged, either directly or indirectly, into the human population. Ongoing surveillance for CoVs in wildlife and increased research efforts to better understand the factors associated with CoV host-switching events are warranted. Technical Appendix: Results of reverse transcription PCR analysis of coronaviruses in bats from Brazil and Mexico. Click here to view.(159K, pdf)

Rabies causes thousands of human and animal deaths worldwide each year. The emergent importance of rabies in wild animals demonstrates the necessity of epidemiologic studies of infection in these species toward the development of better strategies for prevention and control of rabies. We analyzed the circulation of rabies virus among wildlife species from a native rainforest in São Paulo State, Brazil. We used the rapid fluorescent focus inhibition test (RFFIT) to test for rabies virus-neutralizing antibodies in 139 captured terrestrial mammals and the fluorescent antibody test (FAT), mouse inoculation test (MIT), and reverse-transcriptase (RT)-PCR to test for virus in samples from the central nervous system of 53 animals found dead. The percentage of samples positive by RFFIT was 10.8%. All samples tested by FAT, MIT, and RT-PCR were negative. Research should be continued to obtain a better understanding of the role of wildlife in the circulation and transmission of rabies virus.

SUMMARYForty-five human rabies virus isolates from a wide geographical area of Brazil were characterized using an anti-nucleoprotein monoclonal antibody panel and by partial nucleotide sequencing analysis of the nucleoprotein gene. Three major antigenic groups related to the antigenic variants maintained in domestic dogs, vampire bats and marmosets were identified. Phylogenetic analyses revealed that the viruses from dog-related cases segregated into four sister clades: three associated with dog-endemic cycles in Brazil and one with the crab-eating fox cycle in the northeastern region of the country. The vampire bat- and marmoset-related viruses formed two independent groups. The topology of these clades was conserved when these samples were compared to virus representatives of the currently reported rabies endemic cycles in the Americas. These results indicated the presence of multiple endemic transmission cycles maintained in four different reservoirs, domestic dogs, crab-eating foxes, vampire bats and marmosets, which are being transmitted directly to humans and should be considered as a high-risk for rabies infection.

Molecular differences among Mycoplasma hyopneumoniae strains present in pneumonic lungs of swine have been largely studied. However, no comparative studies concerning the strains present in apparently healthy pigs have been carried out. This study aimed to detect, quantify and perform molecular analysis of M. hyopneumoniae strains in pig lungs with and without pneumonic lesions. The detection of M. hyopneumoniae was performed using multiplex PCR (YAMAGUTI, 2008), real-time PCR (STRAIT et al., 2008) and multiple VNTR amplification (VRANCKX et al., 2011). Molecular characterization of the strains was achieved by analysis of the VNTR copy number in P97R1, P146R3, H2R1 and H4. M. hyopneumoniae was detected in samples from healthy and pneumonic pigs and the amount of M. hyopneumoniae positive samples detected varied with the type of assay. The greater number of positive samples was identified by the multiple VNTR amplification combined with capillary electrophoresis. Using real-time PCR, 4.9*104 M. hyopneumoniae genome copies/mL was detected in apparently healthy lungs. A mean quantity of 3.9*106 M. hyopneumoniae genome copies/mL was detected in pneumonic lungs. The analysis of VNTR copy number demonstrated a high genetic variability of the M. hyopneumoniae strains present in apparently healthy and pneumonic lungs. Strains having 3 VNTR copy number in P97R1, were detected only in pneumonic lungs and strains having 40 and 43 VNTR copy number in P146R3 were detected only in apparently healthy lungs. Despite the genetic variability of M. hyopneumoniae, predominant strains in the swine farms could be identified.

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.

Full-length genome sequencing of the rabies virus is not a routine laboratory procedure. To understand fully the epidemiology, genetic variation and evolution of the rabies virus, full-length viral genomes need to be obtained. For rabies virus studies, cDNA synthesis is usually performed using nonspecific oligonucleotides followed by cloning. When specific primers are used, the cDNA obtained is only partial and is limited to the coding regions. Therefore, the development of methods for synthesizing long cDNA using rabies virus-specific primers is of fundamental importance. A new protocol for the synthesis of long cDNA and the development of 19 new primers are described in this study. This procedure allowed the efficient amplification of the full-length genome of the rabies virus variant maintained by hematophagous bat (Desmodus rotundus) populations following the synthesis of a complete long cDNA. Partial sequencing of the rabies virus genome was performed to confirm rabies-specific PCR amplification. Because degenerate primers were employed, this technique can be adapted easily to other variants. Importantly, this new method is faster and less expensive than cloning methods.

Human respiratory syncytial virus (HRSV) strains were isolated from nasopharyngeal aspirates collected from 965 children between 2004 and 2005, yielding 424 positive samples. We sequenced the small hydrophobic protein (SH) gene of 117 strains and compared them with other viruses identified worldwide. Phylogenetic analysis showed a low genetic variability among the isolates but allowed us to classify the viruses into different genotypes for both groups, HRSVA and HRSVB. It is also shown that the novel BA-like genotype was well segregated from the others, indicating that the mutations are not limited to the G gene.

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.

The Human Respiratory Syncytial Virus (HRSV), isolated in 1955, is the main cause of hospitalization of babies and infants with respiratory illness. Several studies have been conducted worldwide aiming the development of a safe and effective vaccine against HRSV. The G2 region of glycoprotein G is used as genotyping default. In the present study, we performed a phylogenetic analysis of G protein and a comparative study between G2 region and ectodomain of attachment glycoprotein. Fifty-three nasal swab samples from children less than 5 years old and presenting symptoms of acute respiratory illness, assisted at the University Hospital (UH) of University of Sao Paulo (USP) in 2004, were submitted to sequencing by PCR and compared with GenBank sequences. We concluded that the G2 region is adequate for HRSV genotyping. O vírus respiratório sincicial humano (HRSV), isolado em 1955, é a principal causa da hospitalização de bebês e crianças pequenas com sintomas de doença respiratória. No mundo inteiro, vários estudos para o desenvolvimento de uma vacina segura e eficiente contra o HRSV têm tido alta prioridade. A região G2 da glicoproteína G é usada como padrão para genotipagem do HRSV. Neste estudo, foi realizada a análise filogenética da glicoproteína G e o estudo comparativo entre a região G2 e o ectodomínio dessa glicoproteína. Cinquenta e três amostras de swab nasal de crianças com menos de cinco anos de idade, apresentando doença respiratória aguda, atendidas no Hospital Universitário (HU) da Universidade de São Paulo durante o ano de 2004, foram submetidas a sequenciamento por PCR e comparadas com seqüências do GenBank. A região G2 mostrou ser adequada para a genotipagem do HRSV.