Rabies surveillance in bats in Northwestern State of São Paulo

Introduction Rabies is an important zoonosis that occurs in mammals, with bats acting as Lyssavirus reservoirs in urban, rural and natural areas. Rabies cases in bats have been recorded primarily in urban areas in Northwestern State of São Paulo since 1998. This study investigated the circulation of rabies virus by seeking to identify the virus in the brain in several species of bats in this region and by measuring rabies-virus neutralizing antibody levels in the hematophagous bat Desmodus rotundus. Methods From 2008 to 2012, 1,490 bat brain samples were sent to the Universidade Estadual Paulista (UNESP) Rabies Laboratory in Araçatuba, and 125 serum samples from vampire bats that were captured in this geographical region were analyzed. Results Rabies virus was detected in the brains of 26 (2%) of 1,314 non-hematophagous bats using the fluorescent antibody test (FAT) and the mouse inoculation test (MIT). None of the 176 hematophagous bat samples were positive for rabies virus when a virus detection test was utilized. Out of 125 vampire bat serum samples, 9 (7%) had levels of rabies virus neutralization antibodies (RVNAs) that were higher than 0.5IU/mL; 65% (81/125) had titers between 0.10IU/mL and 0.5IU/mL; and 28% (35/125) were negative for RVNAs using the simplified fluorescent inhibition microtest (SFIMT) in BHK21 cells. The observed positivity rate (1.7%) was higher than the average positivity rate of 1.3% that was previously found in this region. Conclusions The high percentage of vampire bats with neutralizing antibodies suggests that recent rabies virus exposure has occurred, indicating the necessity of surveillance measures in nearby regions that are at risk to avoid diffusion of the rabies virus and possible rabies occurrences.

Influência do sexo de indivíduos da mesma colônia no controle químico das populações do morcego hematófago Desmodus rotundus (Phyllostomidae) no estado de São Paulo

Os objetivos do presente trabalho foram estudar a fidelidade ao abrigo diurno por indivíduos de ambos os sexos da espécie Desmodus rotundus e verificar a eficácia da ação da pasta vampiricida 2% na redução do tamanho de suas colônias no estado de São Paulo, por meio de estudo experimental de campo. Durante os anos de 1999 e 2000, 626 morcegos distribuídos em 12 abrigos foram capturados com redes-de-espera (armadas durante a noite) e marcados. em seguida, 10% da população previamente estimada recebeu a pasta vampiricida. No Experimento I foram tratados apenas machos, no Experimento II apenas fêmeas e no Experimento III, 5% dos machos e 5% das fêmeas foram tratados. Após 5 e 10 dias, foram feitas contagens dos morcegos que sobreviveram e morreram. As fêmeas mostraram-se mais fiéis aos abrigos (p<0,01) e melhores disseminadoras de pasta vampiricida (p<0,01).

Variabilidade entre genótipos de feijoeiro na eficiência no uso do fósforo

A utilização da variabilidade genética em feijoeiro (Phaseolus vulgaris L.) para eficiência na absorção e utilização de fósforo se mostra como uma alternativa para contornar a deficiência deste elemento em solos de cerrado. Para detectar tais eficiências, foi desenvolvido um experimento em casa de vegetação na Universidade Federal de Uberlândia-MG, entre julho e agosto de 1997. O delineamento experimental foi de blocos casualizados com quatro repetições, em esquema fatorial 8 x 2. Foram avaliados oito genótipos de feijoeiro (BAT 477, Carioca MG, Emgopa 201, Jalo Precoce, Pérola, Rosinha, Roxo e Xamego) e duas doses de P2O5 (24 e 120mg dm-3). Após 45 dias da germinação, foram analisadas a produção de matéria seca da parte aérea e das raízes dos genótipos; conteúdo de fósforo na parte aérea e das raízes; razão raiz:parte aérea e a eficiência no uso do fósforo (valor a), classificando-os em quatro grupos: eficientes e responsivos, não eficientes e responsivos, eficientes e não responsivos e não eficientes e não responsivos. Os genótipos classificados como eficientes na absorção e utilização de fósforo foram BAT 477, Jalo Precoce e Roxo, enquanto que os classificados como responsivos foram os genótipos Carioca MG, Jalo Precoce, Pérola e Roxo.

Antigenic typing of brazilian rabies virus samples isolated from animals and humans, 1989-2000

Amostras de vírus rábico isoladas de animais e humanos no período de 1989 a 2000 foram tipificadas antigenicamente com a utilização de um painel de anticorpos monoclonais contra a nucleoproteína viral, pré-estabelecido para o estudo da epidemiologia molecular do vírus rábico isolado nas Américas. As amostras testadas foram isoladas no laboratório de diagnóstico do Instituto Pasteur e outros centros de diagnóstico de raiva no Brasil. Além das cepas de vírus rábico fixo CVS-31/96-IP, mantida em cérebro de camundongos e a PV-BHK/97, mantida em cultura de células, cepas de vírus rábico isoladas de cães, gatos, bovinos, eqüinos, morcegos, ovinos, caprino, suínos, raposa, sagüí, coatí, guaxinim e humanos, totalizaram 330 amostras. Seis variantes antigênicas foram definidas, compatíveis com perfís observados no painel de anticorpos monoclonais pré-estabelecido utilizado, as de número 2 (cão), 3 (Desmodus rotundus), 4 (Tadarida brasiliensis), 5 (Vampiro da Venezuela), 6 (Lasiurus cinereus) e Lab (reagente a todos os anticorpos utilizados), além de outros seis perfís desconhecidos, não compatíveis com aqueles observados no painel utilizado. A maior variabilidade foi observada entre as amostras isoladas de morcegos insetívoros e a variante mais comum isolada entre as espécies foi a variante 3 (Desmodus rotundus). Estes fatos podem representar a existência de múltiplos ciclos de transmissão independentes, envolvendo diferentes espécies de morcegos.

Genetic and phylogenetic analysis of glycoprotein of rabies virus isolated from several species in Brazil

Genetic and phylogenetic analyses of the region containing the glycoprotein (G) gene, which is related to pathogenicity and antigenicity, and the G-L intergenic region were carried out in 14 Brazilian rabies virus isolates. The isolates were classified as dog-related rabies virus (DRRV) or vampire bat-related rabies virus (VRRV), by nucleoprotein (N) analysis. The nucleotide and amino acid (AA) homologies of the area containing the G protein gene and G-L intergenic region were generally lower than those of the ectodomain. In both regions, nucleotide and deduced AA homologies were lower among VRRVs than among DRRVs. There were AA differences between DRRV and VRRV at 3 antigenic sites and epitopes (IIa, WB+ and III), suggesting that DRRV and VRRV can be distinguished by differences of antigenicity. In a comparison of phylogenetic trees between the ectodomain and the area containing the G protein gene and G-L intergenic region, the branching patterns of the chiropteran and carnivoran rabies virus groups differed, whereas there were clear similarities in patterns within the DRRV and VRRV groups. Additionally, the VRRV isolates were more closely related to chiropteran strains isolated from Latin America than to Brazilian DRRV. These results indicate that Brazilian rabies virus isolates can be classified as DRRV or VRRV by analysis of the G gene and the G-L intergenic region, as well as by N gene analysis.

Genetic analysis of phosphoprotein and matrix protein of rabies viruses isolated in Brazil

To investigate the genetic characteristics of phosphoprotein (P) and matrix protein (M) genes of variable rabies virus (RV) prevalent in Brazil, the authors genetically characterized the P and M genes from 30 Brazilian RV field isolates. Phylogenetic analysis based on the P and M genes revealed the presence of six RV variants that consisted primarily of three insectivorous bats, the vampire bat, dog and fox in Brazil. Specific amino acid substitutions corresponding to these phylogenetic lineages were observed, with ASP(42) and GlU(62) in the P protein found to be characteristic of Brazilian chiroptera- and carnivora-related RVs, respectively. Amino acid sequence motifs predicted to associate with a viral function in the P and M proteins were conserved among Brazilian RV variants.

Características histoquímicas e estruturais do testículo do morcego hematófago (Desmodus rotundus rotundus, Geoffrey, 1810).

The testicular stroma of the vampire bat including the testicular capsula and the lamina propria of the seminiferous tubuli, was strongly PAS-positive. This observation was a possible indication of great amounts of structural glycogen and other glycoconjugates at the level of smooth muscle cells; elongated contractile cells and/or collagen frameworks of the tunica albuginea and tubular lamina propria. In the last the basement membranes of the seminiferous tubules were particularly strongly PAS positive, as an indication of their neutral mucosubstances structural composition, previously described (Malmi et al., 1987). The epithelium lining from the cavitary and surface rete testis complex showed low reactivities to mucosubstances; total proteins and lipids and oxidative enzymes studied. Although the apical granulation at the rete testis epithelium showed an intense PAS reactivity with hypothesis of glycoprotein secretion, through the rete. The PAS, Sudan Black B, NADH, MDH and LDH reactions of the testicular interstitium seem correlate to steroid metabolism (biosynthesis and secretion), at the Leydig cells level. The seminiferous epithelium generally had low reactions to all the histochemical studies realized. Particularly in the adbasal compartment the histochemical localizations of NADH diaphorase and LDH were possible related to glycolytic activities and general carbohydrates metabolism, both enzymes, and hydrogen transport, the NADH. The strong PAS, diastase and PAS, and alcian blue pH 2.5 and PAS reactions observed in the adluminal seminiferous epithelium compartment were directly related to the spermatids acrosomal glycoconjugates structuration. Also the SDH localization at this level seems to be related to the mitochondrial activities at the middle piece level in the late spermatids.

Características ultra-estruturais das células de Sertoli do morecego hematófago (Desmodus rotundus rotundus, Geoffrey, 1810).

This paper deals with the ultrastructural study of mature vampire bat Sertoli cells and their relationships with the different stages of testicular germ cells. In vampire bat seminiferous epithelium there are different types of junctional specializations among Sertoli cells and among Sertoli cells and different germ cells, with special emphasis to tight junctions and to junctions like as desmosomes. Ectoplasmic junctions through the Sertoli cells, including the smooth ER, are observed. These cellular interactions and their cytophysiological roles are discussed. Also are related some ultrastructural peculiarities of the Sertoli cell nucleus, nucleolus, cytoplasmic organelles and lipidic inclusions.

Caracterização antigênica e genotípica de isolados do vírus rábico, de quirópteros da cidade de Botucatu-SP e região

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diversidade da quiropterofauna (Mammalia) no Parque Ecológico de Gunma, Santa Bárbara do Pará

O implemento de Unidades de Conservação constitui um dos meios para se evitar a perda de biodiversidade nas regiões de influência das grandes metrópoles, e o monitoramento de certos grupos zoológicos nessas unidades pode contribuir ao êxito de programas conservacionistas locais. O Parque Ecológico de Gunma (PEG), com área de 540 hectares administrada pela Associação Gunma Kenjin-Kai do Norte do Brasil, protege um remanescente florestal no município de Santa Bárbara do Pará, região da Grande Belém, Estado do Pará, constituído principalmente por fitofisionomias de Terra Firme e Igapó. Este trabalho tem por objetivo oferecer um perfil taxonômico e ecológico da quiropterofauna (Mammalia) nos limites do Parque e seu entorno. A análise quantitativa incluiu medidas de esforço de captura, riqueza e diversidade, assim como uma avaliação da estrutura de comunidades por Escalonamento Multidimensional (MDS) considerando-se as fisionomias vegetais principais e duas áreas selecionadas de Terra Firme. Num conjunto de seis campanhas, de julho a dezembro de 2005, identificaram-se 37 espécies de morcegos, distribuídas em cinco famílias. Contudo, em nenhum ambiente as curvas de rarefação atingiram a assíntota. Os índices de diversidade de Shannon-Wiener (H’) e Simpson (D) foram de 1,86 e 0,69, respectivamente. Carollia perspicillata exibiu a mais alta abundância relativa, abrangendo 50% das capturas, mas de modo distinto entre fisionomias com aproximadamente 59% em Terra Firme e 32% em Igapó. O achado de Neonycteris pusilla representa a segunda ocorrência do táxon para o Estado do Pará. As estimativas de diversidade foram mais elevadas no Igapó (H’ = 1,91; [1-D] = 0,80) do que na Terra Firme (H’ = 1,71; [1-D] = 0,63), apesar da pequena área do primeiro (3,3% da extensão do PEG) e da maior riqueza de espécies da segunda. Isto pode refletir o fato de que a quase totalidade das capturas se deu nos meses mais secos do ano na região, quando em tese haveria tropismo das espécies por espaços de maior oferta hídrica. Assim, parece imperativo complementar o levantamento com amostragens na estação chuvosa para uma descrição fidedigna da quiropterofauna do PEG. Confirmando-se o previsto, a comparação de duas áreas de Terra Firme com distintos graus de alteração apontou maior diversidade de quirópteros na área mais conservada. A importância do PEG como Unidade de Conservação pode ser atestada pela presença de espécies sob risco (vulneráveis ou quase ameaçadas), p.ex. N. pusilla e Tonatia carrikeri. Verificou-se a ameaça de interferências antrópicas sobre os seus ecossistemas devido principalmente à proximidade com áreas rurais e urbanas. O monitoramento da riqueza e dinâmica ecológica dos morcegos no Parque Ecológico de Gunma pode evitar a extinção local de táxons raros, assim como a proliferação de outros, que em situações especiais se tornam ameaçadores à saúde humana, caso do hematófago Desmodus rotundus, potencial hospedeiro e transmissor do vírus rábico.

Rabies surveillance in bats in Northwestern State of Sao Paulo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evaluation of short-interfering RNAs treatment in experimental rabies due to wild-type virus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Valores hematológicos de morcegos hematófagos Desmodus rotundus (E. Geoffroy, 1810) mantidos em cativeiro

Hematological values from captive vampire bats Desmodus rotundus (E. Geoffroy, 1810). Desmodus rotundus is one of the hematophagous bat species that are responsible for significant losses to livestock and has important involvement on public health. The great interest on this bat species made it becomes the target of many investigations a required its maintenance in laboratories. Similarly to others mammals, hematological evaluation has been utilized to assess the health and morbidity state of bats, however there are scarce studies with captive bats. The aim of this study was to investigate the hypothesis that is possible to feed the vampire bat D. rotundus with frozen blood treated with citrate during a long captivity period without hematological dyscrasias. Therefore, complete blood count was performed monthly from 15 adult females kept for 345 days in cages and fed with frozen blood added to citrate. The erythrocyte concentration (9.02 ± 1.43 x1012/L), PCV (0.47 ± 0.08 L/L) and hemoglobin (163.9 ± 31.5 g/L) obtained from free-living bats (immediately after capture) were lower or similar to those obtained after 345 days of captivity, presenting erythrocytes’ count of 11.01 ± 0.82 x1012/L, packed cell volume of 0.50 ± 0.05 L/L and hemoglobin level of 158 ± 10.1 g/L. The total white blood cell (11.09 ± 6.07 x109 /L) and segmented neutrophil counts (9.85 ± 3.5 x109 /L) of free living D. rotundus decreased significantly after 345 days of captivity, with values of 3.98 ± 1.98 and 1.87 ± 978.6 x109 /L respectively, which are similar to bats from temperate regions in hibernation period. This study proved that is possible to feed D. rotundus for long periods of captivity with citrated blood without the occurrence of anemia, erythrocyte or other hematologic dyscrasia

Desmodus rotundus and Artibeus spp. bats might present distinct rabies virus lineages

In Brazil, bats have been assigned an increasing importance in public health as they are important rabies reservoirs. Phylogenetic studies have shown that rabies virus (RABV) strains from frugivorous bats Artibeus spp. are closely associated to those from the vampire bat Desmodus rotundus, but little is known about the molecular diversity of RABV in Artibeus spp. The N and G genes of RABV isolated from Artibeus spp. and cattle infected by D. rotundus were sequenced, and phylogenetic trees were constructed. The N gene nucleotides tree showed three clusters: one for D. rotundus and two for Artibeus spp. Regarding putative N amino acid-trees, two clusters were formed, one for D. rotundus and another for Artibeus spp. RABV G gene phylogeny supported the distinction between D. rotundus and Artibeus spp. strains. These results show the intricate host relationship of RABV's evolutionary history, and are invaluable for the determination of RABV infection sources. (C) 2012 Elsevier Editora Ltda. All rights reserved.

Molecular epidemiology of livestock rabies viruses isolated in the northeastern Brazilian states of Paraíba and Pernambuco from 2003 - 2009

Background Limited or no epidemiological information has been reported for rabies viruses (RABVs) isolated from livestock in the northeastern Brazilian states of Paraíba (PB) and Pernambuco (PE). The aim of this study was to clarify the molecular epidemiology of RABVs circulating in livestock, especially cattle, in these areas between 2003 and 2009. Findings Phylogenetic analysis based on 890 nt of the nucleoprotein (N) gene revealed that the 52 livestock-derived RABV isolates characterized here belonged to a single lineage. These isolates clustered with a vampire bat-related RABV lineage previously identified in other states in Brazil; within PB and PE, this lineage was divided between the previously characterized main lineage and a novel sub-lineage. Conclusions The occurrences of livestock rabies in PB and PE originated from vampire bat RABVs, and the causative RABV lineage has been circulating in this area of northeastern Brazil for at least 7 years. This distribution pattern may correlate to that of a vampire bat population isolated by geographic barriers.