Ocorrência da raiva e de anticorpos antirrábicos em morcegos do noroeste de São Paulo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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

Molecular epidemiology of animal rabies in the semiarid region of Paraíba, Northeastern Brazil

In the semiarid of the state of Paraíba, the anti-rabies vaccination is not common, most of the local inhabitants who deal with the animals do not know the incidence of the disease in the region. In this study, samples of foxes (Pseudalopex vetulus), insectivorous bats (Molossus molossus), raccoons (Procyon cancrivorous) and domestic animals brains were submitted to the diagnosis of rabies, by using the direct fluorescent antibody technique (d-FAT) and mouse inoculation test (MIT). Of the 581 examined materials, 50 (8.60 %) were positive for d-FAT and 47 (8.09 %) for MIT. From the positive samples for rabies, RNAs were extracted and transformed to cDNA, at the Laboratory of Rabies/Faculdade de Medicina Veterinária e Zootecnia/USP, SP. The phylogenetic characterization of the N gene was performed at the Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, Universidade Nihon, Faculdade de Ciências Bioresource, Fujisawa, Kanagawa, Japão. Based on the results of genotyping and phylogenetic analyzes, it is concluded that the epidemiology of rabies is complex in the semiarid of Paraíba, with different viral variants being maintained in domestic dogs, foxes, insectivorous bats and vampire bats. All the isolates examined belong to the genotype I of the genus Lyssavirus and it is possible to state that in the region, foxes are important sylvatic reservoirs of the rabies virus.

A method for simultaneous detection and identification of Brazilian dog- and vampire bat-related rabies virus by reverse transcription loop-mediated isothermal amplification assay

At present, the sporadic occurrence of human rabies in Brazil can be attributed primarily to dog- and vampire bat-related rabies viruses. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) was employed as a simultaneous detection method for both rabies field variants within 60 min. Vampire bat-related rabies viruses could be distinguished from dog variants by digesting amplicons of the RT-LAMP reaction using the restriction enzyme Alwl. Amplification and digestion could both be completed within 120 min after RNA extraction. In addition, the RI-LAMP assay also detected rabies virus in isolates from Brazilian frugivorous bats and Ugandan dog, bovine and goat samples. In contrast, there were false negative results from several Brazilian insectivorous bats and all of Chinese dog, pig, and bovine samples using the RI-LAMP assay. This study showed that the RT-LAMP assay is effective for the rapid detection of rabies virus isolates from the primary reservoir in Brazil. Further improvements are necessary so that the RT-LAMP assay can be employed for the universal detection of genetic variants of rabies virus in the field. (C) 2010 Elsevier B.V. All rights reserved.

Outbreak of vampire bat biting in a Venezuelan village

An outbreak of 154 cases of vampire bat biting in a four-month period in the gold mine of Payapal, a Venezuelan village, is reported. All patients were bitten during the night and the most bites were on their toes. No complication attributed to the bite was reported. Diagnoses of rabies virus made by means of immunofluorescence were negative. A possible reason for this outbreak may been the development of mining areas, with the inhabitants providing an alternative food source for the bats.

A review of complete albinism in bats with five new cases from Brazil

Complete albinism is a rare phenomenon that occurs in all vertebrate groups. In bats, albinism has been recorded in several species but this information has not previously been reviewed in detail nor has there been an analysis of its importance. In this study, cases of albinism from the literature are checked and new cases are presented. Complete albinism in bats is documented in eight families, 38 species, and at least 64 individuals (47.4% female female and 52.6% males; n = 38). of these, 39 individuals were observed and/or captured in sheltered roosts, such as caves (51.3%), mines and galleries (20.5%), buildings (17.9%), and hollow-trees and bird boxes (7.7%). Only one albino bat (2.6%) was captured in an external roost (foliage). This individual is the fruit bat, Artibeus planirostris, which is recorded here for the first time. Information on four additional cases of albino individuals of the common vampire bat, Desmodus rotundus, is also presented. It is suggested that sheltered roosts favour survival of albino bars, offering protection against sunlight, water loss, and visually hunting predators.

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.

A coronavirus detected in the vampire bat Desmodus rotundus

This article reports on the identification of a group 2 coronavirus (BatCoV DR/2007) in a Desmodus rotundus vampire bat in Brazil. Phylogenetic analysis of ORF1b revealed that BatCoV DR/2007 originates from a unique lineage in the archetypical group 2 coronaviruses, as described for bat species elsewhere with putative importance in Public Health.

Serologic Survey for Rickettsiosis in Bats from Sao Paulo City, Brazil

Blood serum samples were collected from 451 bats captured within the Sao Paulo city from April 2007 to November 2008, and individually tested by indirect immunofluorescence assay against antigens derived from five Rickettsia species reported to occur in Brazil: the spotted fever group (SFG) species R. rickettsii, R. parkeri, R. amblyommii, R. rhipicephali, and the ancestral group species R. bellii. For this purpose, an anti-bat immunoglobulin G was produced and used in the present study. Overall, 8.6% (39/451), 9.5% (34/358), 7.8% (28/358), 1.1% (4/358), and 0% (0/358) serum samples were reactive to R. rickettsii, R. parkeri, R. amblyommii, R. rhipicephali, and R. bellii, respectively. Endpoint titers of reactive sera ranged from 64 to 256. From 20 bat species of 3 different families (Molossidae, Vespertilionidae, and Phyllostomidae), 46 animals were shown to be reactive to at least one rickettsial antigen. Seropositivity per bat species ranged from 0% to 33.3%. Most of the serologically positive sera reacted with two or more rickettsial antigens. Seropositivity for SFG rickettsial antigens in the absence of reactivity against R. bellii (ancestral group species) suggests that bats from Sao Paulo city can be infected by SFG rickettsiae. The possible role of soft ticks in serving as vectors of SFG rickettsiae to bats within the Sao Paulo city, associated to its public health risks, is discussed.

Phylogeography of the common vampire bat (Desmodus rotundus): Marked population structure, Neotropical Pleistocene vicariance and incongruence between nuclear and mtDNA markers

Background: The common vampire bat Desmodus rotundus is an excellent model organism for studying ecological vicariance in the Neotropics due to its broad geographic range and its preference for forested areas as roosting sites. With the objective of testing for Pleistocene ecological vicariance, we sequenced a mitocondrial DNA (mtDNA) marker and two nuclear markers (RAG2 and DRB) to try to understand how Pleistocene glaciations affected the distribution of intraspecific lineages in this bat. Results: Five reciprocally monophyletic clades were evident in the mitochondrial gene tree, and in most cases with high bootstrap support: Central America (CA), Amazon and Cerrado (AMC), Pantanal (PAN), Northern Atlantic Forest (NAF) and Southern Atlantic Forest (SAF). The Atlantic forest clades formed a monophyletic clade with high bootstrap support, creating an east/west division for this species in South America. On the one hand, all coalescent and non-coalescent estimates point to a Pleistocene time of divergence between the clades. On the other hand, the nuclear markers showed extensive sharing of haplotypes between distant localities, a result compatible with male-biased gene flow. In order to test if the disparity between the mitochondrial and nuclear markers was due to the difference in mutation rate and effective size, we performed a coalescent simulation to examine the feasibility that, given the time of separation between the observed lineages, even with a gene flow rate close to zero, there would not be reciprocal monophyly for a neutral nuclear marker. We used the observed values of theta and an estimated mutation rate for the nuclear marker gene to perform 1000 iterations of the simulation. The results of this simulation were inconclusive: the number of iterations with and without reciprocal monophyly of one or more clades are similar. Conclusions: We therefore conclude that the pattern exhibited by the common vampire bat, with marked geographical structure for a mitochondrial marker and no phylogeographic structure for nuclear markers is compatible with a historical scenario of complete isolation of refuge-like populations during the Pleistocene. The results on demographic history on this species is compatible with the Carnaval-Moritz model of Pleistocene vicariance, with demographic expansions in the southern Atlantic forest.

A unique substitution at position 333 on the glycoprotein of rabies virus street strains isolated from non-hematophagous bats in Brazil

The amino acid R or K at position 333 on the glycoprotein of the rabies virus is considered necessary for virulence in adult mice. Although some exceptions exist, substitution at this position causes expression of a phenotype that is either less pathogenic or non-virulent. To date, such substitutions have only been found in fixed strains of rabies virus. In this study, the authors found 333H, 333N, and 333Q substitutions at this position in rabies virus street strains isolated from non-hematophagous bats in Brazil. These strains showed pathogenicity and lethality on passage using adult mice with the intracerebral route and were confirmed rabies-positive by immunofluorescent assay. This suggests that these strains maintain virulence. Our findings indicate that rabies virus street strains with these substitutions exist in the field and may result in infection cycles.

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

Insectivorous bats are the main reservoirs of rabies virus (RABV) in various regions of the world. The aims of this study were to (a) establish genealogies for RABV strains from different species of Brazilian insectivorous bats based on the nucleoprotein (N) and glycoprotein (G) genes, (b) investigate specific RABV lineages associated with certain genera of bats and (c) identify molecular markers that can distinguish between these lineages. The genealogic analysis of N and G from 57 RABV strains revealed seven genus-specific clusters related to the insectivorous bats Myotis, Eptesicus, Nyctinomops, Molossus, Tadarida, Histiotus and Lasiurus. Molecular markers in the amino acid sequences were identified which were specific to the seven clusters. These results, which constitute a novel finding for this pathogen, show that there are at least seven independent epidemiological rabies cycles maintained by seven genera of insectivorous bats in Brazil. (C) 2010 Elsevier Inc. All rights reserved.

Comparative analysis of rabies virus isolates from Brazilian canids and bats based on the G gene and G-L intergenic region

Rabies virus (RABV) isolates from two species of canids and three species of bats were analyzed by comparing the C-terminal region of the G gene and the G-L intergenic region of the virus genome. Intercluster identities for the genetic sequences of the isolates showed both regions to be poorly conserved. Phylogenetic trees were generated by the neighbor-joining and maximum parsimony methods, and the results were found to agree between the two methods for both regions. Putative amino acid sequences obtained from the G gene were also analyzed, and genetic markers were identified. Our results suggest that different genetic lineages of RABV are adapted to different animal species in Brazil.

The socio-religious beliefs and nature of the contemporary Vampire Subculture

The contemporary Vampire Subculture can be defined as a multi-faceted, socio-religious movement with its own distinct collective community and network of participants who share a similar belief system and customary lifestyle that reflect their concept of the vampire. The Vampire Subculture consists of individuals who profess to be 'real vampires', vampire communities of like-minded persons, 'blood-donors' who willingly allow vampires to partake of them, occult-based and mystical-orientated groups that appeal to their spirituality, the blood fetishists, and the live-action vampire role-players. In response, a Christian counter-movement of self-proclaimed 'vampire-slayers' has emerged that actively opposes the vampire subculture and its beliefs and practices. The socio-religious nature of the Vampire Subculture can be best described as a Segmented, Polycentric and Integrated Network of participants.