Rabies virus distribution in tissues and molecular characterization of strains from naturally infected non-hematophagous bats

Bats are main reservoirs for Lyssavirus worldwide, which is an important public health issue because it constitutes one of the big challenges in rabies control. Yet, little is known about how the virus is maintained among bats, and the epidemiological relationships remain poorly understood. The aim of the present study was to investigate the distribution of the rabies virus (RABV) in bat tissues and organs and to genetically characterize virus isolates from naturally infected non-hematophagous bats. The heminested reverse transcriptase polymerase chain reaction (hnRT-PCR) and sequencing using primers to the nucleoprotein coding gene were performed. The results showed a dissemination of the RABV in different tissues and organs, particularly in the salivary glands, tongue, lungs, kidneys, bladder, intestine and feces, suggesting other possible forms of RABV elimination and the possibility of transmission among these animals. The phylogenetic analysis confirmed that different variants of RABV are maintained by non-hematophagous bats in nature and have similar tissue distribution irrespective of bat species and phylogenetic characterization. (C) 2012 Elsevier B.V. All rights reserved.

Rabies surveillance in bats in Northwestern State of Sao Paulo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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.

Levantamento da quiropterofauna (Mammalia : Chiroptera) da Ilha de Cotijuba - PA, com observações sobre sua ecologia

Investigações sobre a quiropterofauna da Amazônia revelam a ocorrência de no mínimo 135 espécies regionais, de hábitos alimentares variados- insetívoras, frugívoras, polinívoras, carnívoras e hematófagas. Os morcegos contribuem ao equilíbrio da biota amazônica por diferentes meios, por exemplo, controle populacional de insetos, dispersão de sementes e polinização. Potencialmente perigosos à saúde humana são os hematófagos portadores do vírus rábico. Os objetivos do estudo foram assim definidos: (a) levantamento das espécies da ordem Chiroptera presentes na Ilha de Cotijuba-PA; (b) descrições do período de atividade, alimentação, reprodução e abrigos das diferentes espécies; (c) análise de diagnóstico de raiva dos quirópteros hematófagos coletados. Foram encontradas 31 espécies, duas delas necessitando ainda de estudos mais detalhados para a confirmação taxonômica. A maioria é de espécies frugívoras, isto é, dispersaras potenciais da flora local (por exemplo, Carollia spp, Artibeus spp, Uroderma spp). Duas espécies hematófagas foram observadas: Desmodus rotundus (cerca de 8% da amostra) e Diaemus youngi (menos de 1% da amostra). A pesquisa do vírus rábico foi negativa, mas ainda há registros de agressões de morcegos a ribeirinhos e animais domésticos, exigindo análises laboratoriais de novas amostras, com finalidade de monitoramento sanitário.

Caracterização molecular de cepas do Vírus da raiva (Lyssavirus; Rhabdoviridae) isoladas no Estado do Pará

A raiva é uma das zoonoses mais antigas e temidas pelo homem devido a seu desfecho fatal. Os cães ainda são considerados os principais responsáveis pela manutenção e transmissão da raiva para o homem. Porém, nos últimos anos os morcegos hematófagos têm ganhado destaque como potenciais transmissores de raiva para animais e humanos nas Américas. Recentemente, várias epidemias de raiva humana transmitida por morcegos hematófagos foram relatados no estado do Pará, o que mostra uma grande alteração no ambiente natural destes animais. A amplificação parcial do gene N pela técnica de RT-PCR foi aplicada em 62 amostras positivas para o Vírus da raiva, pela imunofluorescência direta e prova biológica. As seqüências nucleotídicas obtidas foram comparadas entre si e com outras amostras de vírus rábico isoladas no Brasil, utilizando os métodos de análise filogenética máxima verossimilhança e Bayesiano. Estas análises permitiram traçar o perfil epidemiológico molecular das variantes virais circulantes no estado do Pará, observando a emergência da transmissão de casos associados à variante antigênica 3 (VAg3), comumente encontrada em morcegos hematófagos Desmodus rotundus em detrimento dos casos relacionados à variante antigênica 2 (VAg2) associada a cães domésticos, bem como a identificação de três linhagens genéticas relacionadas a VAg3 e uma relacionada a VAg2 e uma possível nova variante isolada de morcego frugívoro Uroderma bilobatum.

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

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Rabies in southeast Brazil: a change in the epidemiological pattern

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.

Serological investigation of rabies virus neutralizing antibodies in bats captured in the eastern Brazilian Amazon

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

First detection of Leishmania spp. DNA in Brazilian bats captured strictly in urban areas

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

The sialotranscriptome of Antricola delacruzi female ticks is compatible with non-hematophagous behavior and an alternative source of food

The hosts for Antricola delacruzi ticks are insectivorous, cave-dwelling bats on which only larvae are found. The mouthparts of nymphal and adult A. delacruzi are compatible with scavenging feeding because the hypostome is small and toothless. How a single blood meal of a larva provides energy for several molts as well as for oviposition by females is not known. Adults of A. delacruzi possibly feed upon an unknown food source in bat guano, a substrate on which nymphal and adult stages are always found. Guano produced by insectivorous bats contains twice the amount of protein and 60 times the amount of iron as beef. In addition, bacteria and chitin-rich fungi proliferate on guano. Comparative data on the transcriptome of the salivary glands of A. delacruzi is nonexistent and would help to understand the physiological adaptations of salivary glands that accompany different sources of food as well as the steps taken by the Acari toward haematophagy, believed to have evolved from scavenging dead animals. Annotation of the transcriptome of salivary glands from female instars of A. delacruzi collected on guano categorized 5.7% of the clusters of expressed genes as putative secreted proteins. They included abundantly expressed TIL-domain-containing proteins (possible anti-microbials), an abundantly expressed protein similar to a serum amyloid found in the sialotranscriptomes of Ornithodoros spp., a savignygrin, a family of mucin/peritrophin/cuticle-like proteins, anti-microbials and an HIV envelope-like glycoprotein also found in soft ticks. When comparing the transcriptome of A. delacruzi with those of blood-feeding female soft and hard ticks some notable differences were observed; they consisted of the following transcripts over- or under-represented or absent in the sialotranscriptome of A. delacruzi that may reflect its source of food: ferritin, mucins with chitin-binding domains and TIL-domain-containing proteins versus lipocalins, basic tail proteins, metalloproteases, glycine-rich proteins and Kunitz protease inhibitors, respectively. (C) 2012 Elsevier Ltd. All rights reserved.

Temporal and spatial distribution and habitat associations of an urban population of New Zealand long-tailed bats (Chalinolobus tuberculatus)

Two ultrasound survey methods were used to determine the presence and activity patterns of New Zealand long-tailed bats (Chalinolobus tuberculatus) in the city of Hamilton. First, 13 monthly surveys conducted at 18 green spaces found C. tuberculatus in only one urban forest reserve, Hammond Bush, where they were found consistently throughout the year. Bat activity was strongly related to temperature. Second, twice-yearly citywide surveys conducted over 2 years determined the distribution and habitat associations of C. tuberculatus. Bats were found only in the southern part of the city and were strongly associated with the Waikato River. Bat activity was negatively correlated with housing and street light density and positively correlated with topographical complexity. In Hamilton, topographical complexity indicates the presence of gullies. Gullies probably provide foraging and roosting opportunities and connect the river to distant forest patches. These results suggest that urban habitats can be useful for bats if gullies can link these to distant habitat fragments.

Is lek breeding rare in bats?

Bats (Mammalia: Chiroptera) are among the most successful mammals and likely display the widest range of mating systems within the Class. One mating system that is underrepresented in the Chiroptera is lek breeding, which is characterized by aggregations of sexually displaying males that are visited by receptive females who appraise male displays and actively choose mates, yet receive no direct benefits such as assistance in parenting. Leks are thought to form when males can defend neither resources nor females, making it more economical to establish small breeding territories and self-advertise through sexual displays. Lekking is rare in mammals, and it has been suggested that a lack in the mobility required by females to economically seek out aggregations of sexually displaying males may explain this rarity. Bats, like birds, do not suffer reduced mobility and yet out of over a thousand described species, only one has been confirmed to breed in leks. We examine the rarity of lekking in bats by providing an overview on the current state of knowledge of their mating systems and discuss the ecological and social determinants for the observed trends, contrasted with the prerequisites of lek-breeding behaviour. We use the breeding behaviour of New Zealand's lesser short-tailed bat Mystacina tuberculata, which is believed to be a lek breeder, as a case study for the examination of potential lekking behaviour in bats, and highlight the importance of such research for the development of effective conservation strategies.

A continental-scale tool for acoustic identification of European bats

Summary 1. Acoustic methods are used increasingly to survey and monitor bat populations. However, the use of acoustic methods at continental scales can be hampered by the lack of standardized and objective methods to identify all species recorded. This makes comparable continent-wide monitoring difficult, impeding progress towards developing biodiversity indicators, transboundary conservation programmes and monitoring species distribution changes. 2. Here we developed a continental-scale classifier for acoustic identification of bats, which can be used throughout Europe to ensure objective, consistent and comparable species identifications. We selected 1350 full-spectrum reference calls from a set of 15 858 calls of 34 European species, from EchoBank, a global echolocation call library. We assessed 24 call parameters to evaluate how well they distinguish between species and used the 12 most useful to train a hierarchy of ensembles of artificial neural networks to distinguish the echolocation calls of these bat species. 3. Calls are first classified to one of five call-type groups, with a median accuracy of 97·6%. The median species-level classification accuracy is 83·7%, providing robust classification for most European species, and an estimate of classification error for each species. 4. These classifiers were packaged into an online tool, iBatsID, which is freely available, enabling anyone to classify European calls in an objective and consistent way, allowing standardized acoustic identification across the continent. 5. Synthesis and applications. iBatsID is the first freely available and easily accessible continental- scale bat call classifier, providing the basis for standardized, continental acoustic bat monitoring in Europe. This method can provide key information to managers and conservation planners on distribution changes and changes in bat species activity through time.

Sex-specific roost selection by bats in clearfell harvested plantation forest : improved knowledge advises management

To maximize energetic savings, female bats often roost communally whilst pregnant or with non-volant dependents, whereas male bats more often roost alone; however, differences in selection of roosts by sex have not often been investigated. Better understanding of female colony locations could focus management to protect the majority of bats. New Zealand's long-tailed bat (Chalinolobus tuberculatus) roost in exotic plantation forest, where sex-specific roost selection has not been investigated, and therefore such management is not possible. We investigated sex-specific roost selection by long-tailed bats for the first time. Roosts and paired nonroosts were characterized testing predictions that males and females select roosts that differ from non-roosts, and males and females select different roosts. Females and males chose Pinus radiata roosts that differed from non-roost trees. Results suggest each sex chose roosts that maximized energetic savings. Female bats used roosts closer to water sources, that warmed earlier in the day, which allowed maintenance of high temperatures. Males appeared to choose roosts that allowed torpor use for long periods of the day. Males may be less selective with their roost locations than females, as they roosted further from water sources. This could allow persistence of male bats in marginal habitat. As all female long-tailed bats chose roosts within 150 m of waterways, management to protect bats could be focused here. To protect bats least able to escape when roosts are harvested, harvest of forest stands selected by female bats as roost sites should be planned when bats are not heavily pregnant nor have non-volant dependents.

Variation in the abundance of ectoparasitic mites of flat-headed bats

We investigated the ectoparasitic mite loads (Macronyssus: Macronyssidae: Acarina) on 2 species of flat-headed bats, Tylonycteris pachypus and T. robustula (Mammalia: Chiroptera: Vespertilionidae) in 2 counties of Guangxi Province, southern China, from 2002 to 2005. In Longzhou County both species of bat occur sympatrically, but only T. pachypus occurs in Ningming County. Individuals of the smaller species (T. pachypus) harbored significantly more mites than did those of T. robustula. In both species males harbored more mites than nonreproductive females, pregnant females had more mites than lactating and nonreproductive females, and juveniles harbored more mites than adults. Mite load on both species of bats showed significant seasonal variation, declining from spring to winter. No correlation was found between mite load and size of the host colony. We discuss our findings in relation to the ecology and biology of both hosts and parasites.