Rabies in Bats (Chiroptera, Mammalia) in Brazil: Prevalence and Potential Risk Factors Based on Twenty Years of Research in the Northwestern Region of São Paulo, Brazil.

The number of rabies cases in bats has increased recently in Brazil and in the state of São Paulo, representing a new epidemiological scenario for this zoonosis. This study aimed to analyze the prevalence of rabies in bats according to food habits, taxonomic classification, sex and season of the year to identify possible risk factors for rabies occurrence in bats. A retrospective analysis of 6389 records of bat samples, from different municipalities of São Paulo, submitted to rabies diagnosis and taxonomic identification was carried out at the Rabies Diagnostic and Chiroptera Laboratories of Unesp Araçatuba, São Paulo, Brazil, from 1998 to 2017. Seventy-six (1.1%) positive rabies cases were detected in bats from ten species and seven genera of three families. The number of rabies-positive cases was higher in the dry season, with a significant association. The prevalence was higher in the Vespertilionidae family (37), especially Myotis nigricans (19) and Eptesicus furinalis (14). Frugivorous bats had a greater association with positivity for rabies, whereas the variable "sex" had no association. We recommend that the surveillance and control of rabies should be undertaken primarily during the dry season, especially in the Vespertilionidae family species and other species with a frugivorous food habit.

Highly Diverse Arenaviruses in Neotropical Bats, Brazil.

We detected arenavirus RNA in 1.6% of 1,047 bats in Brazil that were sampled during 2007-2011. We identified Tacaribe virus in 2 Artibeus sp. bats and a new arenavirus species in Carollia perspicillata bats that we named Tietê mammarenavirus. Our results suggest that bats are an underrecognized arenavirus reservoir.

Biological activity of Morita-Baylis-Hillman adduct homodimers in L. infantum and L. amazonensis: anti-Leishmania activity and cytotoxicity.

This study is a report on the anti-Leishmania activity of Morita-Baylis-Hillman (MBH) homodimers adducts against the promastigote and axenic amastigote forms of Leishmania (Leishmania) infantum and Leishmania (Leishmania) amazonensis and on the cytotoxicity of these adducts to human blood cells. Both studied homodimers, MBH 1 and MBH 2, showed activity against the promastigote forms of L. infantum and L. amazonensis, which are responsible for visceral and cutaneous leishmaniasis, respectively. Additionally, the homodimers presented biological activity against the axenic amastigote forms of these two Leishmania species. The adducts exhibited no hemolytic activity to human peripheral blood mononuclear cells or erythrocytes at the tested concentrations and achieved higher selectivity indices than amphotericin B. Evaluation of cell death by apoptosis revealed that the homodimers had better apoptosis/necrosis profiles than amphotericin B in the promastigote forms of both L. infantum and L. amazonensis. In conclusion, these Morita-Baylis-Hillman adducts had anti-Leishmania activity in an in vitro model and may thus be promising molecules in the search for new drugs to treat leishmaniasis.

Bat Influenza A(HL18NL11) Virus in Fruit Bats, Brazil.

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.

Electrogena brulini sp. nov. and E. vipavensis Zurwerra & Tomka, 1986 syn. nov. revealed by integrative taxonomy of E. gridellii (Grandi, 1953) (Ephemeroptera: Heptageniidae).

A new species belonging to the speciose genus Electrogena Zurwerra & Tomka, 1985 is described based on all stages from material collected in Switzerland, northern Italy and Slovenia. Electrogena brulini Wagner sp. nov. is closely related to E. gridellii (Grandi, 1953). A genetic analysis based on the mitochondrial gene CO1, including 9 specimens from 5 populations, reveals a K2P distance of 22% between both species. Our investigations demonstrate that the description of the nymph of E. gridellii sensu Belfiore (1996) refers in fact to both species. Therefore, E. gridellii is redescribed in all stages. The number of bristles on each mandibular prostheca in the nymphs, the shape of titillators in male genitalia and the shape of the subanal plate in female imagines are the main characters allowing discrimination of these two species. This study also shows that in this case it is possible and helpful to use colour patterns to separate two closely related species. The re-examination of the type material of Electrogena vipavensis Zurwerra & Tomka, 1986 described from western Slovenia indicates that this species is a junior subjective synonym of E. gridellii.

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

Leishmania spp. is a protozoan that maintains its life cycle in domestic and wild animals and it may include bats, a population that has increased in urban environments. This study aimed to investigate the presence of Leishmania spp. in bats captured strictly in urban areas that are endemic for visceral leishmaniasis. The spleen and skin samples of 488 bats from 21 endemic cities in northwestern São Paulo State, Brazil, were tested for the presence of Leishmania kDNA using real-time PCR. Differentiation from Trypanosoma spp. was achieved by amplifying a DNA fragment of the ribosomal RNA gene. The presence of Leishmania spp. kDNA was verified in 23.9% of bats and Trypanosoma spp. DNA was identified in 3.9%. Leishmania species differentiation revealed the presence of Leishmania amazonensis in 78.3% of the bats; L. infantum in 17.4%, and 1 sample (4.3%) showed a mix pattern of L. infantum and L. amazonensis. We also detected, for the first time, L. infantum and L. amazonensis DNA in Desmodus rotundus, the hematophagous bat. The presence of Leishmania spp. DNA in bats strictly from urban areas endemic for visceral leishmaniasis in the State of São Paulo, Brazil indicates that these wild and abundant animals are capable of harboring Leishmania spp. in this new scenario. Due to their longevity, high dispersion capacity and adaptability to synanthropic environments, they may play a role in the maintenance of the life cycle of Leishmania parasites.

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.

Toward a DNA taxonomy of Alpine Rhithrogena (Ephemeroptera: Heptageniidae) using a mixed Yule-coalescent analysis of mitochondrial and nuclear DNA.

Aquatic larvae of many Rhithrogena mayflies (Ephemeroptera) inhabit sensitive Alpine environments. A number of species are on the IUCN Red List and many recognized species have restricted distributions and are of conservation interest. Despite their ecological and conservation importance, ambiguous morphological differences among closely related species suggest that the current taxonomy may not accurately reflect the evolutionary diversity of the group. Here we examined the species status of nearly 50% of European Rhithrogena diversity using a widespread sampling scheme of Alpine species that included 22 type localities, general mixed Yule-coalescent (GMYC) model analysis of one standard mtDNA marker and one newly developed nDNA marker, and morphological identification where possible. Using sequences from 533 individuals from 144 sampling localities, we observed significant clustering of the mitochondrial (cox1) marker into 31 GMYC species. Twenty-one of these could be identified based on the presence of topotypes (expertly identified specimens from the species' type locality) or unambiguous morphology. These results strongly suggest the presence of both cryptic diversity and taxonomic oversplitting in Rhithrogena. Significant clustering was not detected with protein-coding nuclear PEPCK, although nine GMYC species were congruent with well supported terminal clusters of nDNA. Lack of greater congruence in the two data sets may be the result of incomplete sorting of ancestral polymorphism. Bayesian phylogenetic analyses of both gene regions recovered four of the six recognized Rhithrogena species groups in our samples as monophyletic. Future development of more nuclear markers would facilitate multi-locus analysis of unresolved, closely related species pairs. The DNA taxonomy developed here lays the groundwork for a future revision of the important but cryptic Rhithrogena genus in Europe.

Feeding ecology of carnivores (Mammalia, Carnivora) in Atlantic Forest remnants, Southern Brazil / Ecologia alimentar de carnívoros (Mammalia Carnivora) em Fragmentos de floresta Atlantica do Sul do Brasil

The diet of some sympatric carnivore species in three Atlantic Forest remnants of Southern Brazil was studied in order to assess their food niche. We conducted monthly field trips between February 2003 and January 2004 to collect fecal samples that were subsequently examined together with others collected sporadically between November 1994 and January 2003. Of the 416 samples analysed, 198 had the "author" species identified through microscopic analysis guard hairs, which revealed the presence of 10 carnivores and some information about their diet. Puma yagouaroundi had the largest dietary niche breadth, whereas Leopardus tigrinus and Nasua nasua showed the lowest values. Extensive niche overlap was observed between L. tigrinus and N. nasua, L. tigrinus and L. wiedii, and between L. tigrinus and L. pardalis. Data presented here not only increases the understanding of carnivore feeding ecology, but also contributes towards their conservation in the study region and other fragmented landscapes in Brazil and neighboring countries.

Este trabalho apresenta informações sobre a dieta, a amplitude e a sobreposição de nicho alimentar de algumas espécies simpátricas de carnívoros em três fragmentos de Floresta Atlântica do Sul do Brasil. A coleta de material fecal foi realizada com periodicidade mensal de fevereiro de 2003 a janeiro de 2004. No entanto, amostras obtidas esporadicamente entre novembro de 1994 e janeiro de 2003 também foram incluídas nas análises. Das 416 amostras analisadas, 198 possibilitaram a identificação de 10 espécies "autoras" por meio da análise microscópica de pelos-guarda, bem como informações sobre sua dieta. Puma yagouaroundi teve a maior amplitude de nicho, enquanto Leopardus tigrinus e Nasua nasua apresentaram os menores valores. Maiores sobreposições de nicho foram observadas entre L. tigrinus e outras espécies: N. nasua, L. wiedii e L. pardalis. Espera-se que estes dados, além de contribuírem para incrementar o conhecimento sobre a ecologia alimentar de carnívoros neotropicais, possam auxiliar de forma efetiva na conservação das espécies encontradas na região de estudo e em outras paisagens fragmentadas do território brasileiro e países limítrofes.

[Epidemiological profile of rabies in the northwestern region of São Paulo State, from 1993 to 2007]. / Perfil epidemiológico da raiva na região Noroeste do Estado de São Paulo no período de 1993 a 2007.

The epidemiology of animal rabies in the region of Araçatuba, in the northwest of São Paulo State, from 1993 to 2007, is described according to the results from diagnoses made at laboratories in the region, using the fluorescent antibody and mouse inoculation tests. Out of 10,579 samples analyzed, 4.9% were positive (518/10,579). Dogs accounted for 67% of the cases (346/518) and these occurred between 1993 and 1997. Among the other positive samples, 16% (84/518) were in cattle and 9.7% (50/518) were in bats. Among the 42 municipalities in the region, 23 (55%) presented at least one rabies-positive case, while 13 of them had cases in bats. Three distinct cycles of rabies were identified in the northwestern region of the State of São Paulo: the urban cycle characterized predominantly by canine rabies (1993 to 1997); and the aerial and rural cycles starting in 1998, with predominance of cases in bats in urban areas and in herbivores.

Perfil epidemiológico da raiva na região Noroeste do Estado de São Paulo no período de 1993 a 2007 / Epidemiological profile of rabies in the northwestern region of São Paulo State, from 1993 to 2007

Descreve-se a epidemiologia da raiva animal na região de Araçatuba, noroeste do estado de São Paulo, durante o período de 1993 a 2007, com base nos resultados dos diagnósticos realizados em laboratórios da região, utilizando as técnicas de imunofluorescência direta e inoculação intracerebral em camundongos. De 10.579 amostras analisadas, 4,9 por cento foram positivas (518/10.579). Os casos em cães corresponderam a 67 por cento (346/518) do total e ocorreram entre 1993 a 1997. Dentre as demais amostras positivas, 16 por cento do total (84/518) foi detectado em bovinos e 9,7 por cento (50/518) em morcegos. Dos 42 municípios da região, 23 (55 por cento) apresentaram pelo menos um caso positivo da doença, sendo que 13 deles registraram casos em morcegos. Foram identificados três ciclos distintos da raiva na região Noroeste do Estado de São Paulo, o ciclo urbano caracterizado predominantemente pela raiva canina (1993 a 1997) e os ciclos aéreo e rural, a partir de 1998 com predominância de casos em quirópteros nas áreas urbanas e em herbívoros.

The epidemiology of animal rabies in the region of Araçatuba, in the northwest of São Paulo State, from 1993 to 2007, is described according to the results from diagnoses made at laboratories in the region, using the fluorescent antibody and mouse inoculation tests. Out of 10,579 samples analyzed, 4.9 percent were positive (518/10,579). Dogs accounted for 67 percent of the cases (346/518) and these occurred between 1993 and 1997. Among the other positive samples, 16 percent (84/518) were in cattle and 9.7 percent (50/518) were in bats. Among the 42 municipalities in the region, 23 (55 percent) presented at least one rabies-positive case, while 13 of them had cases in bats. Three distinct cycles of rabies were identified in the northwestern region of the State of São Paulo: the urban cycle characterized predominantly by canine rabies (1993 to 1997); and the aerial and rural cycles starting in 1998, with predominance of cases in bats in urban areas and in herbivores.

Bat rabies in the north-northwestern regions of the state of São Paulo, Brazil: 1997-2002.

OBJECTIVE: Reports on bat rabies in Brazil are sporadic and isolated. This study aimed at describing the detection of rabies virus in bats in the state of São Paulo. METHODS: A total of 7,393 bats from 235 municipalities of the north and northwestern areas of the state of São Paulo, Southeastern Brazil, were assessed according to their morphological and morphometric characteristics from 1997 to 2002. Fluorescent antibody test and mice inoculation were used for viral identification. RESULTS: Of all samples examined, 1.3% was rabies virus positive, ranging from 0.2% in 1997 to 1.6% in 2001. There were found 98 bats infected, 87 in the urban area. Fluorescent antibody test was detected in 77 positive samples, whereas 92 produced rabies signs in mice; incubation period ranging from 4 to 23 days. In 43 cities at least one rabid bat was observed. The highest proportion (33.7%) of rabies virus was found in Artibeus lituratus. Eptesicus and Myotis were the most frequent positive species (24.5%) of the Vespertilionidae family. The species Molossus molossus and Molossus rufus showed 14.3% positive bats. There were no differences in the distribution of positive rabies between females (33; 48.5%) and males (35; 51.5%). CONCLUSIONS: Rabies-infected bats were found in environments that pose a risk to both human and domestic animal population and there is a need for actions aiming at the control of these species and public education.

Bat rabies in the north-northwestern regions of the state of São Paulo, Brazil: 1997-2002

OBJECTIVE: Reports on bat rabies in Brazil are sporadic and isolated. This study aimed at describing the detection of rabies virus in bats in the state of São Paulo. METHODS: A total of 7,393 bats from 235 municipalities of the north and northwestern areas of the state of São Paulo, Southeastern Brazil, were assessed according to their morphological and morphometric characteristics from 1997 to 2002. Fluorescent antibody test and mice inoculation were used for viral identification. RESULTS: Of all samples examined, 1.3 percent was rabies virus positive, ranging from 0.2 percent in 1997 to 1.6 percent in 2001. There were found 98 bats infected, 87 in the urban area. Fluorescent antibody test was detected in 77 positive samples, whereas 92 produced rabies signs in mice; incubation period ranging from 4 to 23 days. In 43 cities at least one rabid bat was observed. The highest proportion (33.7 percent) of rabies virus was found in Artibeus lituratus. Eptesicus and Myotis were the most frequent positive species (24.5 percent) of the Vespertilionidae family. The species Molossus molossus and Molossus rufus showed 14.3 percent positive bats. There were no differences in the distribution of positive rabies between females (33; 48.5 percent) and males (35; 51.5 percent). CONCLUSIONS: Rabies-infected bats were found in environments that pose a risk to both human and domestic animal population and there is a need for actions aiming at the control of these species and public education.

OBJETIVO: Os relatos sobre a ocorrência de raiva em morcegos no Brasil são esporádicos e isolados. Assim, o objetivo do estudo foi descrever a detecção do vírus da raiva em morcegos do Estado de São Paulo. MÉTODOS: Foram analisados 7.393 morcegos provenientes de 235 municípios do norte e noroeste do Estado de São Paulo, no período de 1997 a 2002 e identificados por meio de características morfológicas e morfométricas. Para a detecção do antígeno viral foi utilizada a técnica de imunofluorescência direta e o isolamento do vírus foi realizado por inoculação em camundongos. RESULTADOS: Das amostras examinadas, 1,3 por cento foram positivas para raiva, com variação de 0,2 por cento em 1997 a 1,6 por cento em 2001. Foram encontrados 98 morcegos com o vírus, 87 deles em área urbana. O vírus da raiva foi detectado pela imunofluorescência direta em 77 do total de amostras positivas, enquanto 92 produziram doença em camundongos inoculados e o período de incubação variou entre 4-23 dias. Em 43 municípios foi encontrado pelo menos um morcego positivo. Entre as espécies analisadas o vírus da raiva foi detectado com maior freqüência (33,7 por cento) em Artibeus lituratus. Os vespertilionideos do gênero Eptesicus e Myotis totalizaram 24,5 por cento dos morcegos positivos e as espécies do gênero Molossus (Molossus molossus e Molossus rufus), 14,3 por cento. A distribuição do vírus da raiva foi semelhante entre fêmeas (33; 48,5 por cento) e machos (35; 51,5 por cento). CONCLUSÕES: Morcegos positivos para raiva foram encontrados em situações que colocam em risco tanto a população humana como animais de estimação, exigindo medidas voltadas para o manejo destas espécies e de educação da população.

Long-term alpha interferon treatment is effective on anaemia and significantly reduces iron overload in congenital dyserythropoiesis type I.

Interferon has been shown to be an effective treatment of congenital dyserythropoiesis type I (CDA-I), but the optimal dose and the feasibility of this treatment remains to be determined. Here, in a 9-yr follow-up of a single patient, we show that interferon remains active during such a long period. The optimal dose of conventional alpha interferon could be evaluated at 2 million units twice a week. Pegylated interferon could be used as well at a dose of 30 microg/wk. During interferon treatment, serum and erythrocyte ferritin levels decreased progressively, and remained inversely correlated with haemoglobin levels. On repeated liver biopsies, iron overload could be normalized. Low dose interferon is a long-term treatment of CDA-I, and allows a significant decrease in iron overload, that could be interesting even in patients who are only moderately anaemic.

Isolamento do vírus rábico em Molossus ater (Chiroptera: Molossidae) no Estado de Säo Paulo / Rabies in insectivorous bat - Molossus ater in S. Paulo State, Brazil

Descreve-se o isolamento e a identificaçäo do vírus rábico em morcegos insetívoros Molossus ater, no Estado de Säo Paulo, nos municípios de Araçatuba, Penápolis e Säo José do Rio Preto. A maioria dos exemplares foi capturada ainda com vida, näo havendo, porém, contato com pessoas ou animais. O diagnóstico foi realizado pelas provas de imunofluorescência direta e inoculaçäo intracerebral em camundongos