Molecular identification of trypanosomatids in wild animals

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

Pesquisa do Paracoccidioides brasiliensis em aerossóis ambientais: padronização de metodologia e avaliação de amostras relacionadas ao habitat de tatus positivos

P.brasiliensis é o agente etiológico da Paracoccidioidomicose, micose sistêmica mais importante na América Latina. A fase saprofítica produtora dos propágulos infectantes e outros aspectos ecológicos deste patógeno são ainda pouco conhecidos, o que impede a adoção de medidas preventivas. Diversas evidências indicam que o fungo se desenvolve no solo, porém o seu isolamento a partir destes materiais é raro e apenas eventual. O presente projeto visou desenvolver uma metodologia para obtenção de amostras ambientais na forma de aerossóis em locais onde a ocorrência do fungo já foi evidenciada pelo seu isolamento em tatus Dasypus novemcinctus e avaliar a ocorrência do patógeno nestes materiais por cultura direta e por biologia molecular em reações de PCR com “primers” específicos, já desenvolvidos em nosso laboratório. Para tal, avaliamos o padrão de crescimento do P. brasiliensis em quatro diferentes meios ditos como seletivos. Entre estes meios, encontra-se o Meio Sólido de amônia que, por nunca ter sido testado em P. brasiliensis, necessitava de testes quanto à viabilidade de uso e ajuste de concentração. Depois de testado a funcionalidade dos meios e observado o padrão de crescimento que o fungo exibe nesses diferentes meios, partimos para a tentativa de isolamento ambiental do P. brasiliensis através de amostras de solo em forma de aerossóis com o auxílio deles. Tendo em vista os resultados obtidos, optamos, também, por realizar a coleta de teias de aranhas, devido a grande aderência que exerce sobre aerossóis ambientais. Os isolados de P. brasiliensis cultivados apresentaram desenvolvimento semelhante nos quatro meios utilizados, indicando a possibilidade de uso destes meios seletivos para estudos ambientais. P.brasiliensis não foi isolado nestas amostras ambientais... (Resumo completo, clicar acesso eletrônico abaixo)

Abundância e tamanho populacional de mamíferos náo-voadores introduzidos na Ilha Anchieta, SP

Espécies de plantas, animais ou microrganismos introduzidos em ecossistemas do qual não fazem parte originalmente podem ser chamados de exóticos. Esses organismos podem se adaptar e propagar, prejudicando processos naturais e espécies nativas. Os impactos das introduções sobre a biota nativa podem ser imperceptíveis ou mesmo catastróficos, uma vez que estas espécies podem causar profundas alterações na estrutura dos ecossistemas. Em 1983, 15 espécies de mamíferos foram introduzidas na Ilha Anchieta, Ubatuba, SP. Em 2005 a partir de transecções lineares foi possível determinar a abundância relativa, densidade e tamanho populacional de nove espécies introduzidas e uma nativa que mostro que a ilha possui uma alta densidade de mamíferos (480.21 ind/km²). Neste estudo, a partir de dados de coletados em 2008-2009 poderemos comparar os resultados obtidos no mesmo local em 2004-2005. Foram instaladas 42 armadilhas fotográficas para detecção dos mamíferos (sete armadilhas no mês de outubro de 2008 e 35 entre os meses de fevereiro e julho de 2009). Foi utilizado também e como principal ferramenta o censo via transectos lineares onde, foram dispostas 17 trilhas de 400m cada e o levantamento dos mamíferos foi realizado durante o dia, com saídas na parte da manhã às 6:00 e na parte da tarde às 17:00. As trilhas foram percorridas entre os meses de fevereiro e outubro de 2009 e foram percorridos 38,4 km nos transectos lineares. Foram introduzidas duas espécies de cutias (Dasyprocta azarae e D. leporina) entretanto os VIII indivíduos registrados na amostragem por transectos lineares foram identificados apenas ao nível de gênero. Dasyprocta spp. (33%), Dasypus novemcinctus (27,97%) e Callithrix penicillata (19,49%) foram as espécies com maior frequência relativa de ocorrência no censo (vestígios e avistamentos)... (Resumo completo, clicar acesso eletrônico abaixo)

Levantamento rápido de mamíferos terrestres de médio e grande porte no Parque Natural Municipal Cachoeira da Marta, Botucatu/SP

One of the most important strategies for biodiversity conservation is the establishment of protected areas, which protect areas important for the survival of several species. The park “Parque Natural Municipal Cachoeira da Marta”, located in Botucatu, São Paulo, is a unit of strictly protected area and aims to conserve the ecosystems and the biodiversity that exists in the area. The park has a management plan, but this does not include a survey of mammals present in the area, which is relevant as the group has a high degree of threat and a huge ecological importance. Thus, this study aimed to conduct a quick survey of terrestrial mammals of medium and large through the methodology of sand plots, with 25 plots distributed in five areas throughout the park. We recorded 4 orders, 6 families and 9 species (Nasua Nasua, Eira Barbara, Procyon cancrivorous, Leopardus pardalis, Leopardus sp., Dasypus novemcinctus, Tamandua tetradctyla, and Cavia aperea Myrmecophaga tridactyla). Most animals marked are generalist feeding habits, but it was also reported the presence of species more demanding regarding the quality of the habitat

Adesão de Paracoccidioides brasiliensis e outros fungos anemófilos a teias de aranhas: implicações para a identificação de nichos microbianos no ambiente

The dimorphic pathogenic fungus, Paracoccidioides brasiliensis (Pb), is the etiological agent of paracoccidioidomycosis (PCM), the most important systemic mycosis in Latin America. While the yeast phase can be isolated from patients affected by paracoccidioidomycosis, dogs and naturally infected armadillos; several elements related to the ecology of the saprophytic phase of the pathogen, which is responsible for the production of infective propagules, are poorly understood, hampering the adoption of preventive measures. The demonstration of the high incidence of Pb infection in the 9- banded armadillo, Dasypus novemcinctus, has opened new perspectives for the identification of the pathogen’s habitat. At the opening of the armadillos’ burrows, spider webs are commonly found. The objective of this study was to evaluate the presence of Pb in spider webs samples related to the habitat of armadillos. Spider web samples were collected at Lageado Farm, Botucatu/SP and prepared for microscopic, molecular and mycological analyses. Microscopic analysis showed that different fungi were closely attached to spider web samples. Nested-PCR reaction showed positive amplification for Pb in 4 samples, with identity confirmed by amplicon sequencing. Fungal colonies also included members of Aspergillus, Blastobotrys, Penicillium, Candida, and Sporothrix genera, which are related to opportunistic disease and primary infections of great medical importance. In vitro adhesion tests of mycelia and yeast form of Pb to the spider webs were also performed, in order to analyze the possible physical attraction between fungal cells and the spider web protein network. The results showed a clear adherence of fungal particles to spider webs. In the current literature, there are no studies reporting adhesive properties of microorganisms to spider webs... (Complete abstract click electronic access below)

Levantamento da mastofauna e identificação das influências antrópicas em dois fragmentos de Mata Atlântica transição com Cerrado de Rubião Junior – Botucatu - SP

Atlantic Forest and Cerrado are between the biological richest areas in the world, for that are considered top conservation priorities. Those ecosystems are found at Botucatu region, where it´s extremely fragmented by routes, agricultural areas and urban centres. This fragmentation causes restrictions on animals populations, in particular to medium and large mammals, that needs larger areas to live and also are often hunted. Even isolated and suffering anthropic influences the fragments are fundamental for local biodiversity conservation. We realized a mammal survey on two forest fragments of Atlantic forest transition to Cerrado of Rubião Júnior – Botucatu – SP – Brazil: Santo Antônio Church and Parque das Cascatas forests. The Santo Antônio church munt is considered a turistic point and its forest receive many visitants, Parque das Cascatas forest is inside of a residential condominium. Both areas are classify as semidecidual stational forest and are disconected by Domingos Sartori route, that connect Botucatu centre to Rubião Júnior district. Around both fragments can be found residences, plantations and pastures. The utilized method included: recognition of tracks on the forests, interviews with next residents and sand plots mounting. The interviews indicated 29 mammals species, and 19 was validated for animal traces, overruns or visualization. Other 3 species found were not mentioned on interviews. Totally, was found 22 small and medium mammals presence evidences on both forest fragments. Parque das Cascatas forest showed more diversity (19) then Santo Antônio Church forest (11), and species like Eira barbara, Procyon cancrivorus, Mazama americana, Shiggurus villosus, Dasypus novemcinctus, Didelphis albiventris, Lutreolina crassicaudata and Lepus europaeus looked for other areas once that was verifyed overruns of individuals... (Complete abstract click electronic access below)

Molecular detection of Leishmania spp. in road-killed wild mammals in the Central Western area of the State of São Paulo, Brazil

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

Detecção do Paracoccidioides spp. em amostras ambientais e diferenciação do complexo P. brasiliensis da espécie P. lutzii por Nested PCR e hibridização in situ

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Translocation of nine-banded armadillos

During the last 150 years, nine-banded armadillos (Dasypus novemcinctus) have increased their range and abundance in the southeastern United States. When foraging, armadillos cause damage to agricultural crops as, as well as cause structural damage to driveways and foundations. Homeowners frequently use translocation to reduce local armadillo abundance. Despite its popularity with the general public, however, the appropriateness of nuisance wildlife translocation presents concerns for biologists. Our objective was to address some of these concerns by examining survival and movements of translocated armadillos. We translocated 12 armadillos (9 male, 3 female) equipped with radio-transmitters and compared their survival and movements to that of 29 (11 male, 18 female) resident armadillos. Most (92%) of the translocated animals dispersed from their release site within the fi rst few days after release. Resident armadillos generally maintained stable home ranges. We found evidence that translocated animals were abl to return to their original capture sites. We, therefore, recommend against translocating nuisance armadillos.

Evaluation of Attractants for Live-Trapping Nine-Banded Armadillos

In the past 50 years, the range of the nine-banded armadillo (Dasypus novemcinctus) in the south has been rapidly expanding. As their range expands, armadillos increasingly come into conflict with suburban landowners. When foraging, armadillos often uproot ornamental plants. Their rooting also destroys gardens, lawns, and flower beds. Their burrowing can damage tree roots and building foundations. Most armadillo damage is a result of their feeding habits. Armadillos dig shallow holes, 1- 3 inches deep and 3-5 inches long, as they search for soil invertebrates. A recent survey of Georgia county extension agents by scientists at the University of Georgia found that 77.6% of all agents reported receiving complaints or requests for information on armadillos. Armadillo related inquiries made up 10.1 % all inquiries for all agents across the state, surpassing even the white-tail deer (Odocoileus virginianus). Armadillos are often assumed to destroy nests of ground-nesting birds. Armadillo diets have been studied in several states including Alabama, Louisiana, Texas, Georgia, Arkansas, and Florida. According to these studies, vertebrate matter, especially bird eggs, made up an minor portion of their diet. The armadillo’s diet often consists of more than 90% insects, grubs and earthworms. Based on these studies, it seems that claims of armadillos being significant nest predators are unfounded.

Placentation in the anteaters Myrmecophaga tridactyla and Tamandua tetradactyla (Eutheria, Xenarthra)

Background: Since Xenarthra are serious candidates for being basal to Eutheria, their characteristics, e.g. the placental system, influence perceptions of evolution. However, in the subgroup containing the anteaters, data are very limited. The present study aims to elucidate the nature of the feto-maternal interface in the anteater placenta and to interpret these data within an evolutionary context. Methods: Placentas of two species were investigated with histology, immunohistochemistry and transmission electron microscopy. Results: Remnants of the maternal vessel endothelium were absent, resulting in a fully haemochorial barrier throughout the placenta. Two structurally different parts, the villous and trabecular areas were complex and intermingled. In particular, the trabeculae which consisted of cellular, proliferative trophoblast, associated with connective tissue, were attached to the decidua. The villi contained fetal capillaries and hypertrophied mesenchymal cells that occured near the surface near the end of gestation. The surface of the villi consisted of flat, syncytial trophoblast, interspersed with proliferative trophoblast cells. Conclusions: Based on fundamental differences between anteaters and armadillos, we inferred that placental evolution was more complex than previously thought. The haemochorial pattern of anteaters was likely an ancient condition of xenarthrans. Consequently, villous placentation may be attributed, at least in part, by convergent evolution, but was also characterized by some features that were widespread among xenarthrans.

The fetomaternal interface in the placenta of three species of armadillos (Eutheria, Xenarthra, Dasypodidae)

Background: Placental characters vary among Xenarthra, one of four supraordinal clades of Eutheria. Armadillos are known for villous, haemochorial placentas similar to humans. Only the nine-banded armadillo has been well studied so far. Methods: Placentas of three species of armadillos were investigated by means of histology, immunohistochemistry including proliferation marker, and transmission and scanning electron microscopy. Results: The gross anatomy differed: Euphractus sexcinctus and Chaetophractus villosus had extended, zonary placentas, whereas Chaetophractus vellerosus had a disk. All taxa had complex villous areas within the maternal blood sinuses of the endometrium. Immunohistochemistry indicated the validity of former interpretations that the endothelium of the sinuses was largely intact. Tips of the villi and the columns entering the maternal tissue possessed trophoblast cell clusters with proliferation activity. Elsewhere, the feto-maternal barrier was syncytial haemochorial with fetal vessels near the surface. Conclusions: Differences among armadillos occurred in regard to the extension of the placenta, whereas the fine structure was similar. Parallels to the human suggest that armadillos are likely to be useful animal models for human placentation.

Paracoccidioides brasiliensis infection in dogs from Western Brazilian Amazon

The objective of the study was to evaluate Paracoccidioides brasiliensis infection in urban dogs from the municipality of Monte Negro, Rondonia, Western Brazilian Amazon. The serum samples (n=126) were analyzed by indirect ELISA and the immunodiffusion test using P. brasiliensis gp43 and exoantigen as antigens, respectively. A positivity of 54.8% was observed only in the ELISA test and no statistical difference was observed in the seroprevalence in relation to age or sex. This is the first paracoccidioidomycosis survey carried out with dogs from the Western Brazilian Amazon. The higher positivity rates of P. brasiliensis infection observed in this study suggest that veterinarians must be alert to detect new cases of natural disease in dogs living in paracoccidioidomycosis endemic areas.

Importance of xenarthrans in the eco-epidemiology of Paracoccidioides brasiliensis

Abstract Background Several pathogens that cause important zoonotic diseases have been frequently associated with armadillos and other xenarthrans. This mammal group typically has evolved on the South American continent and many of its extant species are seriously threatened with extinction. Natural infection of armadillos with Paracoccidioides brasiliensis in hyperendemic areas has provided a valuable opportunity for understanding the role of this mammal in the eco-epidemiology of Paracoccidioidomycosis (PCM), one of the most important systemic mycoses in Latin America. Findings This study aimed to detect P. brasiliensis in different xenarthran species (Dasypus novemcinctus, Cabassous spp., Euphractus sexcinctus, Tamandua tetradactyla and Myrmecophaga tridactyla), by molecular and mycological approaches, in samples obtained by one of the following strategies: i) from road-killed animals (n = 6); ii) from naturally dead animals (n = 8); iii) from animals that died in captivity (n = 9); and iv) from living animals captured from the wild (n = 2). Specific P. brasiliensis DNA was detected in several organs among 7/20 nine-banded armadillos (D. novemcinctus) and in 2/2 anteaters (M. tridactyla). The fungus was also cultured in tissue samples from one of two armadillos captured from the wild. Conclusion Members of the Xenarthra Order, especially armadillos, have some characteristics, including a weak cellular immune response and low body temperature, which make them suitable models for studying host-pathogen interaction. P. brasiliensis infection in wild animals, from PCM endemic areas, may be more common than initially postulated and reinforces the use of these animals as sentinels for the pathogen in the environment.