Pathogenicities and GP43kDa gene of three Paracoccidioides brasiliensis isolates originated from a nine-banded armadillo (Dasypus novemcinctus)

We studied three different isolates of Paracoccidioides brasiliensis obtained from the mesenteric lymph node (D3LY1), the spleen (D3S1) and the liver (D3LIV1) of the same armadillo (Dasypus novemcinctus ).Pulmonal inflammatory area was evaluated by intravenous inoculation of 10(6) yeast cells of each isolates in young, male, ddY mice. Moreover, the partial sequence of GP43kDa gene of P. brasiliensis was analyzed. The lung inflammatory area was greater in animals inoculated with isolate D3S1. The partial sequence of GP43kDa gene indicated that isolate D3S1 is different from isolates D3LY1 and D3LIV1. This study suggested that the same armadillo might be susceptible to multiple P. brasiliensis isolates simultaneously.

Light and scanning electron microscopic study of the palatine mucosa of nine-banded armadillo (Dasypus novemcinctus)

The three-dimensional structure of the lamina propria of the hard and soft palatine mucosa of the nine-banded armadillo was observed by scanning electron microscopy. Sodium hydroxide cell maceration method was applied to demonstrate the architecture of the connective tissue papillae. The palatine mucosa of the armadillo had a triangular shape and measured appr. 6.5 cm length. The hard palate showed 9 transverse palatine plicae while the soft palate was smooth. In the 10% NaOH treated specimens, the lamina propria of the hard palatine mucosa showed numerous connective tissue papillae with a general finger-like shape. These structures were composed by a meshwork of collagen fibers arranged in several directions. on the other hand, the connective tissue papillae of the soft palate mucosa were scattered and small. Numerous openings of glandular ducts with circular or elliptical shape were located in the interplicae area and in the soft palate.

Cerebriform colonies of Paracoccidioides brasiliensis isolated from nine-banded armadillos (Dasypus novemcinctus) at room temperature

Twelve isolates of Paracoccidioides brasiliensis generated cerebriform colonies at room temperature on potato glucose agar slants (PDA). These isolates contained abundant chlamydospores and yeast-like cells and are a subset of the 65 isolates obtained from nine-banded armadillos (Dasypus novemcinctus). They grew as a yeast form with typical multiple buddings at 37 degreesC on brain heart infusion agar supplemented with 1% glucose. After replating on PDA and culturing at room temperature for 2 months, the mutants appeared as cottonous colonies, which indicated that the morphological characteristics were unstable.

Caracterização molecular de isolados de Paracoccidioides brasiliensis obtidos de Dasypus novemcinctus (tatu) e sua correlação com virulência e origem geográfica

Pós-graduação em Patologia - FMB

Descrição anatômica das estruturas osteoarticulares do esqueleto axial do tatu-galinha (Dasypus novemcinctus Linnaeus, 1758) por meio da radiografia e tomografia computadorizada

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Experimental infections with Paracoccidioides brasiliensis obtained from armadillos: comparison to clinical isolates

Paracoccidioides brasiliensis causes paracoccidioidomycosis (PCM) that is one of the most prevalent systemic human mycoses in Latin America. Armadillos show a high incidence of PCM infection and could, therefore, be a natural reservoir for this fungus. In this study were compared the virulence profiles of isolates obtained from nine-banded armadillos (Dasypus novemcinctus) (PbT1 and PbT4) and isolates from PCM patients (Pb265 and Bt83). Pathogenicity was evaluated by fungal load and analysis of colony morphology. Immunity against the fungus was tested by delayed type hypersensitivity test (DTH) and antibody quantification by ELISA. The higher virulence of PbT1 and PbT4 was suggested by higher fungal load in spleen and lungs. Armadillo isolates and Bt83 presented a cotton-like surface contrasting with the cerebriform appearance of Pb265. All isolates induced cellular and humoral immune responses in infected BALB/c mice. DTH reactions were similarly induced by the four isolates, however, a great variability was observed in specific antibody levels, being the highest ones induced by Bt83 and PbT4. The present work confirms that armadillos harbor P. brasiliensis, whose multiplication and induced immunity in experimentally infected mice are heterogeneous, resembling the behavior of isolates from human PCM. This study reinforces the possibility that armadillos play an important role in the biological cycle of this pathogen.

Molecular detection of Paracoccidioides brasiliensis in road-killed wild animals

Paracoccidioides brasiliensis infections have been little studied in wild and/or domestic animals, which may represent an important indicator of the presence of the pathogen in nature. Road-killed wild animals have been used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies of paracoccidiodomycosis (PCM). The presence of P. brasiliensis infection was evaluated by Nested-PCR in tissue samples collected from 19 road-killed animals; 3 Cavia aperea (guinea pig), 5 Cerdocyon thous (crab-eating-fox), 1 Dasypus novemcinctus (nine-banded armadillo), 1 Dasypus septemcinctus (seven-banded armadillo), 2 Didelphis albiventris (white-eared opossum), 1 Eira barbara (tayra), 2 Gallictis vittata (grison), 2 Procyon cancrivorus (raccoon) and 2 Sphiggurus spinosus (porcupine). Specific P. brasiliensis amplicons were detected in (a) several organs of the two armadillos and one guinea pig, (b) the lung and liver of the porcupine, and (c) the lungs of raccoons and grisons. P. brasiliensis infection in wild animals from endemic areas might be more common than initially postulated. Molecular techniques can be used for detecting new hosts and mapping `hot spot` areas of PCM.

Serological Evidence of Paracoccidioides brasiliensis Infection in Chickens from Parana and Mato Grosso do Sul States, Brazil

The objective of this study was to detect antibodies against Paracoccidioides brasiliensis in free-range and caged chickens Gallus domesticus. Initially, the humoral immune response of two chickens immunized with P. brasiliensis was evaluated. Both animals showed the production of antibodies to gp43, the major P. brasiliensis antigen. The seroepidemiological survey was conducted in chickens from the Pantanal region in Mato Grosso do Sul State (free-range n = 40) and from northern region of Parana State (free-range n = 100, caged n = 43). The serum samples were analyzed by indirect ELISA using gp43 as antigen. The positivity observed in free-range chickens from Mato Grosso do Sul (55%) was significantly higher (P = 0.0001) than in free-range chickens from Parana State (16%). In contrast to the free-range chickens, no positivity was observed in the caged chickens (P = 0.003). This is the first report showing serological evidence of P. brasiliensis infection in chickens. The results suggest that free-range chickens are more frequently infected by P. brasiliensis, probably due to the constant contact with soil than caged chickens and could be useful as epidemiological markers of paracoccidioidomycosis.

Toxoplasmose em animais silvestres e domésticos da região de Botucatu, estado de São Paulo, Brasil

Os Autores analisaram soros de 47 Cannis familiaris, de 9 Felis cattus, de 64 Didelphis marsupialis aurita, de 9 Dasypus novemcinctus, de 4 Cabassous tatouay e de 29 Rattus rattus, através da reação de imunofluorescência indireta, para pesquisar a presença de anticorpos anti Toxoplasma gondii. Estes foram encontrados apenas em C. familiaris (63,8%) e em D. m. aurita (4,7%). Frente aos resultados obtidos, os Autores sugerem que novas pesquisas nesta área sejam realizadas, para que se conheça melhor a importância epidemiológica de várias espécies animais na disseminação da toxoplasmose.

Epidemiologia da doença de Chagas no Ceará VIII - estudo da infecção de animais por T. cruzi no município de Morada Nova

Foram examinados 1.368 mamíferos em ecótopos naturais ou artificiais: 443 cães com 2,5% positivos; 605 gatos com 0,7%, 65 roedores domésticos com 4,6%, 6 coelhos e 10 cobaias negativos. Dentre os animais silvestres, foram examinados 115 preás (Galea s. spixii) com 2,3% positivos, 12 "cassacos" (Didelphis azarae) dos quais 9 positivos; 5 quirópteros, 5 punáres (Cercomys c. laurentius) e 3 pebas (Dasypus sexcinctus) negativos. A amostra apresentada é suficiente para considerada a existência de ciclos doméstico e silvestre de T. cruzi na área estudada e considerada endêmica da Doença de Chagas.

Epidemiological factors related to the transmission risk of Trypanosoma cruzi in a Quilombola community, State of Mato Grosso do Sul, Brazil

INTRODUCTION: This work was an epidemiological investigation of the risk of Trypanosoma cruzi transmission in the rural Quilombola community of Furnas do Dionízio, State of Mato Grosso do Sul, Brazil. METHODS: Of the 71 animals examined, seven were captured (two opossums, Didelphis albiventris; four rats, Rattus rattus; and one nine-banded armadillo, Dasypus novemcinctus) and 64 were domestic (one canine, Canis familiaris; five pigs, Sus scrofa; two bovines, Bos taurus; five caprines, Capra sp.; and 51 ovines, Ovis aries). Parasitological tests were performed to detect parasites in the blood and to identify the morphology of flagellates. These methods included fresh examinations, buffy coat tests and blood cultures. Molecular analysis of DNA for identification of trypanosomatids was performed by polymerase chain reaction (PCR) with primers S35 and S36. RESULTS: The parasitological tests showed flagellates in an opossum and two cattle. The molecular tests showed DNA from T. cruzi in an opossum and a pig. Triatoma sordida was the only triatomine species found in the community, and it colonized households (four specimens) and the surrounding areas (124 specimens). Twenty-three specimens tested positive for flagellates, which were subsequently identified as T. cruzi by PCR. CONCLUSIONS: Data analysis demonstrated that T. cruzi has a peridomestic life cycle that involves both domestic and wild mammals.

Mamíferos de médio e grande porte e sua relação com o mosaico de habitats na cuesta de Botucatu, Estado de São Paulo, Brasil

A região da cuesta de Botucatu caracteriza-se por um gradiente topográfico contendo um mosaico de ambientes com diferentes formações de vegetação natural (floresta estacional semidecidual, cerrado e matas ciliares), além de áreas antropizadas com a predominância de pastagens, plantações de cana-de-açúcar, laranja, e reflorestamentos de eucalipto, com paisagem fragmentada. Inserida nesta região, a Fazenda Experimental Edgardia, pertencente à Universidade Estadual Paulista, Campus de Botucatu, representa uma amostra desta heterogeneidade ambiental, tendo grande importância para a conservação da biodiversidade, tanto de flora como fauna. Entretanto, poucos são os estudos sobre a sua fauna, principalmente de mamíferos. O presente trabalho teve como objetivo conhecer a fauna de mamíferos de médio e grande porte nesta área, e sua relação com o mosaico de habitats. Foram obtidos registros indiretos da presença de mamíferos através de vestígios (pegadas e fezes) em transectos (trilhas pré-existentes), ao longo de um ano. De março de 2004 a março de 2005 foram registradas 18 espécies de mamíferos silvestres de médio e grande porte. Quanto à ocorrência destacou-se Mazama sp., presente em todos os ambientes, com maior abundância relativa no ambiente de transição de floresta/Cerradão. Puma concolor (Linnaeus, 1771), Leopardus pardalis (Linnaeus, 1758), Cerdocyon thous (Linnaeus, 1766), Procyon cancrivorus (Cuvier, 1798) e Dasypus novemcinctus (Linnaeus, 1758) também foram encontradas em praticamente todos os ambientes, e espécies como Chironectes minimus (Zimmermann, 1780), Cuniculus paca Linnaeus, 1766, Eira barbara (Linnaeus, 1758) e uma espécie do gênero Conepatus Gray, 1837 estiveram restritas a ambientes específicos. A análise de correspondência mostrou oito espécies com ocorrência em todos os ambientes: sete mais associadas aos ambientes de várzea, floresta e pastagem e três aos ambientes de cultura de arroz, transição entre floresta/Cerradão e vegetação natural em estádio sucessional secundário. Os resultados sugerem que a fauna de mamíferos de médio e grande porte na Fazenda Experimental Edgardia está sujeita às modificações ambientais que a região vem sofrendo. Embora existam preferências de algumas espécies por determinados hábitats, parece ocorrer certa plasticidade em relação às modificações de seus hábitats originais.

Trypanosomes of the subgenus Megatrypanum from armadillos (Xenarthra: Dasypodidae)

A new species of trypanosome, Trypanosoma (Megatrypanum) peba, is described from the peripheral blood of the armadillo Euphractus sexcinctus setosus from Bahia State, Brazil. Ten out of 29 specimens of the armadillo Dasypus novemcinctus from Pará State were found to have trypanosomes, including epimastigote forms, in impression smears of subcutaneous lymph nodes. The trypanosomes from D. novemcinctus are illustrated and were identified a idenrified as belonging to the subgenus Megatrypanum on the basis of their general appearance, although they failed to multiply is blood-agar culture medium and no bloodstream forms were seen. This is the first published record of trypanosomes of this subgenus from armdillos and the first demonstration of epimastigote trypanosomes in the mammalian host other than in the bloodstream, or in the anal glands of opssums.

Epidemiological and nosological aspects of Leishmania naiffi Lainson & Shaw, 1989

Leismania naiffi was isolated from 10 out of 64 armadillos (Dasypus novemcinctus) examined in Amazonas, Pará and Rondônia States in the Brazilian Amazon Region. The isolates were obtained in culture from samples of liver (3), spleen (3), lymph nodes (2), skin (1) and blood (1) from the infected animals. Heavy infections with the same parasite were detected for the first time in Psychodopygus squamiventris, a common man-biting phlebotomine, in amazonas and Pará. A new case of cutaneous leishmaniasis caused by L. naiffi is described from the Manaus area, making a total of three known cases of human infection by this parasite.