RESUMO
Many microorganisms are able to cause diseases in amphibians, and in the past few years one of the most reported has been Batrachochytrium dendrobatidis. This fungus was first reported in Brazil in 2005; following this, other reports were made in specimens deposited in museum collections, captive and free-living frogs. The aim of this study was to compare singleplex and nested-PCR techniques to detect B. dendrobatidis in free-living and apparently healthy adult frogs from the Brazilian Atlantic Forest. The sample collection area was a protected government park, with no general entrance permitted and no management of the animals there. Swabs were taken from the skin of 107 animals without macroscopic lesions and they were maintained in ethanol p.a. Fungal DNA was extracted and identification of B. dendrobatidis was performed using singleplex and nested-PCR techniques, employing specific primers sequences. B. dendrobatidis was detected in 61/107 (57%) and 18/107 (17%) animals, respectively by nested and singleplex-PCR. Nested-PCR was statistically more sensible than the conventional for the detection of B. dendrobatidis (Chi-square = 37.1; α = 1%) and the agreement between both techniques was considered just fair (Kappa = 0.27). The high prevalence obtained confirms that these fungi occur in free-living frogs from the Brazilian Atlantic Forest with no macroscopic lesions, characterizing the state of asymptomatic carrier. We concluded that the nested-PCR technique, due to its ease of execution and reproducibility, can be recommended as one of the alternatives in epidemiological surveys to detect B. dendrobatidis in healthy free-living frog populations.
Assuntos
Anfíbios/microbiologia , Portador Sadio/veterinária , Quitridiomicetos/isolamento & purificação , Micoses/veterinária , Reação em Cadeia da Polimerase/métodos , Animais , Brasil , Portador Sadio/microbiologia , Quitridiomicetos/genética , Técnicas Microbiológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Micoses/microbiologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Medicina Veterinária/métodosRESUMO
Many microorganisms are able to cause diseases in amphibians, and in the past few years one of the most reported has been Batrachochytrium dendrobatidis. This fungus was first reported in Brazil in 2005; following this, other reports were made in specimens deposited in museum collections, captive and free-living frogs. The aim of this study was to compare singleplex and nested-PCR techniques to detect B. dendrobatidis in free-living and apparently healthy adult frogs from the Brazilian Atlantic Forest. The sample collection area was a protected government park, with no general entrance permitted and no management of the animals there. Swabs were taken from the skin of 107 animals without macroscopic lesions and they were maintained in ethanol p.a. Fungal DNA was extracted and identification of B. dendrobatidis was performed using singleplex and nested-PCR techniques, employing specific primers sequences. B. dendrobatidis was detected in 61/107 (57%) and 18/107 (17%) animals, respectively by nested and singleplex-PCR. Nested-PCR was statistically more sensible than the conventional for the detection of B. dendrobatidis (Chi-square = 37.1; α = 1%) and the agreement between both techniques was considered just fair (Kappa = 0.27). The high prevalence obtained confirms that these fungi occur in free-living frogs from the Brazilian Atlantic Forest with no macroscopic lesions, characterizing the state of asymptomatic carrier. We concluded that the nested-PCR technique, due to its ease of execution and reproducibility, can be recommended as one of the alternatives in epidemiological surveys to detect B. dendrobatidis in healthy free-living frog populations.(AU)
Assuntos
Animais , Anfíbios/microbiologia , Portador Sadio/veterinária , Quitridiomicetos/isolamento & purificação , Micoses/veterinária , Reação em Cadeia da Polimerase/métodos , Brasil , Portador Sadio/microbiologia , /genética , Técnicas Microbiológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Micoses/microbiologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Medicina Veterinária/métodosRESUMO
Many microorganisms are able to cause diseases in amphibians, and in the past few years one of the most reported has been Batrachochytrium dendrobatidis. This fungus was first reported in Brazil in 2005; following this, other reports were made in specimens deposited in museum collections, captive and free-living frogs. The aim of this study was to compare singleplex and nested-PCR techniques to detect B. dendrobatidis in free-living and apparently healthy adult frogs from the Brazilian Atlantic Forest. The sample collection area was a protected government park, with no general entrance permitted and no management of the animals there. Swabs were taken from the skin of 107 animals without macroscopic lesions and they were maintained in ethanol p.a. Fungal DNA was extracted and identification of B. dendrobatidis was performed using singleplex and nested-PCR techniques, employing specific primers sequences. B. dendrobatidis was detected in 61/107 (57%) and 18/107 (17%) animals, respectively by nested and singleplex-PCR. Nested-PCR was statistically more sensible than the conventional for the detection of B. dendrobatidis (Chi-square = 37.1; α = 1%) and the agreement between both techniques was considered just fair (Kappa = 0.27). The high prevalence obtained confirms that these fungi occur in free-living frogs from the Brazilian Atlantic Forest with no macroscopic lesions, characterizing the state of asymptomatic carrier. We concluded that the nested-PCR technique, due to its ease of execution and reproducibility, can be recommended as one of the alternatives in epidemiological surveys to detect B. dendrobatidis in healthy free-living frog populations.
Assuntos
Animais , Anfíbios/microbiologia , Portador Sadio/veterinária , Quitridiomicetos/isolamento & purificação , Micoses/veterinária , Reação em Cadeia da Polimerase/métodos , Brasil , Portador Sadio/microbiologia , Quitridiomicetos/genética , Técnicas Microbiológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Micoses/microbiologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Medicina Veterinária/métodosRESUMO
The aim of this study was to demonstrate the Leptospirosis occurrence in stray dogs found in the campus of Universidade Estadual de Londrina PR., from July to September 2007. The MAT (Microscopic Agglutination Test) was made as diagnostic serological test in 33 serum samples and the direct dark-field microscopy (DFM) in 10 urine samples. In the MAT, seven samples (21,21%) reacted and antibodies against the serovars Canicola, Pyrogenes and Castellonis were detected with titles of 200 to 400. In the DFM, were visualized typical Leptospira spp. morphology and motility structures in one sample. These data indicate the necessity of stray dogs population control because it may represents a public health problem in the university campus population.
O objetivo deste estudo foi demonstrar a ocorrência de anticorpos anti leptospira em cães errantes encontrados entre julho e setembro de 2007, no campus da Universidade Estadual de Londrina PR. Para o diagnóstico da leptospirose foi realizada a soroaglutinação microscópica (SAM) em 33 amostras de soro e o exame direto da urina em microscópio de campo escuro (CE) em 10 amostras. Na SAM, foram reagentes sete (21,21%) amostras e anticorpos contra os sorovares Canicola, Pyrogenes e Castellonis foram detectados com títulos de 200 a 400. No CE foram visualizadas células com morfologia e movimentos compatíveis com Leptospira spp. em uma amostra. Os resultados deste trabalho indicam a necessidade de controle de cães errantes, pois eles podem representar um problema de saúde pública para a população do campus universitário.