Molecular identification and serological profile of Leptospira spp. isolated of white-eared-opossum (Didelphis albiventris) in South of Brazil

A Leptospirose ?? uma antropozoonose de ocorr??ncia mundial, particularmente nos pa??ses em desenvolvimento, causada por bact??rias do g??nero Leptospira. V??rias esp??cies de marsupiais e didelf??deos s??o consideradas suscet??veis a infec????o causada por uma grande variedade de sorovares patog??nicos de Leptospira spp. tendo sido considerados poss??veis hospedeiros deste agente. Neste trabalho foram coletadas amostras de soro e urina de 33 gamb??s-de-orelha-branca (Didelphis albiventris), capturados em diferentes regi??es dos munic??pios do Cap??o do Le??o e Pelotas, no sul do Brasil, com o objetivo de detectar anticorpos e isolar leptospiras. As amostras de soro foram testadas por aglutina????o microsc??pica (MAT), utilizando 58 sorovares de Leptospira spp. e 1 de Leptonema. Para o isolamento, amostras de urina foram inoculadas em meio de cultura EMJH enriquecido com 10% de suplemento Difco??. Um sorovar patog??nico foi obtido da urina, sendo o primeiro isolado do pa??s para esta esp??cie, e foi caracterizado quanto ao g??nero e patogenicidade por PCR utilizando primers para os genes 16S rDNA e Lipl32. A t??cnica de seq??enciamento parcial do gene rpoB foi utilizada para identificar a esp??cie gen??mica. A cepa isolada pertence ?? esp??cie L. borgpertersenii,, que juntamente com L. interrogans s??o as principais causadoras de doen??a em humanos. Anticorpos anti isolado de gamb?? foram avaliados em amostras sorol??gicas de 60 c??es, 60 bovinos, 60 humanos e dos 33 gamb??s capturados atrav??s da MAT. Os soros humanos apresentaram rea????o de 3,3% (2/60), 28,33% (17/60) em soros caninos, 1,67% (1/60) em soros bovinos e 36,36% (12/33) em soros dos gamb??s capturados no estudo. Esses dados sugerem o envolvimento do gamb??-de-orelha-branca na manuten????o de leptospiras no ambiente, uma vez que estando infectados por sorovares patog??nicos podem eliminar o agente pela urina e infectar direta ou indiretamente animais dom??sticos e conseq??entemente o homem.Leptospirosis is a zoonotic disease that occurs all over the world, particularly in developing countries, caused by bacteria of the genus Leptospira. Several marsupial species are considered susceptible to infection caused by a wide variety of Leptospira serovars, acting as reservoirs. In this work were collected serum and urine samples of 33 White-eared opossum, trapped within distinct locations of Cap??o do Le??o and Pelotas cities, in the South of Brazil, aiming to detect antibodies and to isolate leptospires. Serum samples were screened against a panel of 58 Leptospira spp and Leptonema illini serovars using the microscopic agglutination test (MAT). To attempt isolation, urine samples from all animals were inoculated in culture medium.EMJH enriched with 10% of supplement Difco?? One pathogenic serovar was isolated after two months of inoculation, and is first isolate the country for this animal species and was characterized by PCR using primers for 16S and Lipl32 genes. The technique of partial sequencing of the rpoB gene was used to identify the genomic specie. The isolated strain belongs to the specie L. borgpetersenii, which along with L .interrogans are the main cause disease in humans. The isolate, was tested on MAT using dog, cattle and human sera. Cut off titer of 50 was used for domestic animal sera and 25 for human and opossum sera. Both human and animal sera presented agglutination on MAT to isolated strain corresponding to 3.3% (2/60), 28.33% (17/60) and 1.67% (1/60) in human, dog and cattle sera, respectively. The opossum sera presented 36.36% (12/33) of reaction. These findings suggest a probable white-eared opossum role in the maintenance of pathogenic Leptospira on the environment since low titer sera and urine elimination were observed. These animals could be important reservoirs of Leptospira serovars for human and domestic animals