RESUMO
The biological and genetic diversity of Neospora caninum is very limited because of availability of only a few viable isolates worldwide. This study describes the isolation and biological and molecular characterization of a new viable isolate of N. caninum (NC-SP1), from a cattle in Brazil. Approximately 400 g of brain from a naturally infected adult male cattle from an abattoir was fed to a 2-month-old dog. Neospora-like oocysts were observed on day 7 post-inoculation (PI) and the duration of oocyst shedding was 14 days. The DNA obtained from oocysts was characterized molecularly and the final sequence was 99% identical to homologous sequences of N. caninum available in GenBank®. For bioassay, gerbils (Meriones unguiculatus) were orally inoculated with 10 100 and 1000 oocysts; all gerbils remained clinically normal but developed N. caninum antibodies 14 days PI. Cell culture isolation was successful using the brain homogenate from one of the gerbils and tachyzoites were observed 24 days PI. Microsatellite genotyping revealed a unique genetic profile for this new reference isolate.
Assuntos
Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Neospora/isolamento & purificação , Animais , Anticorpos Antiprotozoários/sangue , Bioensaio/veterinária , Encéfalo/parasitologia , Brasil , Bovinos , Coccidiose/parasitologia , DNA de Protozoário/química , Cães , Fezes/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Técnicas de Genotipagem/veterinária , Gerbillinae , Masculino , Repetições de Microssatélites , Neospora/genética , Neospora/imunologia , Oocistos/genética , Oocistos/imunologia , Oocistos/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Soro/parasitologiaRESUMO
Serum samples from 116 horses and 47 dogs from the municipality of Paulicéia, in the state of São Paulo, Brazil, were examined for anti-Toxoplasma gondii, -Neospora spp. and -Sarcocystis neurona antibodies by means of the indirect fluorescent antibody test (IFAT). The results showed that only one horse was seropositive for T. gondii (0.9%) and anti-Neospora spp. antibodies were detected in three out of the 116 horses tested (2.6%). However, 27 horses showed antibodies against S. neurona (23.8%). Amongst the serum samples from the dogs, 10 out of the 47 dogs showed antibodies against T. gondii (21.3%) and three dogs showed antibodies against Neospora caninum (6.4%). This study reports that in the municipality of Paulicéia dogs in both the rural and the urban area were exposed to T. gondii and N. caninum, while horses in the rural area were exposed to all three protozoa studied, with high occurrence of anti-S. neurona antibodies.(AU)
Amostras de soro de 116 equinos e 47 cães provenientes do município de Paulicéia, São Paulo, foram testadas para detecção de anticorpos anti-Toxoplasma gondii, -Neospora spp. e -Sarcocystis neurona por meio da reação de imunofluorescência indireta (RIFI). Apenas um equino, entre 116 equinos testados, teve diagnóstico soropositivo para T. gondii (0,9%), e três deles (2,6%) apresentaram anticorpos anti-Neospora spp. Entretanto, 27 equinos apresentaram anticorpos anti-S. neurona (23,8%). Nas amostras de cães, dez dos 47 animais apresentaram anticorpos anti-T. gondii (21,3%) e três tiveram diagnóstico soropositivo para Neospora caninum (6,4%). Este estudo relata que no município de Paulicéia os cães das áreas urbana e rural foram expostos a T. gondii e N. caninum, enquanto os equinos da área rural foram expostos aos três protozoários estudados, com alta ocorrência de anticorpos anti- S. neurona.(AU)
