RESUMO
In view of the importance of venereal transmission of bovine leptospirosis, the objective of the present study was to apply the polymerase chain reaction (PCR) to 26 serovars of Leptospira interrogans, L. borgpetersenii, L. santarosai, L. noguchii and L. biflexa, to determine the detection threshold in semen samples and to evaluate the possibility of differentiation among serovars using 19 restriction endonucleases. The results showed that all serovars were amplified and the detection threshold in semen samples of a bull was 100 bacteria/ml. Using endonucleases we could classify the 26 serovars into eight groups. The present results show that PCR is a method of great potential for the detection of Leptospira spp. at bovine artificial insemination centers.
Assuntos
Doenças dos Bovinos/microbiologia , Leptospira interrogans/genética , Leptospira/genética , Leptospirose/veterinária , Sêmen/microbiologia , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/prevenção & controle , Primers do DNA/química , DNA Bacteriano/química , DNA Bacteriano/isolamento & purificação , Eletroforese em Gel de Poliacrilamida/veterinária , Inseminação Artificial/veterinária , Leptospira/classificação , Leptospira/isolamento & purificação , Leptospira interrogans/química , Leptospira interrogans/classificação , Leptospirose/diagnóstico , Leptospirose/microbiologia , Leptospirose/prevenção & controle , Masculino , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de RestriçãoRESUMO
Due to the high importance of the venereal transmission of bovine leptospirosis, this study aimed to test the ability of PCR to detect Leptospira interrogans serovar hardjo DNA in experimentally contaminated bovine semen. Employing primers directed to the 16S rRNA gene, 10 bacteria/ml of semen could be detected by PCR. Results achieved in this work show that PCR can have a great potential to detect Leptospira spp. in insemination centers
