Abstract
The aim of the present study was to evaluate a species-specific nested PCR based on a previously described species-specific PCR for detection of B. ovis in semen and urine samples of experimentally infected rams. The performance of the species-specific nested PCR was compared with the results of a genus-specific PCR. Fourteen rams were experimentally infected with the Brucella ovis REO 198 strain and samples of semen and urine were collected every week up to 180 days post infection. Out of 83 semen samples collected, 42 (50.6%) were positive for the species-specific nested PCR, and 23 (27.7%) were positive for the genus-specific PCR. Out of 75 urine samples, 49 (65.3%) were positive for the species-specific nested PCR, whereas 11 (14.6%) were genus-specific PCR positive. Species-specific nested PCR was significantly more sensitive (P<0.001) than the genus-specific PCR in semen and urine from experimentally infected rams. In conclusion, the species-specific nested PCR developed in this study may be used as a diagnostic tool for the detection of B. ovis in semen and urine samples from suspected rams.
Highlights
Ovine brucellosis caused by the Brucella ovis is a contagious disease, with venereal and oral transmission, which causes reproductive failure in rams
Blood samples were collected for serological analysis, as well as semen and urine for culture and PCR, to confirm that the rams were free of B. ovis infection
Urine samples were collected by blocking the breath of the animals for thirty seconds and semen from each animal was collected by eletroejaculation
Summary
Ovine brucellosis caused by the Brucella ovis is a contagious disease, with venereal and oral transmission, which causes reproductive failure in rams. Routine diagnostic tests for ovine brucellosis caused by B. ovis include clinical examination, serological tests, and bacteriology (Alton et al, 1988; Grilló et al, 1999). The bacteriology of B. ovis is time consuming and not always efficient, because bacteria can be excreted intermittently in the semen of infected animals (Worthington et al, 1985; Baigun et al, 2000). Regarding serological tests, they generally fail in the sensitivity displayed. Due to the limitations exposed, the diagnosis of B. ovis infection is usually obtained by the association of two or more of these techniques
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