Infectious pathogens potentially transmitted by semen of the black variety of the Manchega sheep breed: Health constraints for conservation purposes

Conservation of genetic resources from endangered breeds may be conducted through germinal banks. Preservation of healthy samples is paramount to avoid preserving pathogens shed with germinal products. The black variety of Manchega sheep (BMS), and endangered breed endemic to south-central Spain, is the subject of a conservation program; a germinal bank has been recently established. However, several pathogens circulating in BMS flocks may be shed with semen and threaten BMS preservation. Therefore, we investigated the sanitary status of BMS flocks and semen samples from 4 of the 17 flocks in which this variety is bred worldwide. A serological screening for Maedi-Visna virus, bluetongue virus, Pestivirus spp., Brucella spp., Coxiella burnetii, Chlamydophila spp., Mycobacterium tuberculosis complex, Mycobacterium avium paratuberculosis, Anaplasma spp., Mycoplasma agalactiae, Toxoplasma gondii and Neospora caninum was performed to assess for pathogens potentially shed by semen. Semen samples from 11 of the 35 BMS rams and 4 samples from coexisting rams of the white variety (WMS) were analyzed by PCR to detect Maedi-Visna virus, C. burnetii, Anaplasma marginale, Anaplasma phagocytophilum and T. gondii. Maedi-Visna virus RNA was detected in 3 semen samples (2 BMS and 1 WMS) while C. burnetii DNA was detected in 3 samples from WMS rams. Pathogens that can be transmitted by semen were present in BMS flocks, and Maedi-Visna virus and C. burnetii showed the highest potential for transmission by artificial insemination. Our results point to the need of testing semen samples kept for conservation purposes of BMS before using them for artificial insemination.