A RTIFICIAL INSEMINATION using unaccredited frozen semen is a major approach for diseases transmission. This work intended to scan the buffalo bull semen for Br. melitensis, Toxoplasma gondii and Neospora caninum using Polymerase Chain Reaction (PCR). Fifty bulls were used in this study. Semen was assessed for volume, motility, viability, and chromatin damage. Total genomic DNA was extracted from buffalo semen and amplified by PCR. The results showed that, semen samples were positive to Toxoplasma gondii in 6% of the semen samples, while all the semen samples were totally negative to Br. Melitensis and Neospora caninum. No significant differences were found in semen characteristics between bulls positive or negative to Toxoplasma gondii.In conclusion, the present trial suggests the opportunity of venereal transmission of T. gondii among buffaloes. However, further investigations are required using high number of semen samples from different localities of Egypt.