Mycoplasma species are a major cause of mastitis, arthritis, pneumonia and reproductive disorders in small ruminants. Mycoplasmas in reproductive systems of males have been associated with diseases as orchitis, balanoposthitis and abnormal spermatozoa activity. The aim of this study was to detect Mycoplasma species that cause reproductive infections in bucks and rams. Total of 27 preputial swabs was collected from Saanen bucks and Kivircik rams at the Artificial Insemination (Al) Center of Uludag University in Turkey. Bacteriological culture methods, followed by Polymerase Chain Reaction (PCR) and Denaturing Gradient Gel Electrophoresis (DGGE) were used to detect and identify Mycoplasma species. The PCR-DGGE method identified one M. bovigenitalium and one M. arginini from two cases of orchitis in rams, and another M. bovigenitalium was identified from a buck with no clinical signs. The results showed that Mycoplasma species were present in the testicles of rams and bucks that were negative for Brucellosis, and likely causative organisms of orchitis which lead to reduced fertility.