Sheep and goats are one of the important agricultural animals in Egypt. Respiratory affections and mastitis due to mycoplasma in both sheep and goats are regarded as a frequent cause of economic losses. Nasal, conjunctival swabs and milk samples from apparently healthy and diseased sheep and goats were collected from 75 sheep (195 samples) and 50 goat (140 samples) flocks in different areas in El-Fayom governorate. These samples were subjected to Mycoplasma isolation. The results revealed that, the total mycoplasma isolates from sheepwerel35 and from goats wereS4 which identified as follow :(38isolates) M.agalactiae from sheep samples and (18 isolates) from goats. Also M. arginini was isolated from sheep (43 isolates) and goats (17 isolates) and M. ovipneumoniae was isolated from sheep (48 isolates) and goats (16 isolates). Also there 49 were 6 acholeplasma strains from sheep and 4 strains from goat samples. PCR confirm the isolation of M. agalactiae by the presence of 360 bp specific band, PCR amplification of ovipneumoniae strains gave a characteristic band at 1070 bp and PCR amplification of  M. arginine strains gave a gave a characteristic band at 280bp.