A total of 165 cows, 19 buffaloes, 192 sheep and 118 goats were examined for detection of Mycoplasma mastitis. The results revealed that 114 (69.59%) and 6 (31.57%) were clinically mastitic cows and buffaloes respectively while 51 (30.9%) and 13 (68.42%) were apparently healthy cows and buffaloes respectively. On examining the apparently healthy cows and buffaloes, 67 (32.84%) and 18 (34.61%) were subclinically mastitic cows and buffaloes respectively. Mycoplasmas were isolated in percentages of 8.9%, 5.5% from subclinically mastitic cows and buffaloes respectively and in percentages of 12.97%, 12.5% from clinically mastitic cows and buffaloes respectively. M. bovis was isolated from 8 (32%) and M. bovigenitalium from 7 (28%) and 10 (40%) unidentified Mycoplasma. Isolation of Mycoplasma from udder tissue in cows and buffaloes were in a percentage of 28.5% in cows while no Mycoplasma isolates were obtained from buffaloes' udder tissues. Application of PCR technique on these isolates and some of the negative samples was positive 100%. On the other hand, the results revealed that 82 of 192 (42.7%) and 43 of 118 (36.44) of the examined sheep and goats respectively were clinically mastitic. Isolation of Mycoplasma was from 11 (13.41%) and 17 (39.53%) of the examined sheep and goat respectively. Identification of these isolates revealed 8 (29%) M. agalactiae isolates and 20 (71%) unidentified Mycoplasma spp. Application of PCR technique on traditionally identified M. agalactiae isolates revealed negative results on using M. agalactiae specific primer while positive results were obtained for the same 8 isolates (100%) on using M. bovis specific primer.