Buffaloes are considered in the rural regions as a main source for both meat and milk production. Most mycoplasmas are facultative pathogens giving a disease condition only when their hosts succumbed to stress factors or when their body resistance is impaThe present investigation was carried out on 532 samples randomly collected from both apparently healthy and dise-ased buffaloes of both male and female species with different ages and bred in peasantary and governmentalfarms and 75 samples were collected from slaughtered animals.Conjunctival swabs (112), Nasal swabs (1l2), vaginal swabs (133), preputial swabs (59) and preputial wash samples (60) besides (25) lung tissues and (50) tracheal swabs.Moreover (56)milk samples were collected from mastitic cases.Primary isolation revealed that the mycoplasma and aCholeplasma could be isolated from prepuce with an incid-ence of (57.63%), from preputial wash samples with an incidence (23.33%), from vaginal swabs with percentage(56.6%), from vaginal swabs of animal with breeding dis- orders with an incidence (57.5%), from nasal swabs withpercentage (42.86%), from tracheal swabs with an incidence