This study was intended to detect the most common Mycoplasma species involved in 
respiratory diseases in both cattle and buffaloes. In this study, examination of 300 cattle 
and 200 buffaloes was carried out and then the collected samples were subjected to 
isolation of Mycoplasmaspecies followed by PCR confirmation. The results showed that 
the percent of mollicutes isolation from pneumonic lung tissues of cattle& buffaloes was 
22% and 17%, respectively. Moreover, the percent of mollicutes isolation from nasal 
swabs of diseased cattle & buffaloes was 18% and 20%, respectively. PCR was effective 
in the detection of both M. bovis and M. bovigenitalium through successful amplification 
of the Mbo and Mbg gene at 360 and 312 bp, respectively. The minimum inhibitory 
concentration was performed on two field strains of M. bovis against seven different 
antimicrobial agents. The first strain was sensitive to Draxxin® 10 %, Marbocyle® 10%, 
Lincospectin®, Duocycline ® 20%, and Tilmicosin® 30%. The second strain was sensitive 
to Duocycline® 20% and Tilmicosin® 30% and resistant to other antimicrobial agents. 
Regarding the associated risk factors with Mycoplasmosis; the season was significantly 
correlated and the infection was 6.5 times more prevalent in the winter season than 
summer season. As well as the sex revealed a significant effect and 5.08 times in males 
more than females. Additionally, cattle were more susceptible than buffaloes for the 
disease prevalence. Meanwhile, the age was not a significant risk for the disease 
prevalence.