The aim of the present study was to detect the prevalence of Anaplasma marginale in buffaloes in two Iraqi governorates, Al-Qadisiyah and Babylon, by the microscopy as well as the competitive-ELISA that used firstly among the Iraqi buffaloes. A total of 184 buffaloes from both sexes of different age groups of animals were submitted for collection of blood samples to prepare the smears and sera. Overall results were revealed on 10.33% and 36.41% positive animals by microscopy and competitive-ELISA, respectively. In addition, positive rates by both tests were 7.61%; by microscopy only, 2.72%; and by competitive-ELISA only, 28.8%. In Al-Qadisiyah and Babylon governorates, respectively, 8.7% and 11.96% of microscopy samples, and 44.57% and 28.26% of competitive-ELISA were positives with significant differences (P>0.05). Regarding to age factor, the highest prevalence was detected by microscopy in young age group (1-3 years) was 14.15%, whereas by competitive-ELISA, it was 55% in adult buffaloes group (>3 years). Significant increases (P>0.05) in rates of infection were showed in females compared to males, respectively, by microscopy (11.18% and 4.35%) and competitive-ELISA (39.13% and 17.39%).