Vector-borne pathogens have become a major problem since different species of cattle, e.g. indigenous, crossbred dairy, and crossbred beef, are raised together on most farms. Anaplasma spp. are obligate intracellular rickettsial vector-borne pathogens that impact on livestock farmers with major economic constraints and eventually threaten human health in cases of zoonotic species involvement. Therefore, our study aimed to identify Anaplasma spp. infection in a bovine host using both microscopy and molecular techniques. A total of 59 samples were collected from Mingaladon township, Yangon region, and the presence of infection was assessed using a Giemsa-stained thin blood smear and PCR of 16S rRNA gene amplification. Both microscopic and PCR-positive samples were further analyzed for hematobiochemical alteration. As a results, Anaplasma spp. was detected in 3.38% (2/59) of sampled animals. Hematology results revealed severe anemia with a low hemoglobin level together with a low PCV and total platelet count whereas the MCV and total WBC count were shown to be higher. Elevation of some enzymes such as ALT, AST, GGT, total bilirubin, and blood urea nitrogen (BUN) occurred by blood biochemical analysis, whereas total protein and albumin were low. To our best, this study is the first molecular evidence for the presence of Anaplasma marginale infection in cattle in Myanmar, with a sequence similarity range between 98.8 and 100%. By understanding one of the major tick-borne pathogens (TBPs) in Myanmar, possible control measures might be implemented not only to minimize the transmission but also to increase the farm productivity.