The study was designed to investigate the seroprevalence of Anaplasma marginale among cattle from Sargodha, Khushab and Rawalpindi districts of the northern Punjab, Pakistan. A total of 1050 samples were collected from selected small holders and private livestock farms using multistage cluster random sampling technique. The overall seroprevalence of Anapalsma marginale infection was 31.05 percent % using competitive enzyme-linked immunosorbent assay (cELISA). The highest seroprevalence was recorded in Sargodha (37.14%) district followed by Khushab (31.43%) and Rawalpindi (24.57 %) districts. A significant relationship was found among different age groups and breed. The seroprevalence was significantly higher in small holders than private livestock farms in all study districts. It is concluded that small holder’s crossbred cattle of more than four years of age from Sargodha district are more susceptible to Anaplasma marginale infection in summer season.