The current study aimed to investigate the seroprevalence of IgG antibodies to Anaplasma marginale in cattle from Maputo, Gaza and Inhambane provinces, south Mozambique. A total of 809 serum samples from cattle were obtained and tested by indirect enzyme-linked immunosorbent assay (i-ELISA). The chi-square test at 5% significance was used to assess the association between seroprevalence and the variables gender, age and geographic origin of animals. The overall seropositivity was 76.5% (n = 619) and anti-A. marginale antibodies were detected in 89.1% (n = 156), 68.4% (n = 308) and 84.2% (n = 155) of the animals in the provinces of Maputo, Gaza and Inhambane, respectively. A significant association (p < 0.05) was found with the geographic origin of the animals, while sex had no significant relationship. The frequencies of seropositive in the age groups were 63.2% (n = 72), 80.0% (n = 92), 83.1% (n = 98) and 77.3% (n = 357) for animals <12; >12 and ≤24; >24 and ≤36; >36 months, respectively. These results indicate that in southern Mozambique there are areas of enzootic stability to A. marginale. Thus, epidemiological monitoring is required to monitor the immune status of animals in the region.