Haemotropic Mycoplasma species are pathogens that can cause haemolytic anaemia in susceptible mammalian species worldwide. The cause of haemolysis is due to membrane damage through stimulation of IgM cold agglutinins production, which induces autoimmune haemolysis of infected erythrocytes. A study was conducted to establish the prevalence of Mycoplasma haemofelis, ‘Candidatus Mycoplasma haemominutum’ and ‘Candidatus M. turicensis’in ticks and the diversity of tick species that are possible vectors of the pathogens that can transmit the infection to wildlife in Ngorongoro Crater. Three real-time PCR assays were used for the analysis of DNA pools (n = 507) derived from 11 tick species. Mycoplasma haemofelis and ‘Candidatus M. haemominutum’ were detected in Rhipicephalus sanguineus. On average 19.7% and 12.9% of R. sanguineus were PCR-positive for M. haemofelis and ‘Candidatus M. haemominutum’, respectively. This tick species therefore represent an important reservoir for feline haemotropic Mycoplasma species in the crater. These organisms with their known pathological effects are probably one of the factors potentially exacerbating the severity of infection during the disease outbreak in wildlife and can have undesirable outcome to wild cats like lions when under nutritional stress or in case of concurrent infection.