Hemotropic mycoplasmas are small, pleomorphic, gram-negative, epierithrocytic bacteria that infect a wide variety of animals, including domestic cats. These microorganisms cause the disease known as feline hemotropic mycoplasmosis (MHF), also called feline infectious anemia. The objective of the study was to use PCR assays based on amplification of 16S rDNA to detect the species Mycoplasma haemofelis (Mhf) and 'Candidatus Mycoplasma haemominutum' (CMhm) in felines treated at a clinic specialized exclusively in cats in the city of Pelotas, Rio Grande do Sul. Furthermore, the research aimed to identify potential risk factors associated with the infection. A total of 253 blood samples from domestic felines were analyzed, of which 76 samples (30%) were positive for at least one species. The most detected hemoplasma was CMhm, with 51/253 (20.1%), followed by Mhf, with 23/253 (9.1%). Mhf and CMhm co-infection was detected in 2/253 (0.8%) samples. Felines with access to the street, presence of ectoparasites, co-infection with FeLV, FIV and other diseases (unhealthy animals) were more likely to be infected by hemoplasmas.