Mycoplasma suis and Mycoplasma parvum are the two hemotrophic mycoplasmas species described in pigs. M. suis is involved in infectious anemia, while M parvum infection is commonly subclinical. The objectives of this study were twofold: (i) to investigate the prevalence of porcine hemotrophic mycoplasmas in sows from the southern region of Brazil by quantitative real-time PCR (qPCR) and (ii) to genetically characterize a subset of the samples based on the 16S rRNA gene. A total of 429 blood samples were evaluated from 53 different farm sites. Porcine hemoplasmas was detected at all the 53 tested sites and in 79.72% of the samples (342/429). Two sequences were obtained for Mycoplasma spp. The phylogenetic analysis based on the 16S rRNA gene (900 bp) showed that the Mycoplasma sequences were closely related to the M. suis cluster and that one sequence was positioned in the M. parvum cluster. In conclusion, porcine hemoplasmas have a high rate of prevalence in sows from commercial farms in the southern region of Brazil. This study demonstrated the first molecular detection and characterization of partial 16S rRNA gene of M. parvum in Brazil.