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Mycoplasma haemocanis (Mhc) and Candidatus Mycoplasma haematoparvum (CMhp) are two species of canine hemoplasma that may cause hemolytic anemia and chronic disease in canine species. While understanding is limited, blood transfusions, bloodsucking arthropods, biting and fighting are suspected routes of hemoplasma transmission in dogs. The aim of the present survey was to determine the prevalence of hemotropic mycoplasma infections in hunting dogs from Southern Italy and assess related risk factors. Blood samples were collected from 1433 hunting dogs in the Napoli, Avellino and Salerno provinces of Campania region of Southern Italy, and tested by real time polymerase chain reaction (RT-PCR) assays for amplification of Mhc and CMhp DNA. The dogs had no clinical signs at the time of sampling. The overall PCR-positive rates were13.1% for Mhc and 11.4% for CMhp. Coinfection with both hemoplasma species was found in 4% of animals. Statistical analysis revealed living in Salerno province (Mhc: OR=2.94, 95% CI: 2.10–4.11; CMhp: OR=2.27, 95% CI: 1.61–3.20), hound breeds (Mhc: OR=4.0, 95% CI: 2.86–5.59; CMhp: OR=1.61, 95% CI: 1.16–2.24) , pack size more than 10 animals (Mhc: OR=1.67, 95% CI: 1.14–2.47; CMhp: OR=1.61, 95% CI: 1.06–2.44) and wild mammals hunting (Mhc: OR= 3.53, 95% CI: 2.51–4.98; CMhp: OR= 15.7, 95% CI: 8.83–28.0) as associated risk factors for both canine hemoplasma infection. Adult age was a variable significantly associated only to infection by CMhp (OR=1.93, 95% CI: 1.25–2.97). To the authors’ knowledge, this is the first large-scale molecular survey on Mhc and CMhp infections in dogs living in Southern Italy. The obtained data confirm the circulation of these two species of canine hemoplasma within the hunting dog population of Southern Italy, although their pathogenic potential and the possible epidemiological relationships between hunting dogs and sympatric wild animal populations are still unclear.