The aim of this study was to investigate the occurrence of Hepatozoon species infecting dogs in the municipality of Campo Grande, Mato Grosso do Sul (MS), Brazil, using blood samples (n = 165) drawn from dogs. The species Hepatozoon canis was identified in 3.63% of the tested animals using molecular tools. Further studies are needed to determine the clinical relevance of this infection and the main arthropod vectors involved in its transmission.