To determine the prevalence of H. heydorni infection in pet dogs , a total of 315 fresh fecal samples were collected from pet dogs in Anhui and Zhejiang Province. All specimens were screened firstly by nested-PCR targeting H. heydorni/N. caninum ITS1 gene, and the positive amplified products obtained were sequenced and analyzed. Subsequently, the H. heydorni/N. caninum-positive samples were confirmed by H. heydorni ITS1 PCR. PCR and sequence analysis showed that 2 positive samples were found in 315 fresh fecal sample, and the prevalence for H. heydorni was 0.63%. The present study showed that the prevalence of H. heydorni was not high in the pet dogs from Anhui and Zhejiang Provinces.