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Canine and feline heartworm disease caused by Dirofilaria immitis is a serious and sometimes fatal infection transmitted by mosquitos. Little is known about the prevalence or distribution of D. immitis infection in dogs and cats on the island of Hainan island/province or coastal cities of China. The present study examined the occurrence of D. immitis infections in dogs (n ¼ 869) and cats (n ¼ 51) in Hainan island/province and prevalence in dogs from 3 coastal cities (Shenzhen [n ¼ 55], Shanghai [n ¼ 69], and Hangzhou [n ¼ 45]) in southern and eastern China. A commercial antigen detection (AD) test and 2 PCR methods (16S ribosomal RNA and Wolbachia surface protein [WSP] genes) were used to determine the prevalence of D. immitis from animals .6 mo old with no previous history of D. immitis preventive treatment or heartworm infection. Gene sequencing was used to confirm positive PCR samples. The AD test was not used on cat samples. Using the AD test, the prevalence in dogs was 0.5% (4/869) in Hainan island/province, 0% (0/55) in Shenzhen, 1.5% (1/69) in Shanghai, and 0% (0/45) in Hangzhou. Prevalence by 16S rRNA gene PCR was 7.4% (64/869) of dogs from Hainan island/province, 0% (0/55) in dogs from Shenzhen, 1.5% (1/69) in dogs from Shanghai, and 0% (0/45) in dogs from Hangzhou. Prevalence by WSP gene PCR in dogs was 5.3% (46/869) in Hainan island/province, 0% (0/55) in Shenzhen, 1.5% (1/69) in Shanghai, and 0% (0/45) in Hangzhou. Prevalence in the 51 cats from Hainan island/province was 9.8% and 5.9% by 16S rRNA and WSP gene PCR, respectively. The present study demonstrates that canine heartworm exposure occurs in dogs and cats in Hainan island/province and that PCR methods detected a higher prevalence than did the AD method. The 16S rRNA gene PCR detected more positive samples than did the WSP gene PCR in both dogs and cats. The 3 coastal cities had very few dogs that had evidence of D. immitis exposure. [ABSTRACT FROM AUTHOR]