SummaryThe hookworm Ancylostoma caninum is a common nematode of wild and domestic canids worldwide. In Mexico, there are few records of helminths in wild canids, especially in the southeastern region. The aim of the present study was to examine the helminths from a gray fox Urocyon cinereoargenteus in southeastern Mexico. A road-killed female gray fox found in Merida, Yucatan, Mexico, was examined for helminths. Only nematodes were found in the intestine of the gray fox and identified using morphological studies and molecular analysis of 28S rRNA gene fragments. The characteristics exhibited by the nematode specimens were in accordance with descriptions of A. caninum: e. g. oral opening with a pair of prominent chitinous plates bearing three pairs of ventral teeth, lateral rays with a common trunk, dorsal ray divided into two branches with each branch terminating in three digitations. BLAST analysis of the 28S sequence showed similarity and coverage values of 99.8 % and 100 %, respectively, with a sequence of A. caninum from the domestic dog Canis familiaris in Australia. The genetic distance between the Australian specimen and the Yucatan specimen of A. caninum was 0.1 %, that is, they were only different in a single nucleotide. The gray fox examined in this study was found close to a rural community where A. caninum has been recorded from domestic dogs, which could be the source of infection. Our study increases the distribution of this nematode parasitizing the gray fox in Mexico and provides the first nucleotide sequence of A. caninum from the gray fox.