High genetic diversity of

Enterocytozoon bieneusi, a zoonotic pathogen prevalent in both humans and animals, is the most frequently diagnosed microsporidian species in humans and presents significant public health risks. However, data on the prevalence and genotypes of E. bieneusi in farmed minks (Neovison vison) and raccoon dogs (Nyctereutes procyonoides) in China are limited. Therefore, 275 minks (89 from Hebei Province, 57 from Heilongjiang Province, 109 from Liaoning Province, 20 from Shandong Province) and 235 raccoon dogs (114 from Hebei Province, 27 from Heilongjiang Province, 61 from Liaoning Province, 33 from Jilin Province) were examined for the prevalence and genotypes of E. bieneusi through sequence analysis of the internal transcribed spacer (ITS) region of the rRNA gene. The overall prevalence of E. bieneusi was 18.6% (95/510), with 10.5% (29/275) in farmed minks and 28.1% (66/235) in raccoon dogs. Ten genotypes (CHN-F1, genotype D, Type IV, EbpC, NCF2, NCF5, NCF6, Peru8, Henan V, and MJ5) were identified in minks and raccoon dogs. This study is the first to detect the CHN-F1, NCF2, NCF6, Peru8, and Henan V genotypes in minks and the NCF5, NCF6, and MJ5 genotypes in raccoon dogs. Additionally, the D, Type IV, and Peru8 genotypes, previously identified in humans, were also found in minks and raccoon dogs, suggesting that these animals could be potential sources of human microsporidiosis. These findings expand the understanding of E. bieneusi’s host distribution in China and contribute to the prevention of zoonotic E. bieneusi infections among farmed animals.