Molecular Detection and Genotyping of <i>Enterocytozoon bieneusi</i> in Beef Cattle in Shanxi Province, North China

<i>Enterocytozoon bieneusi</i> is an intracellular pathogen that can parasitize humans and a variety of animals. The infection of <i>E. bieneusi</i> in most hosts is asymptomatic, but in immunocompromised individuals, it can lead to serious complications such as acute diarrhea, dehydration, and even death. However, no data on the prevalence and genotyping of <i>E. bieneusi</i> in beef cattle in Shanxi province are currently available. In this study, a total of 401 fecal samples were collected from beef cattle in farms from two representative counties—Qi county and Jishan county—in Shanxi province, north China. Nested PCR was applied to determine the prevalence and genotypes of <i>E. bieneusi</i> by amplifying and sequencing the internal transcribed spacer (ITS) regions of the rRNA gene. A total of 90 out of 401 samples were detected as <i>E. bieneusi</i>-positive, with 22.44% overall prevalence of <i>E. bieneusi</i> in beef cattle in Shanxi province. The highest prevalence of <i>E. bieneusi</i> was detected in calves (28.67%, 41/143) and male beef cattle (28.13%, 54/192). Statistical analysis revealed that the prevalence of <i>E. bieneusi</i> was significantly associated with gender and age factors (<i>p</i> < 0.05), but without any statistical difference among regions. Moreover, six known <i>E. bieneusi</i> genotypes (BEB4, BEB6, BEB8, J, I, and PigSpEb2) and two novel genotypes (designated CSC1 and CSC2) were identified by analysis of ITS sequences, and genotype I was the predominant genotype in these two counties. Phylogenetic analysis showed that five known genotypes and two novel genotypes were clustered into Group 2, but PigSpEb2 belonged to Group 1. To our knowledge, the present study demonstrated the presence and identified genotypes of <i>E. bieneusi</i> in beef cattle in Shanxi province for the first time, extending the data on prevalence and genotypes of <i>E. bieneusi</i> in beef cattle and providing baseline data for executing intervention measures to control it in the study regions.