Enterocytozoon bieneusi in Diarrheal and Asymptomatic Children in Southern Zhejiang Province, China: Prevalence and Genetic Diversity.

Purpose Enterocytozoon bieneusi(E. bieneusi) is a globally prevalent enteric pathogen that poses a major threat to children’s health. This study aimed to investigate the prevalence, genotypic characteristics and zoonotic transmission potential ofE. bieneusi in children under 6 years of age in Wenzhou, Zhejiang, China, where pediatric infection data of this pathogen remains absent.Methods A total of 1,032 stool samples were collected from children at Wenzhou Yuying Children’s Hospital, including 684 from diarrheal individuals and 348 from asymptomatic controls. Nested polymerase chain reaction (PCR) and sequencing, targeting the internal transcribed spacer (ITS) region of the E. bieneusi small subunit(SSU) ribosomal RNA (rRNA) gene, were conducted for pathogen identification and genotyping.Results The overall prevalence of E.bieneusiwas 1.4% (14/1032), with 1.6% in diarrheal children and 0.9% in asymptomatic children. Two zoonotic E. bieneusi genotypes were identified, namely D (n=5) and Peru11 (n=9).Conclusion This study provides the first molecular confirmation of E. bieneusi infection in children in Zhejiang Province, China, despite a relatively low infection rate. The detection of zoonotic genotypes D and Peru11 indicates potential animal to-human transmission of the pathogen, highlighting the need for further epidemiological investigation and strengthened preventive measures.