Enterocytozoon bieneusi is a predominant pathogen that presents significant public health and animal husbandry challenges. Information on the prevalence and genotypes of E. bieneusi in pigeons is scarce. In this study, 207 faecal samples were collected from pigeons in two parks in Zhengzhou, China, and subjected to nested polymerase chain reaction (nested-PCR) analysis for the detection of E. bieneusi. The overall prevalence of E. bieneusi was found to be 15.5% (32/207) across the two parks. ITS sequence analysis identified only the Peru6 genotype (n = 32), which phylogenetic analysis placed within the zoonotic group 1. Among the E. bieneusi isolates, multilocus sequence typing (MLST) at MS1, MS4 and MS7 loci identified 19, 13 and 23 samples, respectively. Of the samples, 11 were successfully amplified and sequenced at all three loci, resulting in nine distinct multilocus genotypes (MLGs). This suggests that pigeons may play a role in the transmission of E. bieneusi to humans and that there is high genetic variability of E. bieneusi in pigeons from Zhengzhou in China.
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