<i>Blastocystis</i>, an enteric protist, has been reported to be an important cause of protozoal gastrointestinal manifestations in humans and animals worldwide. Animals harboring certain <i>Blastocystis</i> subtypes (STs) may serve as a potential source of human infection. However, information about the prevalence and genetic diversity of <i>Blastocystis</i> in alpacas is limited. In the present study, a total of 366 fecal samples from alpacas in Shanxi Province, northern China, were examined for <i>Blastocystis</i> by PCR amplification of the small subunit rRNA gene, followed by sequencing and phylogenetic analysis. The prevalence of <i>Blastocystis</i> in alpacas was 23.8%, and gender difference in the prevalence of <i>Blastocystis</i>was observed. The most predominant <i>Blastocystis</i> ST was ST10, followed by ST14 and ST5. The detection of ST5, a potentially zoonotic genotype, indicates that alpacas harboring ST5 could be a potential source of human infection with <i>Blastocystis</i>. These data provide new insight into the prevalence and genetic diversity of <i>Blastocystis</i> in alpacas.
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