Epidemiology, molecular characterization, and risk factors of Acanthamoeba spp., Blastocystis spp., and Cyclospora spp. infections in snakes in China.

Snakes are widely farmed in China for medicinal purposes and as pets worldwide. Acanthamoeba spp., Blastocystis spp., and Cyclospora spp. are significant zoonotic pathogens frequently discovered in various animals, causing diseases with global public health implications. However, their prevalence and zoonotic potential in snakes remain poorly understood. In this study, 812 snake faecal samples were collected across 28 China provinces. The partial small subunit (SSU) rRNA gene was amplified using polymerase chain reaction (PCR) to assess evolutionary relationships and genetic characterization. Detection rates for Acanthamoeba spp., Blastocystis spp., and Cyclospora spp. were 6.40 %, 3.33 %, and 2.71 %, respectively. Sequencing and phylogenetic analysis revealed that Cyclospora isolates were closely related to those found in humans and cattle. Subtyping for Blastocystis species identified two zoonotic subtypes (ST4, ST6) and four host-specific subtypes (ST10, ST15, ST21, ST42). Multiple Acanthamoeba genotypes were detected, including T4, T11, and T13. Furthermore, species, age, and living conditions are key risk factors. This study provides valuable insights into these infections in snakes and underscores the need for proper hygiene and One Health measures to reduce zoonotic transmission and environmental contamination.