Investigation of Cyclospora spp. in farmed fur animals based on PCR-RFLP

Cyclospora spp., foodborne intestinal protozoa with potential zoonotic risk, are distributed across the globe. However, their presence in animals bred for fur has not been extensively studied, raising concerns about zoonotic transmission. To assess the prevalence of Cyclospora spp. in fur animals in China, we collected fresh fecal samples from 862 animals, including 352 blue foxes, 275 minks, and 235 raccoon dogs, across multiple provinces. The small subunit (SSU) rRNA gene of Cyclospora spp. was amplified using nested PCR, followed by restriction fragment length polymorphism (RFLP) analysis with the Kpn2I enzyme. We also analyzed geographic location and clinical symptoms, such as diarrhea, as potential risk factors. Our findings showed an overall infection rate of 1.3 % for Cyclospora spp. in fur animals, with 2.3 % in blue foxes, 0.4 % in minks, and 0.9 % in raccoon dogs. Geographic location was a significant infection risk factor in blue foxes (P < 0.001), while diarrhea was a risk factor in all fur animals (P < 0.05). A molecular phylogenetic analysis based on SSU rRNA sequences revealed that two isolates clustered with human-derived Cyclospora cayetanensis, while the remaining isolates grouped with previously reported cattle-derived Cyclospora spp. This study is the first to report Cyclospora spp. infections in fur animals worldwide, underscoring the potential for zoonotic transmission. These findings offer crucial insights for controlling infections in fur animals, other animals, and humans.