Sarcocystis species are intracellular coccidian protozoans that can infect a range of animals and humans and cause public health problems and economically significant losses. Sarcocystosis in sheep (Ovis aries) can cause abortion, neurological symptoms, and even death and results in significant economic losses to the livestock industry. To date, however, it is yet unknown whether sheep in Shanxi Province, north China, are infected with Sarcocystis spp. The purpose of this study was to investigate the prevalence of Sarcocystis spp. in sheep in Shanxi Province. Thus, 582 muscle samples of sheep were purchased from farmers’ markets from ten representative counties in Shanxi Province, north China, and examined for the presence and prevalence of Sarcocystis spp. by PCR amplification of the mitochondrial cytochrome c oxidase subunit I (cox1) gene. Of the examined 582 mutton samples, 197 samples (33.85%) were Sarcocystis-positive and were sequenced. Of the obtained 197 cox1 sequences, 196 sequences showed nucleotide similarity of 98.56–99.81% with those of S. tenella, and the remaining one cox1 sequence showed nucleotide similarity of 99.71% with that of S. arieticanis. Two representative cox1 sequences of S. tenella (accession nos. PQ189447 and PQ189448) have 99.52% and 99.61% identity with S. tenalla (KC209725) and S. tenalla (MK419984), respectively. The sequence of S. arieticanis (accession no. PQ165949) obtained in this study has 99.71% identity with S. arieticanis (MK419975). This present study documents the occurrence and prevalence of Sarcocystis spp. in sheep in Shanxi Province for the first time, which enriches the data on the distribution of Sarcocystis spp. in sheep in China and has implications for the control of sheep sarcocystosis.

Entamoeba spp. are common zoonotic intestinal protozoa, which can lead to serious intestinal diseases in both humans and animals through fecal–oral transmission, leading to significant economic losses and public health challenges. To reveal the prevalence of Entamoeba in sheep and cattle in Shanxi Province, North China, fecal samples were collected from 311 sheep, 392 dairy cattle, and 393 beef cattle from three representative counties in the northern, central, and southern regions of Shanxi Province. DNA was extracted from the fecal samples and amplified by PCR with primers targeting the nuclear small subunit ribosomal RNA (SSU rRNA) gene of Entamoeba spp., followed by the sequencing of the positive products. The overall infection rates of Entamoeba were 51.5% (160/311), 82.9% (325/392), and 79.1% (311/393) in sheep, dairy cattle, and beef cattle, respectively. Statistical analysis showed a significant correlation between the infection rate of Entamoeba and the location factor in sheep, dairy cattle, and beef cattle (p < 0.001). According to the obtained SSU rRNA sequences, several Entamoeba species, namely Entamoeba bovis, Entamoeba Ribosomal Lineage (RL) 2, Entamoeba RL4, and Entamoeba RL8, were identified. This study represents the first molecular survey of Entamoeba prevalence in sheep, beef cattle, and dairy cattle in Shanxi Province. The findings extend the geographical distribution of Entamoeba spp. and provide valuable scientific data for the prevention and control of amoebiasis in Shanxi Province.

To date, more than 20 species in the genus Cyclospora have been reported. Among them, Cyclospora cayetanensis has been recognized as the causative agent of human cyclosporiasis, which is characterized by severe intestinal injury and prolonged diarrhea in patients with immune dysfunction. The presence of C. cayetanensis in cattle has been confirmed. To date, however, no surveillance data are available on the occurrence and prevalence of Cyclospora spp. in cattle in Shanxi Province, North China. In the present study, a total of 761 fecal samples collected from cattle in three representative counties (Qi, Jishan, and Shanyin) in this Province were examined for Cyclospora spp. by using a polymerase-chain-reaction–restriction-fragment-length polymorphism (PCR–RFLP) test based on the nuclear small subunit ribosomal RNA (SSU rRNA) gene. The prevalence of Cyclospora spp. in cattle was 2.1%, and region, age, sex, and breed were not identified to be risk factors. Molecular evolutionary analysis based on the SSU rRNA sequences revealed that all 12 of the isolates were relatively distant from the human pathogen C. cayetanensis; seven isolates were grouped with Cyclospora colobi, whereas the others were grouped with cattle Cyclospora spp. reported previously. Though C. cayetanensis was not detected in cattle in the present study, more investigations should be performed in human populations, other animal species, or cattle from other regions of Shanxi Province and other environmental sources from the One Health perspective.