Blastocystis is one of the most common intestinal parasites in humans and animals worldwide. At least 17 subtypes have been identified in mammals and birds. In China, although some studies have reported the occurrence of Blastocystis in humans and animals, our understanding of the role of animals in the transmission of human blastocystosis is only superficial due to a paucity of available molecular data. The aim of the present study was to understand infection rates of Blastocystis and the distribution and genetic diversity of subtypes in various mammal and bird species in northeastern China, as well as to assess the zoonotic potential of Blastocystis isolates. A total of 1265 fresh fecal specimens (1080 from ten mammal species and 185 from eight bird species) were collected in Heilongjiang, Liaoning and Jilin provinces of China. Each specimen was examined for the presence of Blastocystis by PCR amplification and sequence analysis of the partial SSU rRNA gene. Fifty-four specimens (4.3%) were positive for Blastocystis. Birds (7.0%) had a higher infection rate of Blastocystis than mammals (3.8%). Blastocystis was found in seven mammal species, reindeer (6.7%), sika deer (14.6%), racoon dogs (7.5%), Arctic foxes (1.9%), dogs (2.9%), rats (3.7%) and rabbits (3.3%), as well as three bird species, pigeons (2.1%), chickens (13.0%) and red crowned cranes (14.0%). Eight subtypes were identified including ST1 (n = 5), ST3 (n = 3), ST4 (n = 13), ST6 (n = 8), ST7 (n = 6), ST10 (n = 13), ST13 (n = 4) and ST14 (n = 2). 64.8% (35/54) of Blastocystis isolates belonged to potentially zoonotic subtypes. To our knowledge, this is the first report of Blastocystis in reindeer (ST10 and ST13), rabbits (ST4), racoon dogs (ST3) and Arctic foxes (ST1, ST4 and ST7). The findings of potentially zoonotic subtypes suggest that the animals infected with Blastocystis might pose a threat to human health. These data will improve our understanding of the host range and genetic diversity of Blastocystis, and also help develop efficient control strategies to intervene with and prevent the occurrence of human blastocystosis in the investigated areas.

Blastocystis is an enteric protozoan able to infect humans and a large variety of animal hosts worldwide. It exhibits significant genetic diversity, with at least 17 subtypes (STs) identified to date, most of which have low host specificity. In the present study, a cross-sectional survey was conducted on Blastocystis infection among humans and domestic animals sharing habitats in northeastern China's Heilongjiang Province, in order to investigate the frequency and subtype distribution and to evaluate the risk of the zoonotic transmission of Blastocystis sp..Results show that the frequency was 10.5% (6/57), and three subtypes (ST1, ST2, and ST3) were found in humans; in animals, the frequency was 17.9% (46/257), and six subtypes (ST1, ST3, ST5, ST7, ST10, and ST26) were detected. A significant association between Blastocystis sp. infection and eating unwashed vegetables and fruits was found (P = 0.007). We found no effect on gender, age and season on Blastocystis sp. colonization. Phylogenetic analysis showed that Blastocystis ST7 sequences from chicken in two geographical locations formed two distinct clades. Alleles were identified using the Blastocystis 18S database, and a total of 10 different alleles were found in seven STs. Overlap of STs in humans and domestic animals (pig or cattle) was observed in Dongfanghong village. The findings of potentially zoonotic subtypes in domestic animals suggest that these animals may serve as reservoirs of human Blastocystis sp. infections. Multisectoral cooperation is needed to slow down the transmission of Blastocystis in domestic animals, minimize environmental contamination by Blastocystis cysts, and increase molecular epidemiological surveillance of Blastocystis sp. in humans and animals.

Background: Giardia duodenalis is a common parasitic diarrheal agent in humans, especially in developing countries. The aim of this study was to investigate the prevalence and multilocus genetic characterization of G. duodenalis in patients with diarrhea and animals in northeastern China, and to assess the epidemiological role of animals in the transmission of human giardiasis. Methods: A total of 1739 fecal specimens from 413 diarrheal patients and 1326 animals comprising 16 mammal species were collected in Heilongjiang Province of China and screened for G. duodenalis by PCR and sequencing of the SSU rRNA gene. All G. duodenalis-positive specimens were subtyped by PCR and sequencing of the bg, tpi, and gdh genes. To detect additional mixed infections of different assemblages, assemblage A/B/E-specific PCRs were performed to amplify the tpi gene. Results: Sequence analysis of the SSU rRNA gene determined the prevalence of G. duodenalis (5.81%, 24/413) in diarrheal patients, with a peak in minors aged 5–17 years, and identified assemblages A and B. MLG-AII and MLG-B1 were obtained based on concatenated nucleotide sequences of the bg, tpi, and gdh genes, with MLG-AII being identical to a cat-derived isolate reported previously. By sequence analysis of the SSU rRNA gene, G. duodenalis was detected in 214 (16.14%) animals belonging to 11 mammal species, with the prevalence ranging from 1.69 to 53.85%, and assemblages A to G were identified. Sequence analysis of the bg, tpi, and gdh genes from 46 specimens produced 31 MLGs, including MLG-AI (n = 1), MLG-B2–B8 (n = 18), and MLG-E1–E23 (n = 27). Conclusions: The finding of G. duodenalis in diarrheal patients enhances consciousness of detecting G. duodenalis in clinical practice and emphasizes the importance of health education in local inhabitants, especially in the age group of 5–17 years. The identification of seven assemblages (A to G) and 33 MLGs reveals genetic heterogeneity of G. duodenalis in the investigated areas. Due to insufficient homology data on the zoonotic transmission of G. duodenalis, the precise epidemiological role that animals play in the transmission of human giardiasis needs to be assessed by more large-scale molecular epidemiological investigations of local humans and animals. Graphical

Background: Cryptosporidiosis and giardiasis are significant parasitic diseases shared between humans and domestic animals. Due to the close contact between humans and domestic animals in rural areas, it is important to consider the potential transmission of zoonotic parasites from infected domestic animals to humans. This investigation aimed to determine the prevalence and molecular characteristics ofCryptosporidiumspp. andGiardia duodenalisin domestic animals and villagers.Methods: A total of 116 fecal samples from villagers and 686 fecal samples from domestic animals in Heilongjiang Province, China, were analyzed for two parasites using nested polymerase chain reaction (PCR) targeting various genetic loci and DNA sequence analysis of the PCR products.Results: By sequence analysis of theSSUrRNA gene, the prevalence ofCryptosporidiumin humans was 0.9% (1/116), with one species ofC. parvum(n = 1) detected; among domestic animals, the prevalence was 2.6% (18/686), with five species identified:C. suis(n = 7) andC. scrofarum(n = 7) in pigs,C. meleagridis(n = 1) in chickens,C. andersoni(n = 1) in cattle, andC. canis(n = 2) in foxes.C. parvum and C. caniswere further subtyped as IIdA19G1 and XXa4 on the basis ofgp60gene. RegardingG. duodenalis, based on theSSUrRNA,bg,gdh, andtpigenes, the prevalence in domestic animals was 5.1% (31/608), with three assemblages identified: A (n = 1) in pigs, D (n = 1) in foxes, and E (n = 27) in geese, cattle, pigs, ducks, and sheep, along with mixed infection of A + E (n = 1) in one pig and B + E (n = 1) in one sheep. NoG. duodenaliswas detected in humans (0/116).Conclusions: The present results show that no overlap of subtypes between animals and villagers was found inCryptosporidiumspp. andG. duodenalis, indicating a minor role of domestic animals in infecting humans in this population. However, the presence of zoonotic protozoa in domestic animals highlights the need for special attention to high-risk individuals during close contact with domestic animals.Graphical

Rodents constitute the largest and most successful group of mammals worldwide. Brown rats (Rattus norvegicus) are one of the most common rodent species, and they serve as intermediate hosts of Hydatigera taeniaeformis. Although there have been a few studies reporting on the presence of the larval form of H. taeniaeformis (strobilocercus fasciolaris) in brown rats worldwide, little information is available on the genetic characterization of this parasite, with no molecular data from China. Therefore, from April 2014 to March 2016, this study was carried out to understand the prevalence and genetic characters of strobilocercus fasciolaris in brown rats captured in Heilongjiang Province in northeastern China. The livers of brown rats were collected and examined for the presence of cysts. Each cyst was identified based on morphological observation: the larvae with the naked eye and the scolexes under a microscope. The results were confirmed by polymerase chain reaction (PCR) and sequencing of the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 4 (nad4) genes. At the investigated sites, 11.8% (13/110) of the brown rats were infected with strobilocercus fasciolaris. Based on sequence analysis, there were 10 and six haplotypes regarding the cox1 and the nad4 loci, with 24 and 42 polymorphic sites, respectively (degree of intraspecific variation: 0.3%–4.4% and 0.6%–4.7%, respectively). Twelve nucleotide sequences (six of the 10 at the cox1 locus and all six at the nad4 locus) have not previously been described. Base differences in three of the six novel cox1 gene sequences and five of the six novel nad4 gene sequences caused amino acid changes. Phylogenetic analyses of the cox1 and nad4 gene sequences based on neighbor-joining and Bayesian inference trees indicated that all the strobilocercus fasciolaris isolates belonged to Hydatigera taeniaeformis sensu stricto (s.s.). This is the first report on the genetic characterization of strobilocercus fasciolaris in brown rats in China. The findings of novel cox1 and nad4 nucleotide and amino acid sequences may reflect the region-specific genetic characterization of the parasite. The data will be useful to explore the biological and epidemiological significance of the intraspecific variation within H. taeniaeformis s.s.

Introduction: Enterocytozoon bieneusi is a zoonotic parasite causing diarrheal disease in rural settings with frequent human–animal contact. Methods: We conducted a cross-sectional study in Heilongjiang, China, enrolling 69 households across 10 villages. Humans and domestic animals were sampled. Nested PCR targeting the ITS region was performed. Genotypes were determined by sequencing, and genetic diversity was assessed using DnaSP. Results: Human prevalence was 11.0% (13/118), while overall animal prevalence was 23.4% (162/693), with sheep being the highest (66.7%), followed by pigs (46.1%) and geese (20.5%). Prevalence was significantly higher in mammals than poultry (32.9% vs. 13.9%; p < 0.001). Sequence analysis yielded 29 distinct genotypes (10 known, 19 novel), with EbpC (36.0%), BEB6 (13.7%), EbpA (13.7%), and D (11.4%) being the most prevalent, most clustering within zoonotic Group 1. Discussion: Notably, zoonotic genotypes EbpC and EbpA were identified in chickens and geese, extending the known host range of these genotypes. Human–animal genotype sharing was observed in only one household (genotype EbpC between humans, pigs, chickens, and geese), while intraspecies circulation and occasional animal–animal genotype sharing were more common. Phylogenetic and network analyses suggested localized diversification, though single-locus ITS limits definitive evolutionary inference. E. bieneusi is highly prevalent in rural domestic animals in Heilongjiang, with genetically diverse and potentially zoonotic genotypes circulating in livestock and poultry. Although household-level human–animal transmission appears limited in this dataset, the presence of shared zoonotic genotypes highlights the need for expanded surveillance incorporating environmental sampling and multi-locus typing to clarify transmission pathways and inform One Health strategies.