The Molecular Epidemiological Studies Of Four Species Of Intestinal Protozoa In Alpacas(Vicugna Pacos)in Shanxi Province

Alpaca,originated from South America,is an important economic animal prized for its wool fiber and meat.Diarrhea can lead to slow growth,weight loss and low economic benefits,affecting the development of alpaca breeding industry.There are many causative agents of diarrhea,intestinal parasites are one of the most important factors.However,there is limited information on intestinal parasite infection in alpacas in China.Thus,in this study,the prevalence and molecular identification of Enterocytozoon bieneusi,Blastocystis,Entamoeba species and Eimeria species in alpacas were carried out.In the present study,a total of 366 fecal samples from alpacas in Shanxi Province,northern China,were collected to detect the prevalence and genotypes/subtypes/species of E.bieneusi,Blastocystis,Entamoeba spp.and Eimeria spp.by PCR amplification.The results showed that the overall prevalence of E.bieneusi,Blastocystis,Entamoeba spp.and Eimeria spp.in alpacas were 4.4%(16/366),23.8%(87/366),18.0%(66/366)and 1.6%(6/366),respectively.Through the analysis of ITS sequences,four known genotypes of E.bieneusi were identified,namely ALP1,ALP3,P,and SH11,all of which belong to the zoonotic group.ALP3 was the primary genotype of E.bieneusi in alpacas in this study,and this is the first study reporting the presence of SH11 in alpacas.Moreover,ITS-positive samples were further characterized by multilocus sequence typing(MLST),forming eight multilocus genotypes(MLGs).Of which,seven MLGs were observed in the genotype ALP3 and one MLG in the genotype P.The MLGs of E.bieneusi in alpacas showed complicated genetic diversity within the same ITS genotype.SSU r RNA analysis identified three Blastocystis subtypes(namely ST5,ST10 and ST14)among 87 positive samples.ST5 is a genotype of potential zoonotic significance,which indicated that alpacas harbouring Blastocystis ST5 could be a potential source of infection for humans.Six Eimeria-positive samples tested negative for E.macusaniensis.E.bovis was identified as the predominant Entamoeba spp.Nine unique Entamobea sequences were detected and formed a new phylogenetic cluster.In summary,this study analyzed the prevalence,genotype/subtype/species of four important intestinal parasites of alpacas in Shanxi Province,and evaluated their zoonotic risks.These findings not only have important implications for the prevention and control of intestinal protozoa infections,but also provide baseline information for further elucidation of population genetics,zoonotic potential and transmission patterns of these intestinal protozoa.