| Enerocytozoon bieneusi,Giardia duodenalis and Blastocystis are three important zoonotic intestinal protozoans.After the oocysts of the three parasites are excreted from the infected hosts,they are transmitted through water sources and contaminated food,and the oocysts can exist for a long time in the environment.Once humans and animals are infected with E.bieneusi,G.duodenalis and Blastocystis,these parasites can cause diarrhea,weight loss and other gastrointestinal diseases,which cause great harm to humans and significant economic loss to animal production.At present,there are limited reports of the three parasites in goats in China,and there is no report of these three parasites in Yunnan black goats.Therefore,this study investigated the status of infection and genotyping of E.bieneusi,G.duodenalis and Blastocystis in Yunnan black goats in Yunnan province.Object: This study collected fecal samples from 907 Yunnan black goats in five regions in Yunnan,examined the molecular epidemiology and genetic structures of E.bieneusi,G.duodenalis and Blastocystis in Yunnan black goats.The risk that these intestinal parasites may pose to humans and animals in Yunnan was assessed.Meanwhile,this study will enrich host spectra for the three protozoans and provide data support for local parasite prevention and control.Methods: Using molecular biology method,this study amplified the E.bieneusi ITS sequences from 907 fecal samples by nested PCR amplification.Then the positive DNA samples were further amplified for the microsatellite and small satellite loci MS1,MS3,MS4,MS7 using nested PCR,and MLST tools were used to perform multi-genotyping of the ITS and four microsatellites and small satellite loci to construct MLGs.For G.duodenalis: Based on the three loci of bg,gdh and tpi,PCR amplification was performed,and MLGs were constructed by genotypic results of three loci.For Blastocystis: the prevalence and genotypes were determined by PCR amplification of the SSU rRNA gene.Statistical methods were used for the above three intestinal protozoans to assess statistical differences between regions,genders,and ages.Neighbor-Joining trees were constructed to further evaluate the potential public health risks in the study areas.Results: The overall prevalence of E.bieneusi in Yunnan black goats was 10.3%(93/907),the prevalence of G.duodenalis was 4.2%(38/907),and the prevalence of Blastocystis was 41.3%.The Blastocystis prevalence in Yunnan black goats is significantly higher than that of the other two protozoans,so Blastocystis is the dominant intestinal protozoan in Yunnan black goats.E.bieeneusi: The overall prevalence in Yunnan black goats was 10.3%.A total of 7 known genotypes and 15 new genotypes were identified.The analysis of microsatellites and small satellites indicated that CHG1 was the dominant genotype,belonging to the non-zoonotic type;In addition,the eight new zoonotic genotypes have been identified.G.duodenalis: A total of 38 positive samples were detected and identified as cluster E at the three loci of bg,gdh and tpi.Five new genotypes were found in three loci.Three new MLGs genotypes(MLG)MLGE9-MLGE11 were identified.Blastocystis: A total of 375 positive samples were found from 907 samples,3 known subtypes(ST5,ST10,ST14)and 4 novel(ST-novel 1、ST-novel 2、ST-novel 3 and ST-novel 4)subtypes were identified,of which ST5 is the dominant subtype in Yunnan black goats,indicating public health risk.Conclusion: In summary,this study demonstrated that Yunnan black goats were infected with E.bieneusi,G.duodenalis and Blastocystis,and the Blastocystis prevalence is the highest.Through the detection of black goat samples from different regions,the present study elucidated the infection status,genotypes,gene subtype distribution and population structure of three protozoans in each region.This study provided baseline data for the prevention and control of intestinal protozoan infection in Yunnan black goats,and has public health implications. |
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