PCR Detection And Subtype Indentification Of Enterocytozoon Bieneusi In Large-scale Sheep Farms In Southern Xinjiang

Enterocytozoon bieneusi is an intracellular parasitic eukaryotic single-celled microorganism with a wide range of hosts.It was found that sheep from different countries and regions were commonly infected with E.bieneusi,and their genotypes were characterized by genetic diversity and host adaptability.In this study,PCR was used to detect E.bieneusi infection in sheep from some large-scale farms in Southern Xinjiang.Genotypes/subtypes of E.bieneusi were identified and genetic diversity was analyzed by sequence analysis and comparison.1.A total of 892 sheep fecal samples from 7 large-scale farms in 3 regions of Southern Xinjiang were collected.Based on the ITS gene of E.bieneusi,PCR method was used to detect DNA samples from sheep fecal samples,and 309 samples were positive for E.bieneusi,with a total infection rate of 34.6%(309/892).The infection rate of E.bieneusi was 57.1%(60/105)in sheep from Kuqa farms.The infection rate of E.bieneusi in sheep from different farms was significantly different(P<0.01).The highest infection rate was43.1%(103/239)in lambs weaned at 3-6 months of age.The infection rates were 20.5%(42/205),35.5%(75/211)and 21.5%(51/237)in adult sheep,lambs less than 3 months of age and fattening sheep from 6 to12 months of age,respectively.There were significant differences in the infection rates among sheep of different ages(P<0.01).6 known E.bieneusi genotypes were identified by sequence alignment analysis,among which BEB6 accounted for 93.5%(289/309)Is the highest proportion of all genotypes.The other genotypes were COS-I(n = 9),CHS3(n = 5),NESH4(n = 2),CHC8(n = 3)and CHS5(n = 1).Phylogenetic analysis showed that genotype CHS5 belonged to group 1,and the other five genotypes belonged to group2.The results showed that E.bieneusi was prevalent in sheep on large-scale sheep farms in Southern Xinjiang,and genotype BEB6 was the dominant genotype.2.The 289 positive samples of E.bieneusi genotype amplified by ITS locus were selected based on different regions and different ages,and 80 samples were selected for PCR amplification using MS1,MS3,MS4 and MS7 loci.34 and 31 samples were successfully amplified at MS1 and MS7 sites,and 17 samples were amplified at two sites simultaneously.20 t different subtypes were formed at MS1 site,and 3 subtypes with obvious base pair duplication occurred at MS7 site.However,no bright bands were successfully amplified at MS3 and MS4 sites.The results showed that the genetic diversity of E.bieneusi BEB6 subtypes existed in southern Xinjiang.In conclusion,it was found that E.bieneusi was prevalent in sheep of large-scale farms in Southern Xinjiang,and genotype BEB6 was the dominant genotype,and its gene subtypes had diversified differentiation characteristics.These results provide basic reference for the study of the etiology and molecular genetic characteristics of E.bieneusi from sheep in China.