Molecular Epidemiology Of Enterocytozoon Bieneusi And Cryptosporidium In Fox And Raccoon Dog

Cryptosporidium and Enterocytozoon bieneusi are two common zoonotic intestinal protozoa,which are widely prevalent in the world and can infect people and a variety of animals,mainly causing diarrhea.For immunocompromised hosts,persistent watery diarrhea can be caused,and death can be caused in severe cases.It can be spread in a variety of ways.The two pathogens are mainly transmitted by the faecaloral route and can cause infection through contaminated food or water.To understand the infection stuatus of cultured fox and breeding the fox and raccoon dog,the fecal samples of 423 foxes and 415 raccoon dogs were examined by means of Lugo’s iodine solution staining and saturated sucrose floatation.Ball,nematodes and amoeba were found and the infection rates were 1.79%,5.73%and 1.43%,the total infection rate was 8.95%.In order to understand the prevalence stste,species,genotypes and zoonosis risk of Cryptosporidium and Enterocytozoon bieneusi of cultured foxes and raccoon dogs,a total of 1588 fecal samples(1240 and 348,respectively)were collected from 5 farms in 2 districts of Henan Province and 3 farms in 1 district of Hebei Province.Cryptosporidium was amplified by nested PCR based on SSU rRNA gene loci,and 17 Cryptosporidium were found,with a total infection rate of 1.07%.The infection rate of Cryptosporidium in different locations was 0-1.75%,with a statistically significant difference(P<0.05).The highest infection rate was 1.75%in Xinxiang area,and no positive samples were found in Hebi and Changli areas.Three cryptosporidium species were found,and all of them were zoonotic,with the proportions of C.canis(12/17),C.parvum(4/17)and C.andersoni(1/17),respectively.C.canis was the dominant species.A total of 178 Enterocytozoon bieneusi were amplified by nested PCR based on ITS gene locus,with a total infection rate of 11.21%,among which the infection rates of foxes and raccoon dogs were 72.47%(129/178)and 27.53%(49/178),respectively.A total of 10 ITS genotypes were identified by sequence analysis:BEB4,BEB6,NCF2,NCF3,NCF7,D,EBPC,CHN-DC1,SCF2 and CHN-F1,among which CHN-DC1,CHN-F1 and D were common genotypes.NCF2 was the dominant genotype and was found in 6 farms in 2 provinces.Most genotypes belong to the phylogenetic first population and are zoonotic genotypes.A total of 14 MLGs(MLG1-14)were identified by combining ITS and MLST with four microsatellite and microsatellite loci(MS1,MS3,MS7 and MS4)genotypes.In conclusion,based on the epidemiological analysis and zoonotic risk assessment of Cryptosporidium and Enterocytozoon bieneusi of foxes and raccoon dogs in different areas of Henan and Hebei Province,it was found that foxes and raccoon dogs have important public health significance in the prevalence and transmission of Cryptosporidium and Enterocytozoon bieneusi.This study provides scientific basis for the prevention and control of Cryptosporidium and Enterocytozoon bieneusi infection in humans and animals.