Molecular Epidemiology Of Pentatrichomonas Hominis And Enterocytozoon Bieneusi And Intestinal Flora Analysis Of Pet Dogs In Henan Province

Pentatrichomonas hominis and Enterocytozoon bieneusi are two common zoonotic intestinal parasites that can be transmitted through a variety of routes and are widely prevalent worldwide with a wide range of hosts,including domestic animals,wild animals and humans.Infection with these pathogens can cause self-limited diarrhea in individuals with strong immunity,or chronic symptoms in infants,the elderly,and immunocompromised populations.Self-limited diarrhea can occur in hosts with normal immunity,persistent watery diarrhea in immunocompromised hosts,and death in severe cases.The two pathogens are transmitted mainly through the faecal-oral route.Infections can also be caused by contaminated food or water.The pet industry has been booming in China in recent years,with pet ownership continuing to rise.It is well known that dogs,as important companion animals,have a close relationship with humans,which inevitably increases the risk of zoonotic parasites being transmitted from pets to humans,threatening human health.However,the prevalence and potential zoonotic diseases of Pentatrichomonas hominis and Enterocytozoon bieneusi in pet dogs are rarely reported.In order to understand the prevalence of two intestinal parasites in pet dogs,1393 canine feces samples were tested for Pentatrichomonas hominis,569 canine feces samples were tested for Enterocytozoon bieneusi,and the intestinal flora of Pentatrichomonas hominis was analyzed.A total of 1393 fecal samples were collected from 6 regions in Henan Province in order to understand the prevalence,species,genotypes and zoonotic risk of Pentatrichomonas hominis and Enterocytozoon bieneusi in pet dogs.Based on SSU r RNA gene locus,nested PCR was performed for the amplification of Pentatrichomonas hominis.The results showed that 26 Pentatrichomonas hominis samples were detected,and the total infection rate was 1.87%.The infection rate of Pentatrichomonas hominis in different sites range from 0.84% to 2.50%,and the infection rates were significantly different(P<0.01).The infection rate of pet dogs in Zhengzhou was 2.50%.Two Pentatrichomonas hominis genotypes were identified by sequencing of the positive amplification products: CC1(17/26)and CC2(9/26),both of them are zoonotic.Based on ITS gene loci,a total of 198 samples of Enterocytozoon bieneusi were amplified by nested PCR from 569 canine fecal samples,with a total infection rate of 34.80%.Six ITS gene types were identified: Type Ⅳ(24/198),Henan Ⅴ(13/198),Ebp C(19/198),CD 5(34/198),CD 9(25/198)and Pt EbⅨ(83/198),four of which were the common gene types: Type Ⅳ、Henan Ⅴ、Ebp C and Pt EbⅨ,Pt EbⅨ was the dominant genotype.Type Ⅳ,Henan Ⅴ and EbpC are zoonotic.Based on the results of 16 S rRNA sequencing,the effects of infection with Pentatrichomonas hominis on intestinal flora in healthy dogs were analyzed.It was found that the parameters related to species richness in intestinal Alpha diversity of dogs infected with Pentatrichomonas hominis had no significant changes,and species diversity increased significantly at the phylum level.The dominant bacteria in the positive group were Firmicutes and Fusobacteriota.At the genus level,Prevotella,Fusobacterium,Alloprevotella,Negativibacillus,Bibersteinia and Tyzze rella,Uminococcus torques group,Sellimonas and Bacillus were the positive dominant group.PCo A analysis showed that the intestinal microbial communities of the two groups were significantly separated,and the intestinal flora clustering was significantly different,indicating that the intestinal flora structure of healthy dogs infected with Pentatrichomonas hominis had changed.The relative abundance of harmful bacteria(Fusobacterium,Bacillus,Tyzzerella,Bibersteinella)in the intestinal tract of dogs infected with Pentatrichomonas hominis was significantly increased.These results indicate that the diversity and structure of intestinal flora of dogs infected with Pentatrichomonas hominis will be changed,and the relative abundance of harmful bacteria will be increased.In conclusion,the two types of Pentatrichomonas hominis and three types of Enterocytozoon bieneusi infected dogs in this study have the risk of zoonosis,suggesting the possibility of the two intestinal protozoa spreading between pets and humans,which provides a scientific basis for effective clinical prevention and control,and has certain public health significance.The diversity and structure of intestinal flora of dogs infected with Pentatrichomonas hominis will change,and the relative abundance of harmful bacteria will increase,which provides an important reference for the study of the relationship and interaction between the intestinal flora and pet dogs.