Epidemiologic Study And Zoonotic Risk Analyze Of Three Intestinal Parasitic Protozoa In Captive Golden Snub-nosed Monkey (Rhinopithecus Roxcllanae) In China And Chindren Hospitalized In Zhengzhou

Cryptosporidium spp.,G.duodenalis and E.bieneusi are intestinal parasitic protozoa which have been concerned for food-borne transmission causing symptomatic diarrheal disease in humans and animals worldwide.The diarrheal is self-limiting in healthy hosts,but life-threatening in immunosuppressed and young individuals.There is a vast territory in China with abundant wild animal species resources,which complicates the status of both transmission and prevalence of zoonotic parasitic disease.As the closer genetic relationship with human,wild or captive nonhuman primates(NHPs)play a more important role in the transmission of the three protozoa and considered to be the reservoir of many parasites.Meanwhile,some species/Assemblages/genotypes of certain parasites can infect both NHPs and human,so molecular evolution researches on these species/Assemblages/genotypes are conducived to reveal the transmission mechanism between NHPs and human.To a certain extent,the prevalence of zoonotic parasitic diseases is underestimated for the incomplete of routine diagnostic and monitoring as well as reporting system and the lack of the infection reports of these three intestinal parasitic protozoa.To uncover the infection status and epidemiological characteristics of these three parasites in captive golden snub-nosed monkeys in zoos in China,160 fecal specimens were collected from Beijing Zoo,Beijing Wildlife Park,Shanghai Zoo,Shanghai Wild Animal Park,Taiyuan Zoo and Tongling Zoo from June to November 2015.Six parasites were found in 103 fecal specimens by microscopy,with the overall infection rate 64.4%.These parasites are Trichuris spp.,Isospora spp.,Endolimax spp.,C.parvum,G.duodenalis and Strongyloides spp.,and the infection rates are 58.8%,13.8%,10.0%,1.9%,0.6%,0.6%,respectively.There are 30 specimens which co-infected with 2 or more parasites,with the mixed infection rate 29.1%.Additionally,genome DNA of all 160 specimens were extracted in this study,followed by nest PCR based on different genetic loci of these three parasites.The overall infect rate observed was 52.5%(84/160),and the prevalence of Cryptosporidium spp.,G.duodenalis and E.bieneusi is 48.1%(77/160),8.1%(13/160),5.0%(8/160),respectively.Sequencing data revealed the Assemblage of G.duodenalis is B and the species of Cryptosporidium spp.are C.andersoni,C.ubiquitum and C.parvum.To find out the infection status and epidemiological characteristics of these parasites in children hospitalized in Zhengzhou,2284 specimens were collected from 3 hospitals in Zhengzhou from December 2015 to January 2017.Data of molecular biology showed that 47 specimens were positive for at least one of the three parasites and no mixed infection was detected.With the prevalence 1.18%(27/2284),E.bieneusi was proved to be the dominant parasite in this study,and it was also presented as the dominant parasite in each of the three hospitals involved in this study.The infection rate of Cryptosporidium spp.and G.duodenalis is 0.26%(6/2284),0.61%(14/2284),respectively.For children hospitalized in different departments,children hospitalized in Internal Digestive Department have the highest overall infection rate(8.33%,13/156),followed by Renal Rheumatism Department(4.98%,11/221)and Hematology-Oncology Department(4.14%,6/145).To assess the zoonotic risk potential of from animals and human as well as to reveal the role played by different host of E.bieneusi,phylogenetic analysis on different E.bieneusi isolates was implemented based on internal transcribed spacer(ITS).A total of 2284 human samples and 160 golden snub-nosed monkey samples were examined,of which 97 were amplified and sequenced successfully.Ten genotypes were found,inclouding 8 known genotypes: D(n=71),J(n=16),CHG1(n=1),CHG14(n=1),BEB8(n=2),Pig EBITS7(n=2),CM8(n=1)and BEB6(n=1)and 2 novel genotypes: CM20(n=1),CM21(n=1).All the two novel genotypes were found in golden snub-nosed monkey samples.Genotype D,CM8 and Pig EBITS7 were clustered into Group 1 with high zoonotic potential in the phylogenetic analysis.Genotype BEB8,CHG1,CHG14 and J were clustered into Group 2 as well as the two novel genotypes.Prevelance status of intestinal parasites were examined in captive golden snub-nosed monkeys and children hospitalized,with phylogenetic analysis and the zoonotic risk potential assessment on different E.bieneusi isolates from animals and human conducted in this thesis,which have positive significance to protect the golden snub-nosed monkeys better and reduce the threat from human intestinal parasites.Additionally,these datas obtained in this thesis offered a practical and theoretical referrence to the prevention and control of intestinal parasites in captive golden snub-nosed monkeys and children hospitalized.