Molecular Epidemiology Of Three Protozoa In Rabbits In Henan And Genomic Analysis Of Giardia Assemblage B

Giardia duodenalis,Enterocytozoon bieneusi,and Blastocystis sp.are three important zoonotic intestinal parasitic protozoa.Infection in rabbits can cause clinical symptoms such as diarrhea,and even lead to death.Among them,Giardia duodenalis assemblage B shows high host adaptability characteristics for rabbits.To know the situation of intestinal parasitic infections in large-scale rabbit farms in some areas of China,explore and analyze the epidemic characteristics of rabbits in some areas of China and explore the characteristics of the genome of Giardia duodenalis assemblage B.This study investigated the intestinal parasites of rabbits in Henan.Molecular biology methods was used to identify the assemblage of Giardia duodenalis,the genotype of Enterocytozoon bieneusi,and the subtype of Blastocystis sp..Meanwhile,single cyst separation technology and Nanopore sequence was used for obtaining the whole genomic of non human primate derived Giardia duodenalis assemblage B and followed by carry out structural functional analysis and comparative genomic analysis.1 Infection of intestinal parasites in rabbits in some areas of Henan ProvinceThis study conducted a microscopic examination of 1367 fresh rabbit fecal samples from 27 breeding farms and 2 flower markets in 7 cities in Henan Province(Luoyang,Gongyi,Jiyuan,Xinxiang,Zhengzhou,Dengfeng,Kaifeng)to identify the intestinal parasites in rabbits.The results show that the total infection rate of intestinal parasites in rabbits in Henan region was 59.69%(816/1367),and a total of four genotypes of intestinal parasites were detected,Endolimax spp.,Coccidia,Passalurus and Giardia duodenalis.Among them,the infection rate of Coccidia is the highest,with an infection rate of 55.60%(459/1367).The infection rates of Endolimax spp.,Passalurus,and Giardia duodenalis are21.07%(288/1367),23.92%(327/1367),and 0.37%(5/1367).It can be seen that the infection of intestinal parasites in rabbits is relatively widespread.2 Molecular epidemiology of three kinds of zoonotic intestinal protozoa in rabbits in some areas of HenanBased on the bg gene locus,ITS gene locus and SSU r RNA gene locus,a molecular epidemiology survey was carried out on Giardia duodenalis,Enterocytozoon bieneusi and Blastocystis sp.in rabbits.The results show that the total infection rate of the protozoa was5.34%(73/1367).Among them,the overall infection rate of Giardia duodenalis is 0.37%(5/1367),and all of them are assemblage B which are only detected in 60 days age of adult female rabbits from breeding farm of Xinxiang area in spring.The total infection rate of E.bieneusi is 1.76%(24/1367),and the genotypes identified are zoonotic genotype BEB6 and CHG4.Among different areas and age groups,the highest infection rate of E.bieneusi is6.54%(14/214)in Xinxiang area and 2.31%(3/130)in rabbits 30-60 days age.The total infection rate of rabbit Blastocystis sp.is 4.17%(59/1367).The subtype identification shows that it is a zoonotic subtype ST1,ST3,ST4,and ST17.There are differences in infection rates among different areas and age groups,.The highest infection rate is 25%(1/4)in Kaifeng area and 5.33%(38/713)in 30 days age rabbits.3 Sequencing and genomic analysis of Giardia duodenalis assemblage BIn this study,the fecal samples of hamadryas baboon were collected in Zhengzhou Zoo.Single cyst separation technology was used to obtain the single cyst of Giardia duodenalis assemblage B.The whole genome DNA was extracted to Nanopore sequence for genomic data splicing and annotation.The results show that the genome data of Giardia duodenalis assemblage B was 12.8Mb,with 7823 encoded proteins annotated.Compared with the reference genome data of 4 Giardia duodenalis isolates,genomic analysis show that 3059 sequences were clustered among 5 Giardia isolates.More than 95% of the genes obtained from Giardia isolates can be founded as homologous genes in the other four isolates.Therefore,the obtained genome data of Giardia isolates has certain reliability.In summary,this study shows that rabbits raised in large-scale rabbit farms in some areas of Henan Province are generally infected with intestinal parasites.The infection rates in the same region,age and season are all different,and there is a phenomenon of mixed infection.The molecular biological identification results of three genotypes of intestinal parasites in rabbits show the infection rate is relatively low.The zoonotic genotype is the dominant genotype among the three intestinal parasites in rabbits.Meanwhile,this study completed the whole genome sequencing,data splicing and structural analysis of Giardia duodenalis assemblage B.This study lays the foundation for molecular mechanism of variation and pathogenic mechanism of Giardia duodenalis.