A Survey Of Intestinal Parasites In Donkeys Of Animal-exchange Market In Jiaxiang County,Shandong And Multilocus Sequence Typing Of Cryptosporidium Donkey Genotype

Parasitic disease of equine is a most common chronic,covert illness in the modern equine breeding.Infected animals often do not show obvious clinical symptoms,the severe one always manifests as emaciation,anemia,diarrhea and even death.Parasitic disease of equine is mainly caused by Strongylid,Parascaris equorum,Cestode,Coccidium,Cryptosporidium and Giardia.Among these parasites,Cryptosporidium and Giardia are zoonotic protozoiasis and global distribution.In order to understand the infection and epidemic of intestine parasites in donkey,total 508 stool samples of donkey in Livestock Trading Market in Jiaxiang county of Shandong province were collected from March 2013 to May 2014,then the Lug’s iodine staining and Sheather’s sugar flotation technique were uesd to detect intestine parasites.The results indicated that there are 246 positive samples of parasites were detected,the total infection rate was 48.43%,there are 9 species parasites were found,they are Coccidium,Giardia,Cryptosporidium,Strongylid,Parascaris equorum,Cestode,Trichuris,Pinworm,Ancylostoma respectively.Among these parasites,the infection rate of Strongylid was the highest,it is up to 30.71%(156/508),closely followed by Parascaris equorum,reached 20.28%(103/508).The infection rate of Coccidium,Giardia,Cryptosporidium,Cestode,Trichuris,Pinworm,Ancylostoma was 3.74%(19/508),0.39%(2/508),5.91%(30/508),2.76%(14/508),0.59%(3/508),1.77%(9/508),0.20%(1/508),respectively,the mixed infection rate was 15.94%(81/508),most of case were infected by two parasites,and the pathogens is mainly Strongylid and another parasite.In order to know phylogenetic relationship and population genetic structure of 114 Cryptosporidium isolates of donkeys which were kept by our laboratory,HSP70、MSC6-7、MSC6-5、CP47、CP56、ML2、Chom3T、RPGR genetic locus were selected to amplify.Firstly,GP60 genetic locus was used to identify subtype and 69 isolates were amplified successfully,include C.parvum(Ⅱd A19G1,N=18),Horse genotype(N=2)and Donkey genotype(ⅫA16G1,N=49).After screening,CP47 and CP56 genetic locus were abandoned for low amplification efficiency.14 Cryptosporidium Donkey genotype isolates can be amplified in all of those 6 genetic locus(Chom3T,HSP70,ML2,MSC6-5,MSC6-7,RPGR),then MLST was used to analyze the population genetic structure of Cryptosporidium Donkey genotype.The result showed had there are 2,1,3,4,4,3 subtypes of 14 Cryptosporidium Donkey genotype isolates in those 6 genetic locus above-mentioned respectively,and 7 MLST suptypes were founded in these samples.Linkage disequilibrium analysis manifested that Cryptosporidium Donkey genotype in the study was an epidemic population,it implied that the epidemic of donkey Cryptosporidiosis in equine is unstable.