Survey Of Rabbit Intestinal Parasites In Some Areas Of China And Multilocus Sequence Subtyping Of Cryptosporidium Cuniculus

Parasitic disease is a significant chronic, debilitating illness in the rabbit breeding. Mild infected animals often do not show obvious clinical symptoms, the severe one always manifests as diarrhea, weight loss, and inappetence, even life-threatening.Rabbit parasitic disease caused by Coccidia, Giardia, Strongyloides, Passalurus ambiguous and Cryptosporidium. Among these parasites, Giardia and Cryptosporidium are zoonotic agents which can be transmitted by the fecal-oral route and threaten human health.In order to understand epidemiological characteristics of rabbites’ intestine parasites in some areas of China, samples colleceted form Henan, Sichuan, Zhejiang, Guizhou province parasite were detected from March2012to September2012. at the same time, Cryptosporidium cuniculus isolates saved in Hean Agricultural University Parasitology Laboratory were subtyped by a multilocus sequence typing (MLST) and population genetic structure were analyzed too. These studies will help us to understand C. cuniculus dissemination mechanisms and biological characteristics, the results will provide acadmic basis for controlling of Cryptosporidium.To know the prevalence status of in rabbites and epidemiological characteristics in China,2115fecal samples were collected from the partial areas of Henan, Sichuan, Zhejiang, and Guizhou. The samples were examined using the Sheather’s sugar flotation technique and the Lug’s iodine stain method,1559samples were positive.and5parasites were detected, they are Coccidium, Giardia, Strongyloides, Passalurus ambiguous, Amoeba, and Mite. The overage prevalence of intestinal parasites was73.71%(1559/2115), in which, the infection rate of Coccidium was the higehist, reached60.95%(1289/2115), the infection rate of Giardia, Strongyloides, Passalurus ambiguous, Amoeba, and Mite oocysts/eggs/cysts was0.61%(13/2115)、18.39%(389/2115),0.85%(18/2115)、5.67%(120/2115)、3.50%(74/2115), respectively. The results showed that parasite mixed infecition especially two kinds of parasites wer more commonly (mainly Coccidia with other parasite), and two parasites mixed infection rate was10.42%.To assess C.cuniculus phylogenetic relationships and molecular epidemiology, minisatellites, microsatellites and SNP loci, include MSC6-7(serine repeat antigen), RPGR (retinitis pigmentosa GTPase regulator), MSC6-5(hypothetical trans-membrane protein), DZ-HRGP (hydroxyproline-rich glycoprotein), CP56(56kDa trans-membrane protein), Chom3T (T-rich gene fragment), CP47(47kDa protein), Mucinl (mucin-like protein), ML2(possible sporozoite cystein-protein)were amplified and sequenced. The sofeware ClustalX2.1, DnaSP version5.10.01, LIAN version3.5was used to do Sequence alignment, analyze the genetic diversity of the C.cuniculus sequences and testing Linkage disequilibrium (LD) by a parametric method for nine minisatellites, microsatellites and SNP loci, respectively. A total of15isolates were successfully subtyped at all nine loci. For Cryptosporidium cuniculns isolates,1,2,3,1,1,4,1,1, and1haplotypes were identified at the MSC6-7、PRGR、DZ-HRGP、MSC6-5、CP47、CP56、Chom3T、 ML2、Mucin-1loci, respectively.15C.cuniculus isolates were characterized into6MLST subtypes using multilocus sequence typing. Linkage disequilibrium analysis suggested that C.cuniculus population had an epidemic population in Henan province, an epidemic population structure is indicative of recent introduction of the C.cuniculus population into the community and an unstable epidemic of the Cryptosporidiosis.