西藏部分地区马属动物肠道寄生虫的流行病学调查及毕氏肠微孢子虫的基因型鉴定
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摘要
为获悉西藏地区马属动物肠道寄生虫的感染情况,采集西藏4个地区马属动物粪便样品111份,采用卢戈氏碘液法和饱和蔗糖溶液漂浮法,检测到6种寄生虫,感染率分别为:圆线虫82.88%(92/111)、球虫10.81%(12/111)、阿米巴原虫17.12%(19/111)、马副蛔虫12.61%(14/111)、鞭虫2.70%(3/111)和结肠小袋纤毛虫4.50%(5/111),检测到的寄生虫总体感染率为88.29%(98/111)。另对所有样品基于毕氏肠微孢子虫ITS位点进行巢式PCR扩增,检测到毕氏肠微孢子虫的感染率为6.31%(7/111),共鉴定出3种基因型:1个已知基因型Ebp C(5/7)以及2个新基因型,分别命名为XZM1(1/7)和XZM2(1/7)。结果表明,西藏马属动物肠道寄生虫感染情况严重,尤其是圆线虫的感染率较高,检测到毕氏肠微孢子虫的3种基因型,系统发育分析中基因型XZM1归类于具有一定人兽共患风险的第2群,基因型Ebp C和XZM2属于具有高度人兽共患风险的第1群,证实该地区马属动物是毕氏肠微孢子虫的储存宿主,且在其传播过程中扮演着重要的角色。
In order to understand the infection of intestinal parasites in the Equus genus in Tibet,a total of 111 fecal samples of Equus genus were collected from four areas in Tibet.Lugo's iodine solution and saturated sugar flotation method were used,6 species of parasites were detected and their infection rates were Strongylid spp. 82.88%(92/111),coccidium spp.10.81%(12/111),Amoebida spp.17.12%(19/111),Parascaris equorum 12.61%(14/111),Trichuris spp.2.70%(3/111) and Balantidium coli4.50%(5/111),and the intestinal parasites infection rate was 88.29%(98/111).All the samples were amplified using the nested PCR based on the ITS gene of Enterocytozoon bieneusi,and the infection rate of E.bieneusi was 6.31%(7/111),3 genotypes were identified including one known genotype Ebp C(5/7) and two novel genotypes XZM1(1/7) and XZM2(1/7).The results showed that the infection of intestinal para-sites in Equus genus in Tibet was serious,especially the infection rate of Strongylid spp.was higher.Three genotypes of E. bieneusi were detected. In phylogenetic analysis,the novel genotype XZM1 was classified in Group 2,and the genotypes Ebp C and XZM2 belong to Group 1 with zoonotic potential,confirmed that Equus genus may play an important role in the transmission of E. bieneusi to humans.
In order to understand the infection of intestinal parasites in the Equus genus in Tibet,a total of 111 fecal samples of Equus genus were collected from four areas in Tibet.Lugo's iodine solution and saturated sugar flotation method were used,6 species of parasites were detected and their infection rates were Strongylid spp. 82.88%(92/111),coccidium spp.10.81%(12/111),Amoebida spp.17.12%(19/111),Parascaris equorum 12.61%(14/111),Trichuris spp.2.70%(3/111) and Balantidium coli4.50%(5/111),and the intestinal parasites infection rate was 88.29%(98/111).All the samples were amplified using the nested PCR based on the ITS gene of Enterocytozoon bieneusi,and the infection rate of E.bieneusi was 6.31%(7/111),3 genotypes were identified including one known genotype Ebp C(5/7) and two novel genotypes XZM1(1/7) and XZM2(1/7).The results showed that the infection of intestinal para-sites in Equus genus in Tibet was serious,especially the infection rate of Strongylid spp.was higher.Three genotypes of E. bieneusi were detected. In phylogenetic analysis,the novel genotype XZM1 was classified in Group 2,and the genotypes Ebp C and XZM2 belong to Group 1 with zoonotic potential,confirmed that Equus genus may play an important role in the transmission of E. bieneusi to humans.
引文
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[18]KUZMINA T A.Contamination of the environment by strongylid(Nematoda:Strongylidae)infective larvae at horse farms of various types in Ukraine[J].Parasitology Research,2012,110(5):1665-1674.
[19]LAATAMNA A E,WAGNEROVA P,SAK B,et al.Microsporidia and Cryptosporidium in horses and donkeys in Algeria:Detection of a novel Cryptosporidium hominis subtype family(Ik)in a horse[J].Veterinary Parasitology,2015,208(3):135-142.
[20]WAGNEROVA P,SAK B,KVETONOVA D,et al.Enterocytozoon bieneusi and Encephalitozoon cuniculi in horses kept under different management systems in the Czech Republic[J].Veterinary Parasitology,2012,190(3-4):573-577.
[21]QI M,WANG R,WANG H,et al.Enterocytozoon bieneusi genotypes in grazing horses in China and their zoonotic transmission potential[J].Journal of Eukaryotic Microbiology,2016,63(5):591-597.
[22]SANTIN M,VECINO J A C,FAYER R.A zoonotic genotype of Enterocytozoon bieneusi in horses[J].Journal of Parasitology,2010,96(1):157-161.
[2]MARTINS I V F,VEROCAI G G,CORREIA T R,et al.Survey on control and management practices of equine helminthes infection[J].Pesquisa Veterinária Brasileira,2009,29(3):253-257.
[3]KEELING P.Five questions about microsporidia[J].PLo S Pathogens,2009,5(9):e1000489.
[4]SANTIN M,FAYER R.Microsporidiosis:Enterocytozo on bieneusi in domesticated and wild animals[J].Research in Veterinary Science,2011,90(3):363-371.
[5]DIDIER E S,WEISS L M.Microsporidiosis:current status[J].Current Opinion in Infectious Diseases,2006,19(5):485.
[6]SAK B,BRADY D,PELIKANOVA M,et al.Unapparent microsporidial infection among immunocompetent humans in the Czech Republic[J].Journal of Clinical Microbiology,2011,49(3):1064-1070.
[7]MATOS O,LOBO M L,XIAO L.Epidemiology of Enterocytozoon bieneusi infection in humans[J].Journal of Parasitology Research,2012,2012(4):981424.
[8]SANTIN M,FAYER R.Enterocytozoon bieneusi genotype nomenclature based on the internal transcribed spacer sequence:a consensus[J].Journal of Eukaryotic Microbiology,2009,56(1):34-38.
[9]BUCKHOLT M A,LEE J H,TZIPORI S.Prevalence of Enterocytozoon bieneusi in swine:an 18-month survey at a slaughterhouse in Massachusetts[J].Applied&Environmental Microbiology,2002,68(5):2595-2599.
[10]WANG L,XIAO L,DUAN L,et al.Concurrent infections of Giardia duodenalis,Enterocytozoon bieneusi,and Clostridium difficile in children during a cryptosporidiosis outbreak in a pediatric hospital in China[J].PLo S Neglected Tropical Diseases,2013,7(9):e2437.
[11]ZHANG X,WANG Z,SU Y,et al.Identification and genotyping of Enterocytozoon bieneusi in China[J].Journal of Clinical Microbiology,2011,49(5):2006-2008.
[12]缺巴音才次克,达伊力特,钟华,等.巴音布鲁克镇周围散养马消化道寄生虫病感染情况调查[J].兽医导刊,2015(8):124-125.Quebayincaicike,Dayilite,ZHONG Hua,et al.Infection investigation of digestive tract parasite disease of the free-range equine around Bayanbulak Town[J].Veterinary Orientation,2015(8):124-125.(in Chinese)
[13]钱德兴,肖耀南.贵州省马寄生虫区系调查[J].中国兽医科技,1995,25(10):13-15.QIAN De-xing,XIAO Yao-nan.Investigation of the parasitic fauna of horse in Guizhou Province[J].Chinese Journal of Veterinary Science and Technology,1995,25(10):13-15.(in Chinese)
[14]周欢.山东嘉祥县牲畜交易市场驴肠道寄生虫调查及驴隐孢子虫多位点序列分型[D].郑州:河南农业大学,2015.ZHOU Huan.A Survey of Intestinal Parasites in Donkeys of Animal-Exchange Market in Jiaxiang County,Shandong and Multilocus Sequence Typing of Cryptosporidium Donkey Genotype[D].Zhengzhou:Henan Agricultural University,2015.(in Chinese)
[15]菅复春,齐萌,史亚东,等.中国部分地区马属动物肠道寄生虫流行情况调查及隐孢子虫流行种类初步鉴定[C]//中国畜牧兽医学会家畜寄生虫学分会第六次代表大会暨第十一次学术研讨会论文集,2011:256-257.JIAN Fu-chun,QI Meng,SHI Ya-dong,et al.Investigation of intestinal parasites prevalence and identification of Cryptosporidium species in some areas of China[C]//Proceedings of the Sixth Congress and 11th Symposium of Livestock Parasites Branch of Chinese Association of Animal Science and Veterinary Medicine,2011:256-257.(in Chinese)
[16]HINNEY B.Pr覿valenz von Helminthen und Risikofaktoren für ihre Befallsst覿rke bei Pferden in Brandenburg[D].Berlin:Freie Universit覿t,2009.
[17]GRAS L M,USAI F,STANCAMPIANO L.Strongylosis in horses slaughtered in Italy for meat production:Epidemiology,influence of the horse origin and evidence of parasite self-regulation[J].Veterinary Parasitology,2011,179(1):167-174.
[18]KUZMINA T A.Contamination of the environment by strongylid(Nematoda:Strongylidae)infective larvae at horse farms of various types in Ukraine[J].Parasitology Research,2012,110(5):1665-1674.
[19]LAATAMNA A E,WAGNEROVA P,SAK B,et al.Microsporidia and Cryptosporidium in horses and donkeys in Algeria:Detection of a novel Cryptosporidium hominis subtype family(Ik)in a horse[J].Veterinary Parasitology,2015,208(3):135-142.
[20]WAGNEROVA P,SAK B,KVETONOVA D,et al.Enterocytozoon bieneusi and Encephalitozoon cuniculi in horses kept under different management systems in the Czech Republic[J].Veterinary Parasitology,2012,190(3-4):573-577.
[21]QI M,WANG R,WANG H,et al.Enterocytozoon bieneusi genotypes in grazing horses in China and their zoonotic transmission potential[J].Journal of Eukaryotic Microbiology,2016,63(5):591-597.
[22]SANTIN M,VECINO J A C,FAYER R.A zoonotic genotype of Enterocytozoon bieneusi in horses[J].Journal of Parasitology,2010,96(1):157-161.