国内羊胃肠道寄生虫流行病学调查及捻转血矛线虫系统进化分析
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摘要
为更好地掌握我国部分地区羊胃肠道寄生虫病的流行情况以及捻转血矛线虫ZJ株近年来的系统进化关系,从而为捻转血矛线虫病的防治提供科学依据,于2014年9—12月采集了来自山东、吉林、黑龙江、内蒙古、辽宁、河北、江苏、天津和青海9个省(市)的490份羊粪样品,对其进行了寄生虫虫卵分类鉴定,对羊寄生虫病的流行病学特征进行总结分析;此外实验室于2004—2013年采集了来自浙江省嘉兴地区的15份捻转血矛线虫ZJ株成虫样品,设计特异性引物对样品ITS-2和ND4基因进行分子克隆测序鉴定比对,对该虫近10年来的系统进化进行分析。结果表明,全国各地羊体内肠道寄生虫的感染情况均较为严重,其中感染虫种的严重程度依次为原虫(83.27%)、线虫(64.29%)和绦虫(49.80%),感染相对最为严重的3个地区分别为天津、青海和内蒙古,而相对较轻的3个地区为江苏、黑龙江、山东。对捻转血矛线虫ZJ株的15份样品进行系统进化分析显示,ITS-2和ND4基因在10年间均有不同程度的遗传变异发生,变异范围分别为0.0%~3.9%和0.4%~4.8%,而ND4突变程度较大可能是由于虫种固有差异引起的。而2种基因都有部分位点突变后与H.placei(Haemonchus pacei)相同,说明嘉兴地区可能存在捻转血矛线虫和H.placei交叉感染的情况。调查分析结果将为制定更合理的羊寄生虫病防控方案提供参考依据,并为寄生虫系统进化分析研究奠定基础。
To have a better understanding of the sheep gastrointestinal parasites epidemiology in partial areas of China,490 fecal samples were collected from Shandong,Jilin,Heilongjiang,Neimenggu,Liaoning,Hebei,Jiangsu,Tianjin and Qinghai during September to December in 2014 and identified for egg morphology.At the meantime,15 samples of Haemonchus contortus(H.contortus)were collected from Jiangxing area of Zhejiang province since 2004 to perform the phyletic evolution analysis using ITS-2 and ND4 as gene markers.As the results showed,severe infection of gastrointestinal parasites could be observed in all these nine areas with the highest infection rate of protozoans(83.27%),followed by nematodes(64.29%)and tape worms(49.8%).The most severe areas were Tianjin,Qinghai and Neimenggu.The phyletic evolution analysis indicated that both ITS-2 and ND4 genes showed some level of variation among 0.0%-3.9%and 0.4%-4.8%in the last decade.The relatively high variation of ND4 gene may be caused by the inherent difference of ZJ strain compared to the reference strain.Some sites of these two genes of the 15 samples shared identical variations with Haemonchus pacei,this may indicate that there was cross-infection with H.contortus and H.placei in Jiaxing area.This research laid a solid foundation for further study of the control and prevention of sheep gastrointestinal parasites.
To have a better understanding of the sheep gastrointestinal parasites epidemiology in partial areas of China,490 fecal samples were collected from Shandong,Jilin,Heilongjiang,Neimenggu,Liaoning,Hebei,Jiangsu,Tianjin and Qinghai during September to December in 2014 and identified for egg morphology.At the meantime,15 samples of Haemonchus contortus(H.contortus)were collected from Jiangxing area of Zhejiang province since 2004 to perform the phyletic evolution analysis using ITS-2 and ND4 as gene markers.As the results showed,severe infection of gastrointestinal parasites could be observed in all these nine areas with the highest infection rate of protozoans(83.27%),followed by nematodes(64.29%)and tape worms(49.8%).The most severe areas were Tianjin,Qinghai and Neimenggu.The phyletic evolution analysis indicated that both ITS-2 and ND4 genes showed some level of variation among 0.0%-3.9%and 0.4%-4.8%in the last decade.The relatively high variation of ND4 gene may be caused by the inherent difference of ZJ strain compared to the reference strain.Some sites of these two genes of the 15 samples shared identical variations with Haemonchus pacei,this may indicate that there was cross-infection with H.contortus and H.placei in Jiaxing area.This research laid a solid foundation for further study of the control and prevention of sheep gastrointestinal parasites.
引文
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[11]沈杰,叶明忠,陈永军,等.绵羊寄生线虫在体内及牧场上各发育阶段的季节动态研究[J].中国动物传染病学报,1994(1):10-15.
[12]YANG Y,ZHOU Q J,CHEN X Q,et al.Profiling of differentially expressed genes in sheep T lymphocytes response to an artificial primary Haemonchus contortus infection[J].Parasites Vectors,2015,8(1):235.
[13]CABARET J.Relating parasite communities to host environmental conditions using phylogenetic tools[J].Parasite,2003,10(4):287-295.
[14]GASSER R B,BOTT N J,CHILTON N B,et al.Toward practical,DNA-based diagnostic methods for parasitic nematodes of livestock-bionomic and biotechnological implications[J].Biotechnol Adv,2008,26(4):325-334.
[15]O’CONNOR L J,WALKDEN-BROWN S W,KAHN L P.Ecology of the free-living stages of major trichostrongylid parasites of sheep[J].Vet Parasitol,2006,142(1/2):1-15.
[2]姜大昌,张良.农区养羊的现状及应采取的对策[J].草业与畜牧,2012(3):57-58.
[3]龚大春,李助南,何会时.羊捻转血矛线虫病的诊治报告[J].湖北畜牧兽医,2004(1):51.
[4]黄美娜,林振,覃勇.山羊捻转血矛线虫病的诊治[J].中国兽医杂志,2010,46(6):86.
[5]李运兰,陈书民,赵国平.小尾寒羊捻转血矛线虫病的诊治[J].中国兽医杂志,2000,26(7):39.
[6]王晓平,李文艺,周毅,等.花垣与保靖山羊捻转血矛线虫感染情况调查研究[J].现代农业科技,2011(2):341.
[7]魏芝贵,杜乐新.山羊捻转血矛线虫的诊治[J].中国畜牧兽医文摘,2011,27(3):139.
[8]MOHANRAJ K,SUBHADRA S,KALYANASUNDARAM A,et al.Genotyping of benzimidazole resistant and susceptible isolates of Haemonchus contortus,from sheep by allele specific PCR[J].J Parasitic Dis,2017,41(1):282-288.
[9]ZARLENGA D S,BARRY C M,GASBARRE L C,et al.A multiplex PCR assay for differentiating economically important gastrointestinal nematodes of cattle[J].Vet Parasitol,2001,97(3):201-211.
[10]CERUTTI M C,CITTERIO C V,BAZZOCCHI C,et al.Genetic variability of Haemonchus contortus(Nematoda:Trichostrongyloidea)in alpine ruminant host species.[J].J Helminthol,2010,84(3):276-283.
[11]沈杰,叶明忠,陈永军,等.绵羊寄生线虫在体内及牧场上各发育阶段的季节动态研究[J].中国动物传染病学报,1994(1):10-15.
[12]YANG Y,ZHOU Q J,CHEN X Q,et al.Profiling of differentially expressed genes in sheep T lymphocytes response to an artificial primary Haemonchus contortus infection[J].Parasites Vectors,2015,8(1):235.
[13]CABARET J.Relating parasite communities to host environmental conditions using phylogenetic tools[J].Parasite,2003,10(4):287-295.
[14]GASSER R B,BOTT N J,CHILTON N B,et al.Toward practical,DNA-based diagnostic methods for parasitic nematodes of livestock-bionomic and biotechnological implications[J].Biotechnol Adv,2008,26(4):325-334.
[15]O’CONNOR L J,WALKDEN-BROWN S W,KAHN L P.Ecology of the free-living stages of major trichostrongylid parasites of sheep[J].Vet Parasitol,2006,142(1/2):1-15.