陕西绵羊源蛔虫的鉴定
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摘要
为了鉴定分离自陕西咸阳地区绵羊小肠内的蛔虫,本试验对其雌雄虫的形态学指标进行了观察和测定,并提取其基因组DNA,对其内转录间隔区(internal transcribedspacer,ITS)进行了分析。结果显示,形态学符合羊蛔虫的基本特征;雌雄虫的ITS序列长度分别为881,883bp,陕西绵羊源蛔虫与猪蛔虫(GenBank登录号:AB571302)的ITS1序列差异(0.4%)小于绵羊源蛔虫种内差异(0.9%),ITS2在种内和种间均无差异;基于ITS1和ITS2序列构建的遗传进化树显示,绵羊源蛔虫与猪蛔虫、人蛔虫聚于一支。结果表明,绵羊源蛔虫与猪蛔虫可能是同一个种。
To identify Ascaris sp.isolated from small intestines of sheep in Xianyang of Shaanxi province,the present study firstly investigated and measured the morphological features of these worms,and then extracted the genomic DNA and analyzed the internal transcribed spacer(ITS)of them.The results showed that the morphology satisfied characteristic of A.ovis,and the respective sequence lengths of the ITS for male and female worms were 881 bp and 883 bp.The sequence difference(0.4%)of ITS1 between sheep-derived Ascaris from Shaanxi and A.suum was smaller than intra-species difference(0.9%)of sheep-derived Ascaris from Shaanxi,and no interor intravariations were detected in the ITS2.Genetic trees based on respective sequences of ITS1 and ITS2 showed that sheep-derived Ascaris from Shaanxi and A.suum,A.lumbricoides clustered into one clade.These results suggested that sheep-derived Ascaris and A.suum would be the same species.
To identify Ascaris sp.isolated from small intestines of sheep in Xianyang of Shaanxi province,the present study firstly investigated and measured the morphological features of these worms,and then extracted the genomic DNA and analyzed the internal transcribed spacer(ITS)of them.The results showed that the morphology satisfied characteristic of A.ovis,and the respective sequence lengths of the ITS for male and female worms were 881 bp and 883 bp.The sequence difference(0.4%)of ITS1 between sheep-derived Ascaris from Shaanxi and A.suum was smaller than intra-species difference(0.9%)of sheep-derived Ascaris from Shaanxi,and no interor intravariations were detected in the ITS2.Genetic trees based on respective sequences of ITS1 and ITS2 showed that sheep-derived Ascaris from Shaanxi and A.suum,A.lumbricoides clustered into one clade.These results suggested that sheep-derived Ascaris and A.suum would be the same species.
引文
[1]BETSON M,NEJSUM P,BENDALL R P,et al.Molecular epidemiology of ascariasis:aglobal perspective on the transmission dynamics of Ascaris in people and pigs[J].J Infect Dis,2014,210(6):932-941.
[2]HOTEZ P,RAFF S,FENWICK A,et al.Recent progress in integrated neglected tropical disease control[J].Trends Parasitol,2007,23(11):511-514.
[3]苟烈林,姜悦平,晋德普,等.绵羊蛔虫一例[J].中国兽医科学,1985,34(5):61-62.
[4]GOODEY T.On the Ascaris from sheep[J].J Helminthol,1926,4(1):1-6.
[5]VASSILEV I,MUTAFOVA T.Comparative studies on the karyotype of Ascaris suum and“Ascaris ovis”[J].Z Parasitenkd,1974,43(2):115-121.
[6]宋军科,王正皓,王会宝,等.秦岭大熊猫槽盘吸虫的鉴定与遗传进化分析[J].中国兽医学报,2016,36(4):563-566.
[7]PRASAD P K,TANDON V,BISWAL D K,et al.Molecular identification of the Indian liver fluke,Fasciola(Trematoda:Fasciolidae)based on the ribosomal internal transcribed spacer regions[J].Parasitol Res,2008,103(6):1247-1255.
[8]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.
[9]SUBBOTIN S A,MADANI M,KRALL E,et al.Molecular diagnostics,taxonomy,and phylogeny of the stem nematode Ditylenchus dipsaci species complex based on the sequences of the internal transcribed spacer-rDNA[J].Phytopathology,2005,95(11):1308-1315.
[10]HASHEMINASAB S S.Molecular characterization of the first internal transcribed spacer of rDNA of Parabronema skrjabini for the first time in sheep[J].Ann Parasitol,2015,61(4):241-246.
[11]WANG K,XIE H,LI Y,et al.Morphology and molecular analysis of Paratylenchus nanjingensis n.sp.(Nematoda:Paratylenchinae)from the rhizosphere soil of Pinus massoniana in China[J].J Helminthol,2016,90(2):166-173.
[12]ZHU X,GASSER R B,PODOLSKA M,et al.Characterisation of anisakidnematodes with zoonotic potential by nuclear ribosomal DNA sequences[J].Int JParasitol,1998,28(12):1911-1921.
[13]NEJSUM P,BETSON M,BENDALL R P,et al.Assessing the zoonotic potential of Ascaris suum and Trichuris suis:looking to the future from an analysis of the past[J].J Helminthol,2012,86(2):148-155.
[14]ANDERSON T J C.Ascaris infections in humans from North America:molecular evidence for cross-infection[J].Parasitology,1995,110(Pt2):215-219.
[15]BENDALL R P,BARLOW M,BETSON M,et al.Zoonotic ascariasis,United Kingdom[J].Emerg Infect Dis,2011,17(10):1964-1966.
[16]NEJSUM P,PARKER E D,FRYDENBERG J,et al.Ascariasis is a zoonosis in Denmark[J].J Clin Microb,2005,43(3):1142-1148.
[17]CAVALLERO S,SNABEL V,PACELLA F,et al.Phylogeographical studies of Ascaris spp.based on ribosomal and mitochondrial DNA sequences[J].PLoSNegl Trop Dis,2013,7(4):e2170.
[18]LIU G H,WU C Y,SONG H Q,et al.Comparative analyses of the complete mitochondrial genomes of Ascaris lumbricoides and Ascaris suumfrom humans and pigs[J].Gene,2012,492(1):110-116.
[19]BETSON M,HALSTEAD F D,NEJSUM P,et al.Amolecular epidemiological investigation of Ascaris on Unguja,Zanzibar using isoenyzme analysis,DNA barcoding and microsatellite DNA profiling[J].Trans RSoc Trop Med Hyg,2011,105(7):370-379.
[20]BETSON M,STOTHARD J R.Ascaris lumbricoides or Ascaris suum:Whats in a Name[J].J Infect Dis,2016,213(8):1355-1356.
[21]MOTE D C.Ascaris suumin sheep molecular diagnostics,taxonomy,and phylogeny of the stem nematode Ditylenchus dipsaci species complex based on the sequences of the internal transcribed spacer-rDNA[J].Science,1914,40(1023):216.
[22]GUNN A.A case of Ascaris suuminfection in lambs[J].Vet Rec,1980,107(25/26):581.
[23]MCDONALD F E,CHEVIS R A.Ascaris lumbricoides in lambs[J].N Z Vet J,1965,13(2):41.
[24]FITZGERALD P R.The pathogenesis of Ascaris lumbricoides var.suum in lambs[J].Am J Vet Res,1962,23:731-736.
[25]SOE M J,KAPEL C M,NEJSUM P.Ascaris from humans and pigs appear to be reproductively isolated species[J].PLoS Negl Trop Dis,2016,10(9):e0004855.
[2]HOTEZ P,RAFF S,FENWICK A,et al.Recent progress in integrated neglected tropical disease control[J].Trends Parasitol,2007,23(11):511-514.
[3]苟烈林,姜悦平,晋德普,等.绵羊蛔虫一例[J].中国兽医科学,1985,34(5):61-62.
[4]GOODEY T.On the Ascaris from sheep[J].J Helminthol,1926,4(1):1-6.
[5]VASSILEV I,MUTAFOVA T.Comparative studies on the karyotype of Ascaris suum and“Ascaris ovis”[J].Z Parasitenkd,1974,43(2):115-121.
[6]宋军科,王正皓,王会宝,等.秦岭大熊猫槽盘吸虫的鉴定与遗传进化分析[J].中国兽医学报,2016,36(4):563-566.
[7]PRASAD P K,TANDON V,BISWAL D K,et al.Molecular identification of the Indian liver fluke,Fasciola(Trematoda:Fasciolidae)based on the ribosomal internal transcribed spacer regions[J].Parasitol Res,2008,103(6):1247-1255.
[8]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.
[9]SUBBOTIN S A,MADANI M,KRALL E,et al.Molecular diagnostics,taxonomy,and phylogeny of the stem nematode Ditylenchus dipsaci species complex based on the sequences of the internal transcribed spacer-rDNA[J].Phytopathology,2005,95(11):1308-1315.
[10]HASHEMINASAB S S.Molecular characterization of the first internal transcribed spacer of rDNA of Parabronema skrjabini for the first time in sheep[J].Ann Parasitol,2015,61(4):241-246.
[11]WANG K,XIE H,LI Y,et al.Morphology and molecular analysis of Paratylenchus nanjingensis n.sp.(Nematoda:Paratylenchinae)from the rhizosphere soil of Pinus massoniana in China[J].J Helminthol,2016,90(2):166-173.
[12]ZHU X,GASSER R B,PODOLSKA M,et al.Characterisation of anisakidnematodes with zoonotic potential by nuclear ribosomal DNA sequences[J].Int JParasitol,1998,28(12):1911-1921.
[13]NEJSUM P,BETSON M,BENDALL R P,et al.Assessing the zoonotic potential of Ascaris suum and Trichuris suis:looking to the future from an analysis of the past[J].J Helminthol,2012,86(2):148-155.
[14]ANDERSON T J C.Ascaris infections in humans from North America:molecular evidence for cross-infection[J].Parasitology,1995,110(Pt2):215-219.
[15]BENDALL R P,BARLOW M,BETSON M,et al.Zoonotic ascariasis,United Kingdom[J].Emerg Infect Dis,2011,17(10):1964-1966.
[16]NEJSUM P,PARKER E D,FRYDENBERG J,et al.Ascariasis is a zoonosis in Denmark[J].J Clin Microb,2005,43(3):1142-1148.
[17]CAVALLERO S,SNABEL V,PACELLA F,et al.Phylogeographical studies of Ascaris spp.based on ribosomal and mitochondrial DNA sequences[J].PLoSNegl Trop Dis,2013,7(4):e2170.
[18]LIU G H,WU C Y,SONG H Q,et al.Comparative analyses of the complete mitochondrial genomes of Ascaris lumbricoides and Ascaris suumfrom humans and pigs[J].Gene,2012,492(1):110-116.
[19]BETSON M,HALSTEAD F D,NEJSUM P,et al.Amolecular epidemiological investigation of Ascaris on Unguja,Zanzibar using isoenyzme analysis,DNA barcoding and microsatellite DNA profiling[J].Trans RSoc Trop Med Hyg,2011,105(7):370-379.
[20]BETSON M,STOTHARD J R.Ascaris lumbricoides or Ascaris suum:Whats in a Name[J].J Infect Dis,2016,213(8):1355-1356.
[21]MOTE D C.Ascaris suumin sheep molecular diagnostics,taxonomy,and phylogeny of the stem nematode Ditylenchus dipsaci species complex based on the sequences of the internal transcribed spacer-rDNA[J].Science,1914,40(1023):216.
[22]GUNN A.A case of Ascaris suuminfection in lambs[J].Vet Rec,1980,107(25/26):581.
[23]MCDONALD F E,CHEVIS R A.Ascaris lumbricoides in lambs[J].N Z Vet J,1965,13(2):41.
[24]FITZGERALD P R.The pathogenesis of Ascaris lumbricoides var.suum in lambs[J].Am J Vet Res,1962,23:731-736.
[25]SOE M J,KAPEL C M,NEJSUM P.Ascaris from humans and pigs appear to be reproductively isolated species[J].PLoS Negl Trop Dis,2016,10(9):e0004855.