秦岭地区几种珍稀动物毕氏肠微孢子虫基因分型
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摘要
为了评估秦岭地区珍稀野生动物的毕氏肠微孢子虫(Enterocytozoon bieneusi)的人兽共患风险,本研究采用PCR技术和多位点序列分型技术对秦岭地区5种珍稀动物毕氏肠微孢子虫的感染情况及基因型/亚型进行了分析。在采集的22份新鲜粪便中检测到7个阳性分离株,其中斑羚源5个,长颈鹿源1个,麂子源1个。基于ITS位点基因分型发现所有分离株均为人兽共患基因型,包括6个已报道的基因型D和1个尚未报道的D-new。多位点序列分型技术发现秦岭地区珍稀野生动物的毕氏肠微孢子虫存在遗传多样性,7个分离株在MS1、MS3、MS4和MS7位点分别有3、1、2、2个亚型。
In order to determine the genotypes and subtypes of Enterocytozoon bieneusi in wildlife in Qinling Mountains,22 fecal specimens were collected and detected by molecular biological technique based on PCR and multilocus sequence typing.Seven specimens were positive in E.bieneusi in goral(n=5),giraffe(n=1)and barking deer(n=1).Genotyping at ITS locus and phylogenetic analysis indicated one known E.bieneusi genotype D and one novel E.bieneusi genotype D-new,both are zoonotic parasites.Multilocus sequence typing showed 3,1,2,2haplotypes at MS1,MS3,MS4,MS7 loci,respectively.The results in the present study have significant implication in prevention and control of zoonotic E.bieneusi infection.
In order to determine the genotypes and subtypes of Enterocytozoon bieneusi in wildlife in Qinling Mountains,22 fecal specimens were collected and detected by molecular biological technique based on PCR and multilocus sequence typing.Seven specimens were positive in E.bieneusi in goral(n=5),giraffe(n=1)and barking deer(n=1).Genotyping at ITS locus and phylogenetic analysis indicated one known E.bieneusi genotype D and one novel E.bieneusi genotype D-new,both are zoonotic parasites.Multilocus sequence typing showed 3,1,2,2haplotypes at MS1,MS3,MS4,MS7 loci,respectively.The results in the present study have significant implication in prevention and control of zoonotic E.bieneusi infection.
引文
[1]刁瑞南,肖立华,宋铭忻,等.毕氏肠微孢子虫基因分型及公共卫生学重要性[J].中国预防兽医学报,2014,36(12):988-991.
[2]Santín M,Fayer R.Microsporidiosis:Enterocytozoon bieneusi in domesticated and wild animals[J].Res Vet Sci,2011,90(3):363-371.
[3]Mathis A,Weber R,Deplazes P.Zoonotic potential of the microsporidia[J].Clin Microbiol Rev,2005,18(3):423-445.
[4]李梦婕.羊微孢子虫病流行病学调查及分子特征初步研究[D].郑州:河南农业大学,2012.
[5]Anane S,Attouchi H.Microsporidiosis:epidemiology,clinical data and therapy[J].Gastroenterol Clin Biol,2010,34(8-9):450-464.
[6]Ma J,Li P,Zhao X,et al.Occurrence and molecular characterization of Cryptosporidium spp.and Enterocytozoon bieneusi in dairy cattle,beef cattle and water buffaloes in China[J].Vet Parasitol,2015,207(3-4):220-227.
[7]Li N,Xiao L,Wang L,et al.Molecular surveillance of Cryptosporidium spp.,Giardia duodenalis,and Enterocytozoon bieneusi by genotyping and subtyping parasites in wastewater[J].PLoS Negl Trop Dis,2012,6(9):e1809.
[8]Sulaiman I M,Fayer R,Lal A A,et al.Molecular characterization of microsporidia indicates that wild mammals harbor host-adapted Enterocytozoon spp.as well as human-pathogenic Enterocytozoon bieneusi[J].Appl Environ Microbiol,2003,69(8):4495-4501.
[9]Feng Y,Li N,Dearen T,et al.Development of a multilocus sequence typing tool for high-resolution genotyping of Enterocytozoon bieneusi[J].Appl Environ Microbiol,2011,77(14):4822-4828.
[10]Thellier M,Breton J.Enterocytozoon bieneusi in human and animals,focus on laboratory identification and molecular epidemiology[J].Parasite,2008,15(3):349-358.
[11]李巍.肯尼亚狒狒肠道原虫系统进化分析及毕氏肠微孢子虫群体遗传学研究[D].长春:吉林大学,2011.
[12]徐利纳.河南省郑州地区微孢子虫病流行病学调查及分子特征初步研究[D].郑州:河南农业大学,2011.
[13]王永宾,刘吉平.微孢子虫检测技术的研究进展[J].中国寄生虫学与寄生虫病杂志,2009,27(2):161-166.
[14]Wang L,Zhang H,Zhao X,et al.Zoonotic Cryptosporidiumspecies and Enterocytozoon bieneusi genotypes in HIV-positive patients on antiretroviral therapy[J].J Clin Microbiol.2013,51(2):557-563.
[15]Fayer,R.Santin,M.Macarisin,D.Detection of concurrent infection of dairy cattle with Blastocystis,Cryptosporidium,Giardia,and Enterocytozoon by molecular and microscopic methods[J].Parasitol Res,2011,111(3):1349-1355.
[16]Karim MR,Yu F,Li J,et al.First molecular characterization of enteric protozoa and the human pathogenic microsporidian,Enterocytozoon bieneusi,in captive snakes in China[J].Parasitol Res.2014,113(8):3041-3048.
[17]Karim M R,Wang R,He X,et al.Multilocus sequence typing of Enterocytozoon bieneusi in nonhuman primates in China[J].Vet Parasitol,2014,200(1-2):13-23.
[18]Li W,Cama V,Akinbo F O,et al.Multilocus sequence typing of Enterocytozoon bieneusi:Lack of geographic segregation and existence of genetically isolated subpopulations[J].Infect Genet Evol,2013,14:111-119.
[19]Santín M,Fayer R.Enterocytozoon bieneusi genotype nomenclature based on the internal transcribed spacer sequence:a consensus[J].J Eukaryot Microbiol,2009,56(1):34-38.
[2]Santín M,Fayer R.Microsporidiosis:Enterocytozoon bieneusi in domesticated and wild animals[J].Res Vet Sci,2011,90(3):363-371.
[3]Mathis A,Weber R,Deplazes P.Zoonotic potential of the microsporidia[J].Clin Microbiol Rev,2005,18(3):423-445.
[4]李梦婕.羊微孢子虫病流行病学调查及分子特征初步研究[D].郑州:河南农业大学,2012.
[5]Anane S,Attouchi H.Microsporidiosis:epidemiology,clinical data and therapy[J].Gastroenterol Clin Biol,2010,34(8-9):450-464.
[6]Ma J,Li P,Zhao X,et al.Occurrence and molecular characterization of Cryptosporidium spp.and Enterocytozoon bieneusi in dairy cattle,beef cattle and water buffaloes in China[J].Vet Parasitol,2015,207(3-4):220-227.
[7]Li N,Xiao L,Wang L,et al.Molecular surveillance of Cryptosporidium spp.,Giardia duodenalis,and Enterocytozoon bieneusi by genotyping and subtyping parasites in wastewater[J].PLoS Negl Trop Dis,2012,6(9):e1809.
[8]Sulaiman I M,Fayer R,Lal A A,et al.Molecular characterization of microsporidia indicates that wild mammals harbor host-adapted Enterocytozoon spp.as well as human-pathogenic Enterocytozoon bieneusi[J].Appl Environ Microbiol,2003,69(8):4495-4501.
[9]Feng Y,Li N,Dearen T,et al.Development of a multilocus sequence typing tool for high-resolution genotyping of Enterocytozoon bieneusi[J].Appl Environ Microbiol,2011,77(14):4822-4828.
[10]Thellier M,Breton J.Enterocytozoon bieneusi in human and animals,focus on laboratory identification and molecular epidemiology[J].Parasite,2008,15(3):349-358.
[11]李巍.肯尼亚狒狒肠道原虫系统进化分析及毕氏肠微孢子虫群体遗传学研究[D].长春:吉林大学,2011.
[12]徐利纳.河南省郑州地区微孢子虫病流行病学调查及分子特征初步研究[D].郑州:河南农业大学,2011.
[13]王永宾,刘吉平.微孢子虫检测技术的研究进展[J].中国寄生虫学与寄生虫病杂志,2009,27(2):161-166.
[14]Wang L,Zhang H,Zhao X,et al.Zoonotic Cryptosporidiumspecies and Enterocytozoon bieneusi genotypes in HIV-positive patients on antiretroviral therapy[J].J Clin Microbiol.2013,51(2):557-563.
[15]Fayer,R.Santin,M.Macarisin,D.Detection of concurrent infection of dairy cattle with Blastocystis,Cryptosporidium,Giardia,and Enterocytozoon by molecular and microscopic methods[J].Parasitol Res,2011,111(3):1349-1355.
[16]Karim MR,Yu F,Li J,et al.First molecular characterization of enteric protozoa and the human pathogenic microsporidian,Enterocytozoon bieneusi,in captive snakes in China[J].Parasitol Res.2014,113(8):3041-3048.
[17]Karim M R,Wang R,He X,et al.Multilocus sequence typing of Enterocytozoon bieneusi in nonhuman primates in China[J].Vet Parasitol,2014,200(1-2):13-23.
[18]Li W,Cama V,Akinbo F O,et al.Multilocus sequence typing of Enterocytozoon bieneusi:Lack of geographic segregation and existence of genetically isolated subpopulations[J].Infect Genet Evol,2013,14:111-119.
[19]Santín M,Fayer R.Enterocytozoon bieneusi genotype nomenclature based on the internal transcribed spacer sequence:a consensus[J].J Eukaryot Microbiol,2009,56(1):34-38.