长春地区猪十二指肠贾第鞭毛虫感染的分子流行病学调查
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摘要
目的调查长春地区猪十二指肠贾第鞭毛虫(简称贾第虫)感染的分子流行病学特征。方法选择长春地区两个猪场,随机采集60份猪粪便样品,根据贾第虫磷酸丙糖异构酶(triose phosphate isomerase,TPI)基因设计巢式PCR引物,以提取的猪粪便基因组DNA为模板进行巢式PCR扩增,PCR产物经测序及序列比对,并建立贾第虫系统进化树。结果 60份样品中,27份为阳性,阳性率为45%(27/60),基因型均为人兽共患型集聚体A。结论长春地区两个猪场存在较严重的贾第虫感染,基因型均为集聚体A,有较大的散播风险,需加强猪场卫生监督管理,以避免疫病流行。
Objective To investigate the molecular epidemiological characters of Giardia duodenalis infection in pig farms in Changchun region,Jilin Province,China. Methods Primers were designed according to the triose phosphate isomerase(TPI)gene of G. duodenalis. Sixty pig feces samples were collected from two pig farms in Changchun region,from which the genomic DNA was extracted and used as a template for amplification by nested PCR. The PCR product was subjected to sequencing and sequence alignment,based on which a phylogenetic tree of G. duodenalis was constructed. Results Of the 60 samples,27 were positive,indicating a positive rate of 45%(27/60),and the genotype was zoonotic assemblage A. Conclusion Severe G. duodenalis infection was found in the two pig farms in Changchun region,of which the genotype was zoonotic assemblage A with high risk of spread. Therefore,health supervision and management of pig farms should be strengthened to avoid epidemic of the disease.
Objective To investigate the molecular epidemiological characters of Giardia duodenalis infection in pig farms in Changchun region,Jilin Province,China. Methods Primers were designed according to the triose phosphate isomerase(TPI)gene of G. duodenalis. Sixty pig feces samples were collected from two pig farms in Changchun region,from which the genomic DNA was extracted and used as a template for amplification by nested PCR. The PCR product was subjected to sequencing and sequence alignment,based on which a phylogenetic tree of G. duodenalis was constructed. Results Of the 60 samples,27 were positive,indicating a positive rate of 45%(27/60),and the genotype was zoonotic assemblage A. Conclusion Severe G. duodenalis infection was found in the two pig farms in Changchun region,of which the genotype was zoonotic assemblage A with high risk of spread. Therefore,health supervision and management of pig farms should be strengthened to avoid epidemic of the disease.
引文
[1]ZHANG X C,LI J H.Veterinary parasitology[M].Beijing:Science Press,2010:314-315.(in Chinese)张西臣,李建华.动物寄生虫病[M].北京:科学出版社,2010:314-315.
[2]EINARSSON E,MAAYEH S,SVARD S G.An up-date on Giardia and giardiasis[J].Curr Opin Microbiol,2016,34:47-52.
[3]LI Z S.The establishment and application of detection methods for Giardia duodenalis based on RNase P and TPI gene[D].Changchun:Jilin University,2013.(in Chinese)李棕松.犬贾第虫Nested PCR检测方法的建立与应用[D].吉林长春:吉林大学,2013.
[4]LECOVA L,WEISZ F,TUMOVA P,et al.The first multilocus genotype analysis of Giardia intestinalis in humans in the Czech Republic[J].Parasitology,2018,145(12):1577-1587.
[5]HEYWORTH M F.Giardia duodenalis\r,genetic assemblages and hosts[J].Parasite,2016,23(13):1-5.
[6]WANG Z Q,CUI J.Advances in epidemiological studies of Giardia lamblia[J].Foreign Med Sci Parasit Dis,2005,32(3):99-105.(in Chinese)王中全,崔晶.贾第虫病流行病学研究进展[J].国际医学寄生虫病杂志,2005,32:99-105.
[7]VILLALBA-VIZCAíNO V,BUELVAS Y,ARROYO-SALGADOB,et al.Molecular identification of,Giardia intestinalis,in two cities of the Colombian Caribbean Coast[J].Exp Parasitol,2018,189:1-7.
[8]JOHNSTON S P,BALLARD M M,BEACH M J,et al.Evaluation of three commercial assays for detection of Giardia and Cryptosporidium organisms in fecal specimens[J].J Clin Microbiol,2003,41(2):623-626.
[9]SILVA R K N R,PACHECO F T F,MARTINS AS,et al.Perfor mance of microscopy and ELISA for diagnosing Giardia duodenalis infection in different pediatric groups[J].Parasitol Int,2016,65(6):635-640.
[10]HUSSEIN E M,ISMAIL O A,MOKHTAR A B,et al.Nested PCR targeting intergenic spacer(IGS)in genotyping of Giardia duodenalis isolated from symptomatic and asymptomatic infected Egyptian school children[J].Parasitol Res,2017,116(2):763-771.
[11]MONIS P T,ANDREWS R H,MAYRHOFER G,et al.Genetic diversity within the morphological species Giardia intestinalis and its relationship to host origin[J].Infect Genet Evolut,2003,3(1):29-38.
[12]BARUCH A C,ISAACRENTON J,ADAM R D.The molecular epidemiology of Giardia lamblia:a sequence-based approach[J].J Infect Dis,1996,174(1):233-236.
[13]WANG S S,YUAN Y J,YIN Y L,et al.Prevalence and multilocus genotyping of Giardia duodenalis in pigs of Shaanxi Province,northwestern China[J].Parasites&Vectors,2017,10(1):490.
[14]RODRIGUEZ-RIVERA L D,CUMMINGS K J,MCNEELY I,et al.Prevalence and diversity of cryptosporidium and Giardia identified among feral pigs in Texas[J].Vector Borne&Zoonotic Dis,2016,16(12):765-768.
[15]WANG H Y.Molecular epidemiology of three intestinal protozoas on livestock in Henan Province[D].Beijing:China Agricultural University,2015.(in Chinese)王海燕.河南省家畜的三种肠道原虫分子流行病学调查[D].北京:中国农业大学,2015.
[16]LI W,YU X M,ZHONG Z J,et al.Isolation and identification of Giardia and Cryptosporidium genotypes from captive wild boars and Xiang pigs in Sichuan Province[J].Chin Veter Sci,2016(46):1166-1169.(in Chinese)李威,余星明,钟志军,等.四川省圈养野猪源和小香猪源贾第虫及隐孢子虫的分离与鉴定[J].中国兽医科学,2016(46):1166-1169.
[17]SHI L Q,FENG Y,WU J,et al.Molecular detection and analysis of Giardia in some swine farms of Yunnan Province[J].Chin Animal Health Inspect,2018(35):19-21.(in Chinese)石莲琴,冯云,吴杰,等.云南省部分猪场贾第虫分子检测与分析[J].中国动物检疫,2018(35):19-21.
[2]EINARSSON E,MAAYEH S,SVARD S G.An up-date on Giardia and giardiasis[J].Curr Opin Microbiol,2016,34:47-52.
[3]LI Z S.The establishment and application of detection methods for Giardia duodenalis based on RNase P and TPI gene[D].Changchun:Jilin University,2013.(in Chinese)李棕松.犬贾第虫Nested PCR检测方法的建立与应用[D].吉林长春:吉林大学,2013.
[4]LECOVA L,WEISZ F,TUMOVA P,et al.The first multilocus genotype analysis of Giardia intestinalis in humans in the Czech Republic[J].Parasitology,2018,145(12):1577-1587.
[5]HEYWORTH M F.Giardia duodenalis\r,genetic assemblages and hosts[J].Parasite,2016,23(13):1-5.
[6]WANG Z Q,CUI J.Advances in epidemiological studies of Giardia lamblia[J].Foreign Med Sci Parasit Dis,2005,32(3):99-105.(in Chinese)王中全,崔晶.贾第虫病流行病学研究进展[J].国际医学寄生虫病杂志,2005,32:99-105.
[7]VILLALBA-VIZCAíNO V,BUELVAS Y,ARROYO-SALGADOB,et al.Molecular identification of,Giardia intestinalis,in two cities of the Colombian Caribbean Coast[J].Exp Parasitol,2018,189:1-7.
[8]JOHNSTON S P,BALLARD M M,BEACH M J,et al.Evaluation of three commercial assays for detection of Giardia and Cryptosporidium organisms in fecal specimens[J].J Clin Microbiol,2003,41(2):623-626.
[9]SILVA R K N R,PACHECO F T F,MARTINS AS,et al.Perfor mance of microscopy and ELISA for diagnosing Giardia duodenalis infection in different pediatric groups[J].Parasitol Int,2016,65(6):635-640.
[10]HUSSEIN E M,ISMAIL O A,MOKHTAR A B,et al.Nested PCR targeting intergenic spacer(IGS)in genotyping of Giardia duodenalis isolated from symptomatic and asymptomatic infected Egyptian school children[J].Parasitol Res,2017,116(2):763-771.
[11]MONIS P T,ANDREWS R H,MAYRHOFER G,et al.Genetic diversity within the morphological species Giardia intestinalis and its relationship to host origin[J].Infect Genet Evolut,2003,3(1):29-38.
[12]BARUCH A C,ISAACRENTON J,ADAM R D.The molecular epidemiology of Giardia lamblia:a sequence-based approach[J].J Infect Dis,1996,174(1):233-236.
[13]WANG S S,YUAN Y J,YIN Y L,et al.Prevalence and multilocus genotyping of Giardia duodenalis in pigs of Shaanxi Province,northwestern China[J].Parasites&Vectors,2017,10(1):490.
[14]RODRIGUEZ-RIVERA L D,CUMMINGS K J,MCNEELY I,et al.Prevalence and diversity of cryptosporidium and Giardia identified among feral pigs in Texas[J].Vector Borne&Zoonotic Dis,2016,16(12):765-768.
[15]WANG H Y.Molecular epidemiology of three intestinal protozoas on livestock in Henan Province[D].Beijing:China Agricultural University,2015.(in Chinese)王海燕.河南省家畜的三种肠道原虫分子流行病学调查[D].北京:中国农业大学,2015.
[16]LI W,YU X M,ZHONG Z J,et al.Isolation and identification of Giardia and Cryptosporidium genotypes from captive wild boars and Xiang pigs in Sichuan Province[J].Chin Veter Sci,2016(46):1166-1169.(in Chinese)李威,余星明,钟志军,等.四川省圈养野猪源和小香猪源贾第虫及隐孢子虫的分离与鉴定[J].中国兽医科学,2016(46):1166-1169.
[17]SHI L Q,FENG Y,WU J,et al.Molecular detection and analysis of Giardia in some swine farms of Yunnan Province[J].Chin Animal Health Inspect,2018(35):19-21.(in Chinese)石莲琴,冯云,吴杰,等.云南省部分猪场贾第虫分子检测与分析[J].中国动物检疫,2018(35):19-21.