规模羊场环境气载产气荚膜梭菌耐药性及其ERIC-PCR分型
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摘要
为监测不同规模羊场空气中分离的气载产气荚膜梭菌耐药性变迁及不同菌株间的遗传相似性,从而为临床治疗和控制感染提供依据,收集了2016—2017年来自不同规模羊场分离的气载产气荚膜梭菌30株,采用K-B法进行药敏试验,应用基因间重复序列聚合酶链反应(ERIC-PCR),对菌株进行DNA分型及同源性分析。结果显示:30株气载产气荚膜梭菌在监测的9种药物中,对庆大霉素的耐药性最强(86.7%),其次为青霉素(43.3%),对头孢噻肟耐药率最低(1%),对其他药物耐药率在3.3%~36.7%之间;ERIC-PCR谱型表现为14个基因型(Ⅰ~XIV),其中大部分来源于不同的克隆株,且有2个克隆株的相似度为99.0%。本研究结果表明:规模羊场的气载产气荚膜梭菌多重耐药较严重,分子多样性复杂;气载产气荚膜梭菌来源广泛,不同地区、环境和条件下,同一种病原菌的基因型也存在一定差异,可能存在多个基因型;同一个规模羊场的基因型也不同,不同羊场之间也存在相同的基因型。
In order to monitor the drug resistance changes of airborne Clostridium perfringens(C. perfringens)in different goat farms and the genetic similarity of different strains,so as to provide basis for clinical treatment and infection control,30 airborne strains were collected from different scaled goat farms in 2016—2017,the sensitivity of antibiotics was detected by K-B method,and the strains classification and DNA homology were analyzed by ERICPCR. The results showed that,during the detection of the 9 drugs,the resistance of 30 strains of C. perfringens to gentamicin was the highest(86.7%),followed by penicillin(43.3%),and the drug resistance rate of perfringens strains to cefotaxime was the lowest(1%). The rate of drug resistance to other drugs was between 3.3%-36.7%. The ERIC-PCR pattern showed 14 genotypes(Ⅰ-XIV),most of which came from different clones,and the similarity of two clone strains were 99%. The research results showed that the multi drug resistance of airborne C. perfringens in scaled goat farms was serious,and the molecular diversity was complex. The source of airborne C. perfringens was wide,and there was a certain difference in different regions,different environments and different conditions,indicating multiple genotypes may exist. The genotypes in the same goat farms may be different,and the same genotype also may exist in different goat farms.
In order to monitor the drug resistance changes of airborne Clostridium perfringens(C. perfringens)in different goat farms and the genetic similarity of different strains,so as to provide basis for clinical treatment and infection control,30 airborne strains were collected from different scaled goat farms in 2016—2017,the sensitivity of antibiotics was detected by K-B method,and the strains classification and DNA homology were analyzed by ERICPCR. The results showed that,during the detection of the 9 drugs,the resistance of 30 strains of C. perfringens to gentamicin was the highest(86.7%),followed by penicillin(43.3%),and the drug resistance rate of perfringens strains to cefotaxime was the lowest(1%). The rate of drug resistance to other drugs was between 3.3%-36.7%. The ERIC-PCR pattern showed 14 genotypes(Ⅰ-XIV),most of which came from different clones,and the similarity of two clone strains were 99%. The research results showed that the multi drug resistance of airborne C. perfringens in scaled goat farms was serious,and the molecular diversity was complex. The source of airborne C. perfringens was wide,and there was a certain difference in different regions,different environments and different conditions,indicating multiple genotypes may exist. The genotypes in the same goat farms may be different,and the same genotype also may exist in different goat farms.
引文
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[13]MARTAIN G T,SMYTH J A.Prevalence of net B among some clinical isolates of Clostridium perfringens from animals in the United States[J].Veterinary microbiology,2009(136):202-205.
[2]DONALDSON A I,GLOSTER J,HARVEY L D,et al.Use of prediction models to forecast and analyse airborne spread during the foot-and-mouth disease outbreaks in Brittany,Jersey and the Isle of Wight in 1981[J].Veterinary record,1982(110):53-57.
[3]BERRY K,COLVIN S,BLYTHE D,et al.Follow-Up of Deaths Among U.S.Postal Service Workers Potentially Exposed to Bacillus anthracis—District of Columbia,2001—2002[J].Morbidity&mortality weekly report,2003,52(39):937.
[4]ANDERS J,ANNA A,KALDHUSDAL M,et al.Genetic diversity and prevalence of net B in Clostridium perfringens isolated from a broiler flock affected by mild necrotic enteritis[J].Veterinary microbiology,2010(144):87-92.
[5]AIKHALDI S F,VILLANUEVA D,CHIZHIKOV V.Identitication and characterization of Clostridium perfringens using single target DNA microarmy chip[J].International journal of food microbiology,2004,91(3):289-296.
[6]蔡宝祥.家畜传染病学[M].北京:中国农业出版社,1996.
[7]张红英,卢中华,杨霞,等.我国魏氏梭菌病的流行特点[J].中国畜牧兽医,2004,31(1):39-41.
[8]PRAZMO Z,DUTKIEWICZ J,SKORSKA C,et al.Exposure to airborne gram-negative bacteria,dust,and endotoxin in paper factories[J].Annals of agricultural and environmental medicine,2003(52):937-938.
[9]Clinical and Laboratory Standards Institute.Performance standards for antimicrobial susceptibility testing:twentieth informational supplement M100-S20[S].Wayne:NCCLS,2010.
[10]GUIFANG W,LI P,HUIMIN D,et al.ERIC-PCR fingerprinting-based community DNA hybridization to pinpoint genome-specific fragements as molecular markers to identify and track populations common to healthy human guts[J].Journal of microbiological methods,2010(59):91-108.
[11]YUAN W,CHAI T J,MIAO Z M,et al.ERIC-PCR identification of the spread of airborne Escherichia coli in pig houses[J].Science of the total environment,2010(408):1446-1450.
[12]FITTIPALDI M,NOCKER A,CODONY F.Progress in understanding preferential detection of live cells using viability dyes in combination with DNA amplification[J].Microbiology methods,2012,91(2):276-289.
[13]MARTAIN G T,SMYTH J A.Prevalence of net B among some clinical isolates of Clostridium perfringens from animals in the United States[J].Veterinary microbiology,2009(136):202-205.
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