不同宿主来源香港海鸥菌耐药表型和耐药基因分析
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摘要
目的了解不同宿主来源香港海鸥菌的耐药表型及耐药基因的携带情况。方法于2015年1月—2016年8月分离自广东省淡水产品鱼和蛙肠道117株香港海鸥菌,5株病人粪便分离株为本实验室保存。采用K-B纸片法测定菌株对13种抗生素的耐药表型,PCR法扩增磺胺类、四环素类、喹诺酮类和红霉素类等9种耐药基因,应用χ2检验进行率的差异性比较。结果 122株香港海鸥菌主要对利福平、头孢噻吩、氨苄西林、复方新诺明、红霉素、四环素、环丙沙星耐药,耐药率分别为86.9%、78.6%、48.7%、30.8%、18.9%、18.0%、14.5%,多重耐药率达57.4%;对四环素耐药的22株菌中,tet A、tet R检出率分别为72.7%、68.2%;对磺胺类耐药的36株菌中,sul1、sul2检出率分别为86.1%、50.0%;鱼源株对氨苄西林、头孢噻吩、红霉素、利福平的耐药率均高于蛙源株(P均<0.05),而蛙源株对四环素、环丙沙星、复方新诺明和链霉素的耐药率均高于鱼源株(P均<0.05);蛙源株的sul1、sul2,tet A、tet R、aac(6′)-ib和ere A等6种耐药基因携带率均高于鱼源株(P均<0.05);5株人源株耐药率较低,仅有1株表现为多重耐药。结论广东省淡水产品中香港海鸥菌对抗生素的耐药性较为严重,不同宿主来源的菌株具有不同的耐药模式。tet A、tet R和sul1、sul2可能是该菌对磺胺类和四环素类药物耐药的主要原因。
Objective To examine phenotype and genotype distribution of antibiotic-resistant Laribacter hongkongensis(LH) strains isolated from different hosts. Methods Totally 117 LH strains were isolated from intestinal tracts of fresh water fish and frogs in Guangdong province from January 2015 to August 2016, and 5 strains from human faces. Kirby-Bauer(K-B) disk diffusion method was used to test the susceptibility to 13 antibiotics of the strains. Detections of 9 antibioticresistant genes were performed for 122 LH strains with PCR amplification. Chi-square test was adopted to analyze differences in antibiotic resistance of the strains. Results Of the 122 LH strains, 86.89% were resistant to rifampin, 78.6% to cefalotin, 48.7% to ampicillin, 78.6 % to sulfamethoxazole, 18.9% to erythromycin, 18.0% to tetracycline, and 14.5% to ciprofloxacin, respectively, with an overall multiple antibiotic-resistant rate of 57.4%. Among the 22 tetracycline-resistant LH strains, the detection rates of tet A and tet R resistance-related genes were 72.7% and 68.2 %; for the 36 sulfa-resistant LH strains, the detection rates of sul1 and sul2 resistance-related genes were 86.1% and 50.0%. The ampicillin-, cefalotin-,erythromycin-, and rifampicin-resistant rate were significantly higher among LH strains isolated from fresh water fish than among those from frogs(P < 0.05 for all); while the tetracycline-, ciprofloxacin-, sulfamethoxazole-, and streptomycinresistant rate of the LH strains from frogs were significantly higher than those of the strains from fish(P < 0.05 for all).Significantly higher carrying rates of antibiotic resistance-related genes, including sul1, sul2, tet A, tet R, aac(6′)-ib, and ere A,were observed among the LH strains isolated from frogs than among those from fish(all P < 0.05). Of the 5 LH strains from human, only one was multiple antibiotic-resistant. Conclusion The antibiotic-resistant stains are highly prevalent for LH isolated from freshwater products in Guangdong province and the antibiotic resistance is different for the LH strains isolated from various hosts. Carrying tet A, tet R, sul1, and sul2 genes may play a main role for the LH strains′ sulfonamides and tetracyclines resistance.
Objective To examine phenotype and genotype distribution of antibiotic-resistant Laribacter hongkongensis(LH) strains isolated from different hosts. Methods Totally 117 LH strains were isolated from intestinal tracts of fresh water fish and frogs in Guangdong province from January 2015 to August 2016, and 5 strains from human faces. Kirby-Bauer(K-B) disk diffusion method was used to test the susceptibility to 13 antibiotics of the strains. Detections of 9 antibioticresistant genes were performed for 122 LH strains with PCR amplification. Chi-square test was adopted to analyze differences in antibiotic resistance of the strains. Results Of the 122 LH strains, 86.89% were resistant to rifampin, 78.6% to cefalotin, 48.7% to ampicillin, 78.6 % to sulfamethoxazole, 18.9% to erythromycin, 18.0% to tetracycline, and 14.5% to ciprofloxacin, respectively, with an overall multiple antibiotic-resistant rate of 57.4%. Among the 22 tetracycline-resistant LH strains, the detection rates of tet A and tet R resistance-related genes were 72.7% and 68.2 %; for the 36 sulfa-resistant LH strains, the detection rates of sul1 and sul2 resistance-related genes were 86.1% and 50.0%. The ampicillin-, cefalotin-,erythromycin-, and rifampicin-resistant rate were significantly higher among LH strains isolated from fresh water fish than among those from frogs(P < 0.05 for all); while the tetracycline-, ciprofloxacin-, sulfamethoxazole-, and streptomycinresistant rate of the LH strains from frogs were significantly higher than those of the strains from fish(P < 0.05 for all).Significantly higher carrying rates of antibiotic resistance-related genes, including sul1, sul2, tet A, tet R, aac(6′)-ib, and ere A,were observed among the LH strains isolated from frogs than among those from fish(all P < 0.05). Of the 5 LH strains from human, only one was multiple antibiotic-resistant. Conclusion The antibiotic-resistant stains are highly prevalent for LH isolated from freshwater products in Guangdong province and the antibiotic resistance is different for the LH strains isolated from various hosts. Carrying tet A, tet R, sul1, and sul2 genes may play a main role for the LH strains′ sulfonamides and tetracyclines resistance.
引文
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[3]Teng JL,Woo PC,Ma SS,et al.Ecoepidemiology of Laribacter hongkongensis,a novel bacterium associated with gastroenteritis[J].J Clin Microbiol,2005,43(2):919-922.
[4]Feng JL,Yan H,Chowdhury N,et al.Identification andcharacterization of integron-associated antibiotic resistant Laribacter hongkongensis isolated from aquatic products in China[J].International Journal of Food Microbiology,2011,144(3):337-341.
[5]朱江峰,郑南才,廖如燕,等.广东省香港海鸥菌感染性腹泻1例分析[J].中国公共卫生,2011,27(4):403-405.
[6]王祉蕴,冯梅,刘志华,等.广州市香港海鸥菌检测及耐药性分析[J].中国公共卫生,2015,31(4):473-475.
[7]冯梅,刘优招,何雅青,等.深圳市某区淡水产品中香港海鸥菌的检测及耐药特征分析[J].中华疾病控制杂志,2016,20(8):764-767.
[8]Clinical and Laboratory Standards Institute.Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically M7-A7[S].Clinical and Laboratory Standards Institute,2008.
[9]罗毓婷,陈定强,杨羚,等.香港海鸥型菌四环素耐药性及其耐药分子机制研究[J].实用医学杂志,2013,29(4):630-632.
[10]杨锦红,周义正,刘立雪,等.儿童肺炎链球菌红霉素、四环素相关耐药基因的研究[J].中国卫生检验杂志,2008,18(5):866-869.
[11]Chen DQ,Yang L,Luo YT,et al.Prevalence and characterization of quinolone resistance in Laribacter hongkongensis from grass carp and Chinese tiger frog[J].Journal of Medical Microbiology,2013,62(10):1559-1564.
[12]陈冰,冯嘉丽,李美霞,等.耐复方新诺明香港海鸥型菌中I类整合子、磺胺类耐药基因(sul1、sul2)的检测[J].中国人兽共患病杂志,2012,28(3):223-225.
[13]Valeriani F,Giampaoli S,Buggiotti L,et al.Molecular enrichment for detection of S.aureus in recreational waters[J].Water Science and Technology,2012,66(11):2305-2310.
[14]Jiang HX,Tang D,Liu YH,et al.Prevalence and characteristics ofβ-lactamase and plasmid-mediated quinolone resistance genes in Escherichia coli isolated from farmed fish in China[J].Journal of Antimicrobial Chemotherapy,2012,67(10):2350-2353.
[15]林广城,黄烈,刘键,等.葡萄球菌大环内酯类耐药基因检测及其意义[J].中国热带医学,2006,6(11):1926-1927.
[2]Woo PCY,Lau SKP,Teng JLL,et al.Association of Laribacter hongkongensis in community-acquired gastroenteritis with travel and eating fish:a multicentre case-control study[J].The Lancet,2004,363(9425):1941-1947.
[3]Teng JL,Woo PC,Ma SS,et al.Ecoepidemiology of Laribacter hongkongensis,a novel bacterium associated with gastroenteritis[J].J Clin Microbiol,2005,43(2):919-922.
[4]Feng JL,Yan H,Chowdhury N,et al.Identification andcharacterization of integron-associated antibiotic resistant Laribacter hongkongensis isolated from aquatic products in China[J].International Journal of Food Microbiology,2011,144(3):337-341.
[5]朱江峰,郑南才,廖如燕,等.广东省香港海鸥菌感染性腹泻1例分析[J].中国公共卫生,2011,27(4):403-405.
[6]王祉蕴,冯梅,刘志华,等.广州市香港海鸥菌检测及耐药性分析[J].中国公共卫生,2015,31(4):473-475.
[7]冯梅,刘优招,何雅青,等.深圳市某区淡水产品中香港海鸥菌的检测及耐药特征分析[J].中华疾病控制杂志,2016,20(8):764-767.
[8]Clinical and Laboratory Standards Institute.Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically M7-A7[S].Clinical and Laboratory Standards Institute,2008.
[9]罗毓婷,陈定强,杨羚,等.香港海鸥型菌四环素耐药性及其耐药分子机制研究[J].实用医学杂志,2013,29(4):630-632.
[10]杨锦红,周义正,刘立雪,等.儿童肺炎链球菌红霉素、四环素相关耐药基因的研究[J].中国卫生检验杂志,2008,18(5):866-869.
[11]Chen DQ,Yang L,Luo YT,et al.Prevalence and characterization of quinolone resistance in Laribacter hongkongensis from grass carp and Chinese tiger frog[J].Journal of Medical Microbiology,2013,62(10):1559-1564.
[12]陈冰,冯嘉丽,李美霞,等.耐复方新诺明香港海鸥型菌中I类整合子、磺胺类耐药基因(sul1、sul2)的检测[J].中国人兽共患病杂志,2012,28(3):223-225.
[13]Valeriani F,Giampaoli S,Buggiotti L,et al.Molecular enrichment for detection of S.aureus in recreational waters[J].Water Science and Technology,2012,66(11):2305-2310.
[14]Jiang HX,Tang D,Liu YH,et al.Prevalence and characteristics ofβ-lactamase and plasmid-mediated quinolone resistance genes in Escherichia coli isolated from farmed fish in China[J].Journal of Antimicrobial Chemotherapy,2012,67(10):2350-2353.
[15]林广城,黄烈,刘键,等.葡萄球菌大环内酯类耐药基因检测及其意义[J].中国热带医学,2006,6(11):1926-1927.