禽源多杀性巴氏杆菌耐药性分析及氟苯尼考耐药基因floR分布
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摘要
为了解禽源多杀性巴氏杆菌的药物敏感性及氟苯尼考的耐药基因的分布特征,采用琼脂二倍稀释法,对分离自福建、广东和江西等南方数省部分地区113株禽源多杀性巴氏杆菌进行9种抗生素的耐药性分析,并应用PCR方法检测氟苯尼考耐药基因floR的分布情况。结果显示:上述菌株对四环素表现高水平耐药,耐药率为65.49%(74/113);对链霉素、甲氧苄啶/磺胺甲噁唑、氟苯尼考、卡那霉素与恩诺沙星表现中等水平耐药,耐药率分别是43.36%(49/113)、39.82%(45/113)、27.43%(31/113)、25.66%(29/113)与24.78%(28/113);对氨苄西林、头孢噻呋与头孢克洛较为敏感,耐药率分别为4.42%(5/113),2.65%(3/113)与1.77%(2/113);其中52.21%(59/113)菌株能耐受3类及3类以上的抗菌药物;氟苯尼考耐药基因floR的检出率为64.04%(72/113),且所有氟苯尼考耐药菌株均含有floR基因。结论:113株禽源多杀性巴氏杆菌存在多重耐药现象,且floR耐药基因较为流行。本研究结果为上述地区针对禽源多杀性巴氏杆菌临床用药提供科学依据。
In order to understand the antimicrobial sensitivity and distribution of the floR genes of Pasteurella multocida strains isolated from poultry in Fujian,Guangdong,Jiangxi and other provinces in the south of China,113 Pasteurella multocida strains were tested for antimicrobial susceptibility to 9 antibiotics by the agar dilution method.Then,the PCR method was used to detect floR.The results were as follows:The drug sensitivity test results showed that the above strains had high resistance to Tetracycline,at the rate of 65.49%(74/113);intermediate resistance to Streptomycin,Trimethoprim-sulfamethoxazole,Kanamycin,Florfenicol,and Enrofloxacin,at the rates of 43.36%(49/113),39.82%(45/113),25.66%(29/113),27.43%(31/113) and 24.78%(28/113),respectively;and low resistance to Ampicillin,Ceftiofur,Cefaclor,at the rates of 4.42%(5/113),2.65%(3/113) and 1.77%(2/113),and 59 of 113(52.21%),respectively.The Pasteurella multocida strains were of multiple resistances.The detected rates of the resistant floR was 64.04%(72/113),and all the isolates that showed resistance to Florfenicol were positive for the floR gene.The present study indicated that the Pasteurella multocida strains had a multiple drug resistance,and the prevalence of floR was very popular.These data might serve as reference for clinical research.
In order to understand the antimicrobial sensitivity and distribution of the floR genes of Pasteurella multocida strains isolated from poultry in Fujian,Guangdong,Jiangxi and other provinces in the south of China,113 Pasteurella multocida strains were tested for antimicrobial susceptibility to 9 antibiotics by the agar dilution method.Then,the PCR method was used to detect floR.The results were as follows:The drug sensitivity test results showed that the above strains had high resistance to Tetracycline,at the rate of 65.49%(74/113);intermediate resistance to Streptomycin,Trimethoprim-sulfamethoxazole,Kanamycin,Florfenicol,and Enrofloxacin,at the rates of 43.36%(49/113),39.82%(45/113),25.66%(29/113),27.43%(31/113) and 24.78%(28/113),respectively;and low resistance to Ampicillin,Ceftiofur,Cefaclor,at the rates of 4.42%(5/113),2.65%(3/113) and 1.77%(2/113),and 59 of 113(52.21%),respectively.The Pasteurella multocida strains were of multiple resistances.The detected rates of the resistant floR was 64.04%(72/113),and all the isolates that showed resistance to Florfenicol were positive for the floR gene.The present study indicated that the Pasteurella multocida strains had a multiple drug resistance,and the prevalence of floR was very popular.These data might serve as reference for clinical research.
引文
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[11]Furian T Q,Borges K A,Laviniki V,et al.Virulence genes and antimicrobial resistance of Pasteurella multocida isolated from poultry and swine[J].Brazilian Journal of Microbiology,2016,47(1):210-216.
[12]Li Z C,Cheng F J,Lan S M,et al.Investigation of genetic diversity and epidemiological characteristics of Pasteurella multocida isolates from poultry in southwest China by population structure,multi-locus sequence typing and virulence-associated gene profile analysis[J].J Vet Med Sci,2018,80(6):921-929.
[13]Kim E H,Aoki H.Sequence analysis of the florfenicol resistance gene encoded on the transferable R-plasmid of a fish pathogen Pasteurella piscicida[J].Microb Immunol,1996,40(9):665-669.
[14]Bolton L F,Kelley L C,Lee M D,et al.Detection of Salmonella ty phimurium DT104 based on the gene St-flo which confer s resistance to florfenicol an chloramphenicol[J].J Clin Microbiol,1999,37:1348-1351.
[15]Keyes K,Udson C H,Aurer M J J,et al.Detection of florfenicol resistance genes in Escherichia coli isolated from sick chickens[J].Antimicrob Agents Chemother,2000,44:421-424.
[2]Graham R,Palmer D,Pratt B C,et al.In vitro activity of florphenicol[J].European Journal of Clinical Microbiology and Infectious Diseases,1988,7(5):691-694.
[3]Wu B B,Xia C,Du X D,et al.Influence of anti floR antibody on florfenicol accumulation in florfenicol resistant Escherichia coli and enzyme linked immunosorbent assay for detection of florfenicol resistant E.coli isolates[J].J Clin Microb,2006,44(2):378-382.
[4]冯世文,曾芸,李军,等.细菌对氟苯尼考的耐药机制研究[J].黑龙江畜牧兽医,2014(03):52-54.
[5]CLSI(Clinical and Laboratory Standards Institute).Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from Animals;Second Informational SupplementDocument VET01-S2.Wayne,PA:Clinical and Laboratory Standards Institute;2013.
[6]Dai L,Lu L M,Wu C M,et al.Characterization of antimicrobial resistance among Escherichia coli isolates from chickens in China between 2001 and 2006[J].FEMS Microbiol Lett,2008,286(2):178-183.
[7]Tang X,Zhao Z,Hu J,et al.Isolation,Antimicrobial resistance,and virulence genes of Pasteurella multocida strains from swine in china[J].Journal of Clinical Microbobilolgy,2009,47(4):951-958.
[8]Sellyei B,Barqa Z,Szentesi-samu K,et al.Antimicrobial susceptibility of Pasteurella multocida isolated from swine and poultry[J].Acta Veterinaria Hungarica,2009,57(3):357-367.
[9]Katsuda K,Hoshinoo K,Ueno Y,et al.Virulence genes and antimicrobial susceptibility in Pasteurella multocida isolates from calves[J].Veterinary Microbiology,2013,167:737-741.
[10]Faham K,Hassan M,Morteza A M.Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in Iran[J].Frontiers in Microblology,2014,536(5):1-9.
[11]Furian T Q,Borges K A,Laviniki V,et al.Virulence genes and antimicrobial resistance of Pasteurella multocida isolated from poultry and swine[J].Brazilian Journal of Microbiology,2016,47(1):210-216.
[12]Li Z C,Cheng F J,Lan S M,et al.Investigation of genetic diversity and epidemiological characteristics of Pasteurella multocida isolates from poultry in southwest China by population structure,multi-locus sequence typing and virulence-associated gene profile analysis[J].J Vet Med Sci,2018,80(6):921-929.
[13]Kim E H,Aoki H.Sequence analysis of the florfenicol resistance gene encoded on the transferable R-plasmid of a fish pathogen Pasteurella piscicida[J].Microb Immunol,1996,40(9):665-669.
[14]Bolton L F,Kelley L C,Lee M D,et al.Detection of Salmonella ty phimurium DT104 based on the gene St-flo which confer s resistance to florfenicol an chloramphenicol[J].J Clin Microbiol,1999,37:1348-1351.
[15]Keyes K,Udson C H,Aurer M J J,et al.Detection of florfenicol resistance genes in Escherichia coli isolated from sick chickens[J].Antimicrob Agents Chemother,2000,44:421-424.