四川部分区域鸽源沙门菌耐药表型及耐药基因检测
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摘要
为了解四川省成都市及周边区域鸽源沙门菌的耐药表型及耐药基因携带情况,通过常规细菌分离鉴定并结合PCR方法,获得42株鸽源沙门菌,采用K-B纸片法,选取20种抗生素,对42株鸽源沙门菌进行药敏试验,并结合PCR方法对其耐药基因携带情况进行检测。结果显示:受检菌对链霉素、四环素、多西环素耐药率为88. 1%;对阿莫西林耐药率为83. 3%;对头孢拉定、氨苄西林、萘啶酸的耐药率为81. 0%。42株沙门菌皆有不同程度的多重耐药性,其中33株同时耐4种以上抗生素,占检测菌株的78. 6%(33/42),主要的耐药谱为CTX-CRO-FLO-CPIOFL-AKN。42株沙门菌喹诺酮类耐药基因aac (6')-Ib-cr检出率最高,为88. 09%(37/32);氨基糖苷类耐药基因aadA1、aph (3)Ⅱ、aac(3)Ⅱ检出率分别为80. 95%(34/42)、76. 19%(32/42)、16. 67%(7/42);β-内酰胺类耐药基因blaTEM-1、bla CMY-2检出率分别为42. 86%(18/42)、19. 04%(8/42);四环素类耐药基因tet M、tet A、tet B检出率分别为40. 4%(17/42)、38. 09%(16/42)、9. 52%(4/42);磺胺类耐药基因sul2检出率为80. 95%(34/42);喹诺酮类耐药基因qnrA检出率为38. 09%(16/42)。结果表明:四川省成都市及周边区域鸽源沙门菌存在普遍耐药,且多重耐药较为严重,耐药基因携带率高。
This study was to investigate the antibiotic resistance and resistance genes of Salmonella in pigeons in Chengdu and its surrounding areas in Sichuan Province. A total of 42 strains of salmonella were isolated from pigeons using normal method for bacterial isolation combined with PCR. The antimicrobial susceptibility of the Salmonella isolates was examined by the K-B method. Then,PCR was used to detect the resistance genes of the isolates. The results showed that the resistance rates of the 42 isolated strains to streptomycin,tetracycline,doxycycline were high at 88. 1%. Their resistance to amoxicillin was 83. 3%,and to cefradine,ampicillin and nalidixic acid were all 81. 0%. All the 42 strains of Salmonella were of multidrug resistance at different degrees,with 33 of them being resistant to more than four antibiotics and accounting for 78. 6%( 33/42) of the total. The main resistant profile was CTX-CRO-FLO-CPI-OFL-AKN. Detection of resistance determinants revealed that aac( 6')-Ib-cr was the most prevalent gene. Three kinds of aminoglycoside resistance genes aad A1( 80. 95%,34/42),aph( 3) Ⅱ( 76. 19%,32/42),and aac( 3)( 16. 67%,7/42) were all detected in isolates. The detected rates of bla TEM-1 and bla CMY-2 were 42. 86%( 18/42) and 19. 04%( 8/42),respectively. Among the four tetracycline resistance genes,tet M( 40. 4%,17/42),tet A( 38. 09%,16/42) and tet B( 9. 52%,4/42) were the most common resistance genes of the isolated strains. The detection rates of sul2 and qnr A were 80. 95%( 34/42) and 38. 09%( 16/42),respectively. These results indicated that the resistance rate of the isolated strains from pigeons in the investigated areas in Sichuan Province was high with wide a drug resistant spectrum,and the prevalence of drug resistant genes was very common. The present data provide a theoretical basis for treatment of Salmonella in pigeons,and are of significane for public health.
This study was to investigate the antibiotic resistance and resistance genes of Salmonella in pigeons in Chengdu and its surrounding areas in Sichuan Province. A total of 42 strains of salmonella were isolated from pigeons using normal method for bacterial isolation combined with PCR. The antimicrobial susceptibility of the Salmonella isolates was examined by the K-B method. Then,PCR was used to detect the resistance genes of the isolates. The results showed that the resistance rates of the 42 isolated strains to streptomycin,tetracycline,doxycycline were high at 88. 1%. Their resistance to amoxicillin was 83. 3%,and to cefradine,ampicillin and nalidixic acid were all 81. 0%. All the 42 strains of Salmonella were of multidrug resistance at different degrees,with 33 of them being resistant to more than four antibiotics and accounting for 78. 6%( 33/42) of the total. The main resistant profile was CTX-CRO-FLO-CPI-OFL-AKN. Detection of resistance determinants revealed that aac( 6')-Ib-cr was the most prevalent gene. Three kinds of aminoglycoside resistance genes aad A1( 80. 95%,34/42),aph( 3) Ⅱ( 76. 19%,32/42),and aac( 3)( 16. 67%,7/42) were all detected in isolates. The detected rates of bla TEM-1 and bla CMY-2 were 42. 86%( 18/42) and 19. 04%( 8/42),respectively. Among the four tetracycline resistance genes,tet M( 40. 4%,17/42),tet A( 38. 09%,16/42) and tet B( 9. 52%,4/42) were the most common resistance genes of the isolated strains. The detection rates of sul2 and qnr A were 80. 95%( 34/42) and 38. 09%( 16/42),respectively. These results indicated that the resistance rate of the isolated strains from pigeons in the investigated areas in Sichuan Province was high with wide a drug resistant spectrum,and the prevalence of drug resistant genes was very common. The present data provide a theoretical basis for treatment of Salmonella in pigeons,and are of significane for public health.
引文
[1]查华,石火英,吉贞颖,等.华东地区禽源沙门菌的分布及血清型分析[J].畜牧兽医学报,2013,44(2):329-332.
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[7]支威,马海燕,仇永凤,等.猪源沙门氏菌耐药性及耐药基因的分析[J].生物技术通报,2018,34(3):170-176.
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[11]赖海梅,邹立扣,刘书亮,等.肉鸡屠宰生产链中沙门氏菌耐药基因检测与耐药相关性分析[J].食品工业科技,2015,36(7):187-191.
[12] Afema J A,Byarugaba D K,Shah D H,et al. Potential sources and transmission of Salmonella and antimicrobial resistance in Kampala Uganda[J]. PLoS One,2016,11(3),e0152130.
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[14] Noda T,Murakami K,Etoh Y,et al. Increase in resistance to extended-spectrum cephalosporins in Salmonella isolated from retail chicken products in Japan[J]. PLoS One, 2015, 10(2):e0116927.
[15] de Vito D,Monno R,Nuccio F,et al. Diffusion and persistence of multidrug resistant Salmonella typhimurium strains phage type dt120in southern Italy[J]. Biomed Res Int, 2015, doi:10. 1155/2015/265042.
[16] Camarda A,Pupillo A,Pugliese N,et al. Phenotypic and genetic traits of Salmonella enterica subsp. serovar typhimurium strains causing salmonellosis foci in rabbit farms from southern Italy in 1999-2003[J]. Res Vet Sci,2013,94(3):394-398.
[17]王蓓,徐金水,于红,等.解脲脲原体对四环素类药物的耐药及tet M耐药基因研究[J].中国人兽共患病学报,2007,23(1):16.
[18] Folster J P,Tolar B,Pecic G,et al. Characterization of blacmy plasmids and their possible role in source attribution of Salmonella enterica serotype typhimurium infections[J]. Foodborne Pathog Dis,2014,11(4):301-306.
[19]王琼,唐俊妮,汤承,等.一种采用微波炉加热快速提取细菌DNA用于PCR扩增的方法[J].西南民族大学学报(自然科学版),2015,41(2):150-155.
[20] Patrick M E,Adcock P M,Gomez T M,et al. Salmonella enteritidis infections,United States,1985-1999[J]. Emerg Infect Dis,2004,10(1):1-7.
[21]钟传德,程安春,汪铭书,等.规模化鸭场鸭源致性沙门菌的药物敏感性检及耐药性分析[J].中国兽医杂志,2008,44(1):35-37.
[22]陈耀能,梁景涛,陈爱贞,等.佛山市食源性和人源沙门氏菌血清型分布与耐药性研究[J].热带医学杂志,2012,12(8):955-958.
[23]李迎晓,安辉,焦凤超,等.鸭大肠杆菌与沙门氏菌混合感染的分离鉴定及药敏试验[J].江苏农业科学,2015,43(4):224-225.
[24]吴植,郝福星,刘莉,等.禽源沙门菌氨基糖苷类耐药表型及耐药基因研究[J].中国家禽,2016,38(17):56-58.
[25]郭云昌,刘秀梅.市售鸡肉沙菌分离株多重耐药谱测定[J].中国食品卫生杂志,2005,17(2):100-103.
[26]林亚军,郭菲,夏利宁,等.新疆乌鲁木齐市宠物源沙门氏菌耐药性及耐药基因检测[J].中国农业大学学报,2018,23(7):75-83.
[27] Chopra I,Roberts M. Tetracycline antibiotics:mode of action,applications,molecular biology,and epidemiology of bacterial resistance[J]. Microbiol Mol Biolo Rev,2001,65(2):232-260.
[28] Tuckman M,Petersen P J,Howe A Y,et al. Occurrence of tetracycline resistance genes among Escherichia coli isolates from the phase 3 clinical trials for tigecycline[J]. Antimicrob Agents Chemother,2007,51(9):3205-3211.
[29]戴建华,吴植,袁维峰.禽源致病性沙门氏菌耐药表型与耐药基因的分析[J].中国畜牧兽医,2014,41(4):245-248.
[30]宁家宝,陈建红,张济培,等.鸡白痴沙门菌的耐药性及横胺类耐药基因的研究[J].中国家禽,2014,36(3):18-20.
[31]覃春红.动物源沙门氏菌耐药性及质粒介导喹诺酮类耐药基因的流行病学研究[D].成都:四川农业大学,2011.
[2]陆承平.兽医微生物学[M]. 5版.北京:中国农业出版社,2013.
[3] Rahmani M,Peighambari S M,Svendsen C A,et al. Molecular clonality and antimicrobial resistance in Salmonella enterica serovars enteritidis and infantis from broilers in three northern regions of Iran[J]. Bmc Vet Res,2013,9(1):66.
[4] Li D,Wang X,Wang L,et al. The actin-polymerizing activity of sip A is not essential for Salmonella enterica serovar typhimurium-induced mucosal inflammation[J]. Infect Immun,2013,81(5):1541-1549.
[5] Ke B,Sun J,He D,et al. Serovar distribution,antimicrobial resistance profiles and pfge typing of Salmonella enterica strains isolated from 2007-2012 in Guangdong,China[J]. Bmc Infect Dis,2014,14(1),338.
[6]杨丹,贺安祥,邓俊良,等.藏鸡致病性沙门菌的耐药性分析及质粒耐药基因的检测[J].中国兽医科学,2016,46(1):84-90.
[7]支威,马海燕,仇永凤,等.猪源沙门氏菌耐药性及耐药基因的分析[J].生物技术通报,2018,34(3):170-176.
[8] Pereira R V,Siler J D,Cummings K J,et al. Effect of heifer-raising practices on E. coli antimicrobial resistance and Salmonella prevalence in heifer raisers[J]. Epidemiol Infect, 2015, 143(15):3256-3265.
[9]闫雷,徐海.质粒介导的喹诺酮耐药基因qnr的分类、耐药机制及其在国内的流行状况[J].微生物学报,2016,56(2):169-179.
[10]杨保伟,曲东,申进玲,等.陕西食源性沙门氏菌耐药及相关基因[J].微生物学报,2010,50(6):788-796.
[11]赖海梅,邹立扣,刘书亮,等.肉鸡屠宰生产链中沙门氏菌耐药基因检测与耐药相关性分析[J].食品工业科技,2015,36(7):187-191.
[12] Afema J A,Byarugaba D K,Shah D H,et al. Potential sources and transmission of Salmonella and antimicrobial resistance in Kampala Uganda[J]. PLoS One,2016,11(3),e0152130.
[13] Solomon S L,Oliver K B. Antibiotic resistance threats in the United States:stepping back from the brink[J]. Am Fam Physician,2014,89(12):938-941.
[14] Noda T,Murakami K,Etoh Y,et al. Increase in resistance to extended-spectrum cephalosporins in Salmonella isolated from retail chicken products in Japan[J]. PLoS One, 2015, 10(2):e0116927.
[15] de Vito D,Monno R,Nuccio F,et al. Diffusion and persistence of multidrug resistant Salmonella typhimurium strains phage type dt120in southern Italy[J]. Biomed Res Int, 2015, doi:10. 1155/2015/265042.
[16] Camarda A,Pupillo A,Pugliese N,et al. Phenotypic and genetic traits of Salmonella enterica subsp. serovar typhimurium strains causing salmonellosis foci in rabbit farms from southern Italy in 1999-2003[J]. Res Vet Sci,2013,94(3):394-398.
[17]王蓓,徐金水,于红,等.解脲脲原体对四环素类药物的耐药及tet M耐药基因研究[J].中国人兽共患病学报,2007,23(1):16.
[18] Folster J P,Tolar B,Pecic G,et al. Characterization of blacmy plasmids and their possible role in source attribution of Salmonella enterica serotype typhimurium infections[J]. Foodborne Pathog Dis,2014,11(4):301-306.
[19]王琼,唐俊妮,汤承,等.一种采用微波炉加热快速提取细菌DNA用于PCR扩增的方法[J].西南民族大学学报(自然科学版),2015,41(2):150-155.
[20] Patrick M E,Adcock P M,Gomez T M,et al. Salmonella enteritidis infections,United States,1985-1999[J]. Emerg Infect Dis,2004,10(1):1-7.
[21]钟传德,程安春,汪铭书,等.规模化鸭场鸭源致性沙门菌的药物敏感性检及耐药性分析[J].中国兽医杂志,2008,44(1):35-37.
[22]陈耀能,梁景涛,陈爱贞,等.佛山市食源性和人源沙门氏菌血清型分布与耐药性研究[J].热带医学杂志,2012,12(8):955-958.
[23]李迎晓,安辉,焦凤超,等.鸭大肠杆菌与沙门氏菌混合感染的分离鉴定及药敏试验[J].江苏农业科学,2015,43(4):224-225.
[24]吴植,郝福星,刘莉,等.禽源沙门菌氨基糖苷类耐药表型及耐药基因研究[J].中国家禽,2016,38(17):56-58.
[25]郭云昌,刘秀梅.市售鸡肉沙菌分离株多重耐药谱测定[J].中国食品卫生杂志,2005,17(2):100-103.
[26]林亚军,郭菲,夏利宁,等.新疆乌鲁木齐市宠物源沙门氏菌耐药性及耐药基因检测[J].中国农业大学学报,2018,23(7):75-83.
[27] Chopra I,Roberts M. Tetracycline antibiotics:mode of action,applications,molecular biology,and epidemiology of bacterial resistance[J]. Microbiol Mol Biolo Rev,2001,65(2):232-260.
[28] Tuckman M,Petersen P J,Howe A Y,et al. Occurrence of tetracycline resistance genes among Escherichia coli isolates from the phase 3 clinical trials for tigecycline[J]. Antimicrob Agents Chemother,2007,51(9):3205-3211.
[29]戴建华,吴植,袁维峰.禽源致病性沙门氏菌耐药表型与耐药基因的分析[J].中国畜牧兽医,2014,41(4):245-248.
[30]宁家宝,陈建红,张济培,等.鸡白痴沙门菌的耐药性及横胺类耐药基因的研究[J].中国家禽,2014,36(3):18-20.
[31]覃春红.动物源沙门氏菌耐药性及质粒介导喹诺酮类耐药基因的流行病学研究[D].成都:四川农业大学,2011.