豫北地区猪源大肠杆菌耐药性及基因多态性分析
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摘要
为了解豫北地区规模化养殖场猪源大肠杆菌的耐药性及基因多态性,对分离的21株猪源大肠杆菌进行药物敏感实验,采用PCR技术对其耐药基因进行检测,RAPD技术进行基因多态性分析。结果表明:豫北地区规模化养殖场分离的猪源性大肠杆菌菌株对常见16种抗生素存在不同情况的耐药性。其中对四环素、红霉素、麦迪霉素、阿莫西林、磺胺异恶唑、环丙沙星、新生霉素、复方新诺明、恩诺沙星、甲氧苄啶的耐药率均为100%,对庆大霉素、洛美沙星的耐药率为85%以上,对其他药物的耐药率相对较低;PCR检测得到8种耐药基因的条带;对菌株进行RAPD分析得到7种不同的基因型。大肠杆菌极易产生耐药性,耐药基因广泛存在于耐药菌株中,但耐药表型与耐药基因之间无绝对相关性。豫北地区猪源大肠杆菌感染多重耐药十分严重,严重影响了该地区猪源大肠杆菌病的诊治和预防。
To determine the drug resistance and gene polymorphism of Escherichia coli isolated from pigs from north of Henan province,in this study, 21 isolated porcine Escherichia coil were detected with drug sensitivity test, and the drug resistance genes were detected by PCR, the gene polymorphism was analyzed by RAPD. The results showed that the isolated Escherichia coli strains have different resistance to 16 common antibiotics, and the drug resistance rate of strains for tetracycline, erythromycin, medemycin, amoxicillin, sulfamethoxazole, ciprofloxacin, neomycin, sulfamethoxazole, enrofloxacin, trimethoprim were 100%, The drug resistance rate for gentamicin and lomefloxacin were more than 85%, and drug resistance rate for the remained antibiotics had the low resistance rate. At the same time, 8 drug resistance genes was obtained by PCR from the isolated strains; 7 different genotypes were obtained with the RAPD analysis. The above results showed that porcine Escherichia coli was easy to form the resistance to drug, and resistance genes were widely existed in these resistant strains, but there was no correlation between resistant phenotype and resistant genes.This study revealed that porcine Escherichia coli were multidrug resistance in north of Henan, would seriously affect diagnosis and treatment prevention of porcine Escherichia coli.
To determine the drug resistance and gene polymorphism of Escherichia coli isolated from pigs from north of Henan province,in this study, 21 isolated porcine Escherichia coil were detected with drug sensitivity test, and the drug resistance genes were detected by PCR, the gene polymorphism was analyzed by RAPD. The results showed that the isolated Escherichia coli strains have different resistance to 16 common antibiotics, and the drug resistance rate of strains for tetracycline, erythromycin, medemycin, amoxicillin, sulfamethoxazole, ciprofloxacin, neomycin, sulfamethoxazole, enrofloxacin, trimethoprim were 100%, The drug resistance rate for gentamicin and lomefloxacin were more than 85%, and drug resistance rate for the remained antibiotics had the low resistance rate. At the same time, 8 drug resistance genes was obtained by PCR from the isolated strains; 7 different genotypes were obtained with the RAPD analysis. The above results showed that porcine Escherichia coli was easy to form the resistance to drug, and resistance genes were widely existed in these resistant strains, but there was no correlation between resistant phenotype and resistant genes.This study revealed that porcine Escherichia coli were multidrug resistance in north of Henan, would seriously affect diagnosis and treatment prevention of porcine Escherichia coli.
引文
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[13] 王学君,谭艾娟,吕世明,等. 贵州省猪源大肠杆菌对四环素类药物的耐药性及耐药基因检测[J].中国畜牧兽医,2018,45(5):1367-1373. Wang X J, Tan A J, Lv S M, et al. Detection of Resistance and Resistance Genes of Swine Escherichia coli in Guizhou to Tetracyclines[J]. China Animal Husbandry&Veterinary Medicine,2018,45(5):1367-1373.
[14] 颜友荣,方向红,王琳琳. 苏中地区猪大肠杆菌的耐药性与耐药基因的试验[J].中国兽医杂志,2018,52(2) : 90-92. Yan Y R, Fang X H, Wang L L. Drug Resistance and Drug Resistance Genes in Central of Jiangsu [J]. Chinese Journal of Veterinary Medicine,2018,52(2): 90-92.
[15] Wu S1, Dalsgaard A, Hammerum AM, et al. Prevalence and characterization of plasmids carrying sulfonamide resistance genes among Escherichia coli from pigs, pig carcasses and human [J]. Acta Vet Scand. 2010, 30 (6):1-7.
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[17] Hadis Arabi. IrajPakzad. Ayat Nasrollahi. Sulfonamide Resistance Genes (sul) M in Extended Spectrum Beta Lactamase (ESBL) and Non-ESBL Producing Escherichia coli Isolated From Iranian Hospitals [J]. Jundishapur J Microbiol, 2015 , 8(7): e19961.
[18] 陆彦,晏志勋,赵红玉,等. 印第安纳沙门氏菌对氯霉素类药物耐药性分析[J].微生物学通报,2013,40(7):1225-1230. Lu Y, Yan Z X, Zhao H Y , et al. Resistance analysis of Salmonella Indiana to the chlmphenicols[J].Microbiology China, 2013,40(7):1225-1230.
[19] Moore J E, Watabe M, Millar B C, et al. Molecular characterization of verocytoxigenic Escherichia coli O157:H7 by randomamplification of polymorphic DNA (RAPD) typing[J]. Br J Biomed Sci, 2008, 65(3): 161-163.
[20] 陈荣忠,向蓉,莫和国,等. 社区获得性耐甲氧西林金黄色葡萄球菌RAPD基因分型及药敏分析[J].中国抗生素杂志,2018, 43(5): 530-534. Chen R Z, Xiang R, Mo H G, et al. RAPD genotyping and drug sensitivity analysis of community acquired methicillin-resistant Staphylococcus[J]. Chinese Journal of Antibiotics,2018, 43(5): 530-534.
[2] 晏云涛,赵汝,苗淑淑,等. 猪源大肠埃希菌药敏试验及耐药基因检测[J].动物医学进展,2018,39(7):42-46. Yan Y T, Zhao R, Miao S S, et al. Drug Susceptibility Test and Drug Resistant Gene Detection of Pig Source Escherichia coli[J].Progress in Veterinary Medicine, 2018,39(7):42-46.
[3] 艾伟昌,李金磊,董鹏,等. 河南省猪源大肠杆菌的分离鉴定与耐药性分析[J].上海畜牧兽医通讯,2017,(6):29-31. Ai W C, Li J L, Dong P, et al. Isolation, identification and drug resistance of Escherichia coli isolated from pigs in Henan [J]. Shanghai Journal of Animal Husbandry and Veterinary Medicine, 2017, (6):29-31.
[4] 廖成水,程相朝,张春杰,等. 鸡源致病性沙门氏菌新近分离株的耐药性与耐药基因[J].中国兽医科学,2011,41(7): 751-755. Liao C S, Cheng X C, Zhang C J, et al. Antimicrobial resistance and resistance genes of pathogenic Salmonella recently isolated from chicken[J].Chinese Veterinary Science, 2011,41(7): 751-755.
[5] 闫新武,雷连成,张明亮,等. 貂源绿脓杆菌的分离鉴定及生物学特性[J].中国兽医学报,2014,34(10): 1610-1614. Yan X W, Lei L C, Zhang M L, et al. Identification and study on biological characteristics of Pseudomonas aeruginosa isolated from the minks[J]. Chinese Journal of Veterinary Science, 2014, 34(10): 1610-1614.
[6] Clinical and Laboratory Standards Institute (CLSI).Performance standards for antimicrobial susceptibility testing[C]25th informational supplement. Document M100-S25,Wayne,PA: CLSI,2015.
[7] Bonnet R. Growing group of extended-spectrum beta-lactamases the CTX-M enzymes[J].JAntimicrob Chemother, 2012,48(1):1 -14.
[8] 李俊贤,陈鹏,杨晓伟,等. 3种动物源大肠杆菌分离株的耐药性分析[J].黑龙江畜牧兽医,2018,(16) : 106-108. Li J X, Chen P, Yang X W, et al. Drug Resistance Analysis of Escherichia coli Isolated from Three Animal [J].Heilongjiang animal science and veterinary medicine,2018,(16): 106-108.
[9] 王克领,张青娴,徐引弟,等. 河南地区猪源性大肠杆菌血清型鉴定与耐药性调查[J]. 河南农业科学, 2014,(9):164-167. Wang K L, Zhang Q X, Xu Y D, et al. Serotype Identification and Drug Resistance Investigation of Swine Escherichia coli in Henan Area[J].Journal of Henan Agricultural Sciences,2014, (9): 164-167.
[10] 李晴,张泽,曹甸磊,等.养殖废水产ESBLs大肠杆菌多重耐药性及耐药基因检测[J].中国预防兽医学报,2018,40(6):495-499. Li Q, Zhang Z, Cao D L, et al.Detection of multiple drug resistance and the resistance genes of ESBL-producing Escherichia coli from livestock wastewater[J].Chinese Journal of Preventive Veterinary Medicine, 2018, 40(6):495-499.
[11] 毛福超,郁川,韩璐,等. 豫西地区禽源大肠杆菌的分离鉴定与耐药性分析[J].河南农业科学,2016,45( 1) : 127-130. Mao F C, Yu C, Han L, et al. Isolation,Identification and Drug Resistance Analysis of Escherichia coli from Chickens in Western Henan Province[J]. Journal of Henan Agricultural Sciences, 2016,45( 1) : 127-130.
[12] 张炳亮,董发明,王文文,等. 河南地区猪源性大肠杆菌耐药性及耐药基因型分析[J].西北农林科技大学学报(自然科学版),2018,47(4) : 1-7. Zhang B L, Dong F M, Wang W W, et al. Drug resistance and genotype of Escherichia coli in Henan[J]. Journal of Northwest A&F University(Nat. Sci. Ed.), 2018,47(4): 1-7.
[13] 王学君,谭艾娟,吕世明,等. 贵州省猪源大肠杆菌对四环素类药物的耐药性及耐药基因检测[J].中国畜牧兽医,2018,45(5):1367-1373. Wang X J, Tan A J, Lv S M, et al. Detection of Resistance and Resistance Genes of Swine Escherichia coli in Guizhou to Tetracyclines[J]. China Animal Husbandry&Veterinary Medicine,2018,45(5):1367-1373.
[14] 颜友荣,方向红,王琳琳. 苏中地区猪大肠杆菌的耐药性与耐药基因的试验[J].中国兽医杂志,2018,52(2) : 90-92. Yan Y R, Fang X H, Wang L L. Drug Resistance and Drug Resistance Genes in Central of Jiangsu [J]. Chinese Journal of Veterinary Medicine,2018,52(2): 90-92.
[15] Wu S1, Dalsgaard A, Hammerum AM, et al. Prevalence and characterization of plasmids carrying sulfonamide resistance genes among Escherichia coli from pigs, pig carcasses and human [J]. Acta Vet Scand. 2010, 30 (6):1-7.
[16] Margarita Trobos, Henrik Christensen, Marianne Sunde, et al. Characterization of sulphonamide-resistant Escherichia coli using comparison of sul2 gene sequences and multilocus sequence typing[J] Microbiology, 2009,155:831-836.
[17] Hadis Arabi. IrajPakzad. Ayat Nasrollahi. Sulfonamide Resistance Genes (sul) M in Extended Spectrum Beta Lactamase (ESBL) and Non-ESBL Producing Escherichia coli Isolated From Iranian Hospitals [J]. Jundishapur J Microbiol, 2015 , 8(7): e19961.
[18] 陆彦,晏志勋,赵红玉,等. 印第安纳沙门氏菌对氯霉素类药物耐药性分析[J].微生物学通报,2013,40(7):1225-1230. Lu Y, Yan Z X, Zhao H Y , et al. Resistance analysis of Salmonella Indiana to the chlmphenicols[J].Microbiology China, 2013,40(7):1225-1230.
[19] Moore J E, Watabe M, Millar B C, et al. Molecular characterization of verocytoxigenic Escherichia coli O157:H7 by randomamplification of polymorphic DNA (RAPD) typing[J]. Br J Biomed Sci, 2008, 65(3): 161-163.
[20] 陈荣忠,向蓉,莫和国,等. 社区获得性耐甲氧西林金黄色葡萄球菌RAPD基因分型及药敏分析[J].中国抗生素杂志,2018, 43(5): 530-534. Chen R Z, Xiang R, Mo H G, et al. RAPD genotyping and drug sensitivity analysis of community acquired methicillin-resistant Staphylococcus[J]. Chinese Journal of Antibiotics,2018, 43(5): 530-534.
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