中药与抗菌药物联用对猪源大肠杆菌耐药性的抑制作用
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摘要
为了探讨中药与抗菌药物联用对大肠杆菌耐药性的作用效果,本研究对河南部分地区采集的样品(50份)进行分离鉴定、生化试验及致病性试验,对分离鉴定后的致病性猪大肠杆菌选用12种中药采用K-B法进行耐药性检测,将筛选出来的中药与抗生素进行中西联用对致病性大肠杆菌做体外抑菌试验,并分析中西药联用对大肠杆菌的抑制作用。结果显示,采集的50份样品有23株菌符合大肠杆菌的分离培养特性和生化特性,且23株菌均为致病性大肠杆菌;通过耐药性检测,23株试验菌株对测试的中药均有不同程度的耐药性,其中试验菌株对甘草耐药率最高(82.6%),其次为生地(78.2%),而对黄柏(17.3%)和白头翁(21.7%)耐药性较低,具有一定的敏感性;体外抑菌试验结果发现,中药与抗生素联用均有不同程度的抑菌效果,其中黄柏、白头翁与抗菌药物联用的抑菌效果最明显。以上结果说明中药与抗菌药物联用不仅可以抑制大肠杆菌的耐药性,还可以延缓大肠杆菌耐药性的产生。
This study was aimed to discuss the effect of drug resistance of Escherichia coli(E.coli)by combining traditional Chinese medicine and antibiotics drugs.50 samples which collected from Henan province were isolated for identification,biochemical experiments and pathogenicity test.12 kinds of traditional Chinese medicine were selected for isolation and identification of pathogenic porcine E.coli,and the K-B method was used to detect the drug resistance.The screened traditional Chinese medicines and antibiotics were used to conduct in vitro antibacterial tests against pathogenic E.coli,and analyze the inhibitory effect of Chinese and Western drugs on E.coli.The results showed that 23 strains of 50 samples were consistent with the isolation and culture characteristics and biochemical characteristics of E.coli.23 strains of bacteria were pathogenic Escherichia coli.Through the drug resistance test,23 test strains had different degrees of resistance to the tested traditional Chinese medicines.The resistance rate of the tested strains tolicorice was the highest(82.6%),followed by the birthplace(78.2%),and the resistance to cork(17.3%)and pulsatilla(21.7%)were lower,with certain sensitivity.In vitro antibacterial result showed that the combination of traditional Chinese medicine and antibiotics had different degrees of antibacterial effect,and the antibacterial effect of cork,pulsatilla chinensis combined with antibiotics was the most obvious.It showed that the combination of traditional Chinese medicine and antibiotics could not only inhibit the drug resistance of E.coli,but also delay the emergence of drug resistance of E.coli.
This study was aimed to discuss the effect of drug resistance of Escherichia coli(E.coli)by combining traditional Chinese medicine and antibiotics drugs.50 samples which collected from Henan province were isolated for identification,biochemical experiments and pathogenicity test.12 kinds of traditional Chinese medicine were selected for isolation and identification of pathogenic porcine E.coli,and the K-B method was used to detect the drug resistance.The screened traditional Chinese medicines and antibiotics were used to conduct in vitro antibacterial tests against pathogenic E.coli,and analyze the inhibitory effect of Chinese and Western drugs on E.coli.The results showed that 23 strains of 50 samples were consistent with the isolation and culture characteristics and biochemical characteristics of E.coli.23 strains of bacteria were pathogenic Escherichia coli.Through the drug resistance test,23 test strains had different degrees of resistance to the tested traditional Chinese medicines.The resistance rate of the tested strains tolicorice was the highest(82.6%),followed by the birthplace(78.2%),and the resistance to cork(17.3%)and pulsatilla(21.7%)were lower,with certain sensitivity.In vitro antibacterial result showed that the combination of traditional Chinese medicine and antibiotics had different degrees of antibacterial effect,and the antibacterial effect of cork,pulsatilla chinensis combined with antibiotics was the most obvious.It showed that the combination of traditional Chinese medicine and antibiotics could not only inhibit the drug resistance of E.coli,but also delay the emergence of drug resistance of E.coli.
引文
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[18]杜居纯,舒刚,马驰,等.蛇莓水提物对临床菌株质粒消除作用及生物膜形成的影响[J].中国兽医学报,2014,34(2):264-268.DU J C,SHU G,MA C,et al.Effect of Aconitum kusnezoffii on the plasmid elimination and biofilm formation of clinical strain[J].Chinese Journal of Veterinary Science,2014,34(2):264-268.(in Chinese)
[19]李延荣,陈伟.新疆特色植物对金黄色葡萄球菌生物膜形成的影响[J].中国兽医学报,2014,34(8):1253-1256.LI Y R,CHEN W.Effect of Xinjiang species on Staphylococcus aureus biofilm formation[J].Chinese Journal of Veterinary Science,2014,34(8):1253-1256.(in Chinese)
[20]王亚男,柳秉润,邓旭明,等.芦丁对金黄色葡萄球菌Sortase A的抑制作用[J].吉林农业大学学报,2013,35(3):303-307.WANG Y N,LIU B R,DENG X M,et al.Inhibitory effect of rutin on Staphylococcus aureus Sortase A[J].Journal of Jilin Agricultural University,2013,35(3):303-307.(in Chinese)
[21]郭向华,郭润华,宋志军,等.苦参碱对慢性铜绿假单胞菌生物膜肺部感染大鼠的免疫保护作用[J].中国实验方剂学杂志,2010,16(8):185-188.GUO X H,GUO R H,SONG Z J,et al.Immunoprotective effect of matrine on chronic pulmonary Pseudomonas aeruginosa lung infection in rats[J].Chinese Journal of Experimental Traditional Formulae,2010,16(8):185-188.(in Chinese)
[22]王丽华,吕殿红,张苓,等.质粒介导的喹诺酮耐药基因在动物源金黄色葡萄球菌的流行分布[J].中国兽医学报,2014,34(4):606-612.WANG L H,LV D H,ZHANG L,et al.The prevalence distribution of plasmid-mediated quinolone resistance genes in animal origin of Staphylococcus aureus[J].Chinese Journal of Veterinary Science,2014,34(4):606-612.(in Chinese)
[23]张赛奇,王米,杨锐乐,等.15种中药对猪大肠杆菌的体外抑菌试验[J].黑龙江畜牧兽医,2015,3:163-165.ZHANG S Q,WANG M,YANG R L,et al.In vitro bacteriostasis test of 15 kinds of Chinese herbs on Escherichia coli in pigs[J].Heilongjiang Animal Science and Veterinary Medicine,2015,3:163-165.
[24]YANG Z C,WANG B C,YANG X S,et al.The synergtic activity of antibiotics combined with eight traditional Chinese medicines against two different strains of Staphylococcus aureus[J].Colloids and Surfaces B Biointerfaces,2005,41(2-3):79-81.
[25]文英.中西药对大肠埃希菌的体外联合抑菌作用[J].黑龙江畜牧兽医,2014,5:145-146.WEN Y.In vitro combined antibacterial effect of Chinese and Western drugs on Escherichia coli[J].Heilongjiang Animal Science and Veterinary Medicine,2014,5:145-146.(in Chinese)
[26]司红彬,吴永继,黄丽云,等.黄柏与抗菌药联合抗耐药大肠杆菌的作用研究[J].河南农业科学,2015,44(2):123-126.SI H B,WU Y J,HUANG L Y,et al.Study on the effect of cypress and antibacterials on resistance to resistant Escherichia coli[J].Journal of Henan Agricultural Sciences,2015,44(2):123-126.(in Chinese)
[27]时维静,路振香,李立顺.白头翁不同提取物及复方体外抑菌作用的实验研究[J].中国中医药科技,2006,13(3):166-168.SHI W J,LU Z X,LI L S.Experimental study on bacteriostasis of extracts and compounds in vitro[J].Chinese Journal of Traditional Medical Science and Technology,2006,13(3):166-168.(in Chinese)
[2]LOOFT T,JOHNSON T A,ALLEN H K,et al.In-feed antibiotic effects on the swine intestinal microbiome[J].Proceedings of the National Academy of Sciences of the United States of America,2012,109(5):1691-1696.
[3]李晨,张双翔,周碧君,等.腹泻仔猪肠道致病性大肠埃希氏菌和沙门氏菌的分离鉴定与耐药性研究[J].中国畜牧兽医,2017,44(12):3625-3633.LI C,ZHANG S X,ZHOU B J,et al.Isolation,identification and drug resistance of enteric pathogenic Escherichia coli and Salmonellain diarrhoea piglets[J].China Animal Husbandry&Veterinary Medicine,2017,44(12):3625-3633.(in Chinese)
[4]肖莉春,孔祥峰,郭小权.中药抑菌成分及其在养猪生产中的应用前景[J].动物营养学报,2012,24(12):2311-2318.XIAO L C,KONG X F,GUO X Q.Bacteriostasis components of Chinese medicine and its application in pig production[J].Chinese Journal of Animal Nutrition,2012,24(12):2311-2318.(in Chinese)
[5]宁官保,牛艺儒,张鼎,等.鸡源大肠杆菌耐药性分析及中药对大肠杆菌耐药性消除作用的研究[J].畜牧兽医学报,2015,46(6):1018-1025.NING G B,NIU Y R,ZHANG D,et al.Analysis of drug resistance of Escherichia coli from chicken source and study on the effect of traditional Chinese medicine on the elimination of Escherichia coli resistance[J].Chinese Journal of Animal and Veterinary Medicine,2015,46(6):1018-1025.(in Chinese)
[6]HO T Y,LO H Y,LI C C,et al.In vitro and in vivo bioluminescent imaging to evaluate anti-Escherichia coli activity of Galla chinensis[J].BioMedicine,2013,3(4):160-166.
[7]黄名钱,孔祥峰,肖莉春,等.中药水提物对猪源耐四环素大肠杆菌的抑制活性[J].中国农业科学,2013,46(11):2370-2376.HUANG M Q,KONG X F,XIAO L C,et al.Inhibitory activity of Chinese medicine water extract on tetracycline resistant Escherichia coli in pig source[J].Scientia Agricultura Sinica,2013,46(11):2370-2376.(in Chinese)
[8]肖莉春,孔祥峰,黄名钱,等.中药提取物对猪源耐药大肠杆菌分离株的抑制作用[J].江西农业大学学报,2013,35(6):1248-1254.XIAO L C,KONG X F,HUANG M Q,et al.Inhibitory effect of Chinese herbal extracts on porcine derived Escherichia coli isolates[J].Acta Agriculturae Universitatis Jiangxiensis,2013,35(6):1248-1254.(in Chinese)
[9]WALLMANN J.Antimicrobial resistance:Challenges ahead[J].Veterinary Record,2014,175(13):323-324.
[10]黄中利,宋敏训,冯敏燕,等.中药体内抗大肠杆菌效果与药性关系研究[J].山东农业科学,2015,47(7):101-103.HUANG Z L,SONG M X,FENG M Y,et al.Study on the relationship between the effect of anti-Escherichia coli and drug resistance in Chinese medicine[J].Shandong Agricultural Sciences,2015,47(7):101-103.(in Chinese)
[11]王基伟,孙洋,纪雪,等.长春地区猪源大肠杆菌的分离鉴定和耐药性分析[J].中国兽药志,2014,48(11):14-18.WANG J W,SUN Y,JI X,et al.Isolation,identification and drug resistance analysis of Escherichia coli from swine in Changchun region[J].Chinese Journal of Veterinary Drug,2014,48(11):14-18.(in Chinese)
[12]毛福超,郁川,韩璐,等.豫西地区禽源大肠埃希氏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 avian Escherichia coli from poultry in West Henan province[J].Journal of Henan Agricultural Sciences,2016,45(1):127-130.
[13]刘哲,孙洋,纪雪,等.仔猪腹泻大肠埃希氏菌流行株鉴定及毒力与耐药性分析[J].中国畜牧兽医,2017,44(4):1226-1232.LIU Z,SUN Y,JI X,et al.Identification of Escherichia coli strains strain in piglets with diarrhea and analysis of toxicity and drug resistance[J].China Journal of Animal Husbandry&Veterinary Medicine,2017,44(4):1226-1232.(in Chinese)
[14]徐小艳,王国相,印继华,等.1975—2002年大肠杆菌耐药性变化情况调查[J].畜牧与兽医,2003,35(3):16-17.XU X Y,WANG G X,YIN J H,et al.Investigation of the change of Escherichia coli drug resistance in1975-2002[J].Animal Husbanry and Veterinary Medicine,2003,35(3):16-17.(in Chinese)
[15]张文平,刘冬华,闵梦双,等.金银花水浸液对大肠埃希菌R质粒的消除作用[A].中华医学会2014全国微生物学与免疫学学术年会论文汇编[C].成都:中华医学会,2014.ZHANG W P,LIU D H,MIN M S,et al.Elimination effect of Honeysuckle aqueous extract on Escherichia coli R plasmid[A].Chinese Medical Association 2014National Conference on Microbiology and Immunology Annual Meeting Compilation[C].Chengdu:Chinese Medical Association,2014.(in Chinese)
[16]李娟,张学顺,傅春升.中药抗菌作用的研究进展[J].中国药业,2014,23(2):90-92.LI J,ZHANG X S,FU C S.Research progress of antibacterial effect of Chinese medicine[J].China Pharmaceuticals,2014,23(2):90-92.(in Chinese)
[17]ZHOU X Z,JIA F,LIU X M,et al.Total alkaloids of Sophorea alopecuroides induced down regulation of AcrAB-TolC efflux pump reverses susceptibility to ciprofloxacin in clinical multidrug resistant Escherichia coli isolates[J].Phytotherapy Research,2012,26(11):1637-1643.
[18]杜居纯,舒刚,马驰,等.蛇莓水提物对临床菌株质粒消除作用及生物膜形成的影响[J].中国兽医学报,2014,34(2):264-268.DU J C,SHU G,MA C,et al.Effect of Aconitum kusnezoffii on the plasmid elimination and biofilm formation of clinical strain[J].Chinese Journal of Veterinary Science,2014,34(2):264-268.(in Chinese)
[19]李延荣,陈伟.新疆特色植物对金黄色葡萄球菌生物膜形成的影响[J].中国兽医学报,2014,34(8):1253-1256.LI Y R,CHEN W.Effect of Xinjiang species on Staphylococcus aureus biofilm formation[J].Chinese Journal of Veterinary Science,2014,34(8):1253-1256.(in Chinese)
[20]王亚男,柳秉润,邓旭明,等.芦丁对金黄色葡萄球菌Sortase A的抑制作用[J].吉林农业大学学报,2013,35(3):303-307.WANG Y N,LIU B R,DENG X M,et al.Inhibitory effect of rutin on Staphylococcus aureus Sortase A[J].Journal of Jilin Agricultural University,2013,35(3):303-307.(in Chinese)
[21]郭向华,郭润华,宋志军,等.苦参碱对慢性铜绿假单胞菌生物膜肺部感染大鼠的免疫保护作用[J].中国实验方剂学杂志,2010,16(8):185-188.GUO X H,GUO R H,SONG Z J,et al.Immunoprotective effect of matrine on chronic pulmonary Pseudomonas aeruginosa lung infection in rats[J].Chinese Journal of Experimental Traditional Formulae,2010,16(8):185-188.(in Chinese)
[22]王丽华,吕殿红,张苓,等.质粒介导的喹诺酮耐药基因在动物源金黄色葡萄球菌的流行分布[J].中国兽医学报,2014,34(4):606-612.WANG L H,LV D H,ZHANG L,et al.The prevalence distribution of plasmid-mediated quinolone resistance genes in animal origin of Staphylococcus aureus[J].Chinese Journal of Veterinary Science,2014,34(4):606-612.(in Chinese)
[23]张赛奇,王米,杨锐乐,等.15种中药对猪大肠杆菌的体外抑菌试验[J].黑龙江畜牧兽医,2015,3:163-165.ZHANG S Q,WANG M,YANG R L,et al.In vitro bacteriostasis test of 15 kinds of Chinese herbs on Escherichia coli in pigs[J].Heilongjiang Animal Science and Veterinary Medicine,2015,3:163-165.
[24]YANG Z C,WANG B C,YANG X S,et al.The synergtic activity of antibiotics combined with eight traditional Chinese medicines against two different strains of Staphylococcus aureus[J].Colloids and Surfaces B Biointerfaces,2005,41(2-3):79-81.
[25]文英.中西药对大肠埃希菌的体外联合抑菌作用[J].黑龙江畜牧兽医,2014,5:145-146.WEN Y.In vitro combined antibacterial effect of Chinese and Western drugs on Escherichia coli[J].Heilongjiang Animal Science and Veterinary Medicine,2014,5:145-146.(in Chinese)
[26]司红彬,吴永继,黄丽云,等.黄柏与抗菌药联合抗耐药大肠杆菌的作用研究[J].河南农业科学,2015,44(2):123-126.SI H B,WU Y J,HUANG L Y,et al.Study on the effect of cypress and antibacterials on resistance to resistant Escherichia coli[J].Journal of Henan Agricultural Sciences,2015,44(2):123-126.(in Chinese)
[27]时维静,路振香,李立顺.白头翁不同提取物及复方体外抑菌作用的实验研究[J].中国中医药科技,2006,13(3):166-168.SHI W J,LU Z X,LI L S.Experimental study on bacteriostasis of extracts and compounds in vitro[J].Chinese Journal of Traditional Medical Science and Technology,2006,13(3):166-168.(in Chinese)