植物乳杆菌HNU082抗生素抗性与相关基因的关系
|
摘要
以一株分离自海南鱼茶的植物乳杆菌(Lactobacillus plantarum) HNU082为研究对象,经全基因组测序获知该菌株所携带的抗生素抗性基因信息(arlR、emeA、fexA、lmrC、lmrD、macB、mepA、mprF、novA、pbp1a、gyrB、rpoB 9类),并针对11种常用抗生素进行抗性筛选,得到各抗生素对HNU082的最小抑制浓度(Minimum Inhibitory Concentrations,MIC),从而确定其抗性基因与抗生素抗性间的关系。结果表明,并非所有抗性基因都能导致菌株对相应抗生素产生抗性:菌株检出rpoB却未表达利福平抗性;抗性程度与抗性基因所占比例大小无关:与万古霉素和诺氟沙星相关的抗性基因分别占所检测基因总数的6.7%和20.0%,但菌株对二者所表达的抗性均为MIC> 1280μg/mL,而盐酸克林霉素相关基因占所检测基因的20.0%,其MIC(20μg/mL)却与诺氟沙星相差很大;红霉素和罗红霉素的相关抗性基因均为mac B,但菌株对罗红霉素的抗性强于红霉素,即对受同一基因影响的不同药物,菌株所表现的抗性特征也不尽相同。
Taking Lactobacillus plantarum HNU082 isolated from traditional fermented Yucha of Hainan,the antibiotic resistance gene information carried by the strain was interpreted by whole genome sequencing (arlR,emeA,fexA,lmrC,lmrD,macB,mepA,mprF,novA,pbp1a,gyrB,and rpoB). Based on this,resistance screening was carried out for 11 commonly used antibiotics.Thus the minimum inhibitory concentrations (MIC) of the 11 antibiotics against HNU082 were obtained to determine the relationship between the resistance gene and antibiotic resistance under specific experimental conditions.The results showed that not all resistance genes could cause strains to develop resistance to the corresponding antibiotics. For example,rpoB was detected from the strain,but didn't express rifampicin resistance.The degree of resistance was independent on the proportion of resistance genes.Vancomycin and norfloxacin-related genes respectively accounted for 6.7% and 20.0% of the total detected genes,but the resistance of strains to both genes was MIC > 1280 μg/mL. While clindamycin hydrochloride-related genes accounted for 20.0% of the detected genes,the MIC (20 μg/mL) was quite different from that of norfloxacin. The related resistance genes of erythromycin and roxithromycin were mac B,but the resistance of the strain to roxithromycin was stronger than that of erythromycin.So different drugs affected by the same gene would have different resistance characteristics.
Taking Lactobacillus plantarum HNU082 isolated from traditional fermented Yucha of Hainan,the antibiotic resistance gene information carried by the strain was interpreted by whole genome sequencing (arlR,emeA,fexA,lmrC,lmrD,macB,mepA,mprF,novA,pbp1a,gyrB,and rpoB). Based on this,resistance screening was carried out for 11 commonly used antibiotics.Thus the minimum inhibitory concentrations (MIC) of the 11 antibiotics against HNU082 were obtained to determine the relationship between the resistance gene and antibiotic resistance under specific experimental conditions.The results showed that not all resistance genes could cause strains to develop resistance to the corresponding antibiotics. For example,rpoB was detected from the strain,but didn't express rifampicin resistance.The degree of resistance was independent on the proportion of resistance genes.Vancomycin and norfloxacin-related genes respectively accounted for 6.7% and 20.0% of the total detected genes,but the resistance of strains to both genes was MIC > 1280 μg/mL. While clindamycin hydrochloride-related genes accounted for 20.0% of the detected genes,the MIC (20 μg/mL) was quite different from that of norfloxacin. The related resistance genes of erythromycin and roxithromycin were mac B,but the resistance of the strain to roxithromycin was stronger than that of erythromycin.So different drugs affected by the same gene would have different resistance characteristics.
引文
[1]Brandt K K,Amézquita A,Backhaus T,et al.Ecotoxicological assessment of antibiotics:A call for improved consideration of microorganisms[J].Environment International,2015(85):189-205.
[2]方媛瑗,丁惠君.抗生素的生态毒性效应研究进展[J].环境科学与技术,2018,41(5):102-110.
[3]Wellington E M H,Boxall A B A,Cross P,et al.The role of the natural environment in the emergence of antibiotic resistance in gram-negative bacteria[J].The Lancet Infectious Diseases,2013,13(2):155-165.
[4]朱永官,陈青林,苏建强,等.环境中抗生素与抗性基因组的研究[J].科学观察,2017,12(6):60-62.
[5]董祎,张万筠,鲍晨宁,等.抗生素抗性基因在环境中的来源与传播[J].辽宁化工,2018,47(3):253-255.
[6]肖纯凌.益生菌-后抗生素时代的曙光[J].沈阳医学院学报,2017,19(6):461-465.
[7]Levy S B.Antibiotic resistance:An ecological imbalance[J].Ciba Foundation Symposium,1997,207(207):1-9.
[8]Lu J Y,Tian Z,Yu J W,et al.Distribution and abundance of antibiotic resistance genes in sand settling reservoirs and drinking water treatment plants across the Yellow River,China[J].Water,2018,10(3):1-12.
[9]俞慎,王敏,洪有为.环境介质中的抗生素及其微生物生态效应[J].生态学报,2011,31(15):4437-4446.
[10]Hizlisoy H,Onmaz N E,Karadal F,et al.Antibiotic resistance gene profiles of Staphylococcus aureus isolated from foods of animal origin[J].Kafkas Universitesi Veteriner Fakultes Dergisi,2018,24(2):243-249.
[11]李忠玲,李本光,岳淑宁,等.常用抗生素对益生菌存活率影响的研究[J].中国酿造,2012,31(1):61-64.
[12]Zhang Z F,Zhou T X,Ao X,et al.Effects ofβ-glucan and Bacillus subtilis on growth performance,blood profiles,relative organ weight and meat quality in broilers fed maize-soybean mea based diets[J].Livestock Science,2012,150(1-3):419-424.
[13]Johnston B,Supina A,Vohra S.Probiotics for pediatric antibiotic-associated diarrhea:A meta-analysis of randomized placebo-controlled trials[J].Canadian Medical Association Journal,2006,175(4):377-383.
[14]Jicheng W,Xiao D,Wenyi Z,et al.Genome adaptive evolution of Lactobacillus casei under long-term antibiotic selection pressures[J].BMC Genomics,2017(18):1-8.
[15]Netherwood T,Bowden R,Harrison P,et al.Gene transfer in the gastrointestinal tract[J].Applied and Environmenta Microbiology,1999,65(11):5139-5141.
[16]Shalini M,Rameshwar S.Antibiotic resistance in food lactic acid bacteria:A review[J].International Journal of Food Microbiology,2005,105(3):281-295.
[17]Rangasamy K,Murugan A,Natrajan D,et al.Cloning and expression of the organophosphate pesticide-degrading[J].Biomed Research International,2018(17),doi:10.1155/2018/1535209.
[18]Wassenaar T M,Klein G.Safety aspects and implications o regulation of probiotic bacteria in food and food supplements[J].Journal of Food Protection,2008,71(8):1734-1741.
[19]曹承旭,郭晶晶,乌日娜,等.植物乳杆菌的生理功能和组学研究进展[J].乳业科学与技术,2018,41(1):33-39.
[20]Shao Y Y,Huo D X,Peng Q N,et al.Lactobacillus plantarum HNU082-derived improvements in the intestinal microbiome prevent the development of hyperlipidaemia[J].Food&Function,2017,8(12):4508-4516.
[21]Guo X Y,Yan Z,Wang N,et al.Behavior of antibiotic resistance genes under extremely high-level antibiotic selection pressures in pharmaceutical wastewater treatment plants[J].Science of the Total Environment,2018(612):119-128.
[22]王小茹,徐传标,彭湘文,等.海南黎族鱼茶中微生物多样性分析[J].食品科学,2017,38(6):136-141.
[23]李秀银,王月兵,李明川.抗生素的分类及治疗原则[J].临床合理用药杂志,2012,5(28):12.
[24]Gillings M,Paulsen I,Tetu S.Genomics and the evolution of antibiotic resistance[J].Annals of the New York Academy of Sciences,2017,1388(1):92-107.
[25]Liu Y,Liu K,Lai J,et al.Prevalence and antimicrobial resistance of Enterococcus species of food animal origin from Beijing and Shandong province,China[J].Journal of Applied Microbiology,2013,114(2):555-563.
[26]邵玉宇.干酪乳杆菌和植物乳杆菌耐药分子机制的研究[D].呼和浩特:内蒙古农业大学,2015.
[27]杨梅.传统发酵乳制品中乳酸菌的抗生素抗性及相关抗性基因的研究[D].呼和浩特:内蒙古农业大学,2010.
[28]宋晓敏,李少英,马春艳,等.发酵食品中乳酸菌的耐药性现状分析[J].微生物学通报,2015,42(1):207-213.
[29]周宁,张建新,樊明涛,等.细菌药物敏感性实验方法研究进展[J].食品工业科技,2012,33(9):459-464.
[30]韦婉,李少英,王梦姣,等.益生乳酸菌对氟喹诺酮类药物的敏感性检测方法的研究[J].食品科技,2013,38(3):22-26.
[2]方媛瑗,丁惠君.抗生素的生态毒性效应研究进展[J].环境科学与技术,2018,41(5):102-110.
[3]Wellington E M H,Boxall A B A,Cross P,et al.The role of the natural environment in the emergence of antibiotic resistance in gram-negative bacteria[J].The Lancet Infectious Diseases,2013,13(2):155-165.
[4]朱永官,陈青林,苏建强,等.环境中抗生素与抗性基因组的研究[J].科学观察,2017,12(6):60-62.
[5]董祎,张万筠,鲍晨宁,等.抗生素抗性基因在环境中的来源与传播[J].辽宁化工,2018,47(3):253-255.
[6]肖纯凌.益生菌-后抗生素时代的曙光[J].沈阳医学院学报,2017,19(6):461-465.
[7]Levy S B.Antibiotic resistance:An ecological imbalance[J].Ciba Foundation Symposium,1997,207(207):1-9.
[8]Lu J Y,Tian Z,Yu J W,et al.Distribution and abundance of antibiotic resistance genes in sand settling reservoirs and drinking water treatment plants across the Yellow River,China[J].Water,2018,10(3):1-12.
[9]俞慎,王敏,洪有为.环境介质中的抗生素及其微生物生态效应[J].生态学报,2011,31(15):4437-4446.
[10]Hizlisoy H,Onmaz N E,Karadal F,et al.Antibiotic resistance gene profiles of Staphylococcus aureus isolated from foods of animal origin[J].Kafkas Universitesi Veteriner Fakultes Dergisi,2018,24(2):243-249.
[11]李忠玲,李本光,岳淑宁,等.常用抗生素对益生菌存活率影响的研究[J].中国酿造,2012,31(1):61-64.
[12]Zhang Z F,Zhou T X,Ao X,et al.Effects ofβ-glucan and Bacillus subtilis on growth performance,blood profiles,relative organ weight and meat quality in broilers fed maize-soybean mea based diets[J].Livestock Science,2012,150(1-3):419-424.
[13]Johnston B,Supina A,Vohra S.Probiotics for pediatric antibiotic-associated diarrhea:A meta-analysis of randomized placebo-controlled trials[J].Canadian Medical Association Journal,2006,175(4):377-383.
[14]Jicheng W,Xiao D,Wenyi Z,et al.Genome adaptive evolution of Lactobacillus casei under long-term antibiotic selection pressures[J].BMC Genomics,2017(18):1-8.
[15]Netherwood T,Bowden R,Harrison P,et al.Gene transfer in the gastrointestinal tract[J].Applied and Environmenta Microbiology,1999,65(11):5139-5141.
[16]Shalini M,Rameshwar S.Antibiotic resistance in food lactic acid bacteria:A review[J].International Journal of Food Microbiology,2005,105(3):281-295.
[17]Rangasamy K,Murugan A,Natrajan D,et al.Cloning and expression of the organophosphate pesticide-degrading[J].Biomed Research International,2018(17),doi:10.1155/2018/1535209.
[18]Wassenaar T M,Klein G.Safety aspects and implications o regulation of probiotic bacteria in food and food supplements[J].Journal of Food Protection,2008,71(8):1734-1741.
[19]曹承旭,郭晶晶,乌日娜,等.植物乳杆菌的生理功能和组学研究进展[J].乳业科学与技术,2018,41(1):33-39.
[20]Shao Y Y,Huo D X,Peng Q N,et al.Lactobacillus plantarum HNU082-derived improvements in the intestinal microbiome prevent the development of hyperlipidaemia[J].Food&Function,2017,8(12):4508-4516.
[21]Guo X Y,Yan Z,Wang N,et al.Behavior of antibiotic resistance genes under extremely high-level antibiotic selection pressures in pharmaceutical wastewater treatment plants[J].Science of the Total Environment,2018(612):119-128.
[22]王小茹,徐传标,彭湘文,等.海南黎族鱼茶中微生物多样性分析[J].食品科学,2017,38(6):136-141.
[23]李秀银,王月兵,李明川.抗生素的分类及治疗原则[J].临床合理用药杂志,2012,5(28):12.
[24]Gillings M,Paulsen I,Tetu S.Genomics and the evolution of antibiotic resistance[J].Annals of the New York Academy of Sciences,2017,1388(1):92-107.
[25]Liu Y,Liu K,Lai J,et al.Prevalence and antimicrobial resistance of Enterococcus species of food animal origin from Beijing and Shandong province,China[J].Journal of Applied Microbiology,2013,114(2):555-563.
[26]邵玉宇.干酪乳杆菌和植物乳杆菌耐药分子机制的研究[D].呼和浩特:内蒙古农业大学,2015.
[27]杨梅.传统发酵乳制品中乳酸菌的抗生素抗性及相关抗性基因的研究[D].呼和浩特:内蒙古农业大学,2010.
[28]宋晓敏,李少英,马春艳,等.发酵食品中乳酸菌的耐药性现状分析[J].微生物学通报,2015,42(1):207-213.
[29]周宁,张建新,樊明涛,等.细菌药物敏感性实验方法研究进展[J].食品工业科技,2012,33(9):459-464.
[30]韦婉,李少英,王梦姣,等.益生乳酸菌对氟喹诺酮类药物的敏感性检测方法的研究[J].食品科技,2013,38(3):22-26.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |