东江下游典型饮用水源地抗生素抗性基因分布研究
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摘要
抗生素抗性基因(AntibioticsResistanceGenes,ARGs)是环境中的一类新型污染物。为了解东江下游地区水源地中ARGs的污染水平及其影响因素,采用实时荧光定量PCR技术,对东江下游地区9个河流型饮用水源地和5个湖泊型饮用水源地8种ARGs[sul1、sul2、sul3、tet(M)、tet(O)、tet(Q)、tet(G)、ermB]的绝对丰度进行检测分析。结果表明:东江下游饮用水源地总ARGs绝对丰度水平偏低,ARGs总丰度范围为2.37×10~7-4.80×10~8 copies?L~(-1),其中抗性基因sul1的丰度相对较高,这可能与磺胺类药品是常用药品有关。磺胺类ARGs(sul1、sul2)在所有饮用水源地中均有检出,而sul3的检出率为85.7%;四环素类ARGs[tet(M)、tet(O)、tet(Q)、tet(G)]的检出率较高,为64.3%-100%;大环内酯类ARGs检出率最低。河流型水源地上游点位ARGs绝对丰度为1.03×108 copies?L~(-1),下游点位绝对丰度为2.89×10~8 copies?L~(-1),因此河流型饮用水源地的ARGs绝对丰度水平随着河流方向呈现逐渐升高趋势。除了一处曾为水产养殖的备用湖泊型水源地外,河流型水源地除sul1外的抗性基因丰度(3.25×10~5 copies?L~(-1))明显高于湖泊型饮用水源地(1.14×10~5 copies?L~(-1))。通过对东江下游典型饮用水源地ARGs进行研究,为该地区ARGs污染现状提供了基础数据,也为水环境中ARGs污染整治提供依据。
Antibiotics Resistance Genes(ARGs) is a new type of pollutants in the environment. In order to understand the pollution level of ARGs and its influencing factors in the water sources of the lower Dongjiang River, a real-time fluorescent quantitative PCR technique was used in this study. The absolute abundance of 8 kinds of ARGs [sul1, sul2, sul3, tet(M), tet(O), tet(Q), tet(G), ermB] in9 river-type drinking water sources and 5 lake-type drinking water sources in the lower reaches of Dongjiang River were detected and analyzed. The results showed that the absolute abundance of total ARGs was low in drinking water sources of the lower Dongjiang River, and the total abundance of ARGs was in the range of 2.37×10~7-4.80×10~8 copies?L~(-1), in which the abundance of resistant gene sul1 was higher than that of other resistant genes, which may be related to the common use of sulfonamides.Sulfonamides ARGs(sul1, sul2) was detected in all drinking water sources, while sul3 was detected in the rate of 85.7%, the detection rate of tetracycline ARGs [tet(M), tet(O), tet(Q), tet(G)] was high(64.3%-100%) and macrolide ARGs was the lowest. The absolute abundance of ARGs was 1.03×10~8 copies?L~(-1) at upstream and 2.89×10~8 copies?L~(-1) at downstream of river-type water source.Therefore, the absolute abundance level of ARGs in river-type drinking water sources gradually increased with the river's direction.The average ARGs abundance of river-type drinking water source was 3.25×10~5 copies?L~(-1), and that of lake-type drinking water sources was 1.14×10~5 copies?L~(-1) when sul1 was excluded, besides the highest was 1.41×10~6 copies?L~(-1), which was used for aquaculture before becoming a reserve water source. Therefore, ARGs abundance of river-type water source was higher than that of lake-type drinking water source on the whole. This study of ARGs in a typical drinking water source in the lower Dongjiang River not only provides the basic data for the present study of ARGs pollution in this area, but also support the management and regulation of ARGs pollution in the water environment.
Antibiotics Resistance Genes(ARGs) is a new type of pollutants in the environment. In order to understand the pollution level of ARGs and its influencing factors in the water sources of the lower Dongjiang River, a real-time fluorescent quantitative PCR technique was used in this study. The absolute abundance of 8 kinds of ARGs [sul1, sul2, sul3, tet(M), tet(O), tet(Q), tet(G), ermB] in9 river-type drinking water sources and 5 lake-type drinking water sources in the lower reaches of Dongjiang River were detected and analyzed. The results showed that the absolute abundance of total ARGs was low in drinking water sources of the lower Dongjiang River, and the total abundance of ARGs was in the range of 2.37×10~7-4.80×10~8 copies?L~(-1), in which the abundance of resistant gene sul1 was higher than that of other resistant genes, which may be related to the common use of sulfonamides.Sulfonamides ARGs(sul1, sul2) was detected in all drinking water sources, while sul3 was detected in the rate of 85.7%, the detection rate of tetracycline ARGs [tet(M), tet(O), tet(Q), tet(G)] was high(64.3%-100%) and macrolide ARGs was the lowest. The absolute abundance of ARGs was 1.03×10~8 copies?L~(-1) at upstream and 2.89×10~8 copies?L~(-1) at downstream of river-type water source.Therefore, the absolute abundance level of ARGs in river-type drinking water sources gradually increased with the river's direction.The average ARGs abundance of river-type drinking water source was 3.25×10~5 copies?L~(-1), and that of lake-type drinking water sources was 1.14×10~5 copies?L~(-1) when sul1 was excluded, besides the highest was 1.41×10~6 copies?L~(-1), which was used for aquaculture before becoming a reserve water source. Therefore, ARGs abundance of river-type water source was higher than that of lake-type drinking water source on the whole. This study of ARGs in a typical drinking water source in the lower Dongjiang River not only provides the basic data for the present study of ARGs pollution in this area, but also support the management and regulation of ARGs pollution in the water environment.
引文
AMINOV R I,JEANJEAN N G,MACKI R I,2001.Molecular Ecology of Tetracycline Resistance:Development and Validation of Primers for Detection of Tetracycline Resistance Genes Encoding Ribosomal Protection Proteins[J].Applied and Environmenyal Microbiology,67(1):22-32
HOA P T P,NONAKA L,VIET P H,et al.,2008.Detection of the sul1,sul2,and sul3 genes in sulfonamide-resistant bacteria from wastewater and shrimp ponds of North Vietnam[J].Science of the Total environment,405(1-3):377-384.
INYINBOR A A,BELLO O S,FADIJI A E,et al.,2018.Threats from antibiotics:A serious environmental concern[J].Journal of environmental Chemical engineering,6(1):784-793.
JIANG L,HU X L,XU T,et al.,2013.Prevalence of antibiotic resistance genes and their relationship with antibiotics in the Huangpu River and the drinking water sources,Shanghai,China[J].Science of the Total environment,458-460:267-272.
NG L Z,MARTIN I,ALFA M,MULVEY M,2001.Multiplex PCR for the detection of tetracycline resistant gens[J].Molecular and Cellular Probes,15(4):209-215.
PEI R,KIM S C,CARLSON K H,et al.,2006.Effect of River Landscape on the sediment concentrations of antibiotics and corresponding antibiotic resistance genes(ARG)[J].WATER RESEARCH,40(12):2427-2435.
PRUDEN A,PEI R,STORTEBOOM H,et al.,2006.Antibiotic resistance genes as emerging contaminants studies in northern Colorado[J].Environmental Science and Technology,40(23):7445-7450.
SARMAH A K,Meyer M T,BOXALL A B A,2006.A global perspective on the use,sales,exposure pathways,occurrence,fate and effects of veterinary antibiotics(VAs)in the environment[J].ChemospHere,65(5):725-759.
SU H C,LIU Y S,PAN C G,et al.,2018.Persistence of antibiotic resistance genes and bacterial community changes in drinking water treatment system:From drinking water source to tap water[J].Science of the Total Environment,616-617:453-461.
ZHANG A Y,WANG H N,TIAN G B,et al.,2009.Phenotypic and genotypic characterization of antimicrobial resistance in faecal bacteria from 30 Giant pandas[J].International Journal of Antimicrobial Agents,33(5):456-460.
ZHU Y G,JOHNSON T A,SU J Q,et al.,2013.Diverse and abundant antibiotic resistance genes in Chins swine farms[J].Proceedings of the National Academy of Sciences of the United States of America,110(9):3435-3440.
程宪伟,梁银秀,于翔霏,等,2017.水体中抗生素污染及其处理技术研究进展[J].环境科学与技术,40(S1):125-132.CHENG X W,LIANG Y X,YU X F,et al.,2017.The Contamination and Treatment Technologies for Antibiotics in Water:A Review[J].Environmental Science&Technology,40(S1):125-132.
高敏,仇天雷,秦玉成,等,2017.养鸡场空气中抗性基因和条件致病菌污染特征[J].环境科学,38(2):511-516.GAO M,QIU T C,QIN Y C,et al.,2017.Sources and Pollution Characteristics of Antibiotic Resistance Genes and Conditional Pathogenic Bacteria in Concentrated Poultry Feeding Operations[J].Environmental Science,38(2):511-516.
高盼盼,罗义,周启星,等,2009.水产养殖环境中抗生素抗性基因(ARGS)的研究及进展[J].生态毒理学报,4(6):770-779.GAO P P,LUO Y,ZHOU Q X,et al.,2009.Research Advancement of Antibiotics Resistance Genes(ARGs)in Aquaculture Environment[J].Asian Journal of Ecotoxicology,4(6):770-779.
胡亚茹,姜蕾,张天阳,等,2018.华东地区某饮用水源地中磺胺类抗性基因的分布特征[J].环境科学,39(9):4222-4228.HU Y R,JIANG L,ZHANG T Y,et al.,2018.Distribution Characteristics of Sulfonamide Antibiotic Resistance Genes in a Drinking Water Source in East China[J].Environmental Science,39(9):4222-4228.
罗义,周启星,2008.抗生素抗性基因(ARGS):一种新型环境污染物[J].环境科学学报,28(8):1499-1505.LUO Y,ZHOU Q X,2008.Antibiotic resistance genes(ARGs)as emerging pollutants[J].Acta Scientiae Circumstantiae,28(8):1499-1505.
沈群辉,冀秀玲,傅淑珺,等,2012.黄浦江水域抗生素及抗性基因污染初步研究[J].生态环境学报,21(10):1717-1723.SHEN Q H,JI X L,FU S J,et al.,2012.Preliminary studies on the pollution levels of antibiotic and antibiotic resistance genes in Huangpu River,China[J].Ecology and Environmental Sciences,21(10):1717-1723.
苏怀德,1988.青霉素发现及开发简史[J].药学通报,23(8):494-497.SU H D,1988.A brief history of penicillin discovery and development[J].Chinese Pharmaceutical Journal,23(8):494-497.
杨继平,邱志刚,袁兆康,等,2017.天津海河流域抗生素抗性基因分布特征及与指示微生物的关系[J].环境与健康杂志,34(4):313-316.YANG J P,QIN Z G,YUAN Z K,et al.,2017.Distribution of antibiotic resistance genes and their relationship with indicator microorganisms in Haihe River Basin,Tianjin[J].J Environ Health,34(4):313-316.
曾冠军,柳娴,马满英,2017.水体中抗生素污染研究进展[J].安徽农业科学,45(3):72-74.ZENG G J,LIU X,MA M Y,2017.Research Progress of Antibiotic Pollution in Water[J].Journal of Anhui AgriculturaL Sciences,45(3):72-74.
张丹丹,郭亚平,任红云,等,2018.福建省敖江下游抗生素抗性基因分布特征[J].环境科学,39(6):2600-2606.ZHANG D D,GUO Y P,REN H Y,et al.,2018.Characteristics of Antibiotic Resistance Genes in Downstream Areas of the Aojiang River,Fujian Province[J].Environmental Science,39(6):2600-2606.
张俊,杨晓洪,葛峰,等,2014.长期施用四环素残留猪粪对土壤中耐药菌及抗性基因形成的影响[J].环境科学,35(6):2374-2380.ZHANG J,YANG X H,GE F,et al.,2014.Effects of Long-Term Application of Pig Manure Containing Residual Tetracycline on the Formation of Drug-Resistant Bacteria and Resistance Genes[J].Environmental Science,35(6):2374-2380.
张兰河,王佳佳,哈雪姣,等,2016.北京地区菜田土壤抗生素抗性基因的分布特征[J].环境科学,37(11):4395-4401.ZHANG L H,WANG J J,HA X J,et al.,2016.Distribution Characteristics of Antibiotic Resistance Genes in Vegetable Soils in Beijing[J].Environmental Science,37(11):4395-4401.
张瑞全,应光国,丁永祯,等,2013.广东西枝江-东江流域抗生素抗性基因污染特征研究[J].农业环境科学学报,32(12):2471-2479.ZHANG R Q,YING G G,DING Y Z,et al.,2013.Pollution Characteristics of Antibiotic Resistance Genes in Xizhijiang-Dongjiang River Basin,Guangdong Province,China[J].Journal of Agro-Environment Science,32(12):2471-2479.
赵腾辉,陈奕涵,韩巍,等,2016.东江上游典型抗生素污染特征及生态风险评价[J].生态环境学报,25(10):1707-1713.ZHAO T H,CHEN Y H,HAN W,et al.,2016.The Contamination Characteristics and Ecological Risk Assessment of Typical Antibiotics in the Upper Reaches of the Dongjiang River[J].Ecology and Environmental Sciences,25(10):1707-1713.
周婷,李鑫,王晓龙,等,2014.新疆玛纳斯河流域抗生素抗性基因的污染分析[J].石河子大学学报(自然科学版),32(6):755-759.ZHOU T,LI X,WANG X L,et al.,2014.Pollution Analysis of Antibiotic Resistant Genes of Manas River Basin,Xinjiang[J].Journal of Shihezi University(Natural Science),32(6):755-759.
赵晓祥,杨莲,程晨,等,2018.上海水体中抗生素抗性基因的分布特征[J].安全与环境学报,18(3):1191-1197.ZHAO X X,YANG L,CHENG C,et al.,2018.Distributive situation of the antibiotic resistance genes in the surface water bodies in Shanghai[J].Journal of Safety and Environment,18(3):1191-1197.
邹世春,朱春敬,贺竹梅,等,2009.北江河水中抗生素抗性基因污染初步研究[J].生态毒理学报,4(5):655-660.ZOU S C,ZHU J C,HE Z M,et al.,2009.Preliminary Studies on the Pollution Levels of Antibiotic Resistance Genes in the Water of Beijiang River,South China[J].Asian Journal of Ecotoxicology,4(5):655-660.
HOA P T P,NONAKA L,VIET P H,et al.,2008.Detection of the sul1,sul2,and sul3 genes in sulfonamide-resistant bacteria from wastewater and shrimp ponds of North Vietnam[J].Science of the Total environment,405(1-3):377-384.
INYINBOR A A,BELLO O S,FADIJI A E,et al.,2018.Threats from antibiotics:A serious environmental concern[J].Journal of environmental Chemical engineering,6(1):784-793.
JIANG L,HU X L,XU T,et al.,2013.Prevalence of antibiotic resistance genes and their relationship with antibiotics in the Huangpu River and the drinking water sources,Shanghai,China[J].Science of the Total environment,458-460:267-272.
NG L Z,MARTIN I,ALFA M,MULVEY M,2001.Multiplex PCR for the detection of tetracycline resistant gens[J].Molecular and Cellular Probes,15(4):209-215.
PEI R,KIM S C,CARLSON K H,et al.,2006.Effect of River Landscape on the sediment concentrations of antibiotics and corresponding antibiotic resistance genes(ARG)[J].WATER RESEARCH,40(12):2427-2435.
PRUDEN A,PEI R,STORTEBOOM H,et al.,2006.Antibiotic resistance genes as emerging contaminants studies in northern Colorado[J].Environmental Science and Technology,40(23):7445-7450.
SARMAH A K,Meyer M T,BOXALL A B A,2006.A global perspective on the use,sales,exposure pathways,occurrence,fate and effects of veterinary antibiotics(VAs)in the environment[J].ChemospHere,65(5):725-759.
SU H C,LIU Y S,PAN C G,et al.,2018.Persistence of antibiotic resistance genes and bacterial community changes in drinking water treatment system:From drinking water source to tap water[J].Science of the Total Environment,616-617:453-461.
ZHANG A Y,WANG H N,TIAN G B,et al.,2009.Phenotypic and genotypic characterization of antimicrobial resistance in faecal bacteria from 30 Giant pandas[J].International Journal of Antimicrobial Agents,33(5):456-460.
ZHU Y G,JOHNSON T A,SU J Q,et al.,2013.Diverse and abundant antibiotic resistance genes in Chins swine farms[J].Proceedings of the National Academy of Sciences of the United States of America,110(9):3435-3440.
程宪伟,梁银秀,于翔霏,等,2017.水体中抗生素污染及其处理技术研究进展[J].环境科学与技术,40(S1):125-132.CHENG X W,LIANG Y X,YU X F,et al.,2017.The Contamination and Treatment Technologies for Antibiotics in Water:A Review[J].Environmental Science&Technology,40(S1):125-132.
高敏,仇天雷,秦玉成,等,2017.养鸡场空气中抗性基因和条件致病菌污染特征[J].环境科学,38(2):511-516.GAO M,QIU T C,QIN Y C,et al.,2017.Sources and Pollution Characteristics of Antibiotic Resistance Genes and Conditional Pathogenic Bacteria in Concentrated Poultry Feeding Operations[J].Environmental Science,38(2):511-516.
高盼盼,罗义,周启星,等,2009.水产养殖环境中抗生素抗性基因(ARGS)的研究及进展[J].生态毒理学报,4(6):770-779.GAO P P,LUO Y,ZHOU Q X,et al.,2009.Research Advancement of Antibiotics Resistance Genes(ARGs)in Aquaculture Environment[J].Asian Journal of Ecotoxicology,4(6):770-779.
胡亚茹,姜蕾,张天阳,等,2018.华东地区某饮用水源地中磺胺类抗性基因的分布特征[J].环境科学,39(9):4222-4228.HU Y R,JIANG L,ZHANG T Y,et al.,2018.Distribution Characteristics of Sulfonamide Antibiotic Resistance Genes in a Drinking Water Source in East China[J].Environmental Science,39(9):4222-4228.
罗义,周启星,2008.抗生素抗性基因(ARGS):一种新型环境污染物[J].环境科学学报,28(8):1499-1505.LUO Y,ZHOU Q X,2008.Antibiotic resistance genes(ARGs)as emerging pollutants[J].Acta Scientiae Circumstantiae,28(8):1499-1505.
沈群辉,冀秀玲,傅淑珺,等,2012.黄浦江水域抗生素及抗性基因污染初步研究[J].生态环境学报,21(10):1717-1723.SHEN Q H,JI X L,FU S J,et al.,2012.Preliminary studies on the pollution levels of antibiotic and antibiotic resistance genes in Huangpu River,China[J].Ecology and Environmental Sciences,21(10):1717-1723.
苏怀德,1988.青霉素发现及开发简史[J].药学通报,23(8):494-497.SU H D,1988.A brief history of penicillin discovery and development[J].Chinese Pharmaceutical Journal,23(8):494-497.
杨继平,邱志刚,袁兆康,等,2017.天津海河流域抗生素抗性基因分布特征及与指示微生物的关系[J].环境与健康杂志,34(4):313-316.YANG J P,QIN Z G,YUAN Z K,et al.,2017.Distribution of antibiotic resistance genes and their relationship with indicator microorganisms in Haihe River Basin,Tianjin[J].J Environ Health,34(4):313-316.
曾冠军,柳娴,马满英,2017.水体中抗生素污染研究进展[J].安徽农业科学,45(3):72-74.ZENG G J,LIU X,MA M Y,2017.Research Progress of Antibiotic Pollution in Water[J].Journal of Anhui AgriculturaL Sciences,45(3):72-74.
张丹丹,郭亚平,任红云,等,2018.福建省敖江下游抗生素抗性基因分布特征[J].环境科学,39(6):2600-2606.ZHANG D D,GUO Y P,REN H Y,et al.,2018.Characteristics of Antibiotic Resistance Genes in Downstream Areas of the Aojiang River,Fujian Province[J].Environmental Science,39(6):2600-2606.
张俊,杨晓洪,葛峰,等,2014.长期施用四环素残留猪粪对土壤中耐药菌及抗性基因形成的影响[J].环境科学,35(6):2374-2380.ZHANG J,YANG X H,GE F,et al.,2014.Effects of Long-Term Application of Pig Manure Containing Residual Tetracycline on the Formation of Drug-Resistant Bacteria and Resistance Genes[J].Environmental Science,35(6):2374-2380.
张兰河,王佳佳,哈雪姣,等,2016.北京地区菜田土壤抗生素抗性基因的分布特征[J].环境科学,37(11):4395-4401.ZHANG L H,WANG J J,HA X J,et al.,2016.Distribution Characteristics of Antibiotic Resistance Genes in Vegetable Soils in Beijing[J].Environmental Science,37(11):4395-4401.
张瑞全,应光国,丁永祯,等,2013.广东西枝江-东江流域抗生素抗性基因污染特征研究[J].农业环境科学学报,32(12):2471-2479.ZHANG R Q,YING G G,DING Y Z,et al.,2013.Pollution Characteristics of Antibiotic Resistance Genes in Xizhijiang-Dongjiang River Basin,Guangdong Province,China[J].Journal of Agro-Environment Science,32(12):2471-2479.
赵腾辉,陈奕涵,韩巍,等,2016.东江上游典型抗生素污染特征及生态风险评价[J].生态环境学报,25(10):1707-1713.ZHAO T H,CHEN Y H,HAN W,et al.,2016.The Contamination Characteristics and Ecological Risk Assessment of Typical Antibiotics in the Upper Reaches of the Dongjiang River[J].Ecology and Environmental Sciences,25(10):1707-1713.
周婷,李鑫,王晓龙,等,2014.新疆玛纳斯河流域抗生素抗性基因的污染分析[J].石河子大学学报(自然科学版),32(6):755-759.ZHOU T,LI X,WANG X L,et al.,2014.Pollution Analysis of Antibiotic Resistant Genes of Manas River Basin,Xinjiang[J].Journal of Shihezi University(Natural Science),32(6):755-759.
赵晓祥,杨莲,程晨,等,2018.上海水体中抗生素抗性基因的分布特征[J].安全与环境学报,18(3):1191-1197.ZHAO X X,YANG L,CHENG C,et al.,2018.Distributive situation of the antibiotic resistance genes in the surface water bodies in Shanghai[J].Journal of Safety and Environment,18(3):1191-1197.
邹世春,朱春敬,贺竹梅,等,2009.北江河水中抗生素抗性基因污染初步研究[J].生态毒理学报,4(5):655-660.ZOU S C,ZHU J C,HE Z M,et al.,2009.Preliminary Studies on the Pollution Levels of Antibiotic Resistance Genes in the Water of Beijiang River,South China[J].Asian Journal of Ecotoxicology,4(5):655-660.