武汉城市湖泊抗生素及抗性基因的污染特征研究
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摘要
为研究武汉城市湖泊抗生素及抗生素抗性基因的污染水平与分布特征,文章采用超高效液相色谱-质谱法和荧光定量PCR法对南湖、沙湖和东湖水体及底泥中12种抗生素、9种ARGs及Ⅰ类整合子intI1进行定性和定量分析。结果表明,湖泊水体及底泥中均以喹诺酮类抗生素污染为主,浓度范围分别为51.43～105.62 ng/L和9.01～12.93 ng/g。10种目标基因中,tetG、tetM、sul1、sul2、qnrD及intI1的检出率均为100%。磺胺类抗性基因sul1的相对丰度最高(2.39×10~(-2)～3.99×10~(-1)),属于优势抗性基因。intI1含量与4种抗性基因含量(tetG、tetM、sul1及qnrD)、ARGs总量之间均存在显著正相关关系(P<0.05),说明intI1是ARGs在城市湖泊环境中进行水平基因转移的重要媒介。冗余分析表明四环素类、喹诺酮类抗生素污染和intI1含量是影响湖泊水体、底泥中ARGs丰度及分布的重要因素。
In order to investigate the pollution status and distribution characteristics of antibiotics and antibiotic resistance genes(ARGs) in urban lakes of Wuhan, high performance liquid chromatography-mass spectrometry and quantitative PCR were applied to analyze the occurrence and quantities of twelve antibiotics, nine ARGs and class I integron gene intI1 in water and sediment samples of South Lake, Sand Lake and East Lake. The results showed that quinolones were the main contaminants in all samples. The total concentration of quinolones ranged from 51.43 to 105.62 ng/L in water and from 9.01 to 12.93 ng/g in sediments, respectively. Among the ten tested genes, tetG, tetM, sul1, sul2, qnrD as well as intI1 had 100% detection frequency. Sul1 had the highest relative abundance(2.39×10~(-2)～3.99×10~(-1)), and was the predominant resistance gene. There was a significant positive correlation between the relative abundance of intI1 and four resistance genes(tetG, tetM, sul1 and qnrD) as well as total ARGs(P<0.05), indicating that intI1 was a significant media for horizontal gene transfer of ARGs in urban lakes. And redundancy analysis showed that tetracyclines, quinolones and intI1 may play a very important role in the abundance and distribution of ARGs in urban lakes of Wuhan.
In order to investigate the pollution status and distribution characteristics of antibiotics and antibiotic resistance genes(ARGs) in urban lakes of Wuhan, high performance liquid chromatography-mass spectrometry and quantitative PCR were applied to analyze the occurrence and quantities of twelve antibiotics, nine ARGs and class I integron gene intI1 in water and sediment samples of South Lake, Sand Lake and East Lake. The results showed that quinolones were the main contaminants in all samples. The total concentration of quinolones ranged from 51.43 to 105.62 ng/L in water and from 9.01 to 12.93 ng/g in sediments, respectively. Among the ten tested genes, tetG, tetM, sul1, sul2, qnrD as well as intI1 had 100% detection frequency. Sul1 had the highest relative abundance(2.39×10~(-2)～3.99×10~(-1)), and was the predominant resistance gene. There was a significant positive correlation between the relative abundance of intI1 and four resistance genes(tetG, tetM, sul1 and qnrD) as well as total ARGs(P<0.05), indicating that intI1 was a significant media for horizontal gene transfer of ARGs in urban lakes. And redundancy analysis showed that tetracyclines, quinolones and intI1 may play a very important role in the abundance and distribution of ARGs in urban lakes of Wuhan.
引文
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[16]丁惠君,钟家有,吴亦潇,等.鄱阳湖流域南昌市城市湖泊水体抗生素污染特征及生态风险分析[J].湖泊科学,2017,29(4):848-858.Ding Huijun,Zhong Jiayou,Wu Yixiao,et al.Characteristics and ecological risk assessment of antibiotics in five city lakes in Nanchang City,Lake Poyang Catchment[J].Journal of Lake Science,2017,29(4):848-858.
[17]Xu J,Zhang Y,Zhou C,et al.Distribution,sources and composition of antibiotics in sediment,overlying water and pore water from Taihu Lake,China[J].Science of the Total Environment,2014,497/498(3):267-273.
[18]Lei X,Lu J,Liu Z,et al.Concentration and distribution of antibiotics in water-sediment system of Bosten Lake,Xinjiang[J].Environmental Science&Pollution Research International,2015,22(3):1670-8.
[19]章琴琴.北京温榆河流域抗生素污染分布特征及源解析研究[D].重庆:重庆大学,2012.
[20]秦延文,张雷,时瑶,等.大辽河表层水体典型抗生素污染特征与生态风险评价[J].环境科学研究,2015,28(3):361-368.Qin Yanwen,Zhang Lei,Shi Yao,et al.Contamination characteristics and ecological risk assessment of typical antibiotics in surface water of Daliao River,China[J].Research of Environment Science,2015,28(3):361-368.
[21]Li W,Shi Y,Gao L,et al.Occurrence of antibiotics in water,sediments,aquatic plants,and animals from Baiyangdian Lake in North China[J].Chemosphere,2012,89(11):1307-1315.
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[24]Hernando M D,Mezcua M,Fernandezalba A R,et al.Environmental risk assessment of pharmaceutical residues in wastewater effluents,surface waters and sediments[J].Talanta,2006,69(2):334-342.
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[26]Lan L,Ke Y,Liang X,et al.Occurrences and distribution of sulfonamide and tetracycline resistance genes in the Yangtze River Estuary and nearby coastal area[J].Marine Pollution Bulletin,2015,100(1):304-310.
[27]Jiang L,Hu X,Xu T,et al.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,2013,458/459/460(3):267-272.
[28]Yang Y,Liu W,Xu C,et al.Antibiotic resistance genes in lakes from middle and lower reaches of the Yangtze River,China:effect of land use and sediment characteristics[J].Chemosphere,2017,178:19-25.
[29]Stedtfeld R D,Stedtfeld T M,Waseem H,et al.Isothermal assay targeting class 1 integrase gene for environmental surveillance of antibiotic resistance markers[J].Journal of Environmental Management,2017,198(Pt 1):213-220.
[30]Marti E,Variatza E,Balcazar J L.The role of aquatic ecosystems as reservoirs of antibiotic resistance[J].Trends in Microbiology,2014,22(1):36-41.
[31]Hall R M,Collis C M.Mobile gene cassettes and integrons:capture and spread of genes by site-specific recombination[J].Molecular Microbiology,2010,15(4):593-600.
[32]李鲁明,袁晓燕,王明义,等.整合子-基因盒系统与细菌耐药机制的研究进展[J].中国微生态学杂志,2014,26(2).Li Luming,Yuan Xiaoyan,Wang Mingyi,et al.The integron-gene cassette system and the mechanism of bacterial drug resistance:research progress[J].Chinese Journal of Microecology,2014,26(2).
[33]Pei Ruoting,Cha J,And K H C,et al.Response of antibiotic resistance genes(ARG)to biological treatment in dairy lagoon water[J].Environmental Science and Technology,2007,41(14):5108-5113.
[34]Engemann C A,Keen P L,Knapp C W,et al.Fate of tetracycline resistance genes in aquatic systems:migration from the water column to peripheral biofilms[J].Environmental Science and Technology,2008,42(14):5131-5136.
[2]隋倩雯,张俊亚,魏源送,等.畜禽养殖过程抗生素使用与耐药病原菌及其抗性基因赋存的研究进展[J].生态毒理学报,2015,10(5):20-34.Sui Qianwen,Zhang Junya,Wei Yuansong,et al.Veterinary antibiotics use,occurrence of antibiotic resistance pathogen and its antibiotic resistance genes in animal production:an overview[J].Asia Journal of Ecotoxicology,2015,10(5):20-34.
[3]罗义,周启星.抗生素抗性基因(ARGs)——一种新型环境污染物[J].环境科学学报,2008,28(8):1499-1505.Luo Yi,Zhou Qixing.Antibiotic resistance genes(ARGs)as emerging pollutants[J].Acta Scientiae Circumstantiae,2008,28(8):1499-1505.
[4]Pruden A,Pei R,Storteboom H,et al.Antibiotic resistance genes as emerging contaminants:studies in Northern Colorado[J].Environmental Science and Technology,2006,40(23):7445.
[5]苏建强,黄福义,朱永官.环境抗生素抗性基因研究进展[J].生物多样性,2013,21(4):481-487.Su Jianqiang,Huang Fuyi,Zhu Yongguan.Antibiotic resistance genes in the environment[J].Biodiversity Science,2013,21(4):481-487.
[6]Zhang QQ,Ying GG,Pan CG,et al.Comprehensive evaluation of antibiotics emission and fate in the river basins of China:source analysis,multimedia modeling,and linkage to bacterial resistance[J].Environmental Science and Technology,2015,49(11):6772-6782.
[7]Czekalski N,Sigdel R,Birtel J,et al.Does human activity impact the natural antibiotic resistance background?Abundance of antibiotic resistance genes in 21 Swiss lakes[J].Environment International,2015,81:45-55.
[8]Luo Y,Mao DQ,Rysz M,et al.Trends in antibiotic resistance genes occurrence in the Haihe River,China[J].Environmental Science and Technology,2010,44(19):7220-7225.
[9]Ding HJ,Wu YX,Zhang WH,et al.Occurrence,distribution,and risk assessment of antibiotics in the surface water of Poyang Lake,the largest freshwater lake in China[J].Chemosphere,2017,184:137.
[10]Zhang XX,Zhang T,Fang HH.Antibiotic resistance genes in water environment[J].Applied Microbiology&Biotechnology,2009,82(3):397-414.
[11]Mao DQ,Yu S,Rysz M,et al.Prevalence and proliferation of antibiotic resistance genes in two municipal wastewater treatment plants[J].Water Research,2015,85(6):458-466.
[12]冀永般,王雪,曾巾,等.南京市湖泊沉积物中重金属垂直分布及污染状况评价[J].环境科学与技术,2012,35(12):230-233.Ji Yongban,Wang Xue,Zeng Jin,et al.Heavy metal in sediment of urban lakes in Nanjing:vertical distribution and pollution evaluation[J].Environmental Science&Technology,2012,35(12):230-233.
[13]张瑞泉,应光国,丁永祯,等.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,2013,32(12):2471-2479.Zhang Ruiquan,Ying Guangguo,Ding Yongzhen,et al.Pollution characteristics of antibiotic resistance genes in Xizhijiang-Dongjiang river basin,Guangdong Province,China[J].Journal of Agro-environment Science,2013,32(12):2471-2479.
[14]Zhou L J,Ying G G,Liu S,et al.Simultaneous determination of human and veterinary antibiotics in various environmental matrices by rapid resolution liquid chromatographyelectrospray ionization tandem mass spectrometry[J].Journal of Chromatography A,2012,1244(12):123-138.
[15]Managaki S,Murata A,Takada H,et al.Distribution of macrolides,sulfonamides,and trimethoprim in tropical waters:ubiquitous occurrence of veterinary antibiotics in the mekong delta[J].Environmental Science and Technology,2007,41(23):8004-8010.
[16]丁惠君,钟家有,吴亦潇,等.鄱阳湖流域南昌市城市湖泊水体抗生素污染特征及生态风险分析[J].湖泊科学,2017,29(4):848-858.Ding Huijun,Zhong Jiayou,Wu Yixiao,et al.Characteristics and ecological risk assessment of antibiotics in five city lakes in Nanchang City,Lake Poyang Catchment[J].Journal of Lake Science,2017,29(4):848-858.
[17]Xu J,Zhang Y,Zhou C,et al.Distribution,sources and composition of antibiotics in sediment,overlying water and pore water from Taihu Lake,China[J].Science of the Total Environment,2014,497/498(3):267-273.
[18]Lei X,Lu J,Liu Z,et al.Concentration and distribution of antibiotics in water-sediment system of Bosten Lake,Xinjiang[J].Environmental Science&Pollution Research International,2015,22(3):1670-8.
[19]章琴琴.北京温榆河流域抗生素污染分布特征及源解析研究[D].重庆:重庆大学,2012.
[20]秦延文,张雷,时瑶,等.大辽河表层水体典型抗生素污染特征与生态风险评价[J].环境科学研究,2015,28(3):361-368.Qin Yanwen,Zhang Lei,Shi Yao,et al.Contamination characteristics and ecological risk assessment of typical antibiotics in surface water of Daliao River,China[J].Research of Environment Science,2015,28(3):361-368.
[21]Li W,Shi Y,Gao L,et al.Occurrence of antibiotics in water,sediments,aquatic plants,and animals from Baiyangdian Lake in North China[J].Chemosphere,2012,89(11):1307-1315.
[22]Yang Y,Cao X,Lin H,et al.Antibiotics and antibiotic resistance genes in sediment of Honghu Lake and East Dongting Lake,China[J].Microbial Ecology,2016,72(4):791-801.
[23]European Commission.Technical Guidance Document in Support of Commission Directive 93//67/EEC on Risk Assessment for New Notified Substances and Commission Regulation(EC)No.1488/94 on Risk Assessment for Existing Substances[R].Luxembourg:Office for Official Publications of the European Communities,2003.
[24]Hernando M D,Mezcua M,Fernandezalba A R,et al.Environmental risk assessment of pharmaceutical residues in wastewater effluents,surface waters and sediments[J].Talanta,2006,69(2):334-342.
[25]Yang Y,Chen X,Cao X,et al.Antibiotic resistance genes in surface water of eutrophic urban lakes are related to heavy metals,antibiotics,lake morphology and anthropic impact[J].Ecotoxicology,2017,26(6):831-840.
[26]Lan L,Ke Y,Liang X,et al.Occurrences and distribution of sulfonamide and tetracycline resistance genes in the Yangtze River Estuary and nearby coastal area[J].Marine Pollution Bulletin,2015,100(1):304-310.
[27]Jiang L,Hu X,Xu T,et al.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,2013,458/459/460(3):267-272.
[28]Yang Y,Liu W,Xu C,et al.Antibiotic resistance genes in lakes from middle and lower reaches of the Yangtze River,China:effect of land use and sediment characteristics[J].Chemosphere,2017,178:19-25.
[29]Stedtfeld R D,Stedtfeld T M,Waseem H,et al.Isothermal assay targeting class 1 integrase gene for environmental surveillance of antibiotic resistance markers[J].Journal of Environmental Management,2017,198(Pt 1):213-220.
[30]Marti E,Variatza E,Balcazar J L.The role of aquatic ecosystems as reservoirs of antibiotic resistance[J].Trends in Microbiology,2014,22(1):36-41.
[31]Hall R M,Collis C M.Mobile gene cassettes and integrons:capture and spread of genes by site-specific recombination[J].Molecular Microbiology,2010,15(4):593-600.
[32]李鲁明,袁晓燕,王明义,等.整合子-基因盒系统与细菌耐药机制的研究进展[J].中国微生态学杂志,2014,26(2).Li Luming,Yuan Xiaoyan,Wang Mingyi,et al.The integron-gene cassette system and the mechanism of bacterial drug resistance:research progress[J].Chinese Journal of Microecology,2014,26(2).
[33]Pei Ruoting,Cha J,And K H C,et al.Response of antibiotic resistance genes(ARG)to biological treatment in dairy lagoon water[J].Environmental Science and Technology,2007,41(14):5108-5113.
[34]Engemann C A,Keen P L,Knapp C W,et al.Fate of tetracycline resistance genes in aquatic systems:migration from the water column to peripheral biofilms[J].Environmental Science and Technology,2008,42(14):5131-5136.