生活垃圾渗滤液处理过程中抗生素抗性基因的变化特征
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摘要
抗生素抗性基因被认为是一种环境污染物,城市生活垃圾填埋场及其渗滤液中存在着多种类和高丰度的抗生素抗性基因.针对生活垃圾渗滤液处理工艺流程特点,本研究采用超高通量定量PCR技术,分析生活垃圾渗滤液处理过程中抗生素抗性基因的变化特征.结果表明,渗滤液处理使得出水抗生素抗性基因丰度减至9. 2×1010copies·L-1,降低了5个数量级,能有效消减渗滤液中的抗生素抗性基因,是一种有效消减垃圾渗滤液抗生素抗性基因丰度的方法;渗滤液出水中的抗生素抗性基因140种,检出率高达46. 7%,相较于自然环境,出水中抗生素抗性基因种类和丰度仍然维持在较高水平,直接排入市政污水管网可能造成抗生素抗性基因的二次扩散,存在着较高的生态环境风险;渗滤液中抗生素抗性基因与Cr、Cd、Ni、As、MGEs、integron和transposon呈现极显著的正相关关系,表明抗生素抗性基因的赋存和迁移传播可能受重金属元素和可移动遗传元件的共同影响.
Antibiotic resistance genes( ARGs) are considered to be emerging environmental contaminants. ARGs are frequently detected in municipal solid waste landfill and its leachate. In this study,high-throughput quantitative PCR techniques were used to investigate the dynamics of ARGs during the treatment process of landfill leachate. The results showed that ARGs in leachate were diverse and abundant,and leachate treatment could effectively reduce their abundance; the absolute abundance of ARGs was reduced to 9. 2 × 1010 copies·L-1,which was five orders of magnitude less than the abundance of the influent. However,the range and abundance of antibiotic resistance genes in the leachate effluents were still high compared to the natural environment. Therefore,the secondary contamination of ARGs might be caused by leachate being directly discharged into municipal sewage system. In this regard,ecological and environmental risk is an inconvenient truth. Furthermore,strong correlations indicate that heavy metals and mobile genetic elements( MGEs) in landfill leachate might influence the transfer of ARGs.
Antibiotic resistance genes( ARGs) are considered to be emerging environmental contaminants. ARGs are frequently detected in municipal solid waste landfill and its leachate. In this study,high-throughput quantitative PCR techniques were used to investigate the dynamics of ARGs during the treatment process of landfill leachate. The results showed that ARGs in leachate were diverse and abundant,and leachate treatment could effectively reduce their abundance; the absolute abundance of ARGs was reduced to 9. 2 × 1010 copies·L-1,which was five orders of magnitude less than the abundance of the influent. However,the range and abundance of antibiotic resistance genes in the leachate effluents were still high compared to the natural environment. Therefore,the secondary contamination of ARGs might be caused by leachate being directly discharged into municipal sewage system. In this regard,ecological and environmental risk is an inconvenient truth. Furthermore,strong correlations indicate that heavy metals and mobile genetic elements( MGEs) in landfill leachate might influence the transfer of ARGs.
引文
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[13]庄芳芳,苏建强,陈辉煌,等.基于高通量定量PCR研究城市化小流域微生物污染特征[J].生态毒理学报,2017,12(5):141-152.Zhuang F F,Su J Q,Chen H H,et al. Characterization of microbial contaminants using high-throughput quantitative PCR in a small urbanizing catchment[J]. Asian Journal of Ecotoxicology,2017,12(5):141-152.
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[15]黄福义,安新丽,李力,等.生活垃圾填埋场对河流抗生素抗性基因的影响[J].中国环境科学,2017,37(1):203-209.Huang F Y,An X L,Li L,et al. High-throughput profiling of antibiotic resistance genes in river in the vicinity of a landfill[J].China Environmental Science,2017,37(1):203-209.
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[19]王鹤茹,宋永伟,徐峙辉,等.均相/异相催化剂光降解垃圾渗滤液效果的研究[J].环境科学学报,2016,36(9):3243-3250.Wang H R, Song Y W, Xu Z H, et al. Photochemical degradation of landfill leachate by homogeneous/heterogeneous catalyst combinations[J]. Acta Scientiae Circumstantiae,2016,36(9):3243-3250.
[20]李丽,赵成萍,李宏,等.质粒制备绝对定量PCR标准曲线方法的建立[J].农业生物技术学报,2011,19(6):1157-1162.Li L,Zhao C P,Li H,et al. Establishment of the plasmid standard curve generation method for absolute quantification PCR[J]. Journal of Agricultural Biotechnology,2011,19(6):1157-1162.
[21] Su J Q,Wei B,Ouyang W Y,et al. Antibiotic resistome and its association with bacterial communities during sewage sludge composting[J]. Environmental Science&Technology,2015,49(12):7356-7363.
[22] Chen Y H,Su J Q,Zhang J Y,et al. High-throughput profiling of antibiotic resistance gene dynamic in a drinking water riverreservoir system[J]. Water Research,2019,149:179-189.
[23] Chen Q L,An X L,Zhu Y G,et al. Application of Struvite Alters the Antibiotic Resistome in Soil, Rhizosphere, and Phyllosphere[J]. Environmental Science&Technology,2017,51(14):8149-8157.
[24] Klappenbach J A,Saxman P R,Cole J R,et al. rrndb:the ribosomal RNA operon copy number database[J]. Nucleic Acids Research,2001,29(1):181-184.
[25] Aydin S,Ince B,Ince O. Assessment of anaerobic bacterial diversity and its effects on anaerobic system stability and the occurrence of antibiotic resistance genes[J]. Bioresource Technology,2016,207:332-338.
[26] Aydin S,Ince B,Ince O. Development of antibiotic resistance genes in microbial communities during long-term operation of anaerobic reactors in the treatment of pharmaceutical wastewater[J]. Water Research,2015,83:337-344.
[27]张丹丹,郭亚平,任红云,等.福建省敖江下游抗生素抗性基因分布特征[J].环境科学,2018,39(6):2600-2606.Zhang D D,Guo Y P,Ren H Y,et al. Characteristics of antibiotic resistance genes in downstream areas of the Aojiang River,Fujian Province[J]. Environmental Science,2018,39(6):2600-2606.
[28] Zhao Y,Su J Q,An X L,et al. Feed additives shift gut microbiota and enrich antibiotic resistance in swine gut[J].Science of the Total Environment,2018,621:1224-1232.
[2] Pruden A,Pei R T,Storteboom H,et al. Antibiotic resistance genes as emerging contaminants:studies in northern Colorado[J]. Environmental Science&Technology,2006,40(23):7445-7450.
[3]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 J Q,Huang F Y,Zhu Y G. Antibiotic resistance genes in the environment[J]. Biodiversity Science,2013,21(4):481-487.
[4] Martínez J L. Antibiotics and antibiotic resistance genes in natural environments[J]. Science,2008,321(5887):365-367.
[5] Rizzo L,Manaia C,Merlin C,et al. Urban wastewater treatment plants as hotspots for antibiotic resistant bacteria and genes spread into the environment:a review[J]. Science of the Total Environment,2013,447:345-360.
[6]黄福义,李虎,韦蓓,等.长期施用猪粪水稻土抗生素抗性基因污染研究[J].环境科学,2014,35(10):3869-3873.Huang F Y,Li H,Wei B,et al. Long-term manure application induced shift of diversity and abundance of antibiotic resistance genes in paddy soil[J]. Environmental Science,2014,35(10):3869-3873.
[7] Ouyang W Y,Huang F Y,Zhao Y,et al. Increased levels of antibiotic resistance in urban stream of Jiulongjiang River,China[J]. Applied Microbiology and Biotechnology,2015,99(13):5697-5707.
[8] Chen B W,Yang Y,Liang X M,et al. Metagenomic profiles of antibiotic resistance genes(ARGs)between human impacted estuary and deep ocean sediments[J]. Environmental Science&Technology,2013,47(22):12753-12760.
[9]黄福义,杨凯,张子兴,等.微塑料对河口沉积物抗生素抗性基因的影响[J].环境科学,2019,40(5):2234-2239.Huang F Y,Yang K,Zhang Z X,et al. Effects of microplastics on antibiotic resistance genes in estuarine sediments[J].Environmental Science,2019,40(5):2234-2239.
[10] Zhu Y G, Gillings M, Simonet P, et al. Microbial mass movements[J]. Science,2017,357(6356):1099-1100.
[11] Cully M. Public health:the politics of antibiotics[J]. Nature,2014,509(7498):S16-S17.
[12] Smith R A,M'Ikanatha N M,Read A F. Antibiotic resistance:a primer and call to action[J]. Health Communication,2015,30(3):309-314.
[13]庄芳芳,苏建强,陈辉煌,等.基于高通量定量PCR研究城市化小流域微生物污染特征[J].生态毒理学报,2017,12(5):141-152.Zhuang F F,Su J Q,Chen H H,et al. Characterization of microbial contaminants using high-throughput quantitative PCR in a small urbanizing catchment[J]. Asian Journal of Ecotoxicology,2017,12(5):141-152.
[14] Fischbach M A,Walsh C T. Antibiotics for emerging pathogens[J]. Science,2009,325(5944):1089-1093.
[15]黄福义,安新丽,李力,等.生活垃圾填埋场对河流抗生素抗性基因的影响[J].中国环境科学,2017,37(1):203-209.Huang F Y,An X L,Li L,et al. High-throughput profiling of antibiotic resistance genes in river in the vicinity of a landfill[J].China Environmental Science,2017,37(1):203-209.
[16] Chen Q L,Li H,Zhou X Y,et al. An underappreciated hotspot of antibiotic resistance:the groundwater near the municipal solid waste landfill[J]. Science of the Total Environment,2017,609:966-973.
[17] Song L Y,Li L,Yang S,et al. Sulfamethoxazole,tetracycline and oxytetracycline and related antibiotic resistance genes in a large-scale landfill, China[J]. Science of the Total Environment,2016,551-552:9-15.
[18]熊建英,郑正.垃圾填埋场渗滤液溶解性有机质特性及其去除技术综述[J].环境化学,2015,34(1):44-53.Xiong J Y,Zheng Z. Characteristics of the dissolved organic matter in landfill leachate and their removal technology:a review[J]. Environmental Chemistry,2015,34(1):44-53.
[19]王鹤茹,宋永伟,徐峙辉,等.均相/异相催化剂光降解垃圾渗滤液效果的研究[J].环境科学学报,2016,36(9):3243-3250.Wang H R, Song Y W, Xu Z H, et al. Photochemical degradation of landfill leachate by homogeneous/heterogeneous catalyst combinations[J]. Acta Scientiae Circumstantiae,2016,36(9):3243-3250.
[20]李丽,赵成萍,李宏,等.质粒制备绝对定量PCR标准曲线方法的建立[J].农业生物技术学报,2011,19(6):1157-1162.Li L,Zhao C P,Li H,et al. Establishment of the plasmid standard curve generation method for absolute quantification PCR[J]. Journal of Agricultural Biotechnology,2011,19(6):1157-1162.
[21] Su J Q,Wei B,Ouyang W Y,et al. Antibiotic resistome and its association with bacterial communities during sewage sludge composting[J]. Environmental Science&Technology,2015,49(12):7356-7363.
[22] Chen Y H,Su J Q,Zhang J Y,et al. High-throughput profiling of antibiotic resistance gene dynamic in a drinking water riverreservoir system[J]. Water Research,2019,149:179-189.
[23] Chen Q L,An X L,Zhu Y G,et al. Application of Struvite Alters the Antibiotic Resistome in Soil, Rhizosphere, and Phyllosphere[J]. Environmental Science&Technology,2017,51(14):8149-8157.
[24] Klappenbach J A,Saxman P R,Cole J R,et al. rrndb:the ribosomal RNA operon copy number database[J]. Nucleic Acids Research,2001,29(1):181-184.
[25] Aydin S,Ince B,Ince O. Assessment of anaerobic bacterial diversity and its effects on anaerobic system stability and the occurrence of antibiotic resistance genes[J]. Bioresource Technology,2016,207:332-338.
[26] Aydin S,Ince B,Ince O. Development of antibiotic resistance genes in microbial communities during long-term operation of anaerobic reactors in the treatment of pharmaceutical wastewater[J]. Water Research,2015,83:337-344.
[27]张丹丹,郭亚平,任红云,等.福建省敖江下游抗生素抗性基因分布特征[J].环境科学,2018,39(6):2600-2606.Zhang D D,Guo Y P,Ren H Y,et al. Characteristics of antibiotic resistance genes in downstream areas of the Aojiang River,Fujian Province[J]. Environmental Science,2018,39(6):2600-2606.
[28] Zhao Y,Su J Q,An X L,et al. Feed additives shift gut microbiota and enrich antibiotic resistance in swine gut[J].Science of the Total Environment,2018,621:1224-1232.