纳污河流抗性基因和微生物群落相关性
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摘要
通过Miseq高通量测序分析和荧光定量PCR技术,研究某纳污河流中四环素类与磺胺类抗性基因(ARGs)的分布特征、传播情况及微生物群落结构相关性.结果表明:在河流地表水与沉积物中均检测到四环素类抗性基因tetA和tetB与磺胺类抗性基因sul1和sul2,四环素类抗性基因和磺胺类抗性基因的绝对丰度分别为5.09×10~3～1.26×10~7,3.94×10~5～1.32×10~9copics/L,磺胺sul1抗生素抗性基因丰度显著高于其它基因;河流主要优势菌门为Proteobacteria,Bacteroidetes,Firmicutes,Actinobacteria和Cyanobacteria,其平均总相对丰度占总比例的95.62%,且总体差异较小.抗性基因与微生物群落冗余分析显示, Methylotenera菌属是影响tetA抗性基因分布丰度的主要因素, Dechloromonas和Clostridium sensu stricto 1菌属是影响tetB抗性基因分布丰度的主要因素,Dechloromonas、Clostridium sensu stricto 1和Methylotenera菌属是影响sul1、sul2抗性基因分布丰度的主要因素.
The correlation of distribution characteristics, regulations of propagation of tetracyclines and sulfonamide resistance genes(ARGs), and the microbial community structure of the contaminated rivers were studied by using the Miseq sequencing and real-time PCR techniques. The tetracycline resistance genes tetA and tetB and the sulfonamide resistance genes sul1 and sul2 were both detected in water and sediment. And the absolute abundances of tetracycline resistance gene and sulfonamide resistance gene were 5.09×10~3～1.26×10~7 and 3.94×10~5～1.32×10~9 copics/L, respectively. The abundances of sul1 antibiotic resistance gene was significantly higher than other genes. The main dominant bacteria in the river were Proteobacteria, Bacteroidetes, Firmicutes, Actinobacteria and Cyanobacteria. The relative abundance accounted for 95.62% of the total proportion, and there was little difference in overall of the examples. The analysis of resistance gene and microbial community redundancy showed that Methylotenera was the main factor affecting the abundance of tetA resistance gene, Dechloromonas and Clostridium. Sensu stricto 1 were the main factors affecting the abundance of tetB resistance gene, and Dechloromonas, Clostridium sensu stricto 1 and Methylotenera were the main factors affecting of the abundance of sul1 and sul2 resistance gene distribution.
The correlation of distribution characteristics, regulations of propagation of tetracyclines and sulfonamide resistance genes(ARGs), and the microbial community structure of the contaminated rivers were studied by using the Miseq sequencing and real-time PCR techniques. The tetracycline resistance genes tetA and tetB and the sulfonamide resistance genes sul1 and sul2 were both detected in water and sediment. And the absolute abundances of tetracycline resistance gene and sulfonamide resistance gene were 5.09×10~3～1.26×10~7 and 3.94×10~5～1.32×10~9 copics/L, respectively. The abundances of sul1 antibiotic resistance gene was significantly higher than other genes. The main dominant bacteria in the river were Proteobacteria, Bacteroidetes, Firmicutes, Actinobacteria and Cyanobacteria. The relative abundance accounted for 95.62% of the total proportion, and there was little difference in overall of the examples. The analysis of resistance gene and microbial community redundancy showed that Methylotenera was the main factor affecting the abundance of tetA resistance gene, Dechloromonas and Clostridium. Sensu stricto 1 were the main factors affecting the abundance of tetB resistance gene, and Dechloromonas, Clostridium sensu stricto 1 and Methylotenera were the main factors affecting of the abundance of sul1 and sul2 resistance gene distribution.
引文
[1]周启星,罗义,王美娥.抗生素的环境残留、生态毒性及抗性基因污染[J].生态毒理学报,2007,2(3):243-251.Zhou Q X,Luo Y,Wang M E.Environmental residues,ecotoxicity and resistance gene contamination of antibiotics[J].Journal of Ecotoxicology,2007,2(3):243-251.
[2]徐冰洁,罗义,周启星,等.抗生素抗性基因在环境中的来源、传播扩散及生态风险[J].环境化学,2010,29(2):169-178.Xu B J,Luo Y,Zhou Q X,et al.Source,spread and ecological risk of antibiotic resistance genes in the environment[J].Environmental Chemistry,2010,29(2):169-178.
[3]Zhang X X,Tong Z,Fang H H P.Antibiotic resistance genes in water environment[J].Applied Microbiology and Biotechnology,2009,82(3):397-414.
[4]Dzidic S,Bedekovic V.Horizontal gene transfer-emerging multidrug resistance in hospital bacteria[J].Acta Pharmacologica Sinica,2003,24(6):519-526.
[5]Pruden A,Pei R,Heather Storteboom A,et al.Antibiotic resistance genes as emerging contaminants:studies in northern coloradot[J].Environmental Science and Technology,2006,40(23):7445-7450.
[6]Du J,Ren H,Geng J,et al.Occurrence and abundance of tetracycline,sulfonamide resistance genes,and class 1integron in five wastewater treatment plants[J].Environmental Science and Pollution Research International,2014,21(12):72-76.
[7]Lin L,Yuan K,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.
[8]Xu J,Xu Y,Wang H,et al.Occurrence of antibiotics and antibiotic resistance genes in a sewage treatment plant and its effluent-receiving river[J].Chemosphere,2015,119:1379-1385.
[9]Sabri N A,Schmitt H,Van D Z B,et al.Prevalence of antibiotics and antibiotic resistance genes in a wastewater effluent-receiving river in the Netherlands[J].Journal of Environmental Chemical Engineering.DOI:10.1016/j.jece,2018.3:004.
[10]Stoll C,Sidhu J P S,Tiehm A,et al.Prevalence of clinically relevant antibiotic resistance genes in surface water samples collected from Germany and Australia[J].Environmental Science&Technology,2012,46(17):9716-9726.
[11]徐艳,张远,郭昌胜,等.石家庄汪洋沟地区抗生素、抗性细菌和抗性基因污染特征[J].农业环境科学学报,2014,33(6):1174-1182.Xu Y,Zhang Y,Guo C S,et al.Pollution characteristics of antibiotics,resistant bacteria and resistance genes in Wang yang gou area,Shijiazhuang[J].Journal of Agricultural Environmental Science,2014,33(6):1174-1182.
[12]Proia L,Anzil A,Subirats J,et al.Antibiotic resistance along an urban river impacted by treated wastewaters[J].Science of the Total Environment,2018,453:628-629.
[13]张毓森,叶军,苏建强.农田生态系统抗生素抗性研究进展与挑战[J].浙江大学学报(农业与生命科学版),2017,43(6):691-699.Zhang Y S,Ye J,Su J Q.Antibiotic resistance in agroecosystem:Progress and challenges[J].Journal of Zhejiang University(Agriculture and Life Sciences),2017,43(6):691-699.
[14]谭璐.人类活动促进环境中抗生素抗性基因增殖扩散研究[D].天津:天津大学,2017.Tian L.Human Activity promotes proliferation and diffusion of antibiotic resistance genes in the environment[D].Tianjin:Tianjin University,2015.
[15]Huerta B,Marti E,Gros M,et al.Exploring the links between antibiotic occurrence,antibiotic resistance,and bacterial communities in water supply reservoirs[J].Science of the Total Environment,2013,456-457(7):161-170.
[16]Pruden A,Arabi M,Storteboom H N.Correlation between upstream human activities and riverine antibiotic resistance genes[J].Environmental Science and Technology,2012,46(21):11541-11549.
[17]Pei R,Kim S C,Carlson K H,et al.Effect of river landscape on the sediment concentrations of antibiotics and corresponding antibiotic resistance genes(ARG)[J].Water Research,2006,40(12):2427-2435.
[18]Chen H,Zhang M.Effects of advanced treatment systems on the removal of antibiotic resistance genes in wastewater treatment plants from Hangzhou,China[J].Environmental Science and Technology,2013,47(15):8157-8163.
[19]Liu L,Liu C,Zheng J,et al.Elimination of veterinary antibiotics and antibiotic resistance genes from swine wastewater in the vertical flow constructed wetlands[J].Chemosphere,2013,91(8):1088-1093.
[20]张子扬,刘舒巍,张璐,等.人工湿地去除畜禽养殖废水中磺胺类抗生素抗性基因研究[J].环境科学与管理,2016,41(5):89-92.Zhang Z Y,Liu S W,Zhang L,et al.Study on removal of sulfonamide antibiotic resistance genes from livestock and poultry breeding wastewater by constructed wetland[J].Environmental Science and Management,2016,41(5):89-92.
[21]郑加玉,刘琳,高大文,等.四环素抗性基因在人工湿地中的去除及累积[J].环境科学,2013,34(8):3102-3107.Zheng J Y,Liu L,Gao D W,et al.Removal and accumulation of tetracycline resistance gene in constructed wetlands[J].Environmental Science,2013,34(8):3102-3107.
[22]Shu D,He Y,Yue H,et al.Metagenomic insights into the effects of volatile fatty acids on microbial community structures and functional genes in organotrophic anammox process[J].Bioresource Technology,2015,196:621-633.
[23]王智,张志勇,张君倩,等.水葫芦修复富营养化湖泊水体区域内外底栖动物群落特征[J].中国环境科学,2012,32(1):142-149.Wang Z,Zhang Z Y,Zhang J Q,et al.Characteristics of benthos community in the waters of eutrophic lakes repaired by water hyacinth[J].Chinese Environmental Science,2012,32(1):142-149.
[24]刁晓君,李一葳,王曙光.水华生消过程对巢湖沉积物微生物群落结构的影响[J].环境科学,2015,14(1):107-113.Diao X J,Li Y M,Wang S G.Effects of the process of water blooms on the microbial community structure in sediments of Chaohu Lake[J].Environmental Science,2015,14(1):107-113.
[25]曾涛涛,蒋小梅,韩科昌,等.生活污水处理厂微生物群落结构解析[J].安全与环境学报,2018,18(2):697-703.Zeng T T,Jiang X M,Han K C,et al.Analysis of microbial community structure in domestic sewage treatment plant[J].Journal of Safety and Environment,2018,18(2):697-703.
[26]张辉,张小江,徐英春,等.2005~2014年CHINET不动杆菌属细菌耐药性监测[J].中国感染与化疗杂志,2016,16(4):429-436.Zhang H,Zhang X J,Xu Y C,et al.Monitoring of bacterial resistance of Acinetobacter spp.in 2005~2014[J].Chinese Journal of Infection and Chemotherapy,2016,16(4):429-436.
[27]Yang F,Huang L,Li L,et al.Discharge of KPC-2genes from the WWTPs contributed to their enriched abundance in the receiving river[J].Science of the Total Environment,2017,136:581-582.
[28]Teng Z,Zhu Y,Li M,et al.Microbial community composition and activity controls phosphorus transformation in rhizosphere soils of the Yeyahu Wetland in Beijing,China[J].The Science of the Total Environment,2018,628-629:1266-1277.
[29]罗晓,袁立霞,张文丽,等.制药废水厂抗性基因和微生物群落相关性研究[J].中国环境科学,2019,39(2):831-838.Luo X,Yuan L X,Zhang W L,et al.Correlation study between resistance genes and microbial communities in pharmaceutical wastewater treatment plants[J].Chinese Environmental Science,2019,39(2):831-838.
[2]徐冰洁,罗义,周启星,等.抗生素抗性基因在环境中的来源、传播扩散及生态风险[J].环境化学,2010,29(2):169-178.Xu B J,Luo Y,Zhou Q X,et al.Source,spread and ecological risk of antibiotic resistance genes in the environment[J].Environmental Chemistry,2010,29(2):169-178.
[3]Zhang X X,Tong Z,Fang H H P.Antibiotic resistance genes in water environment[J].Applied Microbiology and Biotechnology,2009,82(3):397-414.
[4]Dzidic S,Bedekovic V.Horizontal gene transfer-emerging multidrug resistance in hospital bacteria[J].Acta Pharmacologica Sinica,2003,24(6):519-526.
[5]Pruden A,Pei R,Heather Storteboom A,et al.Antibiotic resistance genes as emerging contaminants:studies in northern coloradot[J].Environmental Science and Technology,2006,40(23):7445-7450.
[6]Du J,Ren H,Geng J,et al.Occurrence and abundance of tetracycline,sulfonamide resistance genes,and class 1integron in five wastewater treatment plants[J].Environmental Science and Pollution Research International,2014,21(12):72-76.
[7]Lin L,Yuan K,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.
[8]Xu J,Xu Y,Wang H,et al.Occurrence of antibiotics and antibiotic resistance genes in a sewage treatment plant and its effluent-receiving river[J].Chemosphere,2015,119:1379-1385.
[9]Sabri N A,Schmitt H,Van D Z B,et al.Prevalence of antibiotics and antibiotic resistance genes in a wastewater effluent-receiving river in the Netherlands[J].Journal of Environmental Chemical Engineering.DOI:10.1016/j.jece,2018.3:004.
[10]Stoll C,Sidhu J P S,Tiehm A,et al.Prevalence of clinically relevant antibiotic resistance genes in surface water samples collected from Germany and Australia[J].Environmental Science&Technology,2012,46(17):9716-9726.
[11]徐艳,张远,郭昌胜,等.石家庄汪洋沟地区抗生素、抗性细菌和抗性基因污染特征[J].农业环境科学学报,2014,33(6):1174-1182.Xu Y,Zhang Y,Guo C S,et al.Pollution characteristics of antibiotics,resistant bacteria and resistance genes in Wang yang gou area,Shijiazhuang[J].Journal of Agricultural Environmental Science,2014,33(6):1174-1182.
[12]Proia L,Anzil A,Subirats J,et al.Antibiotic resistance along an urban river impacted by treated wastewaters[J].Science of the Total Environment,2018,453:628-629.
[13]张毓森,叶军,苏建强.农田生态系统抗生素抗性研究进展与挑战[J].浙江大学学报(农业与生命科学版),2017,43(6):691-699.Zhang Y S,Ye J,Su J Q.Antibiotic resistance in agroecosystem:Progress and challenges[J].Journal of Zhejiang University(Agriculture and Life Sciences),2017,43(6):691-699.
[14]谭璐.人类活动促进环境中抗生素抗性基因增殖扩散研究[D].天津:天津大学,2017.Tian L.Human Activity promotes proliferation and diffusion of antibiotic resistance genes in the environment[D].Tianjin:Tianjin University,2015.
[15]Huerta B,Marti E,Gros M,et al.Exploring the links between antibiotic occurrence,antibiotic resistance,and bacterial communities in water supply reservoirs[J].Science of the Total Environment,2013,456-457(7):161-170.
[16]Pruden A,Arabi M,Storteboom H N.Correlation between upstream human activities and riverine antibiotic resistance genes[J].Environmental Science and Technology,2012,46(21):11541-11549.
[17]Pei R,Kim S C,Carlson K H,et al.Effect of river landscape on the sediment concentrations of antibiotics and corresponding antibiotic resistance genes(ARG)[J].Water Research,2006,40(12):2427-2435.
[18]Chen H,Zhang M.Effects of advanced treatment systems on the removal of antibiotic resistance genes in wastewater treatment plants from Hangzhou,China[J].Environmental Science and Technology,2013,47(15):8157-8163.
[19]Liu L,Liu C,Zheng J,et al.Elimination of veterinary antibiotics and antibiotic resistance genes from swine wastewater in the vertical flow constructed wetlands[J].Chemosphere,2013,91(8):1088-1093.
[20]张子扬,刘舒巍,张璐,等.人工湿地去除畜禽养殖废水中磺胺类抗生素抗性基因研究[J].环境科学与管理,2016,41(5):89-92.Zhang Z Y,Liu S W,Zhang L,et al.Study on removal of sulfonamide antibiotic resistance genes from livestock and poultry breeding wastewater by constructed wetland[J].Environmental Science and Management,2016,41(5):89-92.
[21]郑加玉,刘琳,高大文,等.四环素抗性基因在人工湿地中的去除及累积[J].环境科学,2013,34(8):3102-3107.Zheng J Y,Liu L,Gao D W,et al.Removal and accumulation of tetracycline resistance gene in constructed wetlands[J].Environmental Science,2013,34(8):3102-3107.
[22]Shu D,He Y,Yue H,et al.Metagenomic insights into the effects of volatile fatty acids on microbial community structures and functional genes in organotrophic anammox process[J].Bioresource Technology,2015,196:621-633.
[23]王智,张志勇,张君倩,等.水葫芦修复富营养化湖泊水体区域内外底栖动物群落特征[J].中国环境科学,2012,32(1):142-149.Wang Z,Zhang Z Y,Zhang J Q,et al.Characteristics of benthos community in the waters of eutrophic lakes repaired by water hyacinth[J].Chinese Environmental Science,2012,32(1):142-149.
[24]刁晓君,李一葳,王曙光.水华生消过程对巢湖沉积物微生物群落结构的影响[J].环境科学,2015,14(1):107-113.Diao X J,Li Y M,Wang S G.Effects of the process of water blooms on the microbial community structure in sediments of Chaohu Lake[J].Environmental Science,2015,14(1):107-113.
[25]曾涛涛,蒋小梅,韩科昌,等.生活污水处理厂微生物群落结构解析[J].安全与环境学报,2018,18(2):697-703.Zeng T T,Jiang X M,Han K C,et al.Analysis of microbial community structure in domestic sewage treatment plant[J].Journal of Safety and Environment,2018,18(2):697-703.
[26]张辉,张小江,徐英春,等.2005~2014年CHINET不动杆菌属细菌耐药性监测[J].中国感染与化疗杂志,2016,16(4):429-436.Zhang H,Zhang X J,Xu Y C,et al.Monitoring of bacterial resistance of Acinetobacter spp.in 2005~2014[J].Chinese Journal of Infection and Chemotherapy,2016,16(4):429-436.
[27]Yang F,Huang L,Li L,et al.Discharge of KPC-2genes from the WWTPs contributed to their enriched abundance in the receiving river[J].Science of the Total Environment,2017,136:581-582.
[28]Teng Z,Zhu Y,Li M,et al.Microbial community composition and activity controls phosphorus transformation in rhizosphere soils of the Yeyahu Wetland in Beijing,China[J].The Science of the Total Environment,2018,628-629:1266-1277.
[29]罗晓,袁立霞,张文丽,等.制药废水厂抗性基因和微生物群落相关性研究[J].中国环境科学,2019,39(2):831-838.Luo X,Yuan L X,Zhang W L,et al.Correlation study between resistance genes and microbial communities in pharmaceutical wastewater treatment plants[J].Chinese Environmental Science,2019,39(2):831-838.
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