环丙沙星对膜生物反应器中微生物群落及抗性基因的影响
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摘要
用膜生物反应器(membrane bioreactor,MBR)处理含环丙沙星(ciprofloxacin,CIP)的合成废水,考察了不同CIP进水浓度(0、5、10、15 mg·L-1)下MBR的微生物群落特征和抗性基因丰度的变化.结果表明,随着进水中CIP浓度从0 mg·L-1增加至15 mg·L-1,变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes)仍保持为优势菌门,相对丰度比例分别为57.5%和12.7%;红环菌科(Rhodocyclaceae)、Chitinophagaceae和丛毛单胞菌科(Comamonadaceae)被选择成为优势菌科,比例分别为29.96%、5.44%和6.60%;Methyloversatilis、Ferruginibacter、动胶菌属(Zoogloea)和丛毛单胞菌属(Comamonas)被选择成为优势菌属,比例分别为21.70%、7.56%、5.24%和4.15%;Chao1、ACE、Shannon指数逐渐降低和Simpson指数逐渐升高,表明MBR污泥中微生物丰富度和多样性均降低;亚硝化单胞菌属(Nitrosomonas)、硝化螺旋菌属(Nitrospira)、产碱菌属(Alcaligenes)和硝化杆菌属(Nitrobacter)相对丰度减少,使得氨氮去除率降低.CIP抗性基因(CIP-ARGs)分析表明,当MBR在CIP投加浓度为5 mg·L-1下运行至第33 d时,反应器中的gyr A、gyr B和par C基因相对丰度较CIP投加初期增加,加大了抗药风险.
A membrane bioreactor(MBR) was used to treat ciprofloxacin(CIP)-contaminated artificial wastewater. The microbial community structure and the abundance of antibiotic resistant genes(ARGs) in the MBR were studied at four CIP dosages(0,5 mg·L-1,10 mg·L-1,and 15 mg·L-1). The results showed that Proteobacteria and Bacteroidetes remained the dominant phylum,with relative abundances of 57. 5% and 12. 7%,respectively,as the dosage of CIP was increased from 0 mg·L-1 to 15 mg·L-1.Rhodocyclaceae,Chitinophagaceae,and Comamonadaceae became the dominant family with abundances of 29. 96%,5. 44%,and6. 60%,respectively. Methyloversatilis,Ferruginibacter,Zoogloea,and Comamonas became the dominant genus,with relative abundances of 21. 70%,7. 56%,5. 24%,and 4. 15%,respectively. The decrease of Chao1,ACE,and Shannon and the increase of Simpson indicated a decrease in microbial abundance and diversity. The relative abundances of Nitrosomonas,Nitrospira,Alcaligenes,and Nitrobacter decreased,which caused a decrease in the NH3-N removal rate. A CIP-ARGs analysis revealed that the relative abundances of gyr A,gyr B,and par C were increased,beginning after the sludge was dosed with 5 mg·L-1 of CIP for 33 days,which augmented the risk for microbial drug-resistance.
A membrane bioreactor(MBR) was used to treat ciprofloxacin(CIP)-contaminated artificial wastewater. The microbial community structure and the abundance of antibiotic resistant genes(ARGs) in the MBR were studied at four CIP dosages(0,5 mg·L-1,10 mg·L-1,and 15 mg·L-1). The results showed that Proteobacteria and Bacteroidetes remained the dominant phylum,with relative abundances of 57. 5% and 12. 7%,respectively,as the dosage of CIP was increased from 0 mg·L-1 to 15 mg·L-1.Rhodocyclaceae,Chitinophagaceae,and Comamonadaceae became the dominant family with abundances of 29. 96%,5. 44%,and6. 60%,respectively. Methyloversatilis,Ferruginibacter,Zoogloea,and Comamonas became the dominant genus,with relative abundances of 21. 70%,7. 56%,5. 24%,and 4. 15%,respectively. The decrease of Chao1,ACE,and Shannon and the increase of Simpson indicated a decrease in microbial abundance and diversity. The relative abundances of Nitrosomonas,Nitrospira,Alcaligenes,and Nitrobacter decreased,which caused a decrease in the NH3-N removal rate. A CIP-ARGs analysis revealed that the relative abundances of gyr A,gyr B,and par C were increased,beginning after the sludge was dosed with 5 mg·L-1 of CIP for 33 days,which augmented the risk for microbial drug-resistance.
引文
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[18]Sapkota A R,Curriero F C,Gibson K E,et al.Antibioticresistant enterococci and fecal indicators in surface water and groundwater impacted by a concentrated Swine feeding operation[J].Environmental Health Perspectives,2007,115(7):1040-1045.
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[20]葛伟丽.长期施用鸡粪农田土壤中典型抗生素耐药细菌及其抗性基因污染分析[D].济南:山东农业大学,2014.Ge W L.Typical antibiotic resistant bacteria and resistance gene in the farmland soil fertilized with chicken manure chronically[D].Ji'nan:Shandong Agricultural University,2014.
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[22]Sui Q,Huang J,Deng S B,et al.Occurrence and removal of pharmaceuticals,caffeine and DEET in wastewater treatment plants of Beijing,China[J].Water Research,2010,44(2):417-426.
[23]Liao X B,Li B X,Zou R S,et al.Biodegradation of antibiotic ciprofloxacin:pathways,influential factors,and bacterial community structure[J].Environmental Science and Pollution Research,2016,23(8):7911-7918.
[24]Gonzalez-Martinez A,Rodriguez-Sanchez A,Martinez-Toledo M V,et al.Effect of ciprofloxacin antibiotic on the partialnitritation process and bacterial community structure of a submerged biofilter[J].Science of the Total Environment,2014,476-477:276-287.
[25]Cui H,Wang S P,Fu J,et al.Influence of ciprofloxacin on microbial community structure and function in soils[J].Biology and Fertility of Soils,2014,50(6):939-947.
[26]Xie Y W,Chen L J,Liu R.AOX contamination status and genotoxicity of AOX-bearing pharmaceutical wastewater[J].Journal of Environmental Sciences,2017,52:170-177.
[27]戴琦,刘锐,舒小铭,等.环丙沙星对膜生物反应器运行效能的影响及其去除特性[J].环境科学,2018,39(1):212-218.Dai Q,Liu R,Shu X M,et al.Removal and influence of ciprofloxacin in a membrane bioreactor[J].Environmental Science,2018,39(1):212-218.
[28]魏铭聪.甲烷渗漏环境中自生矿物形成条件的实验研究[D].长春:吉林大学,2016.Wei M C.The formation of authigenic minerals during methane seeping in seafloor:insight from laboratory test[D].Changchun:Jilin University,2016.
[29]Rubin J,Walker R D,Blickenstaff K,et al.Antimicrobial resistance and genetic characterization of fluoroquinolone resistance of Pseudomonas aeruginosa isolated from canine infections[J].Veterinary Microbiology,2008,131(1-2):164-172.
[30]国家环境保护总局.水和废水监测分析方法[M].(第四版).北京:中国环境科学出版社,2002.211-213,277-281.
[31]何势.曝气生物滤池中环丙沙星去除行为及其对降解菌群抗药性的诱导作用[D].上海:东华大学,2016.He S.Removal behavior of ciprofloxacin in the biological aerated filter and its effect on the resistance of bacteria[D].Shanghai:Donghua University,2016.
[32]Jung C M,Heinze T M,Strakosha R,et al.Acetylation of fluoroquinolone antimicrobial agents by an Escherichia coli strain isolated from a municipal wastewater treatment plant[J].Journal of Applied Microbiology,2009,106(2):564-571.
[33]何势,顾超超,魏欣,等.低浓度环丙沙星对曝气生物滤池生物膜硝化过程及硝化微生物的作用影响[J].环境科学,2016,37(4):1485-1491.He S,Gu C C,Wei X,et al.Effect of low-concentration ciprofloxacin on the nitrification and nitrifying microorganisms of biofilms in biological aerated filter[J].Environmental Science,2016,37(4):1485-1491.
[34]Liu L,Liu C X,Zheng J Y,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.
[2]Zhang Q Q,Ying G G,Pan C G,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&Technology,2015,49(11):6772-6782.
[3]Mu1oz I,JoséG M,Molina-Díaz A,et al.Ranking potential impacts of priority and emerging pollutants in urban wastewater through life cycle impact assessment[J].Chemosphere,2008,74(1):37-44.
[4]Guney G,Sponza D T.Comparison of biological and advanced treatment processes for ciprofloxacin removal in a raw hospital wastewater[J].Environmental Technology,2016,37(24):3151-3167.
[5]刘艳萍,刘鸿雁,吴龙华,等.贵阳市某蔬菜地养殖废水污灌土壤重金属、抗生素复合污染研究[J].环境科学学报,2017,37(3):1074-1082.Liu Y P,Liu H Y,Wu L H,et al.Co-contamination of heavy metals and antibiotics in soils under husbandry wastewater irrigation in Guiyang City[J].Acta Scientiae Circumstantiae,2017,37(3):1074-1082.
[6]刘畅,李瑜.环丙沙星在土壤中的吸附及降解研究进展[J].北京农业,2016,(4):195-199.
[7]郭睿,王山军,昌盛,等.嘉兴市饮用水源及城市河网抗生素分布特征[J].环境化学,2016,35(9):1842-1852.Guo R,Wang S J,Chang S,et al.The distribution characteristics of antibiotics in Jiaxing drinking water source and urban river[J].Environmental Chemistry,2016,35(9):1842-1852.
[8]Luo Y,Xu L,Rysz M,et al.Occurrence and transport of tetracycline,sulfonamide,quinolone,and macrolide antibiotics in the Haihe River basin,China[J].Environmental Science&Technology,2011,45(5):1827-1833.
[9]Zhou L J,Ying G G,Zhao J L,et al.Trends in the occurrence of human and veterinary antibiotics in the sediments of the Yellow River,Hai River and Liao River in northern China[J].Environmental Pollution,2011,159(7):1877-1885.
[10]Dorivalgarcía N,Zafra-Gómez A,Navalón A,et al.Removal of quinolone antibiotics from wastewaters by sorption and biological degradation in laboratory-scale membrane bioreactors[J].Science of the Total Environment,2013,442:317-328.
[11]Liu H,Zhang G P,Liu C Q,et al.The occurrence of chloramphenicol and tetracyclines in municipal sewage and the Nanming River,Guiyang City,China[J].Journal of Environmental Monitoring,2009,11(6):1199-1205.
[12]Veldman K,Cavaco L M,Mevius D,et al.International collaborative study on the occurrence of plasmid-mediated quinolone resistance in Salmonella enterica and Escherichia coli isolated from animals,humans,food and the environment in 13European countries[J].Journal of Antimicrobial Chemotherapy,2011,66(6):1278-1286.
[13]王小亮,徐立蒲,曹欢,等.鱼源病原菌对氟喹诺酮类药物的耐药性分析[J].中国畜牧兽医,2013,40(3):195-199.Wang X L,Xu L P,Cao H,et al.Analysis of the fluoroquinolone resistance of pathogens isolated from fish[J].China Animal Husbandry&Veterinary Medicine,2013,40(3):195-199.
[14]Aoike N,Saga T,Sakata R,et al.Molecular characterization of extraintestinal Escherichia coli isolates in Japan:relationship between sequence types and mutation patterns of quinolone resistance-determining regions analyzed by pyrosequencing[J].Journal of Clinical Microbiology,2013,51(6):1692-1698.
[15]马驿,彭金菊,王芸,等.环丙沙星对土壤微生物量碳和土壤微生物群落碳代谢多样性的影响[J].生态学报,2013,33(5):1506-1512.Ma Y,Peng J J,Wang Y,et al.Effects of ciprofloxacin on microbial biomass carbon and carbon metabolism diversity of soil microbial communities[J].Acta Ecologica Sinica,2013,33(5):1506-1512.
[16]Wunder D B,Tan D T,Lapara T M,et al.The effects of antibiotic cocktails at environmentally relevant concentrations on the community composition and acetate biodegradation kinetics of bacterial biofilms[J].Chemosphere,2013,90(8):2261-2266.
[17]Li J,Wang T,Shao B,et al.Plasmid-mediated quinolone resistance genes and antibiotic residues in wastewater and soil adjacent to swine feedlots:potential transfer to agricultural lands[J].Environmental Health Perspectives,2012,120(8):1144-1149.
[18]Sapkota A R,Curriero F C,Gibson K E,et al.Antibioticresistant enterococci and fecal indicators in surface water and groundwater impacted by a concentrated Swine feeding operation[J].Environmental Health Perspectives,2007,115(7):1040-1045.
[19]Rodriguez-Mozaz S,Chamorro S,Marti E,et al.Occurrence of antibiotics and antibiotic resistance genes in hospital and urban wastewaters and their impact on the receiving river[J].Water Research,2015,69:234-242.
[20]葛伟丽.长期施用鸡粪农田土壤中典型抗生素耐药细菌及其抗性基因污染分析[D].济南:山东农业大学,2014.Ge W L.Typical antibiotic resistant bacteria and resistance gene in the farmland soil fertilized with chicken manure chronically[D].Ji'nan:Shandong Agricultural University,2014.
[21]Sharma V K,Johnson N,Cizmas L,et al.A review of the influence of treatment strategies on antibiotic resistant bacteria and antibiotic resistance genes[J].Chemosphere,2016,150:702-714.
[22]Sui Q,Huang J,Deng S B,et al.Occurrence and removal of pharmaceuticals,caffeine and DEET in wastewater treatment plants of Beijing,China[J].Water Research,2010,44(2):417-426.
[23]Liao X B,Li B X,Zou R S,et al.Biodegradation of antibiotic ciprofloxacin:pathways,influential factors,and bacterial community structure[J].Environmental Science and Pollution Research,2016,23(8):7911-7918.
[24]Gonzalez-Martinez A,Rodriguez-Sanchez A,Martinez-Toledo M V,et al.Effect of ciprofloxacin antibiotic on the partialnitritation process and bacterial community structure of a submerged biofilter[J].Science of the Total Environment,2014,476-477:276-287.
[25]Cui H,Wang S P,Fu J,et al.Influence of ciprofloxacin on microbial community structure and function in soils[J].Biology and Fertility of Soils,2014,50(6):939-947.
[26]Xie Y W,Chen L J,Liu R.AOX contamination status and genotoxicity of AOX-bearing pharmaceutical wastewater[J].Journal of Environmental Sciences,2017,52:170-177.
[27]戴琦,刘锐,舒小铭,等.环丙沙星对膜生物反应器运行效能的影响及其去除特性[J].环境科学,2018,39(1):212-218.Dai Q,Liu R,Shu X M,et al.Removal and influence of ciprofloxacin in a membrane bioreactor[J].Environmental Science,2018,39(1):212-218.
[28]魏铭聪.甲烷渗漏环境中自生矿物形成条件的实验研究[D].长春:吉林大学,2016.Wei M C.The formation of authigenic minerals during methane seeping in seafloor:insight from laboratory test[D].Changchun:Jilin University,2016.
[29]Rubin J,Walker R D,Blickenstaff K,et al.Antimicrobial resistance and genetic characterization of fluoroquinolone resistance of Pseudomonas aeruginosa isolated from canine infections[J].Veterinary Microbiology,2008,131(1-2):164-172.
[30]国家环境保护总局.水和废水监测分析方法[M].(第四版).北京:中国环境科学出版社,2002.211-213,277-281.
[31]何势.曝气生物滤池中环丙沙星去除行为及其对降解菌群抗药性的诱导作用[D].上海:东华大学,2016.He S.Removal behavior of ciprofloxacin in the biological aerated filter and its effect on the resistance of bacteria[D].Shanghai:Donghua University,2016.
[32]Jung C M,Heinze T M,Strakosha R,et al.Acetylation of fluoroquinolone antimicrobial agents by an Escherichia coli strain isolated from a municipal wastewater treatment plant[J].Journal of Applied Microbiology,2009,106(2):564-571.
[33]何势,顾超超,魏欣,等.低浓度环丙沙星对曝气生物滤池生物膜硝化过程及硝化微生物的作用影响[J].环境科学,2016,37(4):1485-1491.He S,Gu C C,Wei X,et al.Effect of low-concentration ciprofloxacin on the nitrification and nitrifying microorganisms of biofilms in biological aerated filter[J].Environmental Science,2016,37(4):1485-1491.
[34]Liu L,Liu C X,Zheng J Y,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.