养殖废水处理系统中微生物菌群结构及动态变化
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摘要
采用高通量测序技术对养猪场废水处理系统的细菌群落组成进行分析,同时对细菌群落、污水水质和抗生素含量相关性进行分析.研究结果表明,污水各处理单元中的菌群结构存在显著差异,且细菌菌群结构随处理进程逐步改变,其中集污池与厌氧反应器的细菌菌群优势门类为Firmicutes、Bacteroidetes和Proteobacteria,曝气池的细菌菌群优势门类变为Firmicutes、Verrucomicrobia、Bacteroidetes、Proteobacteria和Candidatus Saccharibacteria,氧化塘的细菌菌群优势门类为Firmicutes、Proteobacteria、Chloroflexi、Euryarchaeota和Bacteroidetes.相关性分析结果表明,污水中的细菌群落与水质环境因子之间存在显著相关(P<0.05).其中,种属Acinetobacter、Bacteroides、Prevotella、Lachnospiracea_incertae_sedis与COD、TN、NH_4~+-N呈显著正相关,Clostridium sensu strict、Methanothrix与TN、NH_4~+-N呈显著负相关,Petrimonas、Lachnospiracea_incertae_sedis、Clostridium Xl Va、Tissierella与磺胺甲恶唑、磺胺间甲氧嘧啶、强力霉素、土霉素呈极显著正相关,Acinetobacter、Bacteroides、Prevotella与恩诺沙星、四环素呈显著正相关,Prevotella、Lachnospiracea_incertae_sedis与环丙沙星呈显著正相关,Clostridium sensu stricto、Petrimonas与氧氟沙星呈显著正相关.
To elucidate the bacterial community dynamics in swine wastewater treatment system, the bacterial communitycharacteristics in different treatment units of swine wastewater was analyzed by Illumina Miseq high-throughput sequencingtechnologies, and the correlation between bacterial community and water quality or antibiotics was investigated. The resultsdemonstrated that microbial community structure varies greatly in different wastewater treatment processes. The dominant bacteria infouling pool and anaerobic reator were Firmicutes, Bacteroidetes and Proteobacteria, the dominant bacteria in aeration tan wereFirmicutes, Verrucomicrobia, Bacteroidetes, Proteobacteria and Candidatus Saccharibacteria, while in oxidation pond, the dominantbacteria were Firmicutes, Proteobacteria, Chloroflexi, Euryarchaeota and Bacteroidetes. The microbial comminity structure inwasterwater treatment system was significantly correlated with both water quality and antibiotics(P<0.05). Acinetobacter,Bacteroides, Prevotella, Lachnospiracea_incertae_sedis correlated closely with COD、TN、NH_4~+-N. Clostridium sensu strict,Methanothrix correlated closely with TN、NH_4~+-N. Petrimonas, Lachnospiracea_incertae_sedis, Clostridium Xl Va, Tissierellaextremely significant positively correlated with sulfamethoxazole, sulfamethoxazole, doxycycline, oxytetracycline. Acinetobacter,Bacteroides, Prevotella significant positively correlated with enrofloxacin and tetracycline. Prevotella,Lachnospiracea_incertae_sedis positively correlated with ciprofloxacin. Clostridium sensu stricto, Petrimonas positively correlatedofloxacin.
To elucidate the bacterial community dynamics in swine wastewater treatment system, the bacterial communitycharacteristics in different treatment units of swine wastewater was analyzed by Illumina Miseq high-throughput sequencingtechnologies, and the correlation between bacterial community and water quality or antibiotics was investigated. The resultsdemonstrated that microbial community structure varies greatly in different wastewater treatment processes. The dominant bacteria infouling pool and anaerobic reator were Firmicutes, Bacteroidetes and Proteobacteria, the dominant bacteria in aeration tan wereFirmicutes, Verrucomicrobia, Bacteroidetes, Proteobacteria and Candidatus Saccharibacteria, while in oxidation pond, the dominantbacteria were Firmicutes, Proteobacteria, Chloroflexi, Euryarchaeota and Bacteroidetes. The microbial comminity structure inwasterwater treatment system was significantly correlated with both water quality and antibiotics(P<0.05). Acinetobacter,Bacteroides, Prevotella, Lachnospiracea_incertae_sedis correlated closely with COD、TN、NH_4~+-N. Clostridium sensu strict,Methanothrix correlated closely with TN、NH_4~+-N. Petrimonas, Lachnospiracea_incertae_sedis, Clostridium Xl Va, Tissierellaextremely significant positively correlated with sulfamethoxazole, sulfamethoxazole, doxycycline, oxytetracycline. Acinetobacter,Bacteroides, Prevotella significant positively correlated with enrofloxacin and tetracycline. Prevotella,Lachnospiracea_incertae_sedis positively correlated with ciprofloxacin. Clostridium sensu stricto, Petrimonas positively correlatedofloxacin.
引文
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[2]翟一帆,袁青彬,胡南.养猪废水处理系统微生物群落结构变化及影响因素研究[J].水资源保护,2018,34(1):88-94.Zhai Yi-fan,Yuan Qing-bin,Hu Nan.Study on structure variation and influencing factors of microbial community in swine wastewater treatment system[J].Water Resources Protection,2018,34(1):88-94.
[3]申童童,袁林江,温丹丹,等.侧流污泥返送到缺氧池对A2/O系统效能影响研究[J].中国环境科学,2018,38(9):3335-3342.Shen Tong-tong,Yuan Lin-jiang,Wen Dan-dan,et al.Effect of sidestream sludge returning to anoxic tank on A2/Osystem effectiveness[J].China Environmental Science,2018,38(9):3335-3342.
[4]杜龑,周北海,袁蓉芳,等.UASB-SBR工艺处理规模化畜禽养殖废水[J].环境工程学报,2018,12(2):497-504.Du Yan,Zhou Bei-hai,Yuan Rong-fang,et al.Treatment of largescale livestock wastewater by UASB-SBR process[J].Chinese Journalof Environmental Engineering,2018,12(2):497-504.
[5]支尧,张光生,钱凯,等.生物吸附/MBR/硫铁自养反硝化组合工艺优化研究[J].中国环境科学,2018,38(6):2097-2104.Zhi Yao,Zhang Guang-sheng,Qian Kai,et al.Research on optimal operation by a combined biological adsorption-MBR-Sulfur/Iron autotrophic denitrification process[J].China Environmental Science,2018,38(6):2097-2104.
[6]陈晴,王毅力,赵丽,等.ABR-MABR耦合工艺处理畜禽养殖废水的同步启动[J].环境科学研究,2017,30(2):298-305.Chen Qing,Wang Yi-li,Zhao Li,et al.Synchronous Start-Up of Anaerobic Baffled Reactor-Membrane Biofilm Bioreactor(ABR-MABR)Coupling Process Treating Synthetic Livestock and Poultry Wastewater[J].Research of Environmental Sciences,2017,30(2):298-305.
[7]赵丽,王毅力.AMBR-MABR耦合工艺处理模拟畜禽养殖废水的启动和运行[J].环境工程学报,2017,11(11):5799-5809.Zhao Li,Wang Yi-li.Start-up and operation of anaerobic migrating blanket reactor-membrane biofilm bioreactor(AMBR-MABR)coupling process treating synthetic livestock wastewater[J].Chinese Journal of Environmental Engineering,2017,11(11):5799-5809.
[8]Cydzikkwiatkowska A,Zielińska M.Bacterial communities in full-scale wastewater treatment systems[J].World Journal of Microbiology&Biotechnology,2016,32(4):66.
[9]王维奇,王秀杰,李军,等.包埋厌氧氨氧化的脱氮特性及其微生物群落结构[J].中国环境科学,2018,38(9):3343-3350.Wang Wei-qi,Wang Xiu-jie,Li Jun,et al.Nitrogen removal characteristics and microbial community structure analysis of entrapped anaerobic ammonium oxidizingbacteria[J].China Environmental Science,2018,38(9):3343-3350.
[10]Fang F,Han H,Zhao Q,et al.Bioaugmentation of biological contact oxidation reactor(BCOR)with phenol-degrading bacteria for coal gasification wastewater(CGW)treatment[J].Bioresource Technology,2013,150:314-320.
[11]Liu X,Chen Y,Zhang X,et al.Aerobic granulation strategy for bioaugmentation of a sequencing batch reactor(SBR)treating high strength pyridine wastewater[J].Journal of Hazardous Materials,2015,295:153-160.
[12]Chen Y,Lan S,Wang L,et al.A review:Driving factors and regulation strategies of microbial community structure and dynamics in wastewater treatment systems[J].Chemosphere,2017,174:173-182.
[13]Renbao L,Cheng M P,Wu M C,et al.Use of metagenomic approaches to isolate lipolytic genes from activated sludge[J].Bioresource Technology,2010,101(21):8323-8329.
[14]Garciaarmisen T,Anzil A,Cornelis P,et al.Identification of antimicrobial resistant bacteria in rivers:insights into the cultivation bias[J].Water Research,2013,47(14):4938-4947.
[15]Li Y,Adams J,Shi Y,et al.Distinct Soil Microbial Communities in habitats of differing soil water balance on the Tibetan Plateau[J].Scientific Reports,2017,7:46407.
[16]Kallenbach C M,Frey S D,Grandy A S.Direct evidence for microbial-derived soil organic matter formation and its ecophysiological controls[J].Nature Communications,2016,7:13630.
[17]Barton W,Penney N C,Cronin O,et al.The microbiome of professional athletes differs from that of more sedentary subjects in composition and particularly at the functional metabolic level[J].Gut,2018,67(4):625-633.
[18]GB 18596-2001畜禽养殖业污染物排放标准[S].GB 18596-2001 Discharge standard of pollutants forlivestock and poultry breeding[S].
[19]GB 12999-1991水质采样样品的保存和管理技术规定[S].GB 12999-1991 Water quality sampling-Technical regulation of the preservation and handling of samples[S].
[20]Mago?T,Salzberg S L.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,2011,27(21):2957-2963.
[21]Martin M.Cutadapt removes adapter sequences from high-throughput sequencing reads[J].Embnet Journal,2011,17(1):10-12.
[22]Schmieder R,Edwards R.Quality control and preprocessing of metagenomic datasets[J].Bioinformatics,2011,27(6):863-864.
[23]Edgar R C.Search and clustering orders of magnitude faster than BLAST[J].Bioinformatics,2010,26(19):2460-2461.
[24]Ye L,Shao M F,Zhang T,et al.Analysis of the bacterial community in a laboratory-scale nitrification reactor and a wastewater treatment plant by 454-pyrosequencing[J].Water Research,2011,45(15):4390-4398.
[25]Schloss P D,Westcott S L,Ryabin T,et al.Introducing mothur:open-source,platform-independent,community-supported software for describing and comparing microbial communities[J].Applied&Environmental Microbiology,2009,75(23):7537-7541.
[26]Caporaso J G,Kuczynski J,Stombaugh J,et al.QIIME allows analysis ofhigh-throughput communitysequencingdata[J].Nat Methods,2010,7(5):335-336.
[27]HJ/T 399-2007水质化学需氧量的测定快速消解分光光度法.HJ/T 399-2007 Water quality-Determination of the chemical oxygen demand-Fast digestion-Spectrophotometric method.
[28]HJ636-2012水质总氮的测定碱性过硫酸钾消解紫外分光光度法.HJ636-2012 Water quality-Determination of total nitrogen-Alkaline potassium persulfate digestion UV spectrophotometric method.
[29]HJ/T 535-2009水质氨氮的测定纳氏试剂分光光度法.HJ/T 535-2009 Waterquality-DeterminationofammonianitrogenNessler’sreagentspectrophotometry.
[30]郭欣妍,王娜,郝利君,等.超高效液相色谱/串联质谱法同时测定水、土壤及粪便中25种抗生素[J].分析化学,2015,(1):13-20.Guo Xin-Yan,Wang Na,Hao Li-Jun,et al.Simultaneous detection of25kinds of veterinary antibiotics in soil,manure and water samples using liquid chromatography_tandem mass spectrometry[J].Chinese journal of analytical chemistry,2015,(1):13-20.
[31]Oksanen J B F,Kindt R,Legendre P,et al.Vegan:community ecology R package,v2.0-10[Z].https://CRAN.R-project.org/package=vegan/2013-12-12.
[32]Warnes G R,Bolker B,Bonebakker L,et al.gplots:Various Rprogramming tools for plotting data.https://CRAN.R-project.org/package=gplots/2016-03-30.
[33]Segata N,Izard J,Waldron L,et al.Metagenomic biomarker discovery and explanation[J].Genome Biology,2011,12(6):R60.
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