不同生物过滤系统铵态氮转化速率及生物膜特性分析
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摘要
实验模拟循环水养殖系统,运行了移动床生物膜反应器(moving bed biofilm reactor,MBBR)和挂帘式生物滴滤池两类生物过滤系统的6种不同填料反应器,对比分析了各反应器的填料挂膜效果、铵态氮转化速率和生物膜微生物群落结构等特征.结果表明,与MBBR相比,挂帘式生物滴滤池挂膜速度快且生物量较多,其中碳纤维挂帘式生物滴滤池的脱膜后生物膜重最大,为45.97 g·m-2,8 h NH+4-N去除率(86.76%)高于其他反应器(61.96%~78.76%),并且NO-2-N累积少,16 h时NO-2-N浓度在0.5 mg·L~(-1)以下.通过Illumina高通量测序技术对生物过滤系统中生物膜微生物群落结构进行解析,结果表明,不同类型反应器的生物膜内的细菌、真核微生物群落构成有明显区别,无论是细菌还是真核微生物,挂帘式生物滴滤池的物种丰度和多样性均高于MBBR,但MBBR的细菌群落物种集中度更高.硝化螺菌属(Nitrospira)及放线菌中的Nakamurella属在两类反应器的生物膜中均占优势,腐螺旋菌科(Saprospiraceae)在挂帘式生物滴滤池更多,而丛毛单胞菌科(Comamonadaceae)更倾向于在MBBR中富集.在真核微生物属水平相对丰度上,挂帘式生物滴滤池中小杆目中的Rhabditida norank占明显优势,而MBBR中绿藻纲中的Chlorophyceae norank占明显优势.以上研究结果为挂帘式生物滴滤池在循环水养殖水质净化中的应用奠定了实验基础.
Three MBBRs and three curtain type trickling filters(CTFs) with different carriers were operated in lab-scale simulated RASs. The characteristics of biofilms,ammonia removal rates and microbial communities in six reactors were compared with each other. Compared with the biofilms of MBBRs,the biofilms of CTFs were heavier and grew faster. The weight of biofilms on CTFs with carbon fiber carriers was the maximum(45. 97 g·m-2),and the ammonia nitrogen removal efficiency(86. 76%) of this CTFs was higher than the other filters or reactors(61. 96% ~ 78. 76%). In addition,the concentration of the accumulated nitrite in the carbon fiber CTFs was the lowest in all the six reactors. The microbial communities of biofilms in the six biofilters were evaluated by the highthroughput Illumina-Mi Seq sequencing technology. The results showed that the microbial(bacteria and eukaryote) community in biofilms of CTFs was different from that in biofilms of MBBRs. At both bacteria and micro-eukaryote level,the species richness and biological diversity of biofilms in the trickling filters were higher than those in the MBBRs. On the contrary,the Simpson index of bacterial community in biofilms of MBBRs was higher than that in the trickling filters. In all the six biofilters,Nitrospira and Nakamurella were the dominated bacterial genera. Saprospiraceae was more abundant in CTFs than in MBBRs,but Comamonadaceae was enriched in the MBBRs. At the micro-eukaryote genus level,Rhabditida norank genus was more abundant in CTFs,while Chlorophyceae norank genus was more abundant in the MBBRs. The results provide useful information about microbial ecology that can be used for the application of CTFs in RAS.
Three MBBRs and three curtain type trickling filters(CTFs) with different carriers were operated in lab-scale simulated RASs. The characteristics of biofilms,ammonia removal rates and microbial communities in six reactors were compared with each other. Compared with the biofilms of MBBRs,the biofilms of CTFs were heavier and grew faster. The weight of biofilms on CTFs with carbon fiber carriers was the maximum(45. 97 g·m-2),and the ammonia nitrogen removal efficiency(86. 76%) of this CTFs was higher than the other filters or reactors(61. 96% ~ 78. 76%). In addition,the concentration of the accumulated nitrite in the carbon fiber CTFs was the lowest in all the six reactors. The microbial communities of biofilms in the six biofilters were evaluated by the highthroughput Illumina-Mi Seq sequencing technology. The results showed that the microbial(bacteria and eukaryote) community in biofilms of CTFs was different from that in biofilms of MBBRs. At both bacteria and micro-eukaryote level,the species richness and biological diversity of biofilms in the trickling filters were higher than those in the MBBRs. On the contrary,the Simpson index of bacterial community in biofilms of MBBRs was higher than that in the trickling filters. In all the six biofilters,Nitrospira and Nakamurella were the dominated bacterial genera. Saprospiraceae was more abundant in CTFs than in MBBRs,but Comamonadaceae was enriched in the MBBRs. At the micro-eukaryote genus level,Rhabditida norank genus was more abundant in CTFs,while Chlorophyceae norank genus was more abundant in the MBBRs. The results provide useful information about microbial ecology that can be used for the application of CTFs in RAS.
引文
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[16]Amato K R,Yeoman C J,Kent A,et al.Habitat degradation impacts black howler monkey(Alouatta pigra)gastrointestinal microbiomes[J].The ISME Journal,2013,7(7):1344-1353.
[17]Zhang S F,Wang Y Y,He W T,et al.Linking nitrifying biofilm characteristics and nitrification performance in moving-bed biofilm reactors for polluted raw water pretreatment[J].Bioresource Technology,2013,146:416-425.
[18]刘雨,赵庆良,郑兴灿.生物膜法污水处理技术[M].北京:中国建筑工业出版社,2000.38-40.
[19]邱立平,马军,张立昕.水力停留时间对曝气生物滤池处理效能及运行特性的影响[J].环境污染与防治,2004,26(6):433-436.Qiu L P,Ma J,Zhang L X.Effect of hydraulic retention time on the treatment efficiency and operational characteristics of biological aerated filter[J].Environmental Pollution&Control,2004,26(6):433-436.
[20]裘钱玲琳,徐如卫,朱建林,等.凡纳滨对虾土塘养殖与循环养殖系统细菌群落结构比较研究[J].生物学杂志,2015,32(6):45-49.Qiu Q L L,Xu R W,Zhu J L,et al.Comparison of bacterial communities in the earthen pond and the recirculating aquaculture system for Litopenaeus vannamei cultivation[J].Journal of Biology,2015,32(6):45-49.
[21]黄志涛,宋协法,李勋,等.基于高通量测序的石斑鱼循环水养殖生物滤池微生物群落分析[J].农业工程学报,2016,32(S1):242-247.Huang Z T,Song X F,Li X,et al.Analysis of microbial diversity of submerged biofilters in recirculating aquaculture system(RAS)for grouper(Epinehelus moara)based on highthroughput DNA sequencing[J].Transactions of the Chinese Society of Agricultural Engineering,2016,32(S1):242-247.
[22]Attramadal K J K,Truong T M H,Bakke I,et al.RAS and microbial maturation as tools for K-selection of microbial communities improve survival in cod larvae[J].Aquaculture,2014,432:483-490.
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[24]Rurangwa E,Verdegem M C J.Microorganisms in recirculating aquaculture systems and their management[J].Reviews in Aquaculture,2015,7(2):117-130.
[25]宋爱红.强化脱氮人工湿地处理分散养猪冲洗水特性研究[D].西安:长安大学,2015.
[26]Kirchman D L.The ecology of Cytophaga-Flavobacteria in aquatic environments[J].FEMS Microbiology Ecology,2002,39(2):91-100.
[27]Itoi S,Niki A,Sugita H.Changes in microbial communities associated with the conditioning of filter material in recirculating aquaculture systems of the pufferfish Takifugu rubripes[J].Aquaculture,2006,256(1-4):287-295.
[28]van Kessel M A,Dutilh B E,Neveling K,et al.Pyrosequencing of 16S rRNA gene amplicons to study the microbiota in the gastrointestinal tract of carp(Cyprinus carpio,L.)[J].AMB Express,2011,1:41.
[29]Hovanec T A,Taylor L T,Blakis A,et al.Nitrospira-Like bacteria associated with nitrite oxidation in freshwater aquaria[J].Applied and Environmental Microbiology,1998,64(1):258-264.
[30]吕福荣,杨海波,李英敏,等.自养条件下小球藻净化氮、磷能力的研究[J].生物技术,2003,13(6):46-47.Lv F R,Yang H B,Li Y M,et al.Study on the N,P purification ability of chlorella under autotrophic condition[J].Biotechnology,2003,13(6):46-47.
[31]孙诗琦,肖文帅,王亮,等.藻类对重金属废水的处理进展[J].广东化工,2014,41(15):201-202,196.Sun S Q,Xiao W S,Wang L,et al.Progress in treatment of heavy metal wastewater by algae[J].Guangdong Chemical Industry,2014,41(15):201-202,196.
[32]李小伟.生物活性炭滤池无脊椎动物群落分析及控制技术[D].广州:暨南大学,2008.10-11.
[33]王家楫.中国淡水轮虫志[M].北京:科学出版社,1961.
[34]Tice H,Mayilraj S,Sims D,et al.Complete genome sequence of Nakamurella multipartita type strain(Y-104T)[J].Standards in Genomic Sciences,2010,2:2.
[35]Schreier H J,Mirzoyan N,Saito K.Microbial diversity of biological filters in recirculating aquaculture systems[J].Current Opinion in Biotechnology,2010,21(3):318-325.
[2]沈加正,史明明,阮贇杰,等.变速流操控对调节循环水养殖系统水质效果的研究[J].农业机械学报,2016,47(7):309-314.Shen J Z,Shi M M,Ruan Y J,et al.Effects of variable flow rates on water qualities in recirculating aquaculture system[J].Transactions of the Chinese Society for Agricultural Machinery,2016,47(7):309-314.
[3]朱建新,刘慧,徐勇,等.循环水养殖系统生物滤器负荷挂膜技术[J].渔业科学进展,2014,35(4):118-124.Zhu J X,Liu H,Xu Y,et al.Dual-Culture techniques for the rapid start-up of recirculating aquaculture system[J].Progress in Fishery Sciences,2014,35(4):118-124.
[4]王威,曲克明,朱建新,等.3种滤料生物滤器的挂膜与黑鲷幼鱼循环水养殖效果[J].中国水产科学,2012,19(5):833-840.Wang W,Qu K M,Zhu J X,et al.Biofilm cultivation with three stuffings and their effect on the growth of young black sea bream,Sparus macrocephalus[J].Journal of Fishery Sciences of China,2012,19(5):833-840.
[5]周子明,李华,刘青松,等.工厂化循环水养殖系统中生物填料的研究现状[J].水处理技术,2015,(12):33-37.Zhou Z M,Li H,Liu Q S,et al.Current situation of the research on biological media in recirculating aquaculture system[J].Technology of Water Treatment,2015,(12):33-37.
[6]sgrd T,Shearer K D.Dietary phosphorus requirement of juvenile Atlantic salmon,Salmo salar L.[J].Aquaculture Nutrition,1997,3(1):17-23.
[7]丁爱中,陈繁忠,雷剑泉,等.光合细菌调控水产养殖业水质的研究[J].农业环境保护,2000,19(6):339-341,344.Ding A Z,Chen F Z,Lei J Q,et al.Water quality for aquiculture improved by photosynthetic bacteria[J].Agroenvironmental Protection,2000,19(6):339-341,344.
[8]Dong B,Tan J,Yang Y,et al.Linking nitrification characteristic and microbial community structures in integrated fixed film activated sludge reactor by high-throughput sequencing[J].Water Science and Technology,2016,74(6):1354-1364.
[9]Leyva-Díaz J C,González-Martínez A,Mu1ío M M,et al.Twostep nitrification in a pure moving bed biofilm reactor-membrane bioreactor for wastewater treatment:nitrifying and denitrifying microbial populations and kinetic modeling[J].Applied Microbiology and Biotechnology,2015,99(23):10333-10343.
[10]于莉芳,杜倩倩,傅学焘,等.城市污水中硝化菌群落结构与性能分析[J].环境科学,2016,37(11):4366-4371.Yu L F,Du Q Q,Fu X T,et al.Community structure and activity analysis of the nitrifiers in raw sewage of wastewater treatment plants[J].Environmental Science,2016,37(11):4366-4371.
[11]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,2002.266-281.
[12]Qiu T L,Liu L L,Gao M,et al.Effects of solid-phase denitrification on the nitrate removal and bacterial community structure in recirculating aquaculture system[J].Biodegradation,2016,27(2-3):165-178.
[13]Brte J,Logares R,Berney C,et al.Freshwater Perkinsea and marine-freshwater colonizations revealed by pyrosequencing and phylogeny of environmental r DNA[J].ISME Journal,2010,4(9):1144-1153.
[14]Luo W,Li H R,Gao S Q,et al.Molecular diversity of microbial eukaryotes in sea water from Fildes Peninsula,King George Island,Antarctica[J].Polar Biology,2016,39(4):605-616.
[15]Wang L,Cheung M K,Kwan H S,et al.Microbial diversity in shallow-water hydrothermal sediments of Kueishan Island,Taiwan as revealed by pyrosequencing[J].Journal of Basic Microbiology,2015,55(11):1308-1318.
[16]Amato K R,Yeoman C J,Kent A,et al.Habitat degradation impacts black howler monkey(Alouatta pigra)gastrointestinal microbiomes[J].The ISME Journal,2013,7(7):1344-1353.
[17]Zhang S F,Wang Y Y,He W T,et al.Linking nitrifying biofilm characteristics and nitrification performance in moving-bed biofilm reactors for polluted raw water pretreatment[J].Bioresource Technology,2013,146:416-425.
[18]刘雨,赵庆良,郑兴灿.生物膜法污水处理技术[M].北京:中国建筑工业出版社,2000.38-40.
[19]邱立平,马军,张立昕.水力停留时间对曝气生物滤池处理效能及运行特性的影响[J].环境污染与防治,2004,26(6):433-436.Qiu L P,Ma J,Zhang L X.Effect of hydraulic retention time on the treatment efficiency and operational characteristics of biological aerated filter[J].Environmental Pollution&Control,2004,26(6):433-436.
[20]裘钱玲琳,徐如卫,朱建林,等.凡纳滨对虾土塘养殖与循环养殖系统细菌群落结构比较研究[J].生物学杂志,2015,32(6):45-49.Qiu Q L L,Xu R W,Zhu J L,et al.Comparison of bacterial communities in the earthen pond and the recirculating aquaculture system for Litopenaeus vannamei cultivation[J].Journal of Biology,2015,32(6):45-49.
[21]黄志涛,宋协法,李勋,等.基于高通量测序的石斑鱼循环水养殖生物滤池微生物群落分析[J].农业工程学报,2016,32(S1):242-247.Huang Z T,Song X F,Li X,et al.Analysis of microbial diversity of submerged biofilters in recirculating aquaculture system(RAS)for grouper(Epinehelus moara)based on highthroughput DNA sequencing[J].Transactions of the Chinese Society of Agricultural Engineering,2016,32(S1):242-247.
[22]Attramadal K J K,Truong T M H,Bakke I,et al.RAS and microbial maturation as tools for K-selection of microbial communities improve survival in cod larvae[J].Aquaculture,2014,432:483-490.
[23]Loreau M,Naeem S,Inchausti P,et al.Biodiversity and ecosystem functioning:current knowledge and future challenges[J].Science,2001,294(5543):804-808.
[24]Rurangwa E,Verdegem M C J.Microorganisms in recirculating aquaculture systems and their management[J].Reviews in Aquaculture,2015,7(2):117-130.
[25]宋爱红.强化脱氮人工湿地处理分散养猪冲洗水特性研究[D].西安:长安大学,2015.
[26]Kirchman D L.The ecology of Cytophaga-Flavobacteria in aquatic environments[J].FEMS Microbiology Ecology,2002,39(2):91-100.
[27]Itoi S,Niki A,Sugita H.Changes in microbial communities associated with the conditioning of filter material in recirculating aquaculture systems of the pufferfish Takifugu rubripes[J].Aquaculture,2006,256(1-4):287-295.
[28]van Kessel M A,Dutilh B E,Neveling K,et al.Pyrosequencing of 16S rRNA gene amplicons to study the microbiota in the gastrointestinal tract of carp(Cyprinus carpio,L.)[J].AMB Express,2011,1:41.
[29]Hovanec T A,Taylor L T,Blakis A,et al.Nitrospira-Like bacteria associated with nitrite oxidation in freshwater aquaria[J].Applied and Environmental Microbiology,1998,64(1):258-264.
[30]吕福荣,杨海波,李英敏,等.自养条件下小球藻净化氮、磷能力的研究[J].生物技术,2003,13(6):46-47.Lv F R,Yang H B,Li Y M,et al.Study on the N,P purification ability of chlorella under autotrophic condition[J].Biotechnology,2003,13(6):46-47.
[31]孙诗琦,肖文帅,王亮,等.藻类对重金属废水的处理进展[J].广东化工,2014,41(15):201-202,196.Sun S Q,Xiao W S,Wang L,et al.Progress in treatment of heavy metal wastewater by algae[J].Guangdong Chemical Industry,2014,41(15):201-202,196.
[32]李小伟.生物活性炭滤池无脊椎动物群落分析及控制技术[D].广州:暨南大学,2008.10-11.
[33]王家楫.中国淡水轮虫志[M].北京:科学出版社,1961.
[34]Tice H,Mayilraj S,Sims D,et al.Complete genome sequence of Nakamurella multipartita type strain(Y-104T)[J].Standards in Genomic Sciences,2010,2:2.
[35]Schreier H J,Mirzoyan N,Saito K.Microbial diversity of biological filters in recirculating aquaculture systems[J].Current Opinion in Biotechnology,2010,21(3):318-325.