冷水鱼循环水养殖系统生物滤池成熟过程及低温氮降解菌的分离与鉴定
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摘要
本文研究了冷水鱼循环水养殖系统生物滤池的成熟过程,同时通过对成熟生物滤料上净化微生物进行了富集,对低温条件下具有有效降解氨氮和亚硝酸氮的细菌进行了分离鉴定。研究表明,在温度17～18℃,pH为7.76±0.28,盐度0的条件下,经过40d的培养,生物滤池可完全成熟。但受较高溶氧条件的限制,成熟后的生物滤池只能将养殖水中高毒害的氨氮和亚硝酸氮转化为低毒害的硝酸氮,而无法通过反硝化作用将氮和磷从系统中移除。同时根据水体中有机碳和无机碳的变化,推测生物滤料上自养菌群的产生滞后于异养菌群。经分离、鉴定得到4株耐低温的氮降解细菌,分别为枯草芽孢杆菌(Bacillus subtilis)、Velezensis芽孢杆菌(Bacillus velezensis)、土芽孢杆菌(Geobacillus sp.)和短小芽孢杆菌(Bacillus pumilus)。
Aiming to explore the maturation of the biofilter in the newly built recirculating aquaculture system rearing cold-water fish,we carried out this study.We also sampled the biofilm to isolate ammonium and nitrite degrading bacteria.The result indicated that after 40 days of cultivation,the biofilm matured at 17～18℃,pH=7.76±0.28 and salinity 0.However,due to the affection of high content dissolved oxygen,the matured biofilter transformed only ammonium and nitrite into nitrate,and it was not able to degrade nitrogen and phosphorus through denitrification.In the biofilter,the degradation of TOC was ahead of TIC in order.We inferred that the production of autotrophic bacteria was later than that of heterotrophic bacteria.Four bacteria were isolated,and identified as Bacillus subtilis,B.velezensis,Geobacillus sp.,and B.pumilus,respectively.
Aiming to explore the maturation of the biofilter in the newly built recirculating aquaculture system rearing cold-water fish,we carried out this study.We also sampled the biofilm to isolate ammonium and nitrite degrading bacteria.The result indicated that after 40 days of cultivation,the biofilm matured at 17～18℃,pH=7.76±0.28 and salinity 0.However,due to the affection of high content dissolved oxygen,the matured biofilter transformed only ammonium and nitrite into nitrate,and it was not able to degrade nitrogen and phosphorus through denitrification.In the biofilter,the degradation of TOC was ahead of TIC in order.We inferred that the production of autotrophic bacteria was later than that of heterotrophic bacteria.Four bacteria were isolated,and identified as Bacillus subtilis,B.velezensis,Geobacillus sp.,and B.pumilus,respectively.
引文
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[3]冯志华,徐军田,李玉,等.封闭循环海水养殖生物滤池生物膜形成过程研究[J].淮海工学院学报(自然科学版),2010,19(4):79-82.Feng Z,Xu J,Li Y,et al.Study on the biofilm formation process of biofilter in a closed seawater recirculating aquaculture system[J].Journal of Huaihai Institute of Technology(Natural Science Edition),2010,19(4):79-82.
[4] Blancheton J P,Attramadal K J K,Michaud L,et al.Insight into bacterial population in aquaculture systems and its implication[J].Aquacult Eng,2012,53:30-39.
[5] Rud I,Kolarevic J,Holan A B,et al.Deep-sequencing of the bacterial microbiota in commercial-scale recirculating and semi-closed aquaculture systems for Atlantic salmon post-smolt production[J].Aquacult Eng,2017,78:50-62.
[6] Pedersen L,Oosterveld R,Pedersen P B.Nitrification performance and robustness of fixed and moving bed biofilters having identical carrier elements[J].Aquacult Eng,2014,10:37-45.
[7] Guerdat T C,Losordo T M,Classen J J,et al.An evaluation of commercially available biological filters for recirculating aquaculture systems[J].Aquacult Eng,2010,42(1):38-49.
[8] Guerdat T C.A Large Scale Evaluation of Commercially Available Biological Filters for Recirculating Aquaculture Systems[D].Raleigh:North Carolina State University,2008.
[9] Badiola M,Mendiola D,Bostock J.Recirculating Aquaculture Systems(RAS)analysis:Main issues on management and future challenges[J].Aquacult Eng,2012,51:26-35.
[10]吴嘉敏,孙大川.循环水养殖系统中浸没式生物滤器的水处理效果[J].上海水产大学学报,2007,16(6):542-548.Wu J,Sun D.Study on the water treatment efficiency of submerged biological filter in the recirculating aquaculture system[J].Journal of Shanghai Fisheries University,2007,16(6):542-548.
[11]管敏,李莎,张建明,等.循环水养殖系统生物滤池自然挂膜启动及效果研究[J].中国给水排水,2015,31(1):80-83.Guan M,Li S,Zhang J,et al.Natural start-up and water treatment effect of biofilter for recirculating aquaculture system[J].China Water&Wastewater,2015,31(1):80-83.
[12]宋协法,李华龙.养殖污水中主要分解氨氮微生物的分离鉴定[J].渔业现代化,2013,40(1):1-6.Song X,Li H.Research of nitrifying bacteria from cultivation wastewater[J].Fishery Modernization,2013,40(1):1-6.
[13]熊焰,刘力源,罗睿,等.1株亚硝酸盐降解菌的筛选-鉴定-降解条件及效果[J].中国水产科学,2010,17(6):1264-1271.Xiong Y,Liu L,Luo R,et al.Screening,identification,degradation condition and effects of anitrite-degradating strain[J].Journal of Fishery Sciences of China,2010,17(6):1264-1271.
[14]罗国芝,陈家捷,于文杰,等.一株新型异养硝化细菌处理养殖水的效果[J].环境工程学报,2016,10(8):4206-4212.Luo G,Chen J,Yu W,et al.A new heterotrophic nitrifying bacteria effect of aquaculture water treatment[J].Chinese Journal of Environmental Engineering,2016,10(8):4206-4212.
[15]李步先.虾蟹养殖池塘益生菌的分离筛选及降解特性初步研究[D].青岛:中国海洋大学,2015.Li B.A Primary Study on Isolation,Screening and Removal Characteristics of Probiotics in Ponds of Shrimp and Crab[D].Qingdao:Ocean University of China,2015.
[16]雷衍之.养殖水环境化学实验[M].北京:中国农业出版社,2006.Lei Y.Waters Enviromental Chemistry for Aquaculture Experiment[M].Beijing:China Agriculture Press,2006.
[17]傅雪军,马绍赛,曲克明,等.循环水养殖系统中自然微生物生物膜形成过程实验[J].海洋环境科学,2010,29(5):713-717.Fu X,Ma S,Qu K,et al.Experiment of biofilm formation process of natural microbed in recirculating aquaculture system[J].Marine Environmental Science,2010,29(5):713-717.
[18]李梅.全封闭式循环水养殖系统中生物滤器生物膜培养[J].农业与技术,2017,37(16):241-242.Li M.Cultivation of the biofilm on the biofilter in enclosed recirculation aquaculture system[J].Agriculture and Technology,2017,37(16):241-242.
[19]鲍鹰,相建海.温度、盐度和pH对生物过滤器去除氨氮效率的影响[J].海洋科学,2001,25(6):42-43.Bao Y,Xiang J.Effect of temperature,salinity and pH on ammonia oxidation in nitrosomonas[J].Marine Sciences,2001,25(6):42-43.
[20]Mora-Gómez J,Freixa A,Perujo N,et al.Aquatic BiofilmsEcology,Water Quality and Wastewater Treatment[M].Norfolk,UK:Caister Academic Press,2016:3-27.
[21] Hatzenpichler R.Diversity,physiology and niche differentiation of ammonia-oxidizing archaea[J].Appl Environ Microbiol,2012,78(21):7501-7510.
[22]刘艳红,罗国芝,朱学宝.海水闭合循环系统生物滤器微生物特性研究[J].农业环境科学学报,2004,23(3):540-544.Liu Y,Luo G,Zhu X.Microbial characteristics of biofilter in a closed marine system[J].Journal of Agro-Environment Science,2004,23(3):540-544.
[23] Van Rijn J,Tal Y,Schreier H J.Denitrification in recirculating systems:Theory and applications[J].Aquacult Eng,2006,34(3):364-376.
[24]王威,王玮,齐继光,等.3种滤料的生物挂膜及硝化反硝化性能[J].应用与环境生物学报,2017,23(2):301-305.Wang W,Wang W,Qi J,et al.Bio-film cultivation and nitrification-denitrification performance of three kinds of fillings[J].Chinese Journal of Applied and Environmental Biology,2017,23(2):301-305.
[25]高霄龙,刘鹰,李贤,等.鲍放养密度对循环水养殖水质的影响及生物滤器净化效果[J].农业工程学报,2017,33(21):244-252.Gao X,Liu Y,Li X,et al.Effects of stocking density on water quality of Haliotis discus hannai Ino in recirculating aquaculture and purification effect of biofilter[J].Transactions of the Chinese Society of Agricultural Engineering,2017,33(21):244-252.
[26]杨世东,刘锦伟,刘志国,等.一种基于脱氮和除磷分开的强化污染物去除组合工艺中试研究[J].环境工程学报,2016,10(4):1609-1614.Yang S,Liu J,Liu Z,et al.Pilot study on a combined process based on separation of nitrogen and phosphorus removal for enhancing pollutant removal[J].Chinese Journal of Environmental Engineering,2016,10(4):1609-1614.
[27]张寒冰,黄凤莲,周艳红,等.生物膜法处理养殖废水的研究[J].生态环境,2005,14(1):26-29.Zhang H,Huang F,Zhou Y,et al.Treatment of aquaculture wastewater by the biological film process[J].Ecology and Environment,2005,14(1):26-29.
[28]张真真,董宏标,段亚飞,等.有机碳驱动的人工基质表面菌群结构组成及其动态变化[J].海洋环境科学,2015,34(5):686-691.Zhang Z,Dong H,Duan Y,et al.Effects of carbohydrate addition on dynamics of bacterial community structures on the biofilms of artificial substrates[J].Marine Environmental Science,2015,34(5):686-691.
[29]李志华,郭强,吴杰,等.自养菌污泥致密过程及其污水处理特性研究[J].环境科学,2010,31(3):738-742.Li Z,Guo Q,Wu J,et al.Densification of autotrophic bacteria sludge and its characteristics for wasterwater treatment[J].Environmental Science,2010,31(3):738-742.
[30]王淑洁.循环水养殖生物滤池中氨氮降解微生物的分离鉴定及其水处理效果研究[D].青岛:中国海洋大学,2015.Wang S.The Study on Isolation and Identification and The Effect of Water Treatment of Microorganism Decomposing Ammonia Nitrogen in the RAS Biological Filter[D].Qingdao:Ocean University of China,2015.
[31]胡修贵,赵培,李玉宏,等.生物絮团中异养亚硝化菌的分离鉴定及其特性[J].渔业科学进展,2013,34(5):97-103.Hu X,Zhao P,Li Y,et al.Isolation and identification of heterotrophic nitrosobacteria from bio-flocs and analysis of their characteristics[J].Progress in Fishery Sciences,2013,34(5):97-103.
[32]刘芳芳,周德平,吴淑杭,等.养殖废水中异养硝化细菌的分离筛选和鉴定[J].农业环境科学学报,2010,29(11):2232-2237.Liu F,Zhou D,Wu S,et al.Isolation and identification of heterotrophic nitrifiers from cultivation wastewater[J].Journal of Agro-Environment Science,2010,29(11):2232-2237.
[33]罗国芝,刘艳红,谭洪新,等.新建海水生物滤器接种培养的研究[J].农业环境保护,2001,20(6):443-445,451.Luo G,Liu Y,Tan H,et al.Inoculation and cultivation in newly established seawater biofilter[J].Agro-environmental Protection,2001,20(6):443-445,451.
[34]罗国芝,孙大川,冯是良,等.闭合循环水产养殖系统生产过程中生物过滤器功能的形成[J].水产学报,2005,29(4):574-577.Luo G,Sun D,Feng S,et al.The function establishment of biofilter in recirculating commercial fish culture system[J].Journal of Fisheries of China,2005,29(4):574-577.
[2]王峰,雷霁霖,高淳仁,等.国内外工厂化循环水养殖模式水质处理研究进展[J].中国工程科学,2013,15(10):16-32.Wang F,Lei Q,Gao C,et al.Research progress of water conditioning in industry recirculating aquaculture mode at home and abroad[J].Engineering Sciences,2013,15(10):16-32.
[3]冯志华,徐军田,李玉,等.封闭循环海水养殖生物滤池生物膜形成过程研究[J].淮海工学院学报(自然科学版),2010,19(4):79-82.Feng Z,Xu J,Li Y,et al.Study on the biofilm formation process of biofilter in a closed seawater recirculating aquaculture system[J].Journal of Huaihai Institute of Technology(Natural Science Edition),2010,19(4):79-82.
[4] Blancheton J P,Attramadal K J K,Michaud L,et al.Insight into bacterial population in aquaculture systems and its implication[J].Aquacult Eng,2012,53:30-39.
[5] Rud I,Kolarevic J,Holan A B,et al.Deep-sequencing of the bacterial microbiota in commercial-scale recirculating and semi-closed aquaculture systems for Atlantic salmon post-smolt production[J].Aquacult Eng,2017,78:50-62.
[6] Pedersen L,Oosterveld R,Pedersen P B.Nitrification performance and robustness of fixed and moving bed biofilters having identical carrier elements[J].Aquacult Eng,2014,10:37-45.
[7] Guerdat T C,Losordo T M,Classen J J,et al.An evaluation of commercially available biological filters for recirculating aquaculture systems[J].Aquacult Eng,2010,42(1):38-49.
[8] Guerdat T C.A Large Scale Evaluation of Commercially Available Biological Filters for Recirculating Aquaculture Systems[D].Raleigh:North Carolina State University,2008.
[9] Badiola M,Mendiola D,Bostock J.Recirculating Aquaculture Systems(RAS)analysis:Main issues on management and future challenges[J].Aquacult Eng,2012,51:26-35.
[10]吴嘉敏,孙大川.循环水养殖系统中浸没式生物滤器的水处理效果[J].上海水产大学学报,2007,16(6):542-548.Wu J,Sun D.Study on the water treatment efficiency of submerged biological filter in the recirculating aquaculture system[J].Journal of Shanghai Fisheries University,2007,16(6):542-548.
[11]管敏,李莎,张建明,等.循环水养殖系统生物滤池自然挂膜启动及效果研究[J].中国给水排水,2015,31(1):80-83.Guan M,Li S,Zhang J,et al.Natural start-up and water treatment effect of biofilter for recirculating aquaculture system[J].China Water&Wastewater,2015,31(1):80-83.
[12]宋协法,李华龙.养殖污水中主要分解氨氮微生物的分离鉴定[J].渔业现代化,2013,40(1):1-6.Song X,Li H.Research of nitrifying bacteria from cultivation wastewater[J].Fishery Modernization,2013,40(1):1-6.
[13]熊焰,刘力源,罗睿,等.1株亚硝酸盐降解菌的筛选-鉴定-降解条件及效果[J].中国水产科学,2010,17(6):1264-1271.Xiong Y,Liu L,Luo R,et al.Screening,identification,degradation condition and effects of anitrite-degradating strain[J].Journal of Fishery Sciences of China,2010,17(6):1264-1271.
[14]罗国芝,陈家捷,于文杰,等.一株新型异养硝化细菌处理养殖水的效果[J].环境工程学报,2016,10(8):4206-4212.Luo G,Chen J,Yu W,et al.A new heterotrophic nitrifying bacteria effect of aquaculture water treatment[J].Chinese Journal of Environmental Engineering,2016,10(8):4206-4212.
[15]李步先.虾蟹养殖池塘益生菌的分离筛选及降解特性初步研究[D].青岛:中国海洋大学,2015.Li B.A Primary Study on Isolation,Screening and Removal Characteristics of Probiotics in Ponds of Shrimp and Crab[D].Qingdao:Ocean University of China,2015.
[16]雷衍之.养殖水环境化学实验[M].北京:中国农业出版社,2006.Lei Y.Waters Enviromental Chemistry for Aquaculture Experiment[M].Beijing:China Agriculture Press,2006.
[17]傅雪军,马绍赛,曲克明,等.循环水养殖系统中自然微生物生物膜形成过程实验[J].海洋环境科学,2010,29(5):713-717.Fu X,Ma S,Qu K,et al.Experiment of biofilm formation process of natural microbed in recirculating aquaculture system[J].Marine Environmental Science,2010,29(5):713-717.
[18]李梅.全封闭式循环水养殖系统中生物滤器生物膜培养[J].农业与技术,2017,37(16):241-242.Li M.Cultivation of the biofilm on the biofilter in enclosed recirculation aquaculture system[J].Agriculture and Technology,2017,37(16):241-242.
[19]鲍鹰,相建海.温度、盐度和pH对生物过滤器去除氨氮效率的影响[J].海洋科学,2001,25(6):42-43.Bao Y,Xiang J.Effect of temperature,salinity and pH on ammonia oxidation in nitrosomonas[J].Marine Sciences,2001,25(6):42-43.
[20]Mora-Gómez J,Freixa A,Perujo N,et al.Aquatic BiofilmsEcology,Water Quality and Wastewater Treatment[M].Norfolk,UK:Caister Academic Press,2016:3-27.
[21] Hatzenpichler R.Diversity,physiology and niche differentiation of ammonia-oxidizing archaea[J].Appl Environ Microbiol,2012,78(21):7501-7510.
[22]刘艳红,罗国芝,朱学宝.海水闭合循环系统生物滤器微生物特性研究[J].农业环境科学学报,2004,23(3):540-544.Liu Y,Luo G,Zhu X.Microbial characteristics of biofilter in a closed marine system[J].Journal of Agro-Environment Science,2004,23(3):540-544.
[23] Van Rijn J,Tal Y,Schreier H J.Denitrification in recirculating systems:Theory and applications[J].Aquacult Eng,2006,34(3):364-376.
[24]王威,王玮,齐继光,等.3种滤料的生物挂膜及硝化反硝化性能[J].应用与环境生物学报,2017,23(2):301-305.Wang W,Wang W,Qi J,et al.Bio-film cultivation and nitrification-denitrification performance of three kinds of fillings[J].Chinese Journal of Applied and Environmental Biology,2017,23(2):301-305.
[25]高霄龙,刘鹰,李贤,等.鲍放养密度对循环水养殖水质的影响及生物滤器净化效果[J].农业工程学报,2017,33(21):244-252.Gao X,Liu Y,Li X,et al.Effects of stocking density on water quality of Haliotis discus hannai Ino in recirculating aquaculture and purification effect of biofilter[J].Transactions of the Chinese Society of Agricultural Engineering,2017,33(21):244-252.
[26]杨世东,刘锦伟,刘志国,等.一种基于脱氮和除磷分开的强化污染物去除组合工艺中试研究[J].环境工程学报,2016,10(4):1609-1614.Yang S,Liu J,Liu Z,et al.Pilot study on a combined process based on separation of nitrogen and phosphorus removal for enhancing pollutant removal[J].Chinese Journal of Environmental Engineering,2016,10(4):1609-1614.
[27]张寒冰,黄凤莲,周艳红,等.生物膜法处理养殖废水的研究[J].生态环境,2005,14(1):26-29.Zhang H,Huang F,Zhou Y,et al.Treatment of aquaculture wastewater by the biological film process[J].Ecology and Environment,2005,14(1):26-29.
[28]张真真,董宏标,段亚飞,等.有机碳驱动的人工基质表面菌群结构组成及其动态变化[J].海洋环境科学,2015,34(5):686-691.Zhang Z,Dong H,Duan Y,et al.Effects of carbohydrate addition on dynamics of bacterial community structures on the biofilms of artificial substrates[J].Marine Environmental Science,2015,34(5):686-691.
[29]李志华,郭强,吴杰,等.自养菌污泥致密过程及其污水处理特性研究[J].环境科学,2010,31(3):738-742.Li Z,Guo Q,Wu J,et al.Densification of autotrophic bacteria sludge and its characteristics for wasterwater treatment[J].Environmental Science,2010,31(3):738-742.
[30]王淑洁.循环水养殖生物滤池中氨氮降解微生物的分离鉴定及其水处理效果研究[D].青岛:中国海洋大学,2015.Wang S.The Study on Isolation and Identification and The Effect of Water Treatment of Microorganism Decomposing Ammonia Nitrogen in the RAS Biological Filter[D].Qingdao:Ocean University of China,2015.
[31]胡修贵,赵培,李玉宏,等.生物絮团中异养亚硝化菌的分离鉴定及其特性[J].渔业科学进展,2013,34(5):97-103.Hu X,Zhao P,Li Y,et al.Isolation and identification of heterotrophic nitrosobacteria from bio-flocs and analysis of their characteristics[J].Progress in Fishery Sciences,2013,34(5):97-103.
[32]刘芳芳,周德平,吴淑杭,等.养殖废水中异养硝化细菌的分离筛选和鉴定[J].农业环境科学学报,2010,29(11):2232-2237.Liu F,Zhou D,Wu S,et al.Isolation and identification of heterotrophic nitrifiers from cultivation wastewater[J].Journal of Agro-Environment Science,2010,29(11):2232-2237.
[33]罗国芝,刘艳红,谭洪新,等.新建海水生物滤器接种培养的研究[J].农业环境保护,2001,20(6):443-445,451.Luo G,Liu Y,Tan H,et al.Inoculation and cultivation in newly established seawater biofilter[J].Agro-environmental Protection,2001,20(6):443-445,451.
[34]罗国芝,孙大川,冯是良,等.闭合循环水产养殖系统生产过程中生物过滤器功能的形成[J].水产学报,2005,29(4):574-577.Luo G,Sun D,Feng S,et al.The function establishment of biofilter in recirculating commercial fish culture system[J].Journal of Fisheries of China,2005,29(4):574-577.