益生菌和复合营养剂对鱼虾混养池塘水质及细菌群落结构的影响
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摘要
通过向鱼虾混养池塘中添加益生菌和复合营养剂对养殖水环境进行生态调控,并评价其水质调控效果;运用Illumina HiSeq2500测序平台对养殖水体细菌多样性进行16S rDNA测序,获得其细菌组成及丰度信息。水质检测结果表明,在该调控方式下,透明度、pH、溶解氧(DO)、氨氮(NH_3-N)及亚硝酸盐(NO_2-N)等参数变化幅度均在养殖鱼、虾适宜范围内,有害弧菌的浓度远低于致病浓度,表明该水质调控方式能够维持养殖水环境稳定,抑制病原菌的繁殖。细菌群落组成及丰度分析结果显示,养殖水体得到的OTU数目为1 878,其中,在界、门、纲、目、科、属和种上的OTU数目分别为4,43,79,91,652,952,16;养殖水体中相对丰度占主要优势的细菌微生物为蓝细菌门(Cyanobacteria)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes),分别占总细菌的38.19%,28.11%,18.85%,12.85%和0.08%。结果表明,添加益生菌和复合营养剂后鱼虾混养池塘细菌群落结构多样性水平较高,含有多种水产养殖中的常用益生菌。本研究结果为鱼虾健康养殖和养殖水体生态环境的改善提供了理论依据。
Probiotics and compound nutrients were added into a fish-shrimp mix-culturing pond to regulate the aquaculture environment in the pond, and their effect on water quality was evaluated. The bacterial diversity in the water of the mix-culturing pond was estimated via a high throughput 16 S rDNA sequencing by using Illumina Instrument HiSeq2500 and its composition and abundance were analyzed. The water quality test showed that the water quality parameters in the mix-culturing ponds, such as transparency, pH, dissolved oxygen(DO), ammonia nitrogen(NH_3-N) and nitrite(NO_2-N), fell within the appropriate range of fish and shrimp farming standard and that the concentration of harmful vibrio bacteria was far lower than the pathogenic concentration, which indicates that this method can maintain stable water environment and inhibit the reproduction of pathogenic bacteria. The analysis of bacterial community composition and abundance showed that the number of bacterial OTU in the aquaculture water was 1878, among which the number of OTU in kingdom, phylum, class, order, family, genus and species was 4, 43, 79, 91, 652, 952 and 16, respectively. The relative abundance of bacteria in the aquaculture water was mainly dominated by Cyanobacteria, Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes, accounting for 38.19%, 28.11%, 18.85%, 12.85% and 0.08% of the total bacteria, respectively. The result indicates that the water in fish-shrimp mix-culturing pond had a higher diversity of bacteria in the bacterial community after the probiotics and compound nutrients were added into the mix-culturing pond, and contained many common probiotics. This study provides a theoretical basis for improving the ecological environment of aquaculture and healthy farming of fish and shrimp.
Probiotics and compound nutrients were added into a fish-shrimp mix-culturing pond to regulate the aquaculture environment in the pond, and their effect on water quality was evaluated. The bacterial diversity in the water of the mix-culturing pond was estimated via a high throughput 16 S rDNA sequencing by using Illumina Instrument HiSeq2500 and its composition and abundance were analyzed. The water quality test showed that the water quality parameters in the mix-culturing ponds, such as transparency, pH, dissolved oxygen(DO), ammonia nitrogen(NH_3-N) and nitrite(NO_2-N), fell within the appropriate range of fish and shrimp farming standard and that the concentration of harmful vibrio bacteria was far lower than the pathogenic concentration, which indicates that this method can maintain stable water environment and inhibit the reproduction of pathogenic bacteria. The analysis of bacterial community composition and abundance showed that the number of bacterial OTU in the aquaculture water was 1878, among which the number of OTU in kingdom, phylum, class, order, family, genus and species was 4, 43, 79, 91, 652, 952 and 16, respectively. The relative abundance of bacteria in the aquaculture water was mainly dominated by Cyanobacteria, Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes, accounting for 38.19%, 28.11%, 18.85%, 12.85% and 0.08% of the total bacteria, respectively. The result indicates that the water in fish-shrimp mix-culturing pond had a higher diversity of bacteria in the bacterial community after the probiotics and compound nutrients were added into the mix-culturing pond, and contained many common probiotics. This study provides a theoretical basis for improving the ecological environment of aquaculture and healthy farming of fish and shrimp.
引文
[1] 姚晓东.抗生素在水产养殖中应用存在的问题及对策[J].农业与技术,2016,36(24):103-103.
[2] 高权新,施兆鸿,彭士明.益生菌在水产养殖中的研究进展[J].海洋渔业,2013,35(3):364-372.
[3] 谢永斌,倪学勤,曾东,等.复合营养剂对浮游生物群落结构和水质调控作用的影响[J].农业生物技术学报,2016,24 (5):738-746.
[4] 何南荣,杨鹏程,金帮宇.养殖塘pH值的变化规律及调控[J].现代畜牧科技,2013(5):229-229.
[5] 卢平克.黄鳍鲷池塘健康养殖技术[J].海洋与渔业,2014(7):56-57.
[6] 杨菁,倪琦,张宇雷,等.对虾工程化循环水养殖系统构建技术[J].农业工程学报,2010,26(8):136-140.
[7] 李斌,张秀珍,马元庆,等.生物絮团对水质的调控作用及仿刺参(Apostichopus japonicus)幼参生长的影响[J].渔业科学进展,2014,35(4):85-90.
[8] 潘腾飞,齐树亭,武洪庆等.影响池塘养殖水体溶解氧的主要因素分析[J].安徽农业科学,2010,38(17):9155-9157.
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[10] 张正光.池塘养殖水体透明度及其在生产上的意义[J].科学养鱼,1994(12):27.
[11] 徐实怀,宋盛宪.对虾养殖中水色和透明度的意义和调节方法[J].江西水产科技,2007(4):22-25.
[12] CHENG S Y,CHEN J C.Accumulation of nitrite in the tissues of Penaeus monodon exposed to elevated ambient nitrite after different time periods[J].Archives of Environmental Contamination and Toxicology,2000,39(2):183-192.
[13] 林小华.南美白对虾鱼虾混养养殖模式[J].水产养殖,2017,38(3):38-39.
[14] 王战蔚,张译丹,李秀颖,等.池塘中氨氮、亚硝酸盐的危害及控制措施[J].吉林水利,2013(3):39-40.
[15] 李健,姜令绪,王文琪,等.氨氮和硫化氢对日本对虾幼体的毒性影响[J].上海水产大学报,2007,16(1):22-27.
[16] CHAND R K,SAHOO P K.Effect of nitrite on the immune response of freshwater prawn Macrobrachium malcolmsonii,and its susceptibility to Aeromonas hydrophila[J].Aquaculture,2006,258(1):150-156.
[17] 宋协法,康萌萌,彭磊,等.益生菌对半滑舌鳎养殖水质及仔稚鱼生长的影响研究[J].渔业现代化,2011,38(6):30-35.
[18] 邹文娟,许晓慧,王国武,等.光合细菌和枯草芽孢杆菌在污水处理中的应用[J].广东农业科学,2010,37(9):199-201.
[19] 聂伟.利用絮凝活性菌株培育生物絮团及其对水质和浮游生物影响的研究[D].武汉:武汉轻工大学,2016.
[20] TRAN L,NUNAN L,REDMAN R M et al.EMS/AHPNS:infectious disease caused by bacteria[J].Global Aquaculture Advocate,2013,18-20.
[21] 倪军.对虾“早期死亡综合征”研究现状[J].海洋与渔业,2014(12):74-75.
[22] 陈爱玲,李秋芬,张立通,等.添加营养物质提高商品水质净化菌剂净化能力的研究[J].水产学报,2010,34(4):581-588.
[23] SCHRYVER P D,VADSTEIN O.Ecological theory as a foundation to control pathogenic invasion in aquaculture.[J].Isme Journal,2014,8(12):2360-2368.
[24] WARNECKE F,AMANN R,PERNTHALER J.Actinobacterial 16S rRNA genes from freshwater habitats cluster in four distinct lineages[J].Environmental Microbiology,2010,6(3):242-253.
[25] KIRCHMAN D L.The ecology of Cytophaga-Flavobacteria in aquatic environments[J].Fems Microbiology,Ecology,2002,39(2):91-100.
[26] 郑佳佳,彭丽莎,张小平,等.复合益生菌对草鱼养殖水体水质和菌群结构的影响[J].水产学报,2013,37(3) :457-464.
[27] 刘树文.海洋聚球藻对铁限制的生理响应[D].武汉:华中师范大学,2012.
[28] 袁军.印度洋深海多环芳烃降解菌的多样性分析及降解菌新种的分类鉴定与降解机理初步研究[D].厦门:厦门大学,2008.
[29] CLATYON R K,SISTRON W R.The photosynthetic Bacteria[M].New York and London:Plenum press:1978:33-44.
[30] SHIPMAN R H,FAN L,KAO I C.Single cell production by photosynthetic bacteria Adv[J].Appl Microb,1977,21:161-183.
[31] BURGESS J G,MIYASHITA H,SUDO H,et al.Antibiotic production by the marine photosynthetic bacterium Chromatium purpuratum NKPB031704:localization of activity to the chromatophores[J].FEMS Microbiology Letters,1991,68(3):301-305.
[32] 曹海鹏,何珊,欧仁建,等.水产用噬菌蛭弧菌研究进展[J].动物医学进展,2013,34(1):86-90.
[33] 袁维道,吴小妹.放线菌在水产养殖中的潜在应用分析[J].自然科学:文摘版,2016,(5):00205-00205.
[34] DAS S,WARD L R,BURKE C.Prospects of using marine Actinobacteria as probiotics in aquaculture[J].Applied Microbiology &Biotechnology,2008,81(3):419-429.
[35] COTTRELL M T,KIRCHMAN D L.Natural assemblages of marine Proteobacteria and members of the Cytophaga-Flavobacter cluster consuming low-and high-molecular-weight dissolved organic matter[J].Applied and Environmental Microbiology,2000,66(4):1692-1697.
[36] 陈琼,李贵阳,罗坤,等.凡纳滨对虾(Litopenaeus vannamei)亲虾繁殖期水体微生物多样性[J].海洋与湖沼,2017,48(1):130-138.
[37] BERGMANN G T,BATES S T,EILERS K G,et al.The under-recognized dominance of Verrucomicrobia in soil bacterial communities [J].Soil Biology & Biochemistry,2011,43(7):1450-1455.
[38] 周恩民.美国大盆地四热泉可培养高温细菌多样性及生态学研究[D].昆明:云南大学,2015.
[39] 王振华,李建臻,王迪,等.益生芽孢杆菌在水产养殖中研究现状及存在问题[J].饲料研究,2018(1):1-8.
[40] 徐亚飞.地衣芽孢杆菌在水产养殖中的应用研究进展[J].渔业研究,2018,40(1):83-88.
[41] 章文明,汪海峰,刘建新.乳酸杆菌益生作用机制的研究进展[J].动物营养学报,2012,24(3):389-396.
[42] 郑瑞珠,何夙旭,杨雅麟,等.益生乳酸菌水产动物消化道黏附机制研究进展[J].中国农业科技导报,2014,16(3):134-142.
[2] 高权新,施兆鸿,彭士明.益生菌在水产养殖中的研究进展[J].海洋渔业,2013,35(3):364-372.
[3] 谢永斌,倪学勤,曾东,等.复合营养剂对浮游生物群落结构和水质调控作用的影响[J].农业生物技术学报,2016,24 (5):738-746.
[4] 何南荣,杨鹏程,金帮宇.养殖塘pH值的变化规律及调控[J].现代畜牧科技,2013(5):229-229.
[5] 卢平克.黄鳍鲷池塘健康养殖技术[J].海洋与渔业,2014(7):56-57.
[6] 杨菁,倪琦,张宇雷,等.对虾工程化循环水养殖系统构建技术[J].农业工程学报,2010,26(8):136-140.
[7] 李斌,张秀珍,马元庆,等.生物絮团对水质的调控作用及仿刺参(Apostichopus japonicus)幼参生长的影响[J].渔业科学进展,2014,35(4):85-90.
[8] 潘腾飞,齐树亭,武洪庆等.影响池塘养殖水体溶解氧的主要因素分析[J].安徽农业科学,2010,38(17):9155-9157.
[9] 宋学林,沈勤.养殖池水质调控技术[J].现代农业科技,2010(7):360-360.
[10] 张正光.池塘养殖水体透明度及其在生产上的意义[J].科学养鱼,1994(12):27.
[11] 徐实怀,宋盛宪.对虾养殖中水色和透明度的意义和调节方法[J].江西水产科技,2007(4):22-25.
[12] CHENG S Y,CHEN J C.Accumulation of nitrite in the tissues of Penaeus monodon exposed to elevated ambient nitrite after different time periods[J].Archives of Environmental Contamination and Toxicology,2000,39(2):183-192.
[13] 林小华.南美白对虾鱼虾混养养殖模式[J].水产养殖,2017,38(3):38-39.
[14] 王战蔚,张译丹,李秀颖,等.池塘中氨氮、亚硝酸盐的危害及控制措施[J].吉林水利,2013(3):39-40.
[15] 李健,姜令绪,王文琪,等.氨氮和硫化氢对日本对虾幼体的毒性影响[J].上海水产大学报,2007,16(1):22-27.
[16] CHAND R K,SAHOO P K.Effect of nitrite on the immune response of freshwater prawn Macrobrachium malcolmsonii,and its susceptibility to Aeromonas hydrophila[J].Aquaculture,2006,258(1):150-156.
[17] 宋协法,康萌萌,彭磊,等.益生菌对半滑舌鳎养殖水质及仔稚鱼生长的影响研究[J].渔业现代化,2011,38(6):30-35.
[18] 邹文娟,许晓慧,王国武,等.光合细菌和枯草芽孢杆菌在污水处理中的应用[J].广东农业科学,2010,37(9):199-201.
[19] 聂伟.利用絮凝活性菌株培育生物絮团及其对水质和浮游生物影响的研究[D].武汉:武汉轻工大学,2016.
[20] TRAN L,NUNAN L,REDMAN R M et al.EMS/AHPNS:infectious disease caused by bacteria[J].Global Aquaculture Advocate,2013,18-20.
[21] 倪军.对虾“早期死亡综合征”研究现状[J].海洋与渔业,2014(12):74-75.
[22] 陈爱玲,李秋芬,张立通,等.添加营养物质提高商品水质净化菌剂净化能力的研究[J].水产学报,2010,34(4):581-588.
[23] SCHRYVER P D,VADSTEIN O.Ecological theory as a foundation to control pathogenic invasion in aquaculture.[J].Isme Journal,2014,8(12):2360-2368.
[24] WARNECKE F,AMANN R,PERNTHALER J.Actinobacterial 16S rRNA genes from freshwater habitats cluster in four distinct lineages[J].Environmental Microbiology,2010,6(3):242-253.
[25] KIRCHMAN D L.The ecology of Cytophaga-Flavobacteria in aquatic environments[J].Fems Microbiology,Ecology,2002,39(2):91-100.
[26] 郑佳佳,彭丽莎,张小平,等.复合益生菌对草鱼养殖水体水质和菌群结构的影响[J].水产学报,2013,37(3) :457-464.
[27] 刘树文.海洋聚球藻对铁限制的生理响应[D].武汉:华中师范大学,2012.
[28] 袁军.印度洋深海多环芳烃降解菌的多样性分析及降解菌新种的分类鉴定与降解机理初步研究[D].厦门:厦门大学,2008.
[29] CLATYON R K,SISTRON W R.The photosynthetic Bacteria[M].New York and London:Plenum press:1978:33-44.
[30] SHIPMAN R H,FAN L,KAO I C.Single cell production by photosynthetic bacteria Adv[J].Appl Microb,1977,21:161-183.
[31] BURGESS J G,MIYASHITA H,SUDO H,et al.Antibiotic production by the marine photosynthetic bacterium Chromatium purpuratum NKPB031704:localization of activity to the chromatophores[J].FEMS Microbiology Letters,1991,68(3):301-305.
[32] 曹海鹏,何珊,欧仁建,等.水产用噬菌蛭弧菌研究进展[J].动物医学进展,2013,34(1):86-90.
[33] 袁维道,吴小妹.放线菌在水产养殖中的潜在应用分析[J].自然科学:文摘版,2016,(5):00205-00205.
[34] DAS S,WARD L R,BURKE C.Prospects of using marine Actinobacteria as probiotics in aquaculture[J].Applied Microbiology &Biotechnology,2008,81(3):419-429.
[35] COTTRELL M T,KIRCHMAN D L.Natural assemblages of marine Proteobacteria and members of the Cytophaga-Flavobacter cluster consuming low-and high-molecular-weight dissolved organic matter[J].Applied and Environmental Microbiology,2000,66(4):1692-1697.
[36] 陈琼,李贵阳,罗坤,等.凡纳滨对虾(Litopenaeus vannamei)亲虾繁殖期水体微生物多样性[J].海洋与湖沼,2017,48(1):130-138.
[37] BERGMANN G T,BATES S T,EILERS K G,et al.The under-recognized dominance of Verrucomicrobia in soil bacterial communities [J].Soil Biology & Biochemistry,2011,43(7):1450-1455.
[38] 周恩民.美国大盆地四热泉可培养高温细菌多样性及生态学研究[D].昆明:云南大学,2015.
[39] 王振华,李建臻,王迪,等.益生芽孢杆菌在水产养殖中研究现状及存在问题[J].饲料研究,2018(1):1-8.
[40] 徐亚飞.地衣芽孢杆菌在水产养殖中的应用研究进展[J].渔业研究,2018,40(1):83-88.
[41] 章文明,汪海峰,刘建新.乳酸杆菌益生作用机制的研究进展[J].动物营养学报,2012,24(3):389-396.
[42] 郑瑞珠,何夙旭,杨雅麟,等.益生乳酸菌水产动物消化道黏附机制研究进展[J].中国农业科技导报,2014,16(3):134-142.