流水和循环水养殖模式对虾养殖系统运行稳定性和细菌群落演替规律  

Operational stability and succession of bacterial community in flow-through and re-circulating shrimp culture system

作　　者：张晋 周灿 王青瑶[1,2] 吴俊敏 申旭东 傅松哲 刘鹰 ZHANG Jin;ZHOU Can;WANG Qingyao;WU Junmin;SHEN Xudong;FU Songzhe;LIU Ying(College of Marine Science and Environment,Dalian Ocean University,Dalian 116023,Liaoning,China;Key Laboratory of Environment Controlled Aquaculture(Dalian Ocean University),Ministry of Education,Dalian 116023,Liaoning,China;Yingkou Enhancement and Experiment Station,Chinese Academy of Fishery Sciences,Yingkou 115004,Liaoning,China)

机构地区：[1]大连海洋大学海洋科技与环境学院,辽宁大连116023 [2]设施渔业教育部重点实验室,辽宁大连116023 [3]中国水产科学研究院营口增殖实验站,辽宁营口115004

出　　处：《渔业现代化》2022年第4期15-25,共11页Fishery Modernization

基　　金：国家自然科学基金“中国北方典型流域水体中致病性弧菌的溯源与传播机制研究(81903372)”;辽参绿色生态养殖关键技术研发与应用(2020ZD23SN009);广东省重点领域研发计划“工厂化水产健康养殖精准调控关键技术研究与示范(2019B020215001)”;辽宁省教育厅项目“毒力拮抗剂对副溶血弧菌毒力表达的抑制作用研究(QL202005)”。

摘　　要：为分析流水和循环水对虾养殖模式的系统运行稳定性和细菌群落演替的差异,对养殖水体中氨氮、亚硝酸盐氮、化学需氧量(COD)、弧菌总数和细菌总数进行了检测。同时采用Illumina Hiseq高通量测序技术,在养殖第1、50、100天对养殖水体的细菌16SrDNA V4区域进行扩增和测序,对基因序列进行OTU聚类,并在此基础上进行细菌群落组成分析。结果显示:随着养殖时间的增加,流水养殖系统(FTS)和循环水养殖系统(RAS)水体中氨氮、亚硝酸盐氮和COD质量浓度逐渐升高;养殖后期,FTS中三者质量浓度明显高于RAS;同时,2种养殖系统水体中弧菌和细菌总数不断增加,其中FTS中弧菌属的细菌丰度显著高于RAS(P<0.05)。细菌门水平分析显示,变形菌纲(Proteobacteria)和拟杆菌门(Bacteroidetes)为两个养殖系统共同的优势类群。在属水平,FTS中主要细菌种群为弧菌属(Vibrio)、红细菌科(Rhodobacteraceae)和交替单胞菌属(Alteromonas),而RAS中核心细菌种群包括红细菌科(Rhodobacteraceae)、交替单胞菌属(Alteromonas)和黄杆菌属(Flavobacterium)。α多样性结果显示,RAS中细菌群落的多样性高于FTS,RAS的Chao1指数平均值也高于FTS(1136.18 vs 1050.82)。最后,两种养殖模式共得到可培养菌株93株,其中FTS中弧菌属(Vibrio)为主要可培养菌株,所占比例为61%,其次为芽孢杆菌(Bacillus),占比为12%。相比之下,弧菌在RAS中仅占9%。研究表明,RAS中系统运行和水体细菌的演替比FTS中更加稳定,突出了RAS在对虾全周期养殖方面的优势。本研究成果为开展高效对虾养殖提供了借鉴和参考。The purpose of this study was to analyze and compare the effects of the flow-through system(FTS)and re-circulating aquaculture systems(RAS)on operational stability and bacterial succession in shrimp farming systems.In this study,the concentrations of ammonia nitrogen,nitrite and COD in rearing water were detected from the above two systems in Yingkou.The total number of Vibrios and bacteria were also measured.Meanwhile,Illumina Hiseq high-throughput sequencing technology was used to amplify and sequence the bacterial 16SrDNA V4 region in the pond and circulating water system on the 1^(st),50^(th) and 100^(th) day of culture.In the beginning,the concentrations of ammonia and COD in FTS and RAS were low.With the increase of aquaculture time,the concentrations of ammonia,nitrite and COD in two types of systems increased.The concentrations of ammonia nitrogen,nitrite and COD in FTS were significantly higher than those in RAS.With the progression of farming,the total number of Vibrio and bacteria increased continuously in both systems,of which FTS has a significant higher abundance of Vibrio spp.and bacteria than those in RAS(P<0.05),indicating that the sequencing depth of the sample is large enough and the number of OTUs is close to the actual number.Then the gene sequences were clustered by the Operational Taxonomic Unit(OTU),and the bacterial community composition was analyzed based on OTU.The phylum level analysis showed that Proteobacteria and Bacteroidetes were the dominant bacteria in both systems.At the genus level,Vibrio,Rhodobacteraceae and Alteromonas were the main genera in FTS,while the core microbiome in RAS consisted of Rhodobacteraceae,Alteromonas and,Flavobacterium.The results ofαdiversity showed that the diversity of the bacterial community in RAS was higher than that of FTS,whereas the mean value of Chao1 index in the RAS was also higher than that of FTS(1136.18 vs 1050.82).Finally,a total of 93 strains were obtained in the two modes,of which Vibrio spp.was the main species in FTS,accounting for 61

关 键 词：种群结构 细菌种群 流水式养殖系统 循环水养殖系统 凡纳滨对虾 高通量测序 

分 类 号：S917.1[农业科学—水产科学]