第38卷第2期大连海洋大学学报Vol.38No.2 2023年4月JOURNAL OF DALIAN OCEAN UNIVERSITY Apr.2023DOI:10.16535/ki.dlhyxb.2022-211文章编号:2095-1388(2023)02-0217-06投喂冰鲜杂鱼和配合饲料对钝吻黄盖鲽生长性能㊁肠道结构及菌群多样性的影响徐晓莹1,王鹤1,李秀梅2,赵延宁1,张宁2,史文凯1,刘蓬1∗(1.烟台市海洋经济研究院,山东烟台264006;2.烟台宗哲海洋科技有限公司,山东烟台265617)摘要:为研究冰鲜杂鱼和配合饲料对钝吻黄盖鲽(Pseudopleuronectes yokohamae)生长及肠道健康的影响,以同一批培育的钝吻黄盖鲽幼鱼(体长为10.16cmʃ0.06cm)为研究对象,经42d的投喂试验,采用体尺性状测量㊁肠道组织石蜡切片制作和高通量测序等方法,研究了冰鲜杂鱼(FF组)㊁配合饲料(CF组)及二者混合投喂(MF组)对钝吻黄盖鲽生长性能㊁肠道结构和肠道菌群的影响㊂结果表明:与配合饲料组相比,冰鲜杂鱼组幼鱼的增重率㊁特定生长率显著提高(P<0.05),饲料系数显著降低(P<0.05);幼鱼肠道及其菌群结构分析显示,投喂冰鲜杂鱼能显著提高幼鱼肠道绒毛长度和基层厚度(P<0.05),提高幼鱼肠道菌群多样性,并促进红杆菌科等有益菌在肠道中的分布;混合投喂组各项指标介于冰鲜杂鱼组与配合饲料组之间㊂研究表明,在钝吻黄盖鲽的养殖过程中,可在配合饲料中适量添加红杆菌科的有益菌以促进钝吻黄盖鲽的生长和营养吸收,而钝吻黄盖鲽人工配合饲料的配方还应进一步优化,以提高其市场优势㊂关键词:钝吻黄盖鲽;生长性能;肠道结构;肠道菌群;配合饲料;冰鲜杂鱼中图分类号:S963.7;Q938.8㊀㊀㊀㊀文献标志码:A㊀㊀钝吻黄盖鲽(Pseudopleuronectes yokohamae)隶属于鲽形目(Pleuronectiformes)鲽科(Pleruonecti-da)黄盖鲽属(Pseudopleuronectes)[1],主要分布于太平洋西部近海,多产于中国的黄海和渤海,是北温带浅海冷水底栖鱼类[2]㊂其肉质细嫩㊁食性温和㊁适应性强并喜好集群[3],是黄海㊁渤海地区拖网作业的重要对象及理想的增殖对象[4]㊂近年来,随着中国海水养殖业的迅速发展,钝吻黄盖鲽的集约化养殖不断扩大㊂钝吻黄盖鲽养殖过程中使用的饲料主要有冰鲜杂鱼和配合饲料㊂随着水产绿色健康养殖 五大行动 中 配合饲料替代幼杂鱼行动 的不断推广,养殖饵料已经开始逐渐向人工配合饲料转变㊂目前,关于冰鲜杂鱼和配合饲料的饲喂对比研究已在大菱鲆(Scoph-thalmus maximus)㊁大黄鱼(Larmichthys crocea)㊁大口黑鲈(Micropterus salmoides)和斜带石斑鱼(Epinephelus coioides)等各种经济鱼类中相继开展,主要涉及生长性能㊁肌肉营养成分㊁免疫功能和肠道微生物结构等方面[5-8],但在钝吻黄盖鲽的相关研究中未见报道㊂不同饲料对鱼类肠道健康状况和菌群组成影响较大,如摄食配合饲料和冰鲜杂鱼的大口黑鲈肠道菌群多样性不同[9],摄食植物和海洋动物的虹鳟(Oncorhynchus mykiss)肠道内优势菌群不同[10]㊂本研究中,通过高通量测序等方法,对投喂不同饲料的钝吻黄盖鲽进行了生长性能㊁肠道结构及肠道菌群分析,从其宏观生长及微观结构㊁肠道菌群等方面探索了钝吻黄盖鲽的营养需求,以期为其饲料的合理配制提供科学参考㊂1㊀材料与方法1.1㊀材料试验用钝吻黄盖鲽为烟台宗哲海洋科技有限公司自繁自育的健康鱼苗,体长为(10.16ʃ0.06)cm㊂配合饲料为赛格林仔稚鱼育苗专用饲料,其常规营养成分为粗蛋白质52%㊁粗脂肪8%㊁粗灰分17%㊁粗纤维8%和水分12%;冰鲜杂鱼捕捞自烟台沿海,其常规营养成分为粗蛋白质21%㊁粗脂肪3%㊁粗灰分4%和水分66%(均为质量分数)㊂㊀收稿日期:2022-07-04㊀基金项目:山东省现代农业产业技术体系项目(SDAIT-12-10)㊀作者简介:徐晓莹(1990 ),女,工程师㊂E-mail:xiaoyingxu810052@ ㊀通信作者:刘蓬(1972 ),男,高级工程师㊂E-mail:135****7477@1.2㊀方法1.2.1㊀试验设计㊀选取健康且规格整齐的钝吻黄盖鲽270尾,随机分为配合饲料投喂组(CF)㊁冰鲜杂鱼与配合饲料混合投喂组(MF)及冰鲜杂鱼投喂组(FF)㊂每组设置3个重复,每个重复30尾鱼,分别放入126L的塑料箱中养殖,水温为(15ʃ1)ħ,盐度为24~30㊂为保证水质,每天定时更换鱼箱中20%的水量㊂试验期间,每天7:30和16:30各投喂一次,混合投喂组上午投喂配合饲料,下午投喂冰鲜杂鱼,冰鲜杂鱼经过粉碎后,每次投饵量为鱼体质量的3%,配合饲料每次投饵量为鱼体质量的1%㊂养殖试验共进行42d㊂1.2.2㊀生长性能指标的测定㊀在试验的第0㊁14㊁28㊁42天时分别称量鱼体质量,第42天时测量鱼体长,每天记录各组鱼的摄食量㊂称量前停食24h,分别从各组随机取10尾鱼进行测量㊂试验结束后,计算各组鱼的增重率(G BW, %)㊁特定生长率(R SG,%/d)㊁饲料系数(R FC)和肥满度(F C,g/cm3)㊂计算公式为㊀㊀G BW=(W t-W0)/W0ˑ100%,(1)㊀㊀R SG=(ln W t-ln W0)/tˑ100%,(2)㊀㊀R FC=F/(W t-W0),(3)㊀㊀F C=W t/L3ˑ100㊂(4)其中:W0㊁W t分别为试验鱼初始体质量(g)和终末体质量(g);t为试验时间(d);F为养殖周期内的摄食量(g);L为试验结束时的鱼体长(cm)㊂1.2.3㊀肠道结构观察㊀养殖试验结束后停食24h,从各组随机取3尾鱼,解剖取其肠道,并立即浸入体积分数为4%的多聚甲醛固定液中固定㊂肠道组织石蜡切片由生工生物工程(上海)股份有限公司制作㊂后肠组织在固定液中固定24h后,用体积分数为70%的乙醇溶液反复冲洗,用叔丁醇脱水,浸蜡包埋,再用冷冻切片机连续切片(厚度6μm),HE染色㊂肠道切片在显微镜下观察拍照,并测量绒毛长度㊁绒毛宽度和基层厚度等指标㊂1.2.4㊀肠道DNA提取与PCR扩增㊀养殖试验结束后,分别从各组随机取9尾鱼,无菌条件下解剖取出完整肠道,每3尾鱼的肠道内容物作为1个混合样品置入冻存管中,将3个混合样品用液氮速冻后保存待测㊂肠道样品基因组DNA的提取和测序由生工生物工程(上海)股份有限公司完成㊂将提取的肠道微生物总DNA用341F(5ᶄCCTAC-GGGNGGCWGCAG3ᶄ)和805R(5ᶄGACTACH-VGGGTATCTAATCC3ᶄ)特定引物扩增16S rDNA 的V3+V4区,构建测序文库后在Illumina平台完成测序㊂1.2.5㊀肠道菌群生物信息学分析㊀下机数据使用FLASH1.2.7[11]㊁QIIME1.7.0[11]对样品原始数据(raw reads)进行拼接㊁过滤及质控,去除嵌合体后获得有效数据(effective tags)㊂应用Uparse 7.0.1001软件[12]在97%的一致性水平上对样品所有effective tags进行聚类分析,获得OTUs㊂基于可操作性分类单元OTUs计算Chao1㊁Shannon㊁Simpson和ACE指数,并进行Alpha多样性分析㊂1.3㊀数据处理试验数据均以平均值ʃ标准差(meanʃS.D.)表示㊂采用SPSS19.0软件进行单因素方差分析,采用Duncan法进行组间多重比较㊂显著性水平设为0.05,极显著性水平设为0.01㊂2㊀结果与分析2.1㊀不同饲料组钝吻黄盖鲽的生长性能从表1可见:与冰鲜杂鱼组相比,配合饲料组钝吻黄盖蝶增重率㊁特定生长率显著降低(P< 0.05),肥满度变化不大(P>0.05);混合投喂组鱼体的增重率㊁特定生长率及肥满度等指标均高于配合饲料组,但低于冰鲜杂鱼组(P>0.05);以湿物质为基础的饲料系数表现为冰鲜杂鱼组显著高于配合饲料组(P<0.05),而以干物质为基础的饲料系数则表现为配合饲料组显著高于冰鲜杂鱼组(P<0.05)㊂2.2㊀不同饲料组钝吻黄盖鲽的肠道结构钝吻黄盖鲽肠道黏膜形态如图1所示,不同饲料影响了钝吻黄盖鲽肠道组织形态,冰鲜杂鱼组绒毛相对致密且高度较高,排列整齐,配合饲料组及混合投喂组的绒毛稀疏且高度较低㊂从表2可见,冰鲜杂鱼组钝吻黄盖鲽肠道的绒图1㊀钝吻黄盖鲽后肠黏膜形态Fig.1㊀Mucosal morphology in distal intestine of Pleu-ronectes yokohama812大连海洋大学学报㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀第38卷表1㊀各组钝吻黄盖鲽的生长性能Tab.1㊀Growth of Pleuronectes yokohama in each group组别group 初体质量/ginitial bodyweight末体质量/gfinal bodyweight增重率/%body weightgain特定生长率/(%㊃d-1)specific growth rate饲料系数(湿物质)food conversionratio(wet matter)饲料系数(干物质)food conversionratio(dry matter)肥满度/(g㊃cm3)condition factorCF13.72ʃ0.4022.38ʃ1.27b64.85ʃ10.00b 1.18ʃ0.15b 1.54ʃ0.01c 1.36ʃ0.01a 1.44ʃ0.14 MF13.97ʃ0.8524.44ʃ1.43a76.25ʃ7.36ab 1.34ʃ0.09ab 2.44ʃ0.03b 1.17ʃ0.03b 1.45ʃ0.13 FF13.69ʃ1.0725.82ʃ0.73a90.78ʃ11.39a 1.53ʃ0.15a 3.14ʃ0.05a 1.07ʃ0.05b 1.55ʃ0.08㊀注:同列中标有不同字母者表示组间有显著性差异(P<0.05),标有相同字母者表示组间无显著性差异(P>0.05),下同㊂Note:The means with different letters within the same column are significantly different in the groups at the0.05probability level,and the means with the same letters within the same column are not significant differences,et sequentia.表2㊀各组肠道黏膜形态参数Tab.2㊀Morphological parameters of intestinal mucosa ineach groupμm组别group绒毛长度villus length绒毛宽度villus width基层厚度muscular thicknessCF410.92ʃ53.39b176.35ʃ32.0941.40ʃ2.30b MF422.56ʃ54.04b178.86ʃ24.9046.40ʃ3.85b FF610.23ʃ62.75a205.63ʃ60.2169.60ʃ7.33a 毛长度和基层厚度显著高于配合饲料组和混合投喂组(P<0.05),配合饲料组最低;3组间绒毛宽度无显著性差异(P>0.05)㊂2.3㊀钝吻黄盖鲽肠道菌群的多样性对MiSeg测序原始数据处理后,3组样品共产生157646条优化序列,有效序列数为49331~ 56251条㊂从图2可见,3组钝吻黄盖鲽肠道样品产生的OTUs数目为429~483个,3组共有OTUs 为107个,CF㊁MF和FF组特有OTUs分别为234㊁225和269个㊂图2㊀各组肠道微生物分类Venn图Fig.2㊀Venn diagram of intestinal microbiome in each group对3组钝吻黄盖鲽的Alpha多样性进行分析,结果如表3所示,3组钝吻黄盖鲽肠道菌群的Chao1和ACE指数依次为FF组>CF组>MF组,说明冰鲜杂鱼组鱼肠道菌群的物种丰富度最高,混合投喂组物种丰富度最低;混合投喂组鱼肠道菌群的Shannon指数最小,Simpson指数最大,说明其多样性较低,其余2组肠道菌群多样性基本相似㊂表3㊀各组肠道菌群多样性指数Tab.3㊀Diversity index of intestinal flora in each group组别groupOUT总数totalOTUsACE指数ACEindexShannon指数ShannonindexChao1指数Chao1indexSimpson指数Simpsonindex CF445448.63 4.14449.70.07 MF429430.18 3.97431.00.08 FF483483.28 4.12483.30.07 2.4㊀钝吻黄盖鲽肠道菌群的结构组成在门水平上,钝吻黄盖鲽肠道微生物中丰度排列前10的菌群如图3所示,冰鲜杂鱼和配合饲料对钝吻黄盖鲽肠道优势菌门的相对丰度影响不同, 3组鱼肠道主要菌群均为变形菌门(Proteobacte-ria)㊁放线菌门(Actinobacteria)㊁厚壁菌门(Fir-micutes)和异常球菌-栖热菌门(Deinococcus-Ther-mus)等,随饲料中冰鲜杂鱼比例的增高,钝吻黄盖鲽肠道菌群中变形菌门相对丰度增大,而厚壁菌图3㊀钝吻黄盖鲽肠道门水平上的物种相对丰度Fig.3㊀Bacterial communities in intestinal flora of Pleu-ronectes yokohama at the phylum level912第2期徐晓莹,等:投喂冰鲜杂鱼和配合饲料对钝吻黄盖鲽生长性能㊁肠道结构及菌群多样性的影响门相对丰度则降低㊂在属水平上,钝吻黄盖鲽肠道菌群中相对丰度较高的10个主要菌群如表4所示,3组鱼肠道的优势菌群均为微杆菌属(Microbacterium ),其中,配合饲料组的次优势菌群为巨型球菌属(Macro-coccus )和异常球菌属(Deinococcus ),混合投喂组的次优势菌群为巨型球菌属(Macrococcus )和发光杆菌属(Photobacterium ),冰鲜杂鱼组的次优势菌群为红杆菌科未分类菌群unclassified_Rhodobacter-aceae 和巨型球菌属(Macrococcus )㊂表4㊀各组钝吻黄盖鲽肠道菌群优势度最高的10个属Tab.4㊀Top ten genera of bacterial communities in intestinal flora of Pleuronectes yokohama in each groupCF 组group CFMF 组group MFFF 组group FF菌种strain species相对丰度/%relative abundance菌种strain species相对丰度/%relative abundance菌种strain species相对丰度/%relative abundance未知菌群other29.41未知菌群other24.96未知菌群other24.49微杆菌属(Microbacterium )20.66微杆菌属(Microbacterium )22.72微杆菌属(Microbacterium )21.69巨型球菌属(Macrococcus )14.77巨型球菌属(Macrococcus )11.07红杆菌科未分类unclassified_Rhodobacteraceae8.60异常球菌属(Deinococcus ) 6.60发光杆菌属(Photobacterium )7.15巨型球菌属(Macrococcus )8.48不动杆菌属(Acinetobacter ) 3.89异常球菌属(Deinococcus ) 5.99异常球菌属(Deinococcus )5.20紫色杆菌属(Janthinobacterium ) 3.85紫色杆菌属(Janthinobacterium ) 3.82不动杆菌属(Acinetobacter ) 3.18细菌未分类unclassified_bacteria2.71不动杆菌属(Acinetobacter )3.78紫色杆菌属(Janthinobacterium ) 3.10鞘脂单胞菌属(Sphingomonas ) 2.61博斯氏菌属(Bosea )3.68发光杆菌属(Photobacterium ) 3.09博斯氏菌属(Bosea ) 2.43细菌未分类unclassified_bacteria 2.68希瓦氏菌属(Shewanella ) 2.45乳球菌属(Lactococcus )2.37鞘脂单胞菌属(Sphingomonas )1.93弧菌属(Vibrio )2.443㊀讨论3.1㊀不同饲料对钝吻黄盖鲽生长的影响集约化养殖鱼类的营养取决于饲料,饲料的质量决定了养殖鱼类的生长发育情况㊂本研究中,冰鲜杂鱼组钝吻黄盖鲽的特定生长率分别比配合饲料组和混合投喂组提高了29.66%和14.18%,说明冰鲜杂鱼更能促进钝吻黄盖鲽的生长,这与佟伟等[13]对大菱鲆㊁丛林梅等[14]对珍珠龙胆石斑鱼(Epinephelus fuscoguttatus ɬˑE .lanceolatus ȶ)㊁牛行健等[7]对斜带石斑鱼及朱择敏等[15]对大口黑鲈的研究结果一致㊂人工配合饲料虽营养丰富,但与天然饵料在营养成分㊁适口性等方面仍存在较大差异㊂本研究中,钝吻黄盖鲽对人工配合饲料还未形成很好的适应性,因而影响了养殖鱼体的生长速度㊂就饲料系数而言,虽然冰鲜杂鱼组的湿物质饲料系数比配合饲料组增加了2倍,但因冰鲜杂鱼水分含量较高,通过计算干物质量饲料系数发现,冰鲜杂鱼组的干物质量饲料系数比配合饲料组更低,加之冰鲜杂鱼的价格优势,配合饲料还应进一步改善质量以提升其饲料利用率,增加市场优势㊂3.2㊀不同饲料对钝吻黄盖鲽肠道结构的影响鱼类肠道健康与肠道绒毛形态密切相关,绒毛的长度和宽度影响着肠道对营养物质的消化和吸收[16]㊂肠道基层厚度影响肠道稳固性,对鱼体健康极为重要㊂本研究中,摄食配合饲料的钝吻黄盖鲽肠道绒毛长度显著低于冰鲜杂鱼组,结合生长数据,配合饲料组的钝吻黄盖鲽生长较慢,这可能是配合饲料适口性差㊁诱食作用弱等原因,导致钝吻黄盖鲽肠道形态发育不如冰鲜杂鱼组好,从而影响肠道对营养物质的吸收,进而影响了鱼的生长㊂3.3㊀不同饲料对钝吻黄盖鲽肠道菌群多样性的影响鱼类肠道中微生物与宿主在长期进化过程中形成稳定的共生关系,直接或间接地影响着宿主的营养吸收㊁代谢免疫等功能[17]㊂鱼类肠道菌群组成与其摄食的饲料也有关[18]㊂本研究中,配合饲料降低了钝吻黄盖鲽肠道菌群的多样性,结合配合饲料高温高压的生产加工方式,这可能与配合饲料中的微生物多样性较低有关㊂本研究中,3组钝吻黄盖鲽肠道的优势菌门均为变形菌门,第二位是放线菌门,第三位是厚壁菌门㊂变形菌门是细菌中最大的门,许多病原菌均位列其中,可导致生物功能失调,增加疾病风险[19]㊂冰鲜杂鱼组变形菌门丰度最高,这可能与冰鲜杂鱼携带的微生物数量高于配合饲料有关㊂放线菌门是水生生态系统中普遍存在的细菌门,是抗生素的主要生产菌,其中许多种类属于潜在的益生菌[20]㊂厚壁菌门中存在许多能够促进碳水化合物转化的细菌,有助于提高宿主对饲料的消化能力[21]㊂研究022大连海洋大学学报㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀第38卷表明,拟杆菌门与厚壁菌门丰度的比值影响养殖鱼类的生长速度[9]㊂本研究中,配合饲料组拟杆菌门与厚壁菌门丰度的比值最低,钝吻黄盖鲽的生长速度也最慢,这与郁二蒙等[9]对摄食不同饲料的大口黑鲈肠道菌群的研究结果一致㊂推测摄食配合饲料的钝吻黄盖鲽生长较慢,可能与其肠道中拟杆菌门与厚壁菌门丰度的比值下降有关㊂研究表明,红杆菌科中的大多数成员具有合成维生素B12的路径,而维生素B12可促进水产动物的生长[22]㊂红杆菌科中的鲁杰氏菌(Ruegeria mo-bilis)对致病菌有抑制作用,具有开发成为益生菌的潜能[23]㊂本研究中,在属水平上对钝吻黄盖鲽肠道进行分析发现,冰鲜杂鱼组未分类的红杆菌科丰度显著高于配合饲料组和混合投喂组㊂由此可见,投喂冰鲜杂鱼相对提高了钝吻黄盖鲽肠道有益菌的含量,这也为钝吻黄盖鲽肠道微生态制剂的开发提供了数据支撑㊂4 结论1)通过分析投喂配合饲料和冰鲜杂鱼在钝吻黄盖鲽生长性能㊁肠道结构和肠道菌群方面的差异,发现冰鲜杂鱼更能促进钝吻黄盖鲽的生长及肠道健康㊂2)配合饲料组钝吻黄盖鲽肠道菌群多样性减少,可能与人工配合饲料的营养成分无法满足鱼体及其肠道内土著细菌的生长需求有关㊂因此,在养殖过程中可在配合饲料中添加有利于钝吻黄盖鲽肠道健康的益生菌,以促进钝吻黄盖鲽的健康生长㊂参考文献:[1]㊀李思忠,王惠民.中国动物志:硬骨鱼纲鲽形目[M].北京:科学出版社,1995:214-216.㊀㊀㊀LI S Z,WANG H M.Zoology of China:Bonefish,Pleuronecti-formes[M].Beijing:Science Press,1995:214-216.(in Chinese) [2]㊀张岩,肖永双,高天翔,等.钝吻黄盖鲽野生群体遗传多样性分析[J].水产学报,2008,32(3):492-496.㊀㊀㊀ZHANG Y,XIAO Y S,GAO T X,et al.Analysis of genetic diver-sity of natural population in Pleuronectes yokohamae[J].Journal of Fisheries of China,2008,32(3):492-496.(in Chinese) [3]㊀窦硕增,杨纪明.黄河口黄盖鲽的食性及摄食的季节性变化[J].海洋学报,1992,14(6):103-112.㊀㊀㊀DOU S Z,YANG J M.Seasonal changes of feeding habits and food intake of Pseudopleuronectes yokohamae in the Yellow River Mouth [J].Haiyang Xuebao,1992,14(6):103-112.(in Chinese) 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[18]㊀MANSFIELD G S,DESAI A R,NILSON S A,et al.Characteriza-tion of rainbow trout(Oncorhynchus mykiss)intestinal microbiotaand inflammatory marker gene expression in a recirculating aqua-culture system[J].Aquaculture,2010,307(1/2):95-104. [19]㊀SHIN N R,WHON T W,BAE J W.Proteobacteria:microbial sig-nature of dysbiosis in gut microbiota[J].Trends in Biotechnolo-gy,2015,33(9):496-503.[20]㊀杜瑞,王梦歌,彭金金,等.环境未培养微生物中新型抗生素的发掘研究进展[J].生物加工过程,2022,20(2):172-181.㊀㊀㊀DU R,WANG M G,PENG J J,et al.Advances in discovery of new antibiotics in uncultured microorganisms[J].Chinese Journalof Bioprocess Engineering,2022,20(2):172-181.(in Chinese) [21]㊀EVANS N J,BROWN J M,MURRAY R D,et al.Characterizationof novel bovine gastrointestinal tract Treponema isolates and com-parison with bovine digital dermatitis treponemes[J].Appliedand Environmental Microbiology,2011,77(1):138-147. [22]㊀吴凡,文华,蒋明,等.维生素B12对草鱼幼鱼生长㊁体组分和造血机能的影响[J].吉林农业大学学报,2007,29(6):695-699.㊀㊀㊀WU F,WEN H,JIANG M,et al.Effects of dietary vitamin B12on growth,body composition and hemopoiesis of juvenile grass carp(Ctenopharyngodon idellus)[J].Journal of Jilin Agricultural U-niversity,2007,29(6):695-699.(in Chinese) [23]㊀SONNENSCHEIN E C,NIELSEN K F,D ALVISE P,et al.Globaloccurrence and heterogeneity of the Roseobacter clade speciesRuegeria mobilis[J].The ISME Journal,2017,11(2):569-583.Effects of iced trash fish and formulated diet on growthperformance,intestinal structure and microflora diversity ofmarbled flounder(Pseudopleuronectes yokohamae)XU Xiaoying1,WANG He1,LI Xiumei2,ZHAO Yanning1,ZHANG Ning2,SHI Wenkai1,LIU Peng1∗(1.Yantai Marine Economic Research Institute,Yantai264006,China;2.Yantai Zongzhe Ocean Technology Company Limited,Yantai265617, China)Abstract:In order to evaluate effects of iced trash fish,formulated diet and their mixture on growth,intestinal structure and microflora of marbled flounder(Pseudopleuronectes yokohamae),the same batch of marbled flounder juveniles with body length of(10.16ʃ0.06)cm were reared in a126L plastic tank and fed iced trash fish,formu-lated diet and their mixture at water temperature of(15ʃ1)ħfor42days.The results showed that the fish fed iced trash had significant increase in the body weight gain and specific growth rate(P<0.05),and decrease in the feed conversion ratio(P<0.05).The intestinal histological structure showed that the length of intestinal villi and the thickness of basal layer were significantly increased in the fish fed iced trash(P<0.05).The intestinal microflora analysis showed that the fish fed iced trash had increase in the intestinal flora diversity,promoting the distribution of beneficial bacteria such as Rhodobacteraceae in the intestinal tract of marbled flounder.It was speculated that the marbled flounder fed formulated diet had slower growth may be involved in the poorly developed intestinal muco-sa and the decline in the ratio of Bacteroides to Firmicutes.The indices in the mixed diet group were mediated be-tween iced trash fish group and formulated feed group.Therefore,beneficial bacteria in Rhodobacteraceae were added in diet to promote the growth and nutrient absorption of marbled flounder during cultivation.The artificial compound feed of marbled flounder should be further optimized to improve market advantage.Key words:Pseudopleuronectes yokohamae;growth performance;intestinal structure;intestinal microflora;formu-lated feed;iced trash fish222大连海洋大学学报㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀㊀第38卷。
