Article(id=1153433739041428450, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250110004, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1736438400000, receivedDateStr=2025-01-10, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752929633150, onlineDateStr=2025-07-19, pubDate=1745510400000, pubDateStr=2025-04-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752929633150, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752929633150, creator=13701087609, updateTime=1752929633150, updator=13701087609, issue=Issue{id=1153433737141412332, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='8', pageStart='1', pageEnd='316', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752929632696, creator=13701087609, updateTime=1757293087150, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1171735391666225233, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1171735391666225234, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=31, endPage=38, ext={EN=ArticleExt(id=1153433739825763315, articleId=1153433739041428450, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Comparison of intestinal microbiota in different aquaculture modes of Opsariichthys bidens, columnId=1151923891565326960, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Processing and Quality Safety of Aquatic Products, runingTitle=null, highlight=null, articleAbstract=

Objective To study the effects of different aquaculture modes on the intestinal microbiota of Opsariichthys bidens. Methods The 16S rRNA gene was amplified by polymerase chain reaction (PCR) using 515F-907R primers using the intestinal microbial DNA of C (monoculture of Opsariichthys bidens), D (mixed culture of Opsariichthys bidens and crayfish), E (mixed culture of Opsariichthys bidens and Macrobrachium nipponense) as templates, and sequenced by Illumina Hiseq2500 sequencing platform. The growth performance of Opsariichthys bidens in different culture modes was simultaneously determined. Results From the perspective of weight and fatness, C>E>D, from the perspective of body length, there was no significant difference between C and E, and there were significant differences between C and D, D and E, which was also C>E>D; from the perspective of hepatosomatic index, there was no significant difference between D and E, but there were significant differences between C and D, C and E, C>E=D; the rule of viscera index and hepatosomatic index was the same, C>D>E; the mixed fish and shrimp farming model significantly increased the diversity and richness of intestinal microbiota in fish compared to the single fish farming model; the mixed farming mode increased the diversity of intestinal microbiota composition in Opsariichthys bidens, and 2 kinds of mixed farming modes had the same effect; the indicator groups for monoculture and 2 kinds of mixed breeding modes of Opsariichthys bidens were different, and the common indicator groups for both mixed breeding modes of Opsariichthys bidens were Microbacteria and Prevotella. Conclusion The comprehensive utilization and quality safety of agricultural products, as well as the single farming mode, result in higher body weight and fat content of Opsariichthys bidens, which is beneficial for increasing the yield of food processing; the mixed farming mode has better microbial diversity and richness, stronger resistance and better quality of Opsariichthys bidens, which is conducive to improving the product quality of food processing. This study can provide theoretical support for the nutritional supply, metabolic homeostasis, epidemic prevention and immunity, food processing, and other aspects of Opsariichthys bidens under different breeding modes.

, correspAuthors=Shui-Rong GUO, Gui-Jie HAO, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Yan-Na CUI, Lei HUANG, Yu-Chen WANG, Jiang-Qi WANG, Ya-Fang SHEN, Ai-Xia HUANG, Yu-Xin WU, Qiu-Yue HU, Shui-Rong GUO, Gui-Jie HAO), CN=ArticleExt(id=1153433756913357104, articleId=1153433739041428450, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=不同养殖模式马口鱼肠道微生物群落比较, columnId=1153433738143851016, journalTitle=食品安全质量检测学报, columnName=本期专题:水产品加工与质量安全, runingTitle=null, highlight=null, articleAbstract=

目的 研究不同养殖模式对马口鱼肠道微生物群落的影响。方法 采取以C(马口鱼单养)、D(马口鱼与小龙虾混养)、E(马口鱼与青虾混养)的马口鱼肠道微生物DNA为模板, 通过聚合酶链式反应(polymerase chain reaction, PCR), 采用515F-907R引物对16S rRNA基因进行扩增, 并用Illumina Hiseq2500测序平台测序, 同步测定了不同养殖模式的马口鱼生长性能。结果 从体重和肥满度来看, C>E>D; 从体长来看, C与E无显著性差异, C与D、D与E均有显著性差异, 也是C>E>D; 从肝体指数来看, D与E无显著性差异, C与D、C与E均有显著性差异, C>E=D; 脏体指数、肝体指数的规律相同, C>D>E; 鱼虾混养模式比单一养鱼模式鱼类肠道菌群的多样性和丰富度均显著增加; 混养模式增加了马口鱼的肠道菌群组成多样性, 两种混养模式效果相同; 单养模式和两种混养模式马口鱼的指示类群不同, 两种混养模式马口鱼的共有指示类群为微杆菌属和普雷沃氏菌属。结论 单养模式马口鱼的体重和肥满度更高, 有利于提升食品加工的产量; 混养模式微生物多样性和丰富度更好, 马口鱼的抵抗力更强, 品质更好, 有利于提升食品加工的产品质量。本研究可对不同养殖模式马口鱼营养供应、代谢稳态和防疫免疫、食品加工等提供了理论支撑。

, correspAuthors=郭水荣, 郝贵杰, authorNote=null, correspAuthorsNote=
* 郭水荣(1966—), 男, 正高级工程师, 主要研究方向为水产养殖。E-mail:
郝贵杰(1979—), 女, 博士, 研究员, 主要研究方向为水产品质量安全与加工。E-mail:
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崔雁娜(1983—), 女, 硕士, 工程师, 主要研究方向为水产品质量安全与加工。E-mail:

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崔雁娜(1983—), 女, 硕士, 工程师, 主要研究方向为水产品质量安全与加工。E-mail:

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注: 不同小写字母表示具有显著性差异(P<0.05)。

, figureFileSmall=5xerdLHmQ7yxyFE+hU5s+w==, figureFileBig=LuRL8nHINWGxuVZtWiB/OA==, tableContent=null), ArticleFig(id=1171733824389038087, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433739041428450, language=EN, label=Fig.3, caption=Relative abundance of bacteria of 3 kinds of farming models of Opsariichthys bidens, figureFileSmall=Th1sD3vr84RkoVM5Bxkfvg==, figureFileBig=3nMV7uo4bocikndpvwSiFQ==, tableContent=null), ArticleFig(id=1171733824451952648, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433739041428450, language=CN, label=图3, caption=3种养殖模式马口鱼细菌相对丰度

注: a. 3种养殖模式马口鱼门水平下细菌相对丰度; b. 3种养殖模式马口鱼种水平下细菌相对丰度。

, figureFileSmall=Th1sD3vr84RkoVM5Bxkfvg==, figureFileBig=3nMV7uo4bocikndpvwSiFQ==, tableContent=null), ArticleFig(id=1171733824514867210, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433739041428450, language=EN, label=Fig.4, caption=Linear discriminant analysis of bacterial intestinal communities of Opsariichthys bidens, figureFileSmall=eprbpaqgaPO5PYOXqhET+A==, figureFileBig=ueozmErpsm37nPNNg0+YNQ==, tableContent=null), ArticleFig(id=1171733824573587468, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433739041428450, language=CN, label=图4, caption=马口鱼肠道细菌群落的线性判别分析, figureFileSmall=eprbpaqgaPO5PYOXqhET+A==, figureFileBig=ueozmErpsm37nPNNg0+YNQ==, tableContent=null), ArticleFig(id=1171733824623919118, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433739041428450, language=EN, label=Table 1, caption=

Growth performance of different culture patterns of Opsariichthys bidens (n≥3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 体重/g 体长/cm 肥满度 肝体指数 脏体指数
C 29.33±7.79a 13.84±1.18a 1.09±0.10a 2.82±0.76a 7.46±4.61a
D 16.77±4.37b 12.70±0.87b 0.80±0.08b 1.66±0.36b 4.22±1.41b
E 23.16±6.88c 13.83±1.37a 0.85±0.07c 1.66±0.39b 4.07±1.39b
), ArticleFig(id=1171733824678445072, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433739041428450, language=CN, label=表1, caption=

马口鱼不同养殖模式生长性能(n≥3)

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组别 体重/g 体长/cm 肥满度 肝体指数 脏体指数
C 29.33±7.79a 13.84±1.18a 1.09±0.10a 2.82±0.76a 7.46±4.61a
D 16.77±4.37b 12.70±0.87b 0.80±0.08b 1.66±0.36b 4.22±1.41b
E 23.16±6.88c 13.83±1.37a 0.85±0.07c 1.66±0.39b 4.07±1.39b
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不同养殖模式马口鱼肠道微生物群落比较
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崔雁娜 1 , 黄雷 1 , 王雨辰 1 , 汪江琦 1 , 沈亚芳 1 , 黄爱霞 1 , 武雨鑫 1 , 胡秋月 1 , 郭水荣 2, * , 郝贵杰 1, *
食品安全质量检测学报 | 本期专题:水产品加工与质量安全 2025,16(8): 31-38
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食品安全质量检测学报 | 本期专题:水产品加工与质量安全 2025, 16(8): 31-38
不同养殖模式马口鱼肠道微生物群落比较
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崔雁娜1 , 黄雷1, 王雨辰1, 汪江琦1, 沈亚芳1, 黄爱霞1, 武雨鑫1, 胡秋月1, 郭水荣2, * , 郝贵杰1, *
作者信息
  • 1.农业农村部淡水渔业健康养殖重点实验室, 浙江省鱼类健康与营养重点实验室, 湖州市水产品品质提升与加工技术重点实验室, 浙江省淡水水产研究所, 湖州 313001
  • 2.杭州市农业技术推广中心, 杭州 310020
  • 崔雁娜(1983—), 女, 硕士, 工程师, 主要研究方向为水产品质量安全与加工。E-mail:

通讯作者:

* 郭水荣(1966—), 男, 正高级工程师, 主要研究方向为水产养殖。E-mail:
郝贵杰(1979—), 女, 博士, 研究员, 主要研究方向为水产品质量安全与加工。E-mail:
Comparison of intestinal microbiota in different aquaculture modes of Opsariichthys bidens
Yan-Na CUI1 , Lei HUANG1, Yu-Chen WANG1, Jiang-Qi WANG1, Ya-Fang SHEN1, Ai-Xia HUANG1, Yu-Xin WU1, Qiu-Yue HU1, Shui-Rong GUO2, * , Gui-Jie HAO1, *
Affiliations
  • 1. Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture and Rural Affairs, Huzhou Key Laboratory of Aquatic Product Quality Improvement and Processing Technology, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China
  • 2. Hangzhou Agricultural Technology Extension Center, Hangzhou 310020, China
出版时间: 2025-04-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250110004
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目的 研究不同养殖模式对马口鱼肠道微生物群落的影响。方法 采取以C(马口鱼单养)、D(马口鱼与小龙虾混养)、E(马口鱼与青虾混养)的马口鱼肠道微生物DNA为模板, 通过聚合酶链式反应(polymerase chain reaction, PCR), 采用515F-907R引物对16S rRNA基因进行扩增, 并用Illumina Hiseq2500测序平台测序, 同步测定了不同养殖模式的马口鱼生长性能。结果 从体重和肥满度来看, C>E>D; 从体长来看, C与E无显著性差异, C与D、D与E均有显著性差异, 也是C>E>D; 从肝体指数来看, D与E无显著性差异, C与D、C与E均有显著性差异, C>E=D; 脏体指数、肝体指数的规律相同, C>D>E; 鱼虾混养模式比单一养鱼模式鱼类肠道菌群的多样性和丰富度均显著增加; 混养模式增加了马口鱼的肠道菌群组成多样性, 两种混养模式效果相同; 单养模式和两种混养模式马口鱼的指示类群不同, 两种混养模式马口鱼的共有指示类群为微杆菌属和普雷沃氏菌属。结论 单养模式马口鱼的体重和肥满度更高, 有利于提升食品加工的产量; 混养模式微生物多样性和丰富度更好, 马口鱼的抵抗力更强, 品质更好, 有利于提升食品加工的产品质量。本研究可对不同养殖模式马口鱼营养供应、代谢稳态和防疫免疫、食品加工等提供了理论支撑。

马口鱼  /  养殖模式  /  肠道微生物  /  生长性能

Objective To study the effects of different aquaculture modes on the intestinal microbiota of Opsariichthys bidens. Methods The 16S rRNA gene was amplified by polymerase chain reaction (PCR) using 515F-907R primers using the intestinal microbial DNA of C (monoculture of Opsariichthys bidens), D (mixed culture of Opsariichthys bidens and crayfish), E (mixed culture of Opsariichthys bidens and Macrobrachium nipponense) as templates, and sequenced by Illumina Hiseq2500 sequencing platform. The growth performance of Opsariichthys bidens in different culture modes was simultaneously determined. Results From the perspective of weight and fatness, C>E>D, from the perspective of body length, there was no significant difference between C and E, and there were significant differences between C and D, D and E, which was also C>E>D; from the perspective of hepatosomatic index, there was no significant difference between D and E, but there were significant differences between C and D, C and E, C>E=D; the rule of viscera index and hepatosomatic index was the same, C>D>E; the mixed fish and shrimp farming model significantly increased the diversity and richness of intestinal microbiota in fish compared to the single fish farming model; the mixed farming mode increased the diversity of intestinal microbiota composition in Opsariichthys bidens, and 2 kinds of mixed farming modes had the same effect; the indicator groups for monoculture and 2 kinds of mixed breeding modes of Opsariichthys bidens were different, and the common indicator groups for both mixed breeding modes of Opsariichthys bidens were Microbacteria and Prevotella. Conclusion The comprehensive utilization and quality safety of agricultural products, as well as the single farming mode, result in higher body weight and fat content of Opsariichthys bidens, which is beneficial for increasing the yield of food processing; the mixed farming mode has better microbial diversity and richness, stronger resistance and better quality of Opsariichthys bidens, which is conducive to improving the product quality of food processing. This study can provide theoretical support for the nutritional supply, metabolic homeostasis, epidemic prevention and immunity, food processing, and other aspects of Opsariichthys bidens under different breeding modes.

Opsariichthys bidens  /  aquaculture modes  /  intestinal microbiota  /  growth properties
崔雁娜, 黄雷, 王雨辰, 汪江琦, 沈亚芳, 黄爱霞, 武雨鑫, 胡秋月, 郭水荣, 郝贵杰. 不同养殖模式马口鱼肠道微生物群落比较. 食品安全质量检测学报, 2025 , 16 (8) : 31 -38 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250110004
Yan-Na CUI, Lei HUANG, Yu-Chen WANG, Jiang-Qi WANG, Ya-Fang SHEN, Ai-Xia HUANG, Yu-Xin WU, Qiu-Yue HU, Shui-Rong GUO, Gui-Jie HAO. Comparison of intestinal microbiota in different aquaculture modes of Opsariichthys bidens[J]. Journal of Food Safety & Quality, 2025 , 16 (8) : 31 -38 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250110004
马口鱼(Opsariichthys bidens)隶属鲤形目(Cypriniformes), 鲤科(Cyprinidae), 马口鱼属(Opsariichthys), 俗名花杈鱼、桃花鱼、山鳡等, 杂食偏肉食性淡水鱼类, 溪流中多见, 肉质鲜美, 营养价值高[1]。马口鱼可以产生非常好的经济效益, 市场价为50~120元/kg[2-3]。不同的养殖模式, 利润差别较大。例如翦晓红[4]采用土池微流水方式养殖马口鱼, 利润5945元/亩; 张乐乐等[5]在江西赣南地区池塘单养马口鱼, 产值达17952元/亩, 利润达8810元/亩; 吴燕琴等[6]采取马口鱼单养模式, 实现利润9754元/亩; 采取马口鱼搭养鲢、鳙和土鲫夏花的模式, 马口鱼实现利润10406.6元/亩; 采取马口鱼搭养鲢鱼和唇䱻夏花的模式, 马口鱼实现利润8682.06元/亩[7]; 王燕波等[8]采用砖砌水泥池、塑料膜铺底土池、一般土池、稻田轮养4种模式进行养殖, 最高产量632 kg/亩, 净效益14914元/亩, 平均产量214 kg/亩, 产值15215元/亩, 效益3892元/亩, 比当地同类养殖水域的养殖效益提高1倍以上; 林建忠[9]在丘陵山坳池塘养殖实验, 马口鱼产量达506.1 kg/亩, 平均利润为7788.8元/亩。
马口鱼具有良好的生态效益, 是我国乃至世界鱼类物种多样性保护资源, 是贵州省黔东南州道雷公山自然保护区鱼类之一[10]。近年来, 由于过度捕捞、严重的水环境污染和河流筑坝, 马口鱼这种自然资源迅速下降, 影响了多地的自然水生态系统[11-13]。从多重生态维度来讲, 马口鱼具有优先保护的价值[14]
然而, 关于马口鱼肠道微生物群落的研究还比较少。鱼类的肠道微生物群落在鱼类生长过程中的营养供应、代谢稳态和免疫防疫中都会起到重要的作用[15]。鱼类的肠道微生物群包括原生动物、真菌、酵母、病毒以及细菌和古菌[16]。细菌是鱼类肠道的主要微生物群[17]。关于鱼类不同养殖模式的研究, 马明星[18]比较了循环水和流水模式下许氏平鲉的肠道菌群差异性, 发现Alpha多样性指数间没有显著性差异, 具体菌群存在显著性差异; LUO等[19]研究了尼罗罗非鱼和鳙鱼混养、鲤鱼和鳙鱼混养两种养殖模式中鳙鱼肠道菌群的差异, 尽管饲喂相同的饲料配方, 但鳙鱼在两种养殖模式下的肠道菌群β多样性具有显著差异; 严雪瑜等[20]研究了稻田和池塘两种模式下金边鲤与建鲤肠道菌群差异分析, 发现稻田养殖的金边鲤和建鲤肠道菌群多样性高于相应的池塘养殖模式; 王金林等[21]对野生和养殖的异齿裂腹鱼肠道菌群进行了结构分析, 发现野生和人工养殖两种模式下异齿裂腹鱼肠道菌群的丰富度和多样性差异不显著, 但两种养殖模式的优势菌群不同; 邓智明[22]进行了野生与养殖圆口铜鱼肠道微生物研究, 发现无论是养殖、野生, 还是不同的摄食强度圆口铜鱼肠道微生物的优势门均为变形菌门, 养殖和野生的前、中肠优势菌不同, 后肠优势菌相同。
综合上述, 同一种鱼处于循环水和流水两种养殖模式, 同一种鱼分别与两种不同鱼混养, 同一种鱼在稻田和池塘两种不同养殖环境下, 同一种野生和养殖鱼类的肠道微生物均有显著性差异, 有的是肠道菌群多样性存在差异性, 有的是优势菌群存在差异性。然而, 对于同一种鱼类和不同的虾混养过程中, 鱼的肠道微生物变化规律还少有报道。因此, 本研究基于16S rRNA检测对不同养殖模式(小龙虾与马口鱼混养、青虾与马口鱼混养)的马口鱼体内肠道微生物群落进行了研究, 以期为不同养殖模式马口鱼营养供应、代谢稳态和防疫免疫、食品加工等提供参考依据。
2022年4月底在浙江湖州某家庭农场放养马口鱼水花苗, 放养密度70000尾/亩。放苗之前池塘已经清塘、晒干、消毒。2022年6月底将养成后的马口鱼夏花分成3个塘, 分别为马口鱼单养塘(C), 马口鱼夏花放养密度为7000尾/亩; 马口鱼和小龙虾混养塘(D), 马口鱼夏花放养密度为7000尾/亩, 小龙虾苗放养密度60000尾/亩; 马口鱼和青虾混养塘(E), 马口鱼夏花放养密度为7000尾/亩, 青虾苗放养密度60000尾/亩。3种养殖模式马口鱼的饲料均是万盛恒通的鱼膨化配合饲料, 小龙虾和青虾使用的饲料均是万盛恒通的虾颗粒配合饲料。3种养殖模式养殖过程全程不使用兽药。2023年1月进行采样分析。
选取活力好、无外伤, 平行样品大小相近的C(平均体重为25.1 g±0.1 g)、D(平均体重为24.8 g±0.1 g)、E的马口鱼(平均体重为25.0 g±0.1 g)各25尾, 对取样工具消毒后, 分别取样后测体重、体长、肠重、肝重, 并将马口鱼肠道放置于-80 ℃保存待测肠道微生物。
E.Z.N.A. Soil DNA Kit DNA抽提试剂盒(美国Omega公司); FastPfu Polymerase(北京全式金生物技术有限公司)。
ABI GeneAmp® 9700型聚合酶链式反应(polymerase chain reaction, PCR)仪(美国ABI公司); QuantiFluor™-ST蓝色荧光定量系统(美国Promega公司); SECURA125-1CN/SQP十万分之一分析天平(德国Sartorius科学仪器有限公司)。
取C、D、E马口鱼样品各25尾, 测量其体重、体长, 解剖后取内脏和肝脏, 称重, 计算各组肥满度(condition factor, CF)、肝体指数(hepatopancreas somatic indices, HSI)、脏体指数(visceral sensitivity index, VSI)。各指标按照公式(1)~(3)计算:
CF/%=W/L3×100%
HSI/%=W肝脏/W×100%
VSI/%=W内脏/W×100%
式中: W肝脏表示肝脏重, g; W内脏表示内脏重, g; W表示对应的体重, g; L表示体长, cm。
以C、D、E马口鱼的肠道微生物DNA为模板, 通过PCR, 采用515F-907R引物对16S rRNA基因进行扩增, 并用Illumina Hiseq2500测序平台测序获得原始数据。所选引物515F和907R的序列分别为: 515F (GTGCCAGCMGCCGCGG)、907R (CCGTCAATTCMTT TRAGTTT)。PCR正式实验采用2×Phanta Max Master Mix, 20 μL反应体系: 2×Phanta Max Master Mix 10 µL, 引物515F和907R各0.8 µL, DNA模板10 ng, 补ddH2O至20 µL。扩增条件: 95 ℃ 3 min; 95 ℃ 30 s, 53 ℃ 30 s, 72 ℃ 45 s, 29个循环; 72 ℃延伸10 min。
将同一样本的PCR产物混合后切胶回收, Tris_HCl洗脱; 2%琼脂糖电泳检测。参照电泳初步定量结果进行检测定量, 按相应比例混合。采用Hieff NGS®Ultima Pro DNA Library Prep Kit试剂盒文库构建, 在Novaseq 6000 PE250平台进行高通量测序。
首先需要根据barcode得到所有样品的有效序列; 对序列进行质控; 用Usearch 11.0软件和gold数据库去除嵌合体, 对高通量测序数据进行分析。用SPSS 19.0软件分析其他数据的显著性差异。数据的表现形式为平均值±标准偏差。
从体重来看, 3种养殖模式均有显著性差异, C>E>D; 肥满度也有显著性差异, 也是C>E>D; 从体长来看, C与E无显著性差异, C与D、D与E均有显著性差异, 也是C>E>D; 从肝体指数来看, D与E无显著性差异, C与D、C与E均有显著性差异, C>E=D; 脏体指数与肝体指数的规律相同, C>D>E。具体差异数值见表1
对C、D、E 3种养殖末端的马口鱼肠道微生物16S rRNA基因测序数据进行双端拼接、质控以及嵌合体过滤后, 共获得有效序列1367623条。主要序列长度集中在361~380 bp, 占比达99.5%。3组样品的覆盖度均达到0.999以上, 表明检测的样本量具有代表性。物种累计曲线(图1)可以看出抽样充分。
马口鱼微生物菌群α多样性分析采用两种分析方法—Kruskal-Wallis函数分析和Anova函数分析, 两者均显示: 3种养殖模式的总体ace指数、chao1指数、shannon指数、simpson指数均有显著性差异(P<0.05)(图2), 且D、E的α多样性显著高于C, D与E之间无显著性差异。
其中ace指数是一种通过已知物种估计未知物种丰富度的指标, 一般来说, ace指数越高, 表示物种丰富度越大; chao1指数是在ace指数基础上发展起来的, 与ace指数类似, chao1指数越高, 物种丰富度越大; shannon指数是用于衡量生物群落物种多样性的一种指标, 它综合了物种丰富度和物种均匀度两个方面, shannon指数越高, 表示物种多样性越高; simpson指数也是衡量生物群落多样性的一种指标, 与shannon指数相似, 但更加注重物种均匀度的影响, 其值越小表示物种多样性越高。
图2来看, D、E的物种丰富度和多样性都显著高于C。
对3种养殖模式的马口鱼肠道菌群进行了菌群组成分析(图3)。
在门水平, C模式下的肠道菌群主要包括硬壁菌门(84.40%)、变形菌门(13.60%)、拟杆菌门(<1.00%); D模式下的肠道菌群主要包括硬壁菌门(10.10%)、变形菌门(62.90%)、拟杆菌属(13.40%)、放线菌属(8.20%)、蓝细菌(1.65%)等; E模式下的肠道菌群主要包括硬壁菌门(35.60%)、变形菌门48.80%、拟杆菌门8.78%、放线菌门4.61%、其他1.32%。
可见, 硬壁菌门、变形菌门和拟杆菌门是3种养殖模式马口鱼肠道菌群的共有菌群。D和E两种模式共有菌群相对丰度达90%以上。
在属水平, C模式下的肠道菌群主要包括阿尔卡利根科(1.82%)、支原体属(84.40%)、气单胞菌属(9.56%)、希瓦氏菌属(1.75%)等; D模式下的肠道菌群主要包括阿尔卡利根科(46.10%)、波波杆菌(10.70%)、假单胞菌属(6.62%)、未分类的细菌(3.84%)、不动杆菌(4.16%)、丙酸杆菌科(3.56%)、毛球菌(3.25%)等; E模式下的肠道菌群主要包括阿尔卡利根科(41.90%)、波波杆菌(8.68%)、乳酸杆菌(18.70%)、假单胞菌属(4.32%)、未分类的细菌(2.99%)、罗伊氏乳杆菌(2.70%)、唾液乳酸杆菌(2.70%)、芽孢杆菌科未培养的杆菌(2.12%)、鞘氨醇单胞菌(1.01%)、乳酸杆菌属(1.01%)等。可见, 3种养殖模式共有菌群是阿尔卡利根科。D、E两种模式相同菌群的丰度达50%以上。
利用不同线性判别分析筛选了3种养殖模式下马口鱼肠道中的细菌主要指示类群。
C模式下的指示类群为金抗菌属; D模式的指示类群为双杆菌目, 嗜酸杆菌纲, Aminicenantales, 戈登氏属, 红球菌属, 诺卡氏科, 棒状杆菌目, 微杆菌属, 谷氨酸杆菌属, 丙酸杆菌属、科、目, 链球菌科, 放线菌纲, 未分类的细菌属、科、目、纲, 普雷沃氏菌属、科, 文肯菌科, Blvii28_wastewater_sludge, 威廉.惠特曼尼科, Lentimicrobium, Pedobacte, 鞘氨醇杆菌科、目, 拟杆菌纲、目、属、科; E模式的指示类群为未分类的杆菌属、科、目, 维氏菌属纲, 未分类的酸性微生物属、科、目、纲, Iamia, Iamiaceae, Microtrichales, Microtrichales, Microtrichales, Flaviflexus, Tomitella, Corynebacteriales_f_ Corynebacteriales_ Incertae_Sedis, 尤泽比氏菌属、科、目, 间孢囊菌科属, 微杆菌属, 微球藻属、科, 小单胞菌属、科、目, PeM15属、科、目, 裂孢菌属, 链球菌目, 柯林斯菌属, Enorma, CHKCI002, Eggerthellaceae, 未分类的OPB41属、科、目, 白毛杆菌纲, 放线菌目, 康乃斯氏杆菌属, 红杆菌科、目, 嗜热油菌纲, Dysgonomonadaceae, 发酵单胞菌属, 普雷沃氏菌属, WCHB1_32, Flavitalea, Ohtaekwangia, Microscillaceae, 寒冷微菌属, Myroides, Ulvibacter(图4)。
影响马口鱼生长性能的因素很多, 其中食物的可得性为其中之一[23], 但本研究的马口鱼养殖密度比较低, 显然不存在这样的问题。实验证明, 低放养密度对鱼的体重、体长、CF、HSI、VSI无显著影响[24]。从体重和CF来看, C>E>D, 且有显著性差异。这说明单养马口鱼的出肉率更高[25], 有利于提升食品加工的产量。且可推断是因为混养模式造成虾为了躲避敌人, 活动量增大, 导致体重降低[26]
前人研究证明: 与单独养虾相比, 鱼虾混养模式中后期弧菌数量显著低于单独养虾模式[27]; 稻鱼共作模式和池塘单养模式在黄颡鱼肠道微生物方面无显著差异, 但稻鱼共作模式下, 黄颡鱼拥有更丰富的肠道微生物多样性[28]; 不同地区鱼类养殖的水质细菌群落多样性不存在显著差异, 但ace指数与Chao1指数均表现为混养鱼类池塘水质>单养池塘(P<0.01)[29]; 与稻虾共作模式相比, 稻虾鱼共作模式降低了水体中的微生物多样性[30]
本研究发现马口鱼肠道微生物多样性与马口鱼虾养殖环境变化规律基本相同: 马口鱼虾混养模式比马口鱼单养模式鱼类肠道菌群的多样性和丰富度均有显著增加。
在门水平, D和E两种模式共有菌群相对丰度达90%以上且包含了C模式, 说明了混养模式增加了马口鱼的肠道菌群组成多样性, 两种混养模式效果相同。有研究表明, 硬壁菌门和变形菌门是淡水鱼最常见的菌门, 且同一品种的鱼在不同的环境中肠道微生物优势菌门不会变化[31], 这与本研究结论相同。
在种水平, 3种养殖模式共有菌群是阿尔卡利根科。D、E两种模式相同菌群的丰度达50%以上。和门水平变化规律相似, 但共有菌群发生了变化, 且D、E两种模式的相同菌群丰度明显减少, 可推断, 在门水平下, 对3种养殖模式的菌群组成进行比较更为合适。
C模式仅有一种指示菌属—金抗菌属, 且与混养模式不同。两种混养模式(D和E)的指示菌很多, 共发现两种相同的指示菌属, 分别为微杆菌属和普雷沃氏菌属。金抗菌属较少见文献报道; 微杆菌属是一类重要的产蛋白酶菌属[32], 是在鲫鱼多地养殖池塘底泥中出现的共同菌属, 被认为是益生菌[33]; 普雷沃氏菌属属于拟杆菌门, 是瘤胃中降解植物纤维素的主要微生物[34]。通过已有文献可推断, 两种混养模式马口鱼共有指示类群均为有益微生物。有研究证明: 有益微生物会影响鱼体内消化酶的活性, 从而提高鱼对营养物质的吸收和生长[35], 会改善鱼体中的脂肪和蛋白质含量[36]
本研究比较了不同养殖模式下马口鱼的生长性能、微生物多样性、肠道菌群组成及指示类群的差异。结果表明, 从体重和CF来看, C(马口鱼单养)>E(马口鱼与青虾混养)>D(马口鱼与小龙虾混养); 鱼虾混养模式比单一养鱼模式鱼类肠道菌群的多样性和丰富度都有显著增加; 混养模式增加了马口鱼的肠道菌群组成多样性, 两种混养模式效果相同; 单养模式和两种混养模式马口鱼的指示类群不同, 两种混养模式马口鱼的共有指示类群为微杆菌属和普雷沃氏菌属。由于实验时间和精力所限, 本研究未能深入研究马口鱼肠道菌群和马口鱼肉品质的直接关联性, 建议后续可以开展相关方面的研究。
  • 湖州市科技特派员项目(2021KT40)
  • 江西省渔业种业联合育种攻关项目(2023YYZYGG-08)
  • 杭州市农业科技协作与创新攻关项目(202209SX08)
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2025年第16卷第8期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250110004
  • 接收时间:2025-01-10
  • 首发时间:2025-07-19
  • 出版时间:2025-04-25
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  • 收稿日期:2025-01-10
基金
湖州市科技特派员项目(2021KT40)
江西省渔业种业联合育种攻关项目(2023YYZYGG-08)
杭州市农业科技协作与创新攻关项目(202209SX08)
作者信息
    1.农业农村部淡水渔业健康养殖重点实验室, 浙江省鱼类健康与营养重点实验室, 湖州市水产品品质提升与加工技术重点实验室, 浙江省淡水水产研究所, 湖州 313001
    2.杭州市农业技术推广中心, 杭州 310020

通讯作者:

* 郭水荣(1966—), 男, 正高级工程师, 主要研究方向为水产养殖。E-mail:
郝贵杰(1979—), 女, 博士, 研究员, 主要研究方向为水产品质量安全与加工。E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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