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牛磺酸对卵形鲳鲹肠道微生物及免疫功能的影响

马启伟 郭梁 刘波 刘宝锁 朱克诚 郭华阳 张楠 杨静文 张殿昌

马启伟, 郭梁, 刘波, 刘宝锁, 朱克诚, 郭华阳, 张楠, 杨静文, 张殿昌. 牛磺酸对卵形鲳鲹肠道微生物及免疫功能的影响[J]. 南方水产科学, 2021, 17(2): 87-96. doi: 10.12131/20200193
引用本文: 马启伟, 郭梁, 刘波, 刘宝锁, 朱克诚, 郭华阳, 张楠, 杨静文, 张殿昌. 牛磺酸对卵形鲳鲹肠道微生物及免疫功能的影响[J]. 南方水产科学, 2021, 17(2): 87-96. doi: 10.12131/20200193
Qiwei MA, Liang GUO, Bo LIU, Baosuo LIU, Kecheng ZHU, Huayang GUO, Nan ZHANG, Jingwen YANG, Dianchang ZHANG. Effect of taurine on intestinal microbes and immune function in golden pompano (Trachinotus ovatus)[J]. South China Fisheries Science, 2021, 17(2): 87-96. doi: 10.12131/20200193
Citation: Qiwei MA, Liang GUO, Bo LIU, Baosuo LIU, Kecheng ZHU, Huayang GUO, Nan ZHANG, Jingwen YANG, Dianchang ZHANG. Effect of taurine on intestinal microbes and immune function in golden pompano (Trachinotus ovatus)[J]. South China Fisheries Science, 2021, 17(2): 87-96. doi: 10.12131/20200193

牛磺酸对卵形鲳鲹肠道微生物及免疫功能的影响

doi: 10.12131/20200193
基金项目: 现代农业产业技术体系建设专项资金 (CARS-47-G07);中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金资助 (2020TD29);广东省现代农业产业技术体系海水鱼产业创新项目 (2019KJ143);国家自然科学基金项目 (U20A2064)
详细信息
    作者简介:

    马启伟 (1995—),男,硕士研究生,研究方向为水产动物营养和遗传育种。E-mail: sdmaqw@163.com

    通讯作者:

    张殿昌 (1977—),男,博士,研究员,从事水产种质资源与遗传育种研究。E-mail: zhangdch@scsfri.ac.cn

  • 中图分类号: S963.73

Effect of taurine on intestinal microbes and immune function in golden pompano (Trachinotus ovatus)

  • 摘要: 为了探讨牛磺酸对卵形鲳鲹 (Trachinotus ovatus) 肠道微生物菌群结构和免疫功能的影响,试验以鱼粉、发酵豆粕和玉米蛋白粉为基础蛋白源配制了牛磺酸质量分数分别为1.3 g·kg−1 (T0)、4.4 g·kg−1 (T1)、7.4 g·kg−1 (T2)、10.5 g·kg−1 (T3)、12.7 g·kg−1 (T4)的等氮等脂饲料。将750尾卵形鲳鲹 (81.0±0.5) g随机分为5组 (每组150尾,分为3个平行),试验为期56 d。高通量测序结果显示在卵形鲳鲹肠道内共获得87 707条序列,能够注释到数据库的OTUs数目为5 130 (95.32%),在门水平上,变形菌门、软壁菌门、螺旋体门为优势菌群。Alpha和Beta多样性分析表明,T2组中的肠道微生物丰富度和多样性最低,T1和T4组间物种组成差异较大 (P<0.05)。随外源性牛磺酸的添加,血清溶菌酶活性显著提高,补体C4和免疫球蛋白含量显著增加 (P<0.05);各试验组ToTLR-1、ToTLR-2、ToTNF-αToIL-1β基因表达量显著降低 (P<0.05);T1、T2和T4组的ToNFkB P65基因表达量显著低于对照组 (P<0.05);T3和T4组ToIL-10基因表达量显著高于对照组 (P<0.05)。该试验结果显示,外源牛磺酸对卵形鲳鲹肠道微生物菌群和免疫功能具有显著影响。
  • 图  1  卵形鲳鲹肠道微生物稀释曲线 (a) 和Rank abundance曲线 (b) (n=9)

    Figure  1.  Rarefaction (a) and Rank abundance (b) curve of intestinal microbial diversity of T. ovatus

    图  2  牛磺酸对卵形鲳鲹肠道微生物Alpha多样性分析 (n=9)

    *. 差异显著 (P<0.05);**. 差异极显著 (P<0.01)

    Figure  2.  Alpha diversity analysis of intestinal microbial diversity of T. ovatus

    *. Significant difference (P<0.05); **. Very significant difference (P<0.01)

    图  3  卵形鲳鲹肠道微生物Beta多样性分析分析 (n=9)

    Figure  3.  Beta diversity analysis of intestinal microbiota of T. ovatus

    图  4  基于门水平上的卵形鲳鲹肠道菌群物种相对丰度柱形图 (n=9)

    Figure  4.  Bacterial relative abundance in intestine of T. ovatus at phylum level

    图  5  基于属水平上的卵形鲳鲹肠道菌群物种相对丰度柱形图 (n=9)

    Figure  5.  Bacterial relative abundance in intestine of T. ovatus at genus level

    图  6  免疫相关基因在卵形鲳鲹肠道中mRNA相对表达量

    Figure  6.  Relative expression levels of immune-related genes in intestine of T. ovatus

    表  1  细菌16S rDNA V3—V4可变区引物

    Table  1.   Bacterial 16S rDNA V3–V4 corresponding primers

    引物名称
    Primer name
    引物序列 (5'−3')
    Primer sequence
    341F CCTAYGGGRBGCASCAG
    806R GGACTACNNGGGTATCTAAT
    下载: 导出CSV

    表  2  卵形鲳鲹肠道免疫基因引物序列

    Table  2.   Real-time quantitative PCR primers for immune related genes in T.ovatus

    引物名称
    Primer name
    引物序列 (5'−3')
    Primer sequence
    登录号
    Accession No.
    用途
    Amplification target
    ToTLR-1 F: GAACCTTCTGATGCTGAATCTG
    R: TGCTCCAAGTGCTAATCTCT
    MG762971.1 qRT-PCR
    ToTLR-2 F: CTCCACCTTGCGATACCT
    R: TCCAACACCTCCAGAGATG
    MG762972.1 qRT-PCR
    ToNFkB P65 F: GTTCGCATCTCGCTCGTAA
    R: TGGCCTCATTCACATCCTT
    MN233364.1 qRT-PCR
    ToIL-1β F: GGAGACTGTGGAGGACAAGAGC
    R: GCGGGCAGACATGAAGGTG
    MK224504.1 qRT-PCR
    ToIL-10 F: AGTCAGTCTCCACCCCCATCTT
    R: GCCCACTGGAGTTCAGATGCT
    KY231908 qRT-PCR
    ToTNF-α F: GGCGTCGTTCAGAGTCTCCT
    R: TCCTCCTGGGCAGTGGTTT
    [24] qRT-PCR
    EF-1α F: CCCCTTGGTCGTTTTGCC
    R: GCCTTGGTTGTCTTTCCGCTA
    [22] 内参
    下载: 导出CSV

    表  3  牛磺酸对卵形鲳鲹血清免疫指标的影响

    Table  3.   Effect of taurine on serum immune parameters of T. ovatus

    指标
    Index
    组别 Group
    T0T1T2T3T4
    补体C3 C3/(g·L−1) 0.13±0.45 0.14±0.02 0.15±0.21 0.16±0.48 0.16±0.13
    补体C4 C4/(g·L−1) 0.17±0.25b 0.23±0.14ab 0.24±0.81ab 0.26±0.60a 0.24±0.64ab
    溶菌酶 LZM/(U·mL−1) 175.91±21.08c 194.30±17.42bc 204.30±15.72b 253.18±18.79a 211.29±12.96b
    免疫球蛋白A IgA/(g·L−1) 0.39±0.74b 0.51±0.16ab 0.46±0.16ab 0.64±0.21a 0.48±0.18ab
    免疫球蛋白G IgG/(g·L−1) 1.16±0.19b 1.27±0.46ab 1.44±0.68ab 2.01±0.78a 1.59±0.71ab
    免疫球蛋白M IgM/(g·L−1) 1.94±0.63 2.51±0.66 2.61±0.68 3.07±1.21 2.78±0.91
    注:同行数据不同小写字母表示差异显著 (P<0.05) Note: Values in the same column with different letters are significantly different (P<0.05)
    下载: 导出CSV
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出版历程
  • 收稿日期:  2020-09-17
  • 修回日期:  2020-12-15
  • 网络出版日期:  2021-01-20
  • 刊出日期:  2021-04-05

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