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

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

马启伟^{1, 2,},
郭梁^{2, 3},
刘波^{2, 3},
刘宝锁^{2, 3},
朱克诚^{2, 3},
郭华阳^{2, 3},
张楠^{2, 3},
杨静文^{2, 3},
张殿昌^{2, 3, ,}

1.
上海海洋大学水产与生命学院，上海 201306
2.
中国水产科学研究院南海水产研究所/农业农村部南海渔业资源开发利用重点实验室，广东广州 510300
3.
广东省海洋生物种业工程技术研究中心，广东广州 510300

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
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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

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

Qiwei MA^{1, 2
,},
Liang GUO^{2, 3},
Bo LIU^{2, 3},
Baosuo LIU^{2, 3},
Kecheng ZHU^{2, 3},
Huayang GUO^{2, 3},
Nan ZHANG^{2, 3},
Jingwen YANG^{2, 3},
Dianchang ZHANG^{2, 3
, ,}

1.
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
2.
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences/Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, Guangzhou 510300, China
3.
Guangdong Provincial Engineer Technology Research Center of Marine Biological Seed Industry, Guangzhou 510300, China

摘要

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-α和ToIL-1β基因表达显著降低 (P<0.05)；T1、T2和T4组的ToNFkB P65基因表达显著低于对照组 (P<0.05)；T3和T4组ToIL-10基因表达显著高于对照组 (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。
- 卵形鲳鲹 /
- 牛磺酸 /
- 肠道微生物 /
- 免疫功能
Abstract: In order to investigate the effects of taurine on the intestinal microflora structure and immune function of Trachinotus ovatus, we used fish meal, fermented soybean meal and corn gluten meal as the basic protein sources to prepare the nitrogen and fat feed with taurine contents of 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), respectively. Seven hundred and fifty T. olvatus with an average body mass of (80.0±0.5) g were randomly devided into five groups with three replicates of fifty fish, and the experiment lasted 56 d. Sequencing results show that a total of 87 707 sequences had been obtained in the intestine of T. ovatus. The number of OTUs that can be annotated to the database was 5 130 (95.32%). At the phylum level, Proteobacteria, Tenericutes and Spirochaetes were the predominant phyla. Alpha diversity and beta diversity analyses show that the richness and diversity of intestinal microbial in T2 group were the lowest and the species composition was significantly different between T1 and T4 groups (P<0.05). The serum lysozyme activity, C4, and Ig increased significantly with the addition of exogenous taurine (P<0.05). The expression of ToTLR-1, ToTLR-2, ToTNF-α and ToIL-1β in each group decreased significantly (P<0.05). The expression of ToNFkB P65 of T1, T2 and T4 groups was significantly lower than that of the control group (P<0.05), while the expression of ToIL-10 of T3 and T4 groups was significantly higher than that of control group (P<0.05). The results indicate that exogenous taurine has a significant impact on the intestinal microflora and immune function.
- Trachinotus ovatus /
- Taurine /
- Intestinal microbes /
- Immune function

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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

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图 2 牛磺酸对卵形鲳鲹肠道微生物Alpha多样性分析 (n=9)

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

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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

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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

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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

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图 6 免疫相关基因在卵形鲳鲹肠道中mRNA表达量

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

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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]	reference genes

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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
指标 Index	T0	T1	T2	T3	T4
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±0.25^b	0.23±0.14^ab	0.24±0.81^ab	0.26±0.60^a	0.24±0.64^ab
溶菌酶 LZM/(U·ml⁻¹)	175.91±21.08^c	194.30±17.42^bc	204.30±15.72^b	253.18±18.79^a	211.29±12.96^b
免疫球蛋白A IgA/(g·L⁻¹)	0.39±0.74^b	0.51±0.16^ab	0.46±0.16^ab	0.64±0.21^a	0.48±0.18^ab
免疫球蛋白G IgG/(g·L⁻¹)	1.16±0.19^b	1.27±0.46^ab	1.44±0.68^ab	2.01±0.78^a	1.59±0.71^ab
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)

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