草鱼肠道微生物谱及其发育性变化

doi:10.3969/j.issn.1004-1524.20230519

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (4): 780-789.DOI: 10.3969/j.issn.1004-1524.20230519

草鱼肠道微生物谱及其发育性变化

张红芳¹^,²(), 钱涛³, 金婷³, 谢小玲¹, 吴酬飞¹, 肖英平², 马灵燕²^,^*()

1.湖州师范学院生命科学学院,浙江湖州 313000
2.浙江省农业科学院农产品质量安全与营养研究所,省部共建农产品质量安全危害因子与风险防控国家重点实验室, 浙江杭州 310021
3.开化县农业农村局,浙江开化 324300

收稿日期:2023-04-21 出版日期:2024-04-25 发布日期:2024-04-29
作者简介:张红芳(1996—),女,甘肃甘谷人,硕士研究生,研究方向为鱼肠道微生物与健康养殖。E-mail:hfzhangcth@126.com
通讯作者: *马灵燕,E-mail:maly1124@163.com
基金资助:
农产品标准化生产示范县建设(“一县一品一策”)重大专项(ZJNY2022001-007)

Gut microbial profiles and its developmental changes of grass carp (Ctenopharyngodon idella)

ZHANG Hongfang¹^,²(), QIAN Tao³, JIN Ting³, XIE Xiaoling¹, WU Choufei¹, XIAO Yingping², MA Lingyan²^,^*()

1. College of Life Sciences, Huzhou University, Huzhou 313000, Zhejiang, China
2. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Agro-Product Safety and Nutrition, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. Agriculture and Rural Affairs Bureau of Kaihua County, Kaihua 324300, Zhejiang, China

Received:2023-04-21 Online:2024-04-25 Published:2024-04-29
Contact: MA Lingyan

摘要/Abstract

摘要：

为了研究草鱼不同肠段微生物结构以及草鱼不同发育阶段肠道菌群的分布和变化。试验分2个部分进行, 第一部分:试验选取24月龄生理状况良好的草鱼8尾,用 MS-222对其深度麻醉,收集草鱼前肠、中肠、后肠段内容物进行DNA提取和16S rRNA基因测序;第二部分:从浙江省衢州市开化县一典型的开化清水鱼养殖基地购买不同月龄(1、7、15、24月龄)的健康草鱼各25尾,为了保证足够的样本量和最大限度降低个体差异,每5尾鱼的后肠段内容物混合为一个样本。用16S rRNA基因高通量测序方法,分析不同月龄草鱼肠道菌群的分布和变化。结果显示:1)Alpha多样性表明,草鱼的后肠段微生物群落多样性和丰富度显著高于前肠和中肠段(P<0.05);Beta 多样性表明,后肠段的微生物群落与前肠段和中肠段存在显著差异(P<0.05)。2)热图(Heatmap)显示,罗斯氏菌属(Ralstonia)是前肠段的主要优势菌属;砷氧化菌(Aliihoeflea)是中肠段的主要优势菌属;纤发鞘丝蓝细菌属(Leptolyngbya)和鲸杆菌属(Cetobacterium) 是后肠段的主要优势菌属。3)从 1到24月龄,草鱼后肠段微生物群落多样性和丰富度显著降低(P<0.05);Beta 多样性表明,不同月龄的草鱼后肠段微生物结构存在显著差异(P<0.05)。4)热图(Heatmap)显示,Rubellimicrobium和Synechococcus是1月龄后肠段的主要优势菌属;Aestuariimicrobium和Streptomyces是7月龄后肠段的主要优势菌属;Kineosporia和Prevotella是15月龄后肠段的主要优势菌属;Mesonia是24月龄后肠段的主要优势菌属。这表明不同肠段的草鱼微生物组成和结构存在差异,以及草鱼肠道微生物结构在生长的不同阶段呈发育性变化。

关键词: 草鱼, 肠道菌群, 发育性变化

Abstract:

The study was aim to expolar the diversity of the microbial structure in different intestinal segments during the growth and development processes of grass carp. The experiment was conducted in two parts, Part I: Eight grass carp at 24 months were anesthetized with MS-222 for DNA extraction and 16S rRNA sequencing analysis; Part Ⅱ: 25 grass carp with different ages, including 1 month, 7 months, 15 months and 24 months, were purchased from a typical Kaihua clear water fish breeding base in Kaihua County, Quzhou City, Zhejiang Province. To ensure sufficient sample size and minimize individual differences, the contents of the hindgut segments of each five fish were mixed into a single sample. The contents of grass carp at different months were extracted for DNA extraction and 16S rRNA sequencing analysis. The results indicated that: 1) Alpha diversity analysis showed that the diversity and richness of microbial community in hindgut were significantly higher than those in foregut and midgut (P<0.05); Beta diversity analysis showed that the microbial communities in foregut and midgut were similar, but significantly different from those in hindgut (P<0.05). 2) LEfSE analysis showed that Streptococcus and Ralstonia were dominant bacteria in the foregut. Escherichia-Shigella and Aliihoeflea were dominant bacteria in the midgut segment. Leptolyngbya and Cetobacterium are the main dominant genera in the hindgut. 3) The diversity and richness of microbial community in the hindgut of grass carp significantly decreased from 1 month to 24 months of age (P<0.05); Beta diversity analysis showed that there were significant differences in the microbial structure of the hindgut of spring water grass carp at different months (P<0.05). 4) Heatmap shows that Rubellimicrobium and Synechococcus are the main dominant bacteria in the hindgut at the age of 1 month. Aestuariimicrobium and Streptomyces were the dominant genera in the hindgut intestine of the 7-month-old. Kineosporia and Prevotella were dominant genera in the 15-month-old hindgut, while Vibrio and Mesonia were dominant genera in the 24-month-old hindgut. This indicates that there are differences in the composition and structure of microbia communities in different intestinal segments of grass carp, as well as developmental changes in the structure of gut microbiota during different stages of growth.

Key words: grass carp, microbial community structure, developmental change

中图分类号:

S917.1

张红芳, 钱涛, 金婷, 谢小玲, 吴酬飞, 肖英平, 马灵燕. 草鱼肠道微生物谱及其发育性变化[J]. 浙江农业学报, 2024, 36(4): 780-789.

ZHANG Hongfang, QIAN Tao, JIN Ting, XIE Xiaoling, WU Choufei, XIAO Yingping, MA Lingyan. Gut microbial profiles and its developmental changes of grass carp (Ctenopharyngodon idella)[J]. Acta Agriculturae Zhejiangensis, 2024, 36(4): 780-789.

图/表 10

表1 草鱼肠道微生物Alpha多样性分析(n=8)

Table 1 Alpha diversity analysis of the intestinal microflora in grass carp (n=8)

样本名称 Sample name	操作分类单元数 OTUs	香农指数 Shannon index	辛普森指数 Simpson index	丰度指数 Chao index
前肠Foregut	272± 27 c	3.23±0.26 b	0.17± 0.04	311.13± 41.68 c
中肠Midgut	524± 12 b	3.51±0.16 ab	0.11± 0.02	603.13± 20.90 b
后肠Hindgut	694± 23 a	3.93±0.21 a	0.09± 0.03	917.88± 29.44 a

图1 草鱼肠道微生物结构主坐标分析(n=8)

Fig.1 Principal comporent analysis of microbial community structure in grass carp (n=8)

图2 草鱼肠道微生物结构的分析

Fig.2 Analysis of the intestinal microflora structure in grass carp

图3 草鱼肠道微生物结构进化树

Fig.3 Phylogenetic tree of intestinal microflora structure in grass carp

图4 草鱼肠道微生物结构热图(前35种属)

Fig.4 Heatmap of microbial community structure in grass carp (Top 35 genus)

表2 草鱼后肠微生物Alpha多样性分析(n=5)

Table 2 Alpha diversity analysis of hindgut microbial of grass carp at different months (n=5)

样本名称 Sample name	操作分类单元数 OTUs	香农指数 Shannon index	辛普森指数 Simpson index	丰度指数 Chao index
1月龄1-month-old	562±106 a	3.74±0.61 a	0.11±0.09 ab	776.00±131.87 a
7月龄7-month-old	475±50 ab	3.60±0.17 a	0.11±0.02 ab	611.20±54.03 b
15月龄15-month-old	470±57 abc	3.54±0.29 a	0.12±0.04 ab	551.00±71.25 bc
24月龄24-month-old	270±85 c	2.30±0.16 b	0.27±0.04 a	292.60±43.90 cd

图5 不同月龄的草鱼后肠微生物结构主坐标分析

Fig.5 Principal component analysis of hindgut microbial community structure of grass carp at different months

图6 不同月龄的草鱼后肠微生物结构

Fig.6 Hindgut microbial community structure of grass carp at different months

图7 不同月龄的草鱼后肠微生物结构热图(前35种属)

Fig.7 Heatmap of hindgut microbial community structure of grass carp at different months (Top 35 genus)

图8 不同月龄水环境中微生物组成

Fig.8 Microbial composition in water environment at different months

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草鱼肠道微生物谱及其发育性变化

Gut microbial profiles and its developmental changes of grass carp (Ctenopharyngodon idella)

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