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不同生理状态中华鲟体表黏液与生境微生物组成差异分析
郑跃平,刘杰豪,金审,孙若凌,徐嘉楠,范厚勇,王有基,胡梦红
1.长江口水生生物监测与保护联合实验室 上海 200092;2.上海市水生野生动植物保护研究中心 上海 200092;3.上海海洋大学 水产种质资源发掘与利用教育部重点实验室 上海 201306;4.上海海洋大学 上海水产养殖工程技术研究中心 上海 201306;5.上海市水生野生动植物保护研究中心 上海 200093
摘要:
为揭示不同生理状态下中华鲟(Acipenser sinensis)体表黏液与生境微生物组成差异,本研究比较了健康和亚健康中华鲟体表黏液菌群的群落结构差异,探讨细菌群落结构与中华鲟生理状态的相关性。本研究采集健康与亚健康中华鲟的体表黏液和养殖水体,利用高通量测序技术分析其菌群结构。结果表明,亚健康组微生物多样性与丰富度显著高于健康组(P<0.05)。序列的统计分析表明,相较于水体,黏液样本存在大量的独有操作分类单元(operational taxonomic unit, OTU),且与水体的共有OTU数目因中华鲟生理状况而发生变化。根据组间的相似性与差异分析,不同组之间的主成分与主坐标分析显示出较强的分离趋势,且组内样本相对距离较小。在门和属水平下,中华鲟体表黏液和水体群落组成和优势种有显著差异。水体中优势菌门为变形菌门(Proteobacteria) (31.84%)和厚壁菌门(Firmicutes) (24.37%);健康组优势菌门为变形菌门(55.23%)和拟杆菌门(Bacteroidetes) (21.00%);亚健康组的变形菌门(40.23%)占比相较于健康组有所下降,但酸杆菌门(Acidobacteria) (18.29%)、芽单胞菌门(Gemmatimonadetes) (10.08%)、放线菌门(Actinobacteria)、疣微菌门(Verrucomicrobia)的比例均有不同程度的增加。水体中优势菌群包括球形发丝菌属(Sphaerochaeta) (10.56%)和下水道菌属(Cloacibacillus) (7.95%)。健康组中的优势菌群包括罗尔斯通氏菌属(Ralstonia) (19.83%)和黏液杆菌属(Mucilaginibacter) (8.11%)。亚健康组的优势菌群则相较于健康组发生了变化,主要由鞘氨醇单胞菌属(Sphingomonas) (16.71%)和WPS-1_genera_incertae_sedis (5.05%)组成。功能预测发现,亚健康组的菌群功能丰度相较于健康组显示出下降的趋势,包括环境适应、生物代谢、信号转导等。通过对不同生理状态中华鲟黏液微生物组与水体的相似性及差异分析发现,随着中华鲟的生理状态的改变,体表黏液核心菌群产生差异,亚健康组中鞘氨醇单胞菌属是黏液菌群的优势属,该属可作为健康检测的标志物。通过对健康和亚健康中华鲟体表黏液微生物群落的比较,发现不同健康状态存在不同的微生物群落结构,为中华鲟的健康养殖与监测提供了新的视角。
关键词:  中华鲟  生理状态  黏液  微生物组  养殖水体
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Skin mucus flora and habitat microbial composition of Chinese sturgeon (Acipenser sinensis) under different physiological states
ZHENG Yueping1,2, LIU Jiehao3,4, JIN Shen3,5, SUN Ruoling3,6, XU Jianan1,2, FAN Houyong1,2, WANG Youji3,4, HU Menghong3,5
1.Joint Laboratory for Monitoring and Conservation of Aquatic Living Resources in the Yangtze Estuary, Shanghai 200092, China;2.Shanghai Aquatic Wildlife Conservation Research Center, Shanghai 200092, China;3.Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University, Shanghai 201306, China;4.Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201306, China;5.Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201307, China;6.Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai 201308, China
Abstract:
To reveal the differences in the microbial composition of skin mucus and the habitat of Chinese sturgeon (Acipenser sinensis) in different physiological states, we compared the community structure of the skin mucus flora of healthy and sub-healthy Chinese sturgeons and explored the correlation between the structure of the bacterial community and physiological state of Chinese sturgeons. Skin mucus and aquaculture water of healthy and sub-healthy Chinese sturgeons were collected, and the bacterial flora composition was analyzed using high-throughput sequencing technology. The results showed that microbial diversity and richness were significantly higher in the sub-healthy group than in the healthy group (P<0.05). Statistical analysis of the sequences showed that many unique operational taxonomic units (OTUs) were present in the mucus samples compared with the aquaculture water, and the number of shared OTUs with the aquaculture water changed according to the health status. The principal component and principal coordinate analyses between different groups showed a strong separation tendency, and the relative distance between samples within groups was small. There were significant differences in the composition and dominant species of skin mucus and aquaculture water communities of Chinese sturgeons at the phylum and genus levels. The dominant phyla in the water were Proteobacteria (31.84%) and Firmicutes (24.37%); the dominant phyla in the healthy group were Proteobacteria (55.23%) and Bacteroidetes (21.00%); and the percentage of Proteobacteria (40.23%) in the sub-healthy group, but the proportion of Acidobacteria (18.29%), Gemmatimonadetes (10.08%), Actinobacteria, and Verrucomicrobia increased. The dominant flora in water samples were Sphaerochaeta (10.56%) and Cloacibacillus (7.95%). The dominant flora in the healthy group were Ralstonia (19.83%) and Mucilaginibacter (8.11%). The dominant flora in the sub-healthy group, compared to the healthy group, consisted mainly of Sphingomonas (16.71%) and WPS-1_genera_incertae_sedis (5.05%). Functional prediction revealed that the functional abundance of flora in the sub-healthy group showed a decreasing trend compared with that in the healthy group, including environmental adaptation, biological metabolism, and signal transduction. Similarity and difference analysis of the mucus microbiome of Chinese sturgeons in different physiological states and aquaculture water showed that it changed with the physiological state, and the genus Sphingomonas was the dominant genus of mucus flora in the sub-healthy group, which can be used as a health marker. By comparing the microbial communities in the skin mucus of healthy and sub-healthy Chinese sturgeons, different microbial community structures were observed in different health states, which provides a new perspective for their healthy culture and monitoring.
Key words:  Chinese sturgeon  Physiological states  Mucus  Microbiome  Aquacultural water