仿刺参夏眠期间肠道菌群结构特征研究

THE INTESTINAL MICROBIOME OF SEA CUCUMBER APOSTICHOPUS JAPONICUS DURING THE AESTIVATION PERIOD

  • 摘要: 为了解析仿刺参(Apostichopus japonicus)夏眠期间肠道菌群结构特征, 以仿刺参为研究对象, 设置正常生长组(对照组, 15℃)和夏眠组(26℃), 经过近3个月的养殖实验和先后7次采样, 通过16S rRNA基因测序技术分析了夏眠期间仿刺参肠道菌群多样性和结构动态变化。研究结果表明, 夏眠组和正常组仿刺参肠道菌群丰度和多样性随采样时间均发生动态变化; 仿刺参夏眠初期肠道菌群丰度和多样性较夏眠之前显著下降(P<0.05)。NMDS和Anosim分析表明正常组和夏眠组刺参肠道菌群结构存在显著性差异(P<0.05)。变形菌门(Proteobacteria)、拟杆菌门(Bacteroidota)、厚壁菌门(Firmicutes)、疣微菌门(Verrucomicrobiota)、放线菌门(Actinobacteriota)和弯曲杆菌门(Campylobacterota)为主要优势菌门, 相对丰度为90.01%—99.80%。在属水平上, 夏眠组和正常组的仿刺参肠道菌群结构组成存在明显差异, 且优势菌属随采样时间动态变化。LEfSe分析共发现9个Biomarkers, 其中正常组和夏眠组分别有7个和2个。共发生网络表明正常组仿刺参肠道菌群相互作用比夏眠组更加紧密和复杂。中性群落模型(NCM)分析表明仿刺参微生物群落的构建受到随机过程的影响较小, 受确定性过程的影响较大。研究为阐明仿刺参夏眠提供了肠道菌群层面的解释, 为仿刺参夏季健康养殖提供了理论依据。

     

    Abstract: Sea cucumber Apostichopus japonicus is an economically important marine echinoderm in China. The optimal temperature for feeding and growth of A. japonicus is around 14—16℃, while it enters an aestivation state when the temperature exceeds 25℃. Among marine invertebrates, A. japonicus has emerged as a suitable model organism for investigating aestivation induced by environmental factors, primarily due to increased temperature. The studies on elucidating the aestivation mechanism A. japonicus is increasing in recent years, which focused on physiology, metabolism, gene expression, and epigenetic regulation. The intestinal microbiota plays an essential role in the growth, development, and immune regulation of the host, while the knowledge regarding the succession of intestinal microbial community in A. japonicus during the aestivation period is still unclear. In this study, sea cucumbers with an average weight of (54.39±2.94) g were collected and acclimated for two weeks in a recirculating aquaculture system at 15℃. After acclimation, a total of 300sea cucumbers were randomly allocated into 10 tanks with five replicates of non-aestivation group (NAT) and aestivation group (AT). In the NAT group, the temperature was maintained at 15℃, while in the AT group, the water temperature increased from 15℃ to 26℃. The aestivation period lasted for one month, then the temperature decreased from 26℃ to 16—17℃ until the AT group recovered and began feeding activities. Seven times were collected according the temperature in the AT group, including after acclimation (1 time, labeled NAT0 and AT0), during aestivation (five times, NAT1-5 and AT1-5), and arousal from aestivation (1 time, labeled NAT6 and AT6). The intestinal microbial communities were investigated using high-throughput sequencing. The results showed that a total of 11552 ASVs, with only 14.57% shared between the NAT and AT groups. The intestinal and richness of microbial diversity were assessed using the Shannon and Chao 1 indices, showing distinct temporal dynamics both in the NAT and AT groups. The richness and diversity in AT1 and AT2samples were significantly lower than that in AT0 (P<0.05). The NMDS analysis based on Bray-Curtis distances showed that samples in the NAT and AT groups clustered separately, with significant differences confirmed by Anosim analysis. Proteobacteria, Bacteroidota, Firmicutes, Verrucomicrobiota, Actinobacteriota and Campylobacterota were the dominant phyla, with relative abundances ranging from 90.01% to 99.80%. The composition of the dominant genera differed significantly during the aestivation period. LEfSe analysis identified 9 biomarkers, including 7 of NAT and 2 of AT. Co-occurrence network analysis revealed that the NAT group had a more complex network structure than that of AT. The neutral model analysis (NCM) revealed that both NAT and AT microbial communities were predominantly influenced by deterministic processes. These findings provide valuable insights into the mechanisms of sea cucumber aestivation and offer a theoretical basis for the healthy breeding of A. japonicus in summer.

     

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