患“红斑综合征”的海胆体腔液中菌群结构特征

Structural Characteristics of Bacterial Flora in Coelomatic Fluid of Sea Urchin With"Red Spot Syndrome Disease"

为探究患“红斑综合征”海胆的细菌群落结构、功能,以及该病在海胆种间的传播性,采用16S rDNA高通量测序技术人工回归感染,从患病马粪海胆(Hemicentrotus pulcherrimus)体腔内分离、鉴定出一株灿烂弧菌(Vibrio splendidus),之后配置其菌悬液注射到光棘球海胆(Strongylocentrotus nudus)、中间球海胆(Strongylocentrotus intermedius)和海刺猬(Glyptocidaris crenularis)体腔内,构建海胆感染“红斑综合征”后体腔液菌群16S rRNA基因测序文库,分析不同种类海胆感染“红斑综合征”后体腔内菌群结构。结果显示:光棘球海胆、中间球海胆、海刺猬均可被灿烂弧菌感染,出现“红斑”病征,在10 d内均死亡,其中中间球海胆表现出围口膜周围变黑,体壁出现红色斑块;光棘球海胆体腔液变为紫黑色等。3种海胆注射感染后体腔液内优势菌门均为变形菌门(Proteobacteria),所占比例分别为99.94%、60.18%和92.38%;弧菌属(Vibrio)为优势菌属,所占比例分别上升到99.90%、48.96%和97.47%。COG及KEGG数据库分析表明,体腔内菌群共注释到24组COG功能分类,主要集中在氨基酸转运与代谢、转录、信号转导机制;41条KEGG信号通路,集中分布在膜运输(Membrane Transport)、氨基酸的运输和代谢(Amino acid transport and metabolism)、转录(Transcription)、信号传导机制(Signal transduction mechanisms)。本研究结果可为海胆“红斑综合征”的预防治疗提供理论基础。

In order to explore the causes of sea urchin suffering from"red spot disease"and whether"red spot disease"is transmissible among sea urchin species,16S rDNA high-throughput sequencing technology and artificial regression infection were used to isolate and identify Vibrio splendidus from the coelomic cavity of sick sea urchin Hemicentrotus pulcherrimus,and the bacterial suspension was injected into the coelomic cavity of sea urchin Strongylocentrotus nudus,S.intermedius and Glyptocidaris crenularis.The 16S rRNA Gene Sequencing Library of the coelomic fluid flora was constructed in sea urchins challenged with erythema,and the structure of the coelomic flora of different sea urchins infected with"red spot disease"was analyzed.The results showed that Strongylocentrotus nudus,S.intermedius and Glyptocidaris crenularis were infected by Vibrio splendidus,all sea urchins died or showed the symptoms of"red spotting"within 10 days.The S.intermedius infected by Vibrio splendidus showed the blackening around the perioral membrane,the color of coelomic fluid changed to purple black,with red patches on the body wall of sea urchin.The dominant bacteria in the coelomic fluid of the three species of sea urchin challenged were Bacteroidetes,accounting for 99.94%,60.18%and 92.38%respectively.The proportion of Vibrio as the dominant genus increased to 99.90%,48.96%and 97.47%respectively.The coelomic flora was analyzed by COG and KEGG databases,and 24 groups of COG functional classifications were annotated,mainly focusing on the mechanisms of amino acid transport and metabolism,transcription and signal transduction;41 KEGG signal pathways are concentrated in membrane transport,amino acid transport and metabolism,transcription and signal transmission mechanisms.The findings provide a theoretical basis for the prevention and treatment of"red spot disease"of sea urchin,and provide a reference basis for the construction of a healthy breeding model of sea urchin.