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| 福建吊笼养殖刺参肠炎病的发病特征及其菌群结构分析 |
| Analysis on the pathogenic characteristics and bacterial community structure of enteritis disease of Apostichopus japonicus in cage culture areas in the Fujian Dongwuyang Bay |
| 投稿时间：2024-04-14 修订日期：2024-05-06 |
| DOI： |
| 中文关键词: 刺参，肠炎病，混合饲料，菌群结构，高通量测序 |
| 英文关键词: Apostichopus japonicus, enteritis disease, mixed feed, flora structure, high-throughput sequencing |
| 基金项目:农业农村部东海海水健康养殖重点实验室开放课题（2022ESHML03）、青岛市重点研发计划课题（22-3-3-hygg-1-hy）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(2020TD40)和霞浦县政府采购项目（350921MJHDY2021002）共同资助 |
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| 中文摘要: |
| 本研究以福建东吾洋海域吊笼养殖区暴发的刺参肠炎病为研究对象，对其发病特征、病原病理以及刺参肠道菌群结构进行解析，以期掌握该养殖模式下相关疾病的发病原因，为该区域刺参病害防控提供科学依据。流行病学调查发现，患病刺参肠道变细且肠道内含有大量积水，组织病理学结果显示，病变肠道结缔组织细胞肿大且部分崩解，排布较为松散，结缔组织与肌肉层出现分离现象。采用涂布培养法对刺参肠道及环境样品进行可培养细菌培养和占比分析，结果显示患病刺参肠道总菌数和弧菌数均显著高于健康刺参。通过对16S rDNA可变区V3~V4区域进行高通量测序，菌群结构分析结果表明检测到的细菌归属于25门，311科，健康组肠道排名前三的为肠杆菌科（Enterobacteriaceae）、杆菌科（Bacillaceae）和毛螺菌科（Lachnospiraceae），相对丰度分别为8.69%、5.53%和5.12%，而患病组肠道排名前三的优势菌科依次为立克次体科（Rickettsiaceae）、弧菌科（Vibrionaceae）和肠杆菌科，相对丰度分别为 12.66%、8.22% 和 5.71%，患病组的弧菌科丰度显著高于健康组，环境样品中混合饲料的弧菌科相对丰度最高，达到了34.99%。PCoA分析及UPGMA聚类分析显示，各组样品聚类良好，且混合饲料菌群类别与肠道菌群较为接近。在差异菌群的高丰度OTU定位分析中,相对丰度高于0.5%的OTU主要定位到了泥单胞菌属（Lutimonas）、另类弧菌属（Aliivibrio）和华美菌属（Formosa），且弧菌属多为致病菌，华美菌属多为益生菌，高丰度OTU在健康组和患病组的肠道中差异显著。综合上述结果表明，患病刺参肠道组织结构发生改变，且肠道中的大量弧菌主要来源于混合饲料，刺参饵料中的大量菌群对其健康具有一定的威胁性，益生菌在健康刺参肠道中的含量高于患病刺参，致病菌在患病刺参中的含量高于健康刺参，相关研究结果将有助于为潜在益生菌的开发利用和潜在病原菌的筛查提供科学依据，并进一步推动刺参饵料的优化，为科学养殖及病害防控提供理论依据。 |
| 英文摘要: |
| In recent years, the sea cucumber industry has developed rapidly, and the breeding area has gradually expanded to the southern region. By utilizing the warmer water found in the southern sea area during the winter, Fujian Province set up cage culture. With less than 1% of the nation's breeding area, the Fujian Province produced 18% of the nation's sea cucumber output, establishing a new paradigm known as "north sea cucumber and south cultivation," which has become a major force in the growth of the sea cucumber industry. However, with the rapid development of the sea cucumber breeding industry in this area, due to the short development time of sea cucumber breeding industry and the extensive use of breeding technology in Fujian Province, breeding diseases have become more common in recent years, significantly affecting both the survival and weight gain rates of breeding. In order to understand the causes of related diseases under this culture mode and provide a scientific basis for the prevention and control of sea cucumber disease in this area, we investigated an outbreak of sea cucumber enteritis in the cage culture area of the Dongwuyang sea area of Fujian Province. We analyzed the epidemiological characteristics, pathogenic pathology, environmental microbial groups, and the structure of the intestinal flora of sea cucumber. The epidemiological investigation found that the intestinal tract of the diseased sea cucumber became smaller and contained a large amount of water. The histopathological results show that the connective tissue and muscle layer were separated, the connective tissue cells in the sick intestinal tract were swollen and partially destroyed, and the arrangement was generally loose in the diseased intestinal tract. Utilizing microbial culture data, analysis of cultivable bacterial culture and fraction of intestinal and ambient samples from sea cucumbers. The total quantity and Vibrio quantity of bacteria in the intestinal samples of the diseased sea cucumber were found to be significantly greater than those in the intestinal samples of the healthy sea cucumber. Based on high-throughput sequencing of the 16S rDNA variable region V3-V4, 25 phylums and 311 families were identified. Enterobacteriaceae, Bacillaceae, and Lachnospiraceae were the top three bacteria families in the healthy group, with relative abundances of 8.69%, 5.53%, and 5.12%, respectively, while Rickettsiaceae, Vibrionaceae, and Enterobacteriaceae were the top three prominent bacterial families in the diseased group, with relative abundances of 12.66%, 8.22%, and 5.71%, respectively. Vibriaceae were more prevalent in the diseased group than in the healthy group. With the relative abundance of Vibriococcaceae was 34.99%, the mixed diet had the highest abundance in the environmental samples. According to PCoA and UPGMA cluster analyses, the samples of each group were well grouped, and the mixed diet's flora types were similar to the intestinal flora. In conclusion, the intestinal tissue structure of the diseased sea cucumber was changed, and a significant amount of Vibrio bacteria in the intestinal tract primarily derived from the mixed diet, and a significant amount of bacteria in the diet were posed a threat to the health of the sea cucumber. In the analysis of high abundance OTUs with differential microbial communities, OTUs with relative abundance of 0.5% were mainly located in the genera Lutimonas, Aliivibrio, and Formosa. Vibrio was mostly pathogenic, while Formosa was mostly probiotic. There is a significant difference in gut microbiota between the healthy and diseased groups in terms of high abundance OTU. The above results indicate that the intestinal tissue structure of diseased sea cucumber has changed, and the large amount of Vibrio in the intestine mainly comes from mixed feed. The large microbial community in sea cucumber feed poses a certain threat to its health. The content of probiotics in the intestinal tract of healthy sea cucumber is higher than that of diseased sea cucumber, and the content of pathogenic bacteria in diseased sea cucumber is higher than that of healthy sea cucumber. The relevant research results will help provide scientific basis for the development and utilization of potential probiotics and the screening of potential pathogenic bacteria. The results will support the development of sea cucumber bait optimization and offer a theoretical foundation for both scientific breeding and disease management. |
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