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草鱼感染维氏气单胞菌后肠道微生物和免疫因子的变化及拮抗菌株(黄棕色链霉菌)的筛选。

Alteration of the gut microbiome and immune factors of grass carp infected with Aeromonas veronii and screening of an antagonistic bacterial strain (Streptomyces flavotricini).

机构信息

State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Provincial Key Laboratory of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha, 410081, China.

出版信息

Microb Pathog. 2020 Jun;143:104092. doi: 10.1016/j.micpath.2020.104092. Epub 2020 Mar 4.

Abstract

Aeromonas veronii is a widely distributed novel pathogen that can affect humans and animals, it can cause sepsis in fish with high mortality and serious economic losses to aquaculture. In the study, the gut microbiome of the infected and uninfected grass carp with Aeromonas veronii were analyzed probiotics and pathogenic bacteria by the Miseq high-throughput sequencing, the results showed that the infected fish were significantly higher in Proteobacteria, Firmicutes, Fusobacteria, and the immune factors in liver and kidney were up-regulated by qRT-PCR. In order to effectively inhibit the pathogen, we screened an actinomycete strain and had good antibacterial effect on Aeromonas veronii. The new antagonistic bacteria was named as Streptomyces flavotricini X101, the whole genome sequencing revealed that the metabolic process was most active. After grass carp was inoculated with the minimum inhibitory concentration of 900 μg/mL of the strain's fermentation supernatant, then Aeromonas veronii was injected, we found that the pathological symptoms such as body surface, anus and abdominal congestion were alleviated by H&E staining. Cellular experiments showed that it wasn't toxic to liver cells of grass carp. Overall, this is the first study of changes in intestinal flora, phenotype, and immune factors in grass crap infected with Aeromonas veronii, it had important theoretical significance and application value for immunization and prevention.

摘要

维氏气单胞菌是一种广泛分布的新型病原体,可感染人类和动物,可引起鱼类败血症,死亡率高,给水产养殖业造成严重经济损失。本研究采用 Miseq 高通量测序技术分析了感染和未感染维氏气单胞菌的草鱼的肠道微生物群,结果表明,感染组鱼类的变形菌门、厚壁菌门、梭杆菌门显著升高,qRT-PCR 检测到肝脏和肾脏的免疫因子上调。为了有效抑制病原菌,我们筛选了一株放线菌,对维氏气单胞菌具有良好的抑菌作用。新的拮抗细菌被命名为链霉菌 X101,全基因组测序显示其代谢过程最为活跃。将最小抑菌浓度为 900μg/mL 的菌株发酵上清液接种草鱼后,再注射维氏气单胞菌,通过 H&E 染色发现草鱼体表、肛门和腹部充血等病理症状得到缓解。细胞实验表明,该菌株对草鱼肝细胞没有毒性。总之,本研究首次研究了维氏气单胞菌感染草鱼后肠道菌群、表型和免疫因子的变化,对免疫和预防具有重要的理论意义和应用价值。

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