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地衣芽孢杆菌 T-1 作为群体感应淬灭细菌保护斑马鱼抵抗嗜水气单胞菌感染。

The Quorum Quenching Bacterium Bacillus licheniformis T-1 Protects Zebrafish against Aeromonas hydrophila Infection.

机构信息

National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, 201306, Shanghai, People's Republic of China.

National Pathogen Collection for Aquatic Animals, Shanghai Ocean University, Shanghai, 201306, China.

出版信息

Probiotics Antimicrob Proteins. 2020 Mar;12(1):160-171. doi: 10.1007/s12602-018-9495-7.

Abstract

Quorum sensing, bacterial cell-to-cell communication via small signaling molecules regulates virulence in many bacterial pathogens, and is a promising target for antivirulence therapy, which may inhibit virulence rather than cell growth and division. Herein, Bacillus strains capable of degrading QS molecules from freshwater environments were screened as potential aquaculture probiotics. A total of 34 Bacillus strains were isolated. Strain T-1 was selected with "H" streaking and double layer agar plate methods using Chromabacterium violaceum ATCC12472 as reporter, and eventually identified as Bacillus licheniformis based on biochemical and molecular identification. Quorum quenching by T-1 was confirmed using C. violaceum CV026. T-1 was non-hemolytic in vitro. In biocontrol experiments, T-1 reduced the pathogenicity of Aeromonas hydrophila cb15 in zebrafish co-injected intraperitoneally with both strains, achieving a relative percentage survival of 70%. Determination and analysis of the T-1 draft genome using the Illumina Hiseq 2500 platform identified the quorum quenching gene ytnP, encoding an acyl-homoserine lactone metallo-β-lactamase, as a potential QS quencher in T-1. In conclusion, B. licheniformis T-1 could be a safe and effective quorum quenching bacterium for protecting hosts against pathogenic bacterial infections in aquaculture.

摘要

群体感应,通过小分子信号分子进行的细菌细胞间通讯调节许多细菌病原体的毒力,是抗毒力治疗的有前途的靶点,它可能抑制毒力而不是细胞生长和分裂。在此,筛选了能够从淡水环境中降解 QS 分子的芽孢杆菌菌株作为潜在的水产养殖益生菌。共分离出 34 株芽孢杆菌。使用 Chromabacterium violaceum ATCC12472 作为报告菌,采用划线法和双层琼脂平板法,通过“H”划线和双层琼脂平板法,选择菌株 T-1,并最终根据生化和分子鉴定鉴定为地衣芽孢杆菌。使用 C. violaceum CV026 证实了 T-1 的群体感应猝灭作用。T-1 在体外无溶血作用。在生物防治实验中,T-1 降低了两种菌株同时腹腔注射斑马鱼时嗜水气单胞菌 cb15 的致病性,相对存活率达到 70%。使用 Illumina Hiseq 2500 平台对 T-1 的草图基因组进行测定和分析,鉴定出群体感应淬灭基因 ytnP,编码酰基高丝氨酸内酯金属β-内酰胺酶,是 T-1 中潜在的 QS 淬灭剂。总之,地衣芽孢杆菌 T-1 可以作为一种安全有效的群体感应猝灭细菌,用于保护水产养殖宿主免受致病性细菌感染。

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