Study about the combination strategy of Bacillus subtilis wt55 with AiiO-AIO6 to improve the resistance of zebrafish to Aeromonas veronii infection

文献类型： 外文期刊

第一作者： Xia, Rui

作者： Xia, Rui;Hao, Qiang;Zhou, Zhi-Gang;Yao, Yuan-Yuan;Yang, Ya-Lin;Ran, Chao;Zhang, Zhen

作者机构：

关键词： B. subtilis; Quenching enzyme AiiO-AIO6; Intestinal microbiota; Immune regulation; Germ-free zebrafish

期刊名称：FISH & SHELLFISH IMMUNOLOGY （ 影响因子：4.622； 五年影响因子：4.799 ）

ISSN： 1050-4648

年卷期： 2022 年 128 卷

页码：

收录情况： SCI

摘要： Disease problems will seriously restrict the sustainable development of aquaculture, and the environmental-friendly prevention strategies are urgently needed. Probiotics and quorum-quenching enzyme are innovative strategies to control bacterial diseases. Firstly, the bacteriostatic activity of Bacillus subtilis wt55 strain and quenching enzyme AiiO-AIO6 on the growth of Aeromonas veronii were tested in vitro, and the results showed wt55 inhibit the growth of A. veronii, but AiiO-AIO6 did not. Then, the synergistic effects of simple combination of B. subtilis wt55 and AiiO-AIO6 were evaluated next. The results showed this combination could improve the survival rate and significantly reduce the number of invasive A. veronii in gut after challenge compared to the other groups, corresponding to the lower intestinal alkaline phosphatase activity. One of its effect mechanisms is the combination could inhibit the growth of A. veronii in vitro; the other is direct immersion of germ-free zebrafish proved AiiO-AIO6 did not directly regulate the innate immune response of the host, but wt55 did it, and the simple combination group could significantly reduce the expression of nuclear factor kappa-B (NF-kappa B) and proinflammatory cytokine interleukin-1 beta (IL-1 beta), increase the expression of lysozyme gene; and the third is intestinal microbiota also plays a regulatory role: the gut microbiota from combination group could significantly inhibit the expression of IL-1 beta and NF-kappa B, and increased the expression of transforming growth factor-beta (TGF-beta) and lysozyme. Given the effectiveness of this simple combination, a B. subtilis quorum-quenching recombinant expression strain in which AiiO-AIO6 was surface displayed on the spores and secreted by vegetative cells was built. The results showed that the survival rate after challenge was lower than that of the group treated with AiiO-AIO6 or wt55 alone, and the expression of proinflammatory cytokine IL-1 beta and NF-kappa B were significantly higher. Our study demonstrated the effectiveness of B. subtilis and AiiO-AIO6 simple combination and established an efficient B. subtilis expression system.

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