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Published on: September 9, 2015
Diet-Supplemented Probiotic Bacillus velezensis Protects Against Viral Infection Via the Lipopeptide Surfactin in
Wenhao Zhou1, Hui Liang2, Delong Meng1
1Sino-Norway Joint Lab on Fish Gastrointestinal Microbiota, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, China.
Background:
Probiotic nutrition has the potential to improve health and prevents diseases, and probiotics such as Bacillus spp may protect against viral infection. However, the in vivo antiviral mechanism has not yet been fully determined.
Objectives:
We aimed to elucidate the antiviral mechanism of Bacillus velezensis strain by comparing the effect of wild type and isogenic mutant in a zebrafish model.
Methods:
Zebrafish (60 ± 3 mg) were fed control diet or diet-supplemented B. velelzensis T23 and were challenged with spring viremia of carp virus (SVCV). T23 mutants deficient in the production of lipopeptides and/or polyketides were constructed by 2-step replacement recombination. Zebrafish were fed diet supplemented with wild type or mutant followed by SVCV infection. Germ-free zebrafish were monocolonized with T23 or its mutant and were infected by SVCV. When appropriate, type I interferon (IFN) receptors or myeloid differentiation factor (Myd)88 were knocked down by vivo-morpholino. ZF4 (zebrafish embryonic fibroblast like cell line) were treated with bacterial fractions of T23 or surfactin and were infected with SVCV.
Results:
B. velezensis T23 improved survival of zebrafish after SVCV challenge (P < 0.05) by 250% and inhibited viral replication in germ-free zebrafish (P < 0.05). Genes involved in biosynthesis of secondary metabolites were knocked out in T23, and the antiviral function of Δsfp and ΔsrfAA mutants deficient in the production of surfactin was abrogated both in vitro and in vivo. T23 enhanced type I IFN antiviral immunity after SVCV infection (P < 0.05). Knockdown of type I IFN receptors or Myd88 in gnotobiotic zebrafish did not influence the antiviral effect of T23. Finally, surfactin can exert antiviral effect by directly inhibiting the infectivity of SVCV.
Conclusions:
Probiotic B. velezensis T23 protects against viral infection in zebrafish. The antiviral effect of B. velezensis T23 is mainly mediated by direct inhibition of surfactin on virus and is not attributable to type I IFN antiviral immunity.
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