Applied Microbiology and Biotechnology

, Volume 103, Issue 21–22, pp 8987–8999 | Cite as

Interaction of a novel Bacillus velezensis (BvL03) against Aeromonas hydrophila in vitro and in vivo in grass carp

  • Lina Cao
  • Lifei Pan
  • Liang Gong
  • Yahui Yang
  • Haocheng He
  • Yanping Li
  • Yanan Peng
  • Dongjie Li
  • Liang Yan
  • Xuezhi Ding
  • Shengbiao Hu
  • Ziquan Yu
  • Yunjun Sun
  • Weitao Huang
  • Yibo Hu
  • Ganfeng YiEmail author
  • Liqiu XiaEmail author
Applied microbial and cell physiology


This study evaluated the inhibition and interaction of Bacillus velezensis BvL03 as a probiotic agent against Aeromonas hydrophila. Strain BvL03 isolated from sediment samples of fish ponds had excellent antimicrobial activity against several fish pathogenic bacteria, especially Aeromonas, including A. hydrophila, A. veronii, A. caviae, and A. sobria. The successful amplification of lipopeptide antimicrobial chemical biosynthetic genes, including iturin family (ituA, ituB, and ituD), bacillomycin family (bacA, bacD, and bacAB), surfactin family (srfAB, srfC, and srfAA), and subtilosin family (albF and sunT) from the genome of BvL03 strain, confirmed its predominant antimicrobial activity. The challenge test suggested that BvL03 significantly decreased fish mortality when challenged with A. hydrophila, which had a cumulative mortality of 12.5% in the treatment group. Toxicity and hemolytic activity of A. hydrophila after co-cultured with BvL03 were relieved as confirmed by the cell experiments, when the initial inoculated concentration of BvL03 was 109 cfu/mL or higher. Moreover, the BvL03 strain labeled with GFP protein (BvL03-GFP) and AhX040 strain labeled with mCherry protein (AhX040-mCherry) were injected into grass carps. The fluorescence levels were monitored by using In Vivo Imaging System (IVIS), in which the green color was steadily increasing, whereas the red color was gradually weakening. Whole genome sequencing revealed that strain BvL03 possesses 15 gene clusters related to antibacterial compounds, including 5 NRPS gene clusters and 3 PKS gene clusters. These results suggested that B. velezensis BvL03 has the potential to be developed as a probiotic candidate against A. hydrophila infection in aquaculture.


Bacillus velezensis Aeromonas hydrophila Antibacterial activity Grass carps Fluorescence labeling Interaction 


Author contributions

L.N.C. and L.F.P. designed the experiments. L.N.C., L.F.P., Y.H.Y., H.C.H., Y.P.L., Y.N.P., D.J.L., and S.B.H. contributed in performing the experiments. L.N.C., L.G., X.Z.D., Z.Q.Y., Y.J.S., and Y.B.H. analyzed the data. L.N.C., L.G., and W.T.H. wrote a draft of the manuscript. G.F.Y. and L.Q.X. supervised the research.

Funding information

This study was funded by the National Natural Science Foundation of China (31770106), the National Basic Research Program of China (“973” program; 2012CB722301), the Major Research Projects in Hunan Province (2017NK1030), and “Hunan Province Biological Development Engineering and New Product Development Collaborative Innovation Center project” (20134486).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

Ethical approval. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the Animal Care Committee of Hunan Normal University at which the studies were conducted.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Developmental Biology of Freshwater Fish, Hunan Provincial Key Laboratory of Microbial Molecular Biology, College of Life ScienceHunan Normal UniversityChangshaPeople’s Republic of China

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