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Isolation and characterisation of Bacillus spp. antagonistic to Vibrio parahaemolyticus for use as probiotics in aquaculture

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Abstract

Acute gastroenteritis caused by pathogenic Vibrio parahaemolyticus is one of the major factors affecting the development of aquaculture and the safety of seafood. Using the antagonism of probiotics against pathogens is an alternative strategy to antibiotics and a common trend to control food-borne pathogenic bacteria. In this study, a total of 249 isolates were isolated from four types of seafood (Litopenaeus vannamei, Oratosquilla oratoria, Mactra veneriformis and Portunus trituberculatus) and coastal sediment from Liaodong Bay in the Bohai Sea, China with five different separation agars. The most isolates came from the sample of coastal sediment and on agar of 2216E, which accounted for 36.14 and 54.62 % respectively. Twenty-four among 249 isolates displayed direct antimicrobial activity to V. parahaemolyticus with spot inoculation. Sixteen active isolates were selected for extracellular antimicrobial activity using the Oxford cup method. Only strains of B16 and J7 showed extracellular antimicrobial activity and were identified as Bacillus pumilus and Bacillus mojavensis respectively based on the physiological identification and 16S rRNA sequence analysis. Both of the strains B16 and J7 exhibited extracellular hydrolytic enzyme activity and antagonism against more than one indicator bacteria in vitro, which indicates that the two strains have broad potential application as suitable probiotic candidates in aquaculture while B. mojavensis was first reported to inhibit pathogenic Vibrio spp. in vitro. There is no particular trait as to antagonism of B. pumilus B16 or B. mojavensis J7 to Gram-positive or Gram-negative indicator bacteria.

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Acknowledgments

This work was supported financially by the National Key Technologies R & D Programme of China during the 12th Five-Year Plan Period (2012BAD29B06), Food Safety Key Laboratory of Liaoning Province, and Engineering and Technology Research Centre for Food Preservation, Processing, and Safety Control of Liaoning Province (LNSAKF2011031).

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Authors and Affiliations

  1. Food College, Shenyang Agriculture University, Shenyang, 110866, China

    Xue-Fei Liu & Shu-Xia Lyu

  2. Food Science Research Institute of Bohai University, Jinzhou, 121013, China

    Xue-Fei Liu, Ya Li, Jian-Rong Li, Lu-Yun Cai & Xiu-Xia Li

  3. Department of Food Science and Technology, The University of Georgia, Athens, GA, 30602-1661, USA

    Jian-Rong Li & Jin-Ru Chen

  4. College of Biological Science and Technology, Shenyang Agriculture University, Shenyang, 110866, China

    Shu-Xia Lyu

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  1. Xue-Fei Liu
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  2. Ya Li
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  3. Jian-Rong Li
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  4. Lu-Yun Cai
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  5. Xiu-Xia Li
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Correspondence to Jian-Rong Li or Shu-Xia Lyu.

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Liu, XF., Li, Y., Li, JR. et al. Isolation and characterisation of Bacillus spp. antagonistic to Vibrio parahaemolyticus for use as probiotics in aquaculture. World J Microbiol Biotechnol 31, 795–803 (2015). https://doi.org/10.1007/s11274-015-1833-2

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