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Biological Characterization of the Biocontrol Agent Bacillus amyloliquefaciens CPA-8: The Effect of Temperature, pH and Water Activity on Growth, Susceptibility to Antibiotics and Detection of Enterotoxic Genes

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Abstract

This work focuses on the biological understanding of the biocontrol agent Bacillus amyloliquefaciens CPA-8 in order to accomplish the characterization required in the registration process for the development of a microorganism-based product. The tolerance of CPA-8 to grow under different pH–temperature and water activity (a w)–temperature conditions was widely demonstrated. Regarding the pH results, optimum growth at the evaluated conditions was observed at 37 °C and pH between 7 and 5. On the contrary, the slowest growth was recorded at 20 °C and pH 4.5. Moreover, the type of solute used to reduce a w had a great influence on the minimum a w at which the bacterium was able to grow. The lowest a w values for CPA-8 growth in media modified with glycerol and glucose were 0.950 and 0.960, respectively. Besides, the lowest a w for CPA-8 growth increased when the temperature decreased to 20 °C, at which CPA-8 was not able to grow at less than 0.990 a w, regardless of the type of solute. Antibiotic susceptibility tests were carried out to determine which antibiotic could affect the behavior of the bacteria and revealed that CPA-8 was clearly resistant to hygromycin. Finally, a PCR amplification assay to detect the presence of enterotoxic genes from Bacillus cereus in CPA-8 was also performed. CPA-8 gave negative results for all the genes tested except for nheA gene, which is not enough for the toxicity expression, suggesting that fruit treated with this antagonist will not be a potential vehicle for foodborne illnesses.

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Acknowledgments

This research was supported by the European project BIOCOMES FP7-612713 and by the Secretaria d’Universitats i Recerca del Departament d’Economia i Coneixement de la Generalitat de Catalunya for the PhD Grant 2014-FI-B00367 (Amparo M. Gotor Vila). The authors also thank CERCA Program (Generalitat de Catalunya).

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Authors and Affiliations

  1. IRTA, XaRTA-Postharvest, Edifici Fruitcentre, Parc Científic i Tecnològic Agroalimentari de Lleida, 25003, Lleida, Catalonia, Spain

    Amparo Gotor-Vila, Neus Teixidó, María Sisquella, Rosario Torres & Josep Usall

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  1. Amparo Gotor-Vila
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  2. Neus Teixidó
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  3. María Sisquella
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  4. Rosario Torres
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Correspondence to Neus Teixidó.

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Gotor-Vila, A., Teixidó, N., Sisquella, M. et al. Biological Characterization of the Biocontrol Agent Bacillus amyloliquefaciens CPA-8: The Effect of Temperature, pH and Water Activity on Growth, Susceptibility to Antibiotics and Detection of Enterotoxic Genes. Curr Microbiol 74, 1089–1099 (2017). https://doi.org/10.1007/s00284-017-1289-8

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  • DOI: https://doi.org/10.1007/s00284-017-1289-8

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