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Prevalence and Characteristics of Bacillus cereus Group Isolated from Raw and Pasteurised Milk

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Abstract

The elimination of spore-forming bacteria is not guaranteed by current pasteurisation processes and is a challenging problem for the dairy industry. Given that Bacillus cereus sensu lato (B. cereus group) is an important foodborne pathogen and spoiler in the dairy industry, this study aimed at evaluating the prevalence and characteristics of B. cereus group in raw and pasteurised milk samples collected in Victoria, Australia. Isolated B. cereus group were tested for antimicrobial susceptibility, biofilm formation and virulence properties. Genetic diversity was assessed using ERIC-PCR. Proteomic profiling using MALDI-TOF MS and chemical profiling using Fourier-transform infrared (FTIR) spectroscopy were also applied for clustering of the isolates. Results showed 42.3% of milk samples contained B. cereus group, with a higher contamination level for pasteurised milk. Virulence studies identified genes nheA, nheB, hblA and nheC in most isolates and cyk gene in 46% of all isolates. Antimicrobial susceptibility testing showed a high prevalence of resistance towards ampicillin, ceftriaxone and penicillin. The biofilm-forming capacity of our isolates showed that most (53.7%) had the ability to form a biofilm. Genetic profiling using ERIC-PCR placed most B. cereus group isolates from pasteurised milk in the same cluster, indicating that they probably originated from a similar source. Raw milk isolates showed greater diversity indicating various sources. FTIR spectroscopy showed high agreement with genetic profiling. In contrast, low agreement between proteomic (MALDI-TOF MS) and genetic typing was observed. The present study showed that the FTIR spectroscopy could be adopted as a rapid tool for the typing of B. cereus group. Overall, the virulence and antimicrobial resistance characteristics, together with the ability of isolates to produce biofilm, indicate the importance of B. cereus group in the Australian dairy industry.

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Acknowledgements

This work was supported by Dairy Food Safety Victoria (DFSV). The authors would like to acknowledge Doug Eddy for his help during the project and all technical staff in the Department of Chemistry and Biotechnology, Swinburne University of Technology.

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Authors and Affiliations

  1. Department of Chemistry and Biotechnology, Swinburne University of Technology, Hawthorn, VIC, 3122, Australia

    Behrad Radmehr, Bita Zaferanloo & Enzo A. Palombo

  2. Bruker Pty Ltd, Preston, VIC, 3072, Australia

    Thien Tran

  3. Land and Water, Commonwealth Scientific and Industrial Research Organisation (CSIRO), Ecosciences Precinct, Dutton Park, QLD, 4102, Australia

    David J. Beale

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  1. Behrad Radmehr
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All authors designed the experiments. BR and TT performed the experimentation and analysed the data. All authors wrote the manuscript.

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Correspondence to Enzo A. Palombo.

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The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Thien Tran is an employee of Bruker Pty. Ltd. All other authors declare no conflict of interest.

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Radmehr, B., Zaferanloo, B., Tran, T. et al. Prevalence and Characteristics of Bacillus cereus Group Isolated from Raw and Pasteurised Milk. Curr Microbiol 77, 3065–3075 (2020). https://doi.org/10.1007/s00284-020-02129-6

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