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Rapid detection and differentiation of Staphylococcus colonies using an optical scattering technology

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Abstract

Staphylococcus species are a major pathogen responsible for nosocomial infections and foodborne illnesses. We applied a laser-based BARDOT (bacterial rapid detection using optical scattering technology) for rapid colony screening and detection of Staphylococcus on an agar plate and differentiate these from non-Staphylococcus spp. Among the six growth media tested, phenol red mannitol agar (PRMA) was found most suitable for building the Staphylococcus species scatter image libraries. Scatter image library for Staphylococcus species gave a high positive predictive value (PPV 87.5–100%) when tested against known laboratory strains of Staphylococcus spp., while the PPV against non-Staphylococcus spp. was 0–38%. A total of nine naturally contaminated bovine raw milk and ready-to-eat chicken salad samples were tested, and BARDOT detected Staphylococcus including Staphylococcus aureus with 80–100% PPV. Forty-five BARDOT-identified bacterial isolates from naturally contaminated foods were further confirmed by tuf and nuc gene-specific PCR and 16S rRNA gene sequence. This label-free, non-invasive on-plate colony screening technology can be adopted by the food industries, biotechnology companies, and public health laboratories for Staphylococcus species detection including S. aureus from various samples for food safety and public health management.

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References

  1. Baird-Parker A. An improved diagnostic and selective medium for isolating coagulase positive staphylococci. J Appl Microbiol. 1962;25(1):12–9.

    Google Scholar 

  2. Bhunia AK. One day to one hour: how quickly can foodborne pathogens be detected? Future Microbiol. 2014;9(8):935–46.

    Article  CAS  PubMed  Google Scholar 

  3. Banada PP, Guo S, Bayraktar B, Bae E, Rajwa B, Robinson JP, et al. Optical forward-scattering for detection of Listeria monocytogenes and other Listeria species. Biosens Bioelectron. 2007;22(8):1664–71.

    Article  CAS  PubMed  Google Scholar 

  4. Bae E, Banada PP, Huff K, Bhunia AK, Robinson JP, Hirleman ED. Biophysical modeling of forward scattering from bacterial colonies using scalar diffraction theory. Appl Opt. 2007;46(17):3639–48.

    Article  PubMed  Google Scholar 

  5. Singh AK, Bettasso AM, Bae E, Rajwa B, Dundar MM, Forster MD, et al. Laser optical sensor, a label-free on-plate Salmonella enterica colony detection tool. MBio. 2014;5(1):e01019–3.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Ahmed WM, Bayraktar B, Bhunia AK, Hirleman ED, Robinson JP, Rajwa B. Classification of bacterial contamination using image processing and distributed computing. IEEE J Biomed Health Inform. 2013;17(1):232–9.

    Article  CAS  PubMed  Google Scholar 

  7. Sousa AM, Pereira MO, Lourenço A. MorphoCol: an ontology-based knowledgebase for the characterisation of clinically significant bacterial colony morphologies. J Biomed Inform. 2015;55:55–63.

    Article  PubMed  Google Scholar 

  8. Zieliński B, Plichta A, Misztal K, Spurek P, Brzychczy-Włoch M, Ochońska D. Deep learning approach to bacterial colony classification. PLoS One. 2017;12(9):e0184554.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Banada PP, Huff K, Bae E, Rajwa B, Aroonnual A, Bayraktar B, et al. Label-free detection of multiple bacterial pathogens using light-scattering sensor. Biosens Bioelectron. 2009;24:1685–92.

    Article  CAS  PubMed  Google Scholar 

  10. Abdelhaseib MU, Singh AK, Bailey M, Singh M, El-Khateib T, Bhunia AK. Fiber optic and light scattering sensors: complimentary approaches to rapid detection of Salmonella enterica in food samples. Food Control. 2016;61:135–45.

    Article  CAS  Google Scholar 

  11. Tang Y, Kim H, Singh AK, Aroonnual A, Bae E, Rajwa B, et al. Light scattering sensor for direct identification of colonies of Escherichia coli serogroups O26, O45, O103, O111, O121, O145 and O157. PLoS One. 2014;9(8):e105272.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Singh AK, Sun XL, Bai XJ, Kim H, Abdalhaseib MU, Bae E, et al. Label-free, non-invasive light scattering sensor for rapid screening of Bacillus colonies. J Microbiol Methods. 2015;109:56–66.

    Article  CAS  PubMed  Google Scholar 

  13. Singh AK, Drolia R, Bai X, Bhunia AK. Streptomycin induced stress response in Salmonella enterica serovar Typhimurium shows distinct colony scatter signature. PLoS One. 2015;10(8):e0135035.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Singh AK, Leprun L, Drolia R, Bai X, Kim H, Aroonnual A, et al. Virulence gene-associated mutant bacterial colonies generate differentiating two-dimensional laser scatter fingerprints. Appl Environ Microbiol. 2016;82(11):3256–68.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Andrews WH, Hammack TS. Food sampling and preparation of sample homogenate. In: Bacteriological analytical manual online vol 3. 8th ed. Silver Spring: Center for Food Safety & Applied Nutrition, US Food and Drug Administration; 2003. p. 2004.

    Google Scholar 

  16. Morot-Bizot SC, Talon R, Leroy S. Development of a multiplex PCR for the identification of Staphylococcus genus and four staphylococcal species isolated from food. J Appl Microbiol. 2004;97(5):1087–94.

    Article  CAS  PubMed  Google Scholar 

  17. Sunagar R, Deore S, Deshpande P, Rizwan A, Sannejal A, Sundareshan S, et al. Differentiation of Staphylococcus aureus and Staphylococcus epidermidis by PCR for the fibrinogen binding protein gene. J Dairy Sci. 2013;96(5):2857–65.

    Article  CAS  PubMed  Google Scholar 

  18. Singh AK, Ramesh A. Evaluation of a facile method of template DNA preparation for PCR-based detection and typing of lactic acid bacteria. Food Microbiol. 2009;26(5):504–13.

    Article  CAS  PubMed  Google Scholar 

  19. Bae E, Bai N, Aroonnual A, Robinson JP, Bhunia AK, Hirleman ED. Modeling light propagation through bacterial colonies and its correlation with forward scattering patterns. J Biomed Opt. 2010;15(4):045001.

    Article  PubMed  Google Scholar 

  20. Kim H, Doh I-J, Sturgis J, Bhunia AK, Robinson JP, Bae E. Reflected scatterometry for noninvasive interrogation of bacterial colonies. J Biomed Opt. 2016;21(10):107004.

    Article  PubMed  Google Scholar 

  21. Cooper A, Dean A, Hinshelwood C. Factors affecting the growth of bacterial colonies on agar plates. Proc R Soc Lond B Biol Sci. 1968;171(1023):175–99.

    Article  CAS  PubMed  Google Scholar 

  22. Mialon M, Tang Y, Singh AK, Bae E, Bhunia AK. Effects of preparation and storage of agar media on the sensitivity of bacterial forward scattering patterns. Open J Appl Biosens. 2012;1(03):26.

    Article  CAS  Google Scholar 

  23. Suchwałko A, Buzalewicz I, Podbielska H. Bacteria identification in an optical system with optimized diffraction pattern registration condition supported by enhanced statistical analysis. Opt Express. 2014;22(21):26312–27.

    Article  PubMed  Google Scholar 

  24. Buzalewicz I, Kujawińska M, Krauze W, Podbielska H. Novel perspectives on the characterization of species-dependent optical signatures of bacterial colonies by digital holography. PLoS One. 2016;11(3):e0150449.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Skovgaard N. Scientific criteria to ensure safe food: Institute of Medicine, Natural Research Council of the National Academies. The National Academy Press, Elsevier, Washington DC. 2004.

  26. NACMCF. Response to questions posed by the Department of Defense Regarding Microbiological Criteria as indicators of process control or insanitary conditions, National Advisory Committee on Microbiological Criteria for Foods. USDA-FSIS, Washington, DC. 2015.

  27. Chye FY, Abdullah A, Ayob MK. Bacteriological quality and safety of raw milk in Malaysia. Food Microbiol. 2004;21(5):535–41.

    Article  Google Scholar 

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Acknowledgments

T.S.A acknowledges King Saud University, the Kingdom of Saudi Arabia, for a graduate student fellowship. X.Z., a visiting scholar at Purdue University from College of Science, Nanjing University of Aeronautics and Astronautics, Nanjing, China, acknowledges the scholarship support from China Scholarship Council, Beijing, China. We also acknowledge the help of Bruce M. Applegate and Marcela M. Chavez in procuring raw milk samples.

Funding

This research was supported through a cooperative agreement with the Agricultural Research Service of the US Department of Agriculture project number 8072-42000-077, USDA National Institute of Food and Agriculture (Hatch) project, and the Center for Food Safety Engineering at Purdue University.

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Correspondence to Atul K. Singh or Arun K. Bhunia.

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The authors declare that there were no ethical implications or research involving human or animals.

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The authors declare that they have no conflict of interest.

Additional information

Published in the topical collection Food Safety Analysis with guest editor Steven J. Lehotay.

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Alsulami, T.S., Zhu, X., Abdelhaseib, M.U. et al. Rapid detection and differentiation of Staphylococcus colonies using an optical scattering technology. Anal Bioanal Chem 410, 5445–5454 (2018). https://doi.org/10.1007/s00216-018-1133-4

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  • DOI: https://doi.org/10.1007/s00216-018-1133-4

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