Skip to main content
SpringerLink
Log in

Optimization and evaluation of heat-shock condition for spore enumeration being used in thermal-process verification: Differential responses of spores and vegetative cells of Clostridium sporogenes to heat shock

  • Research Article
  • Published:
Food Science and Biotechnology Aims and scope Submit manuscript

Abstract

To evaluate a heat-shock condition for the enumeration of Clostridium sporogenes spores, a surrogate for C. botulinum spores, we examined the heat tolerance of C. sporogenes spores and vegetative cells exposed to a heat shock at 90°C. From the D values of the spores determined in the temperature range of 113–121°C, z value (±SD) and D90°C value were estimated to be 10.16±0.90°C and 1,071.52 min, respectively, and the inactivation rates were predicted to be only approximately 2% at 90°C for up to 10 min. Meanwhile, the viable count of spores was significantly higher when activated under a heat-shock condition of 90°C for over 9 min than those activated for shorter time periods. The heat tolerance of vegetative cells was extremely low, showing a D90°C value (±SD) of 0.21±0.01 min. Finally, 3 different heat-shock conditions were compared: 70°C for 30 min, 80°C for 20 min, and 90°C for 10 min, and the experimental comparative data showed no significant differences in viable spore counts. Consequently, these results support that the heat-shock treatment at 90°C for 10 min is suitable to activate spores and to inactivate vegetative cells of C. sporogenes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Ocio MJ, Sánchez T, Fernández PS, Rodrigo M, Martínez A. Thermal resistance characteristics of PA 3679 in the temperature range of 110–121°C as affected by pH, type of acidulant, and substrate. Int. J. Food Microbiol. 22: 239–247 (1994)

    Article  CAS  Google Scholar 

  2. Welt BA, Tong CH, Rossen JL, Lund DB. Effect of microwave radiation on inactivation of Clostridium sporogenes (PA 3679) spores. Appl. Environ. Microb. 60: 482–488 (1994)

    CAS  Google Scholar 

  3. Guan D, Gray P, Kang D-H, Tang J, Shafer B, Ito K, Younce F, Yang TCS. Microbiological validation of microwave-circulated water combination heating technology by inoculated pack studies. J. Food Sci. 68: 1428–1432 (2003)

    Article  CAS  Google Scholar 

  4. McGlynn WG, Davis DR, Johnson MG, Crandall PG. Modified spore inoculation method for thermal-process verification of pinto beans and green beans canned in two large reusable containers. J. Food Sci. 68: 988–991 (2003)

    Article  CAS  Google Scholar 

  5. IFT (Institute of Food Technologists). Overarching principles: Kinetics and pathogens of concern for all technologies. J. Food Sci. 65(suppl.): 16–31 (2000)

    Google Scholar 

  6. Luechapattanaporn K, Wang Y, Wang J, Al-Holy M, Kang D-H, Tang J, Hallberg LM. Microbial safety in radio-frequency processing of packaged foods. J. Food Sci. 69: M201–M206 (2004)

    Article  CAS  Google Scholar 

  7. Luechapattanaporn K, Wang Y, Wang J, Tang J, Hallberg LM, Dunne CP. Sterilization of scrambled eggs in military polymeric trays by radio frequency energy. J. Food Sci. 70: E288–E294 (2005)

    Article  CAS  Google Scholar 

  8. Kalchayanand N, Dunne CP, Sikes A, Ray B. Inactivation of bacterial spores by combined action of hydrostatic pressure and bacteriocins in roast beef. J. Food Safety 23: 219–231 (2003)

    Article  CAS  Google Scholar 

  9. Kalchayanand N, Dunne CP, Sikes A, Ray B. Germination induction and inactivation of Clostridium spores at medium-range hydrostatic pressure treatment. Innov. Food Sci. Emerg. 5: 277–283 (2004)

    Article  CAS  Google Scholar 

  10. AOAC. Official Methods of Analysis of AOAC Int. 18th ed. Method 966.04. Association of Official Analytical Chemists International, Gaithersburg, MD, USA (2005)

    Google Scholar 

  11. Tomasino SF, Hamilton MA. Comparative evaluation of two quantitative test methods for determining the efficacy of liquid sporicides and sterilants on a hard surface: A precollaborative study. J. AOAC Int. 90: 456–464 (2007)

    CAS  Google Scholar 

  12. Tomasino SF, Samalot-Freire LC. AOAC method 966.04: Preliminary evaluation of cooked meat medium with manganese sulfate for the cultivation of Clostridium sporogenes: Precollaborative study. J. AOAC Int. 90: 825–933 (2007)

    CAS  Google Scholar 

  13. Keynan A, Evenchick Z. Activation. pp. 359–396. In: The Bacterial Spore. Gould GW, Hurst A (eds). Academic Press, Inc., New York, NY, USA (1969)

    Google Scholar 

  14. Turnbull PCB, Frawley DA, Bull RL. Heat activation/shock temperatures for Bacillus anthracis spores and the issue of spore plate counts versus true number of spores. J. Microbiol. Meth. 68: 353–357 (2007)

    Article  CAS  Google Scholar 

  15. Uehara M, Fujioka RS, Frank HA. Method for obtaining cleaned putrefactive anaerobe 3679 spores. J. Bacteriol. 89: 929–930 (1965)

    CAS  Google Scholar 

  16. USFDA (US Food and Drug Administration). Clostridium botulinum (Ch. 17). In: Bacteriological Analytical Manual. 8th ed. Rev. A. Available from: http://www.fda.gov/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070879.htm. Accessed Apr. 18, 2011.

  17. Chung H-J, Wang S, Tang J. Influence of heat transfer with tube methods on measured thermal inactivation parameters for Escherichia coli. J. Food Protect. 70: 851–859 (2007)

    Google Scholar 

  18. Bolton DJ, Pearce R, Sheridan JJ, McDowell DA, Blair IS. Decontamination of pork carcasses during scalding and the prevention of Salmonella cross-contamination. J. Appl. Microbiol. 94: 1036–1042 (2003)

    Article  CAS  Google Scholar 

  19. Byrne B, Dunne G, Bolton DJ. Thermal inactivation of Bacillus cereus and Clostridium perfringens vegetative cells and spores in pork luncheon roll. Food Microbiol. 23: 803–808 (2006)

    Article  CAS  Google Scholar 

  20. Licciardello JJ, Nickerson JTR. Effect of radiation environment on the thermal resistance of irradiated spores of Clostridium sporogenes P.A. 3679. J. Food Sci. 27: 211–218 (1962)

    Article  CAS  Google Scholar 

  21. Blank G, Akomas G, Henderson M, Zawistowski J. Heat sensitization of Bacillus subtilis spores by selected spices. J. Food Safety 9: 83–96 (1988)

    Article  Google Scholar 

  22. Augustin JAL, Pflug IJ. Recovery patterns of spores of putrefactive anaerobe 3679 in various subculture media after heat treatment. Appl. Microbiol. 15: 266–276 (1967)

    CAS  Google Scholar 

  23. Juneja VK, Marmer BS. Thermal inactivation of Clostridium perfringens vegetative cells in ground beef and turkey as affected by sodium pyrophosphate. Food Microbiol. 15: 281–287 (1998)

    Article  CAS  Google Scholar 

  24. Juneja VK, Novak JS, Eblen BS, Mcclane BA. Heat resistance of Clostridium perfringens vegetative cells as affected by prior heat shock. J. Food Safety 21: 127–139 (2001)

    Article  CAS  Google Scholar 

  25. Abraham G, Debray E, Candau Y, Piar G. Mathematical model of thermal destruction of Bacillus stearothermophilus spores. Appl. Environ. Microb. 56: 3073–3080 (1990)

    CAS  Google Scholar 

  26. Mathys A, Heinz V, Schwartz FH, Knorr D. Impact of agglomeration on the quantitative assessment of Bacillus stearothermophilus heat inactivation. J. Food Eng. 81: 380–387 (2007)

    Article  Google Scholar 

  27. Powell JF, Hunter JR. Spore germination in the genus Bacillus: The modification of germination requirements as a result of preheating. J. Gen. Microbiol. 13: 59–67 (1955)

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Food and Biotechnology, Korea University, Jochiwon, Chungnam, 339-700, Korea

    Bo Young Byun, Hong-Yon Cho, Han-Joon Hwang & Jae-Hyung Mah

  2. College of Engineering, China Agricultural University, Beijing, 100083, China

    Yanhong Liu

  3. Department of Biological Systems Engineering, Washington State University, Pullman, WA, 99164-6120, USA

    Juming Tang

  4. Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea

    Dong-Hyun Kang

Authors
  1. Bo Young Byun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hong-Yon Cho
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Han-Joon Hwang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jae-Hyung Mah
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yanhong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Juming Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Dong-Hyun Kang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jae-Hyung Mah.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Byun, B.Y., Cho, HY., Hwang, HJ. et al. Optimization and evaluation of heat-shock condition for spore enumeration being used in thermal-process verification: Differential responses of spores and vegetative cells of Clostridium sporogenes to heat shock. Food Sci Biotechnol 20, 751–757 (2011). https://doi.org/10.1007/s10068-011-0105-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10068-011-0105-7

Keywords

Search

Navigation