Skip to main content
SpringerLink
Log in

PCR Detection of Thermophilic Spore-Forming Bacteria Involved in Canned Food Spoilage

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

Thermophilic bacteria that form highly heat-resistant spores constitute an important group of spoilage bacteria of low-acid canned food. A PCR assay was developed in order to rapidly trace these bacteria. Three PCR primer pairs were designed from rRNA gene sequences. These primers were evaluated for the specificity and the sensitivity of detection. Two primer pairs allowed detection at the species level of Geobacillus stearothermophilus and Moorella thermoacetica/thermoautrophica. The other pair allowed group-specific detection of anaerobic thermophilic bacteria of the genera Thermoanaerobacterium, Thermoanaerobacter, Caldanerobium and Caldanaerobacter. After a single enrichment step, these PCR assays allowed the detection of 28 thermophiles from 34 cans of spoiled low-acid food. In addition, 13 ingredients were screened for the presence of these bacteria. This PCR assay serves as a detection method for strains able to spoil low-acid canned food treated at 55°C. It will lead to better reactivity in the canning industry. Raw materials and ingredients might be qualified not only for quantitative spore contamination, but also for qualitative contamination by highly heat-resistant spores.

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

Fig. 1

Similar content being viewed by others

References

  1. Anonymous (1997) Contrôle de la stabilité des produits appertisés et assimilés—Méthode de routine, in Microbiologie des aliments. AFNOR (Association Française de Normalisation), Paris

  2. Ashton D (1981) Thermophilic microorganisms involved in food spoilage: thermophilic anaerobes not producing hydrogen sulfide. J Food Protect 44(2):146–148

    Google Scholar 

  3. Ashton D, Bernard D (1992) Thermophilic anaerobic sporeformers. In: Vanderzantz C, Splittstoesser DF (eds) Compendium of methods for the microbiological examination of foods, 3rd edn. American Public Health Association, Washington, DC, pp 309–316

  4. Byrer DE, Rainey FA, Wiegel J (2000) Novel strains of Moorella thermoacetica form unusually heat-resistant spores. Arch Microbiol 174(5):334–339

    Article  CAS  PubMed  Google Scholar 

  5. Carlier JP, Bedora-Faure M (2006) Phenotypic and genotypic characterization of some Moorella sp. strains isolated from canned foods. Syst Appl Microbiol 29(7):581–588

    Article  CAS  PubMed  Google Scholar 

  6. Cooper RM, McKillip JL (2006) Enterotoxigenic Bacillus spp. DNA fingerprint revealed in naturally contaminated nonfat dry milk powder using rep-PCR. J Basic Microbiol 46(5):358–364

    Article  CAS  PubMed  Google Scholar 

  7. De Vos P, Garrity GM, Jones D et al (eds) (2009) Bergey’s manual of systematic bacteriology, vol 3. The Firmicutes, 2nd edn. Springer, Dordrecht, p 1450

  8. Denny CB, Corlett DAJ (1992) Canned foods—tests for cause of spoilage. In: Vanderzantz C, Splittstoesser DF (eds) Compendium of methods for the microbiological examination of foods, 3rd edn. American Public Health Association, Washington, DC, pp 1051–1092

  9. Dotzauer C, Ehrmann M, Vogel R (2002) Occurence and detection of Thermoanaerobacterium and Thermoanaerobacter in canned food. Food Technol Biotechnol 40(1):21–26

    Google Scholar 

  10. Esty JR, Meyer KF (1922) The heat resistance of the spores of B. botulinus and allied anaerobes XI. J Infect Dis 31:650–663

    Google Scholar 

  11. Feeherry FE, Munsey DT, Rowley DB (1987) Thermal inactivation and injury of Bacillus stearothermophilus spores. Appl Environ Microbiol 53(2):365–370

    CAS  PubMed  Google Scholar 

  12. Herman LM, Vaerewijck MJ, Moermans RJ et al (1997) Identification and detection of Bacillus sporothermodurans spores in 1, 10, and 100 milliliters of raw milk by PCR. Appl Environ Microbiol 63(8):3139–3143

    CAS  PubMed  Google Scholar 

  13. Hornstra LM, Ter Beek A, Smelt JP et al (2009) On the origin of heterogeneity in (preservation) resistance of Bacillus spores: input for a ‘systems’ analysis approach of bacterial spore outgrowth. Int J Food Microbiol 134(1–2):9–15

    Article  PubMed  Google Scholar 

  14. Karnholz A, Kusel K, Gossner A et al (2002) Tolerance and metabolic response of acetogenic bacteria toward oxygen. Appl Environ Microbiol 68(2):1005–1009

    Article  CAS  PubMed  Google Scholar 

  15. Kotzekidou P (1996) A microtitre tray procedure for a simplified identification of Bacillus spp. in spoiled canned foods. Food Microbiol 13:35–40

    Article  Google Scholar 

  16. Liu D (2008) Preparation of Listeria monocytogenes specimens for molecular detection and identification. Int J Food Microbiol 122(3):229–242

    Article  CAS  PubMed  Google Scholar 

  17. Mafart P (2000) Taking injuries of surviving bacteria into account for optimising heat treatments. Int J Food Microbiol 55:175–179

    Article  CAS  PubMed  Google Scholar 

  18. Olson KE, Sorrells KM (1992) Thermophilic flat sour sporeformers. In: Vanderzantz C, Splittstoesser DF (eds) Compendium of methods for the microbiological examination of foods, 3rd edn. American Public Health Association, Washington, DC, pp 299–308

  19. Oomes SJ, van Zuijlen AC, Hehenkamp JO et al (2007) The characterisation of Bacillus spores occurring in the manufacturing of (low acid) canned products. Int J Food Microbiol 120(1–2):85–94

    Article  CAS  PubMed  Google Scholar 

  20. Penna TC, Machoshvili IA, Taqueda ME et al (2000) The effect of media composition on the thermal resistance of Bacillus stearothermophilus. PDA J Pharm Sci Technol 54(5):398–412

    CAS  PubMed  Google Scholar 

  21. Robert H, Gabriel V, Fontagne-Faucher C (2009) Biodiversity of lactic acid bacteria in French wheat sourdough as determined by molecular characterization using species-specific PCR. Int J Food Microbiol 135(1):53–59

    Article  CAS  PubMed  Google Scholar 

  22. Ruckert A, Ronimus RS, Morgan HW (2004) A RAPD-based survey of thermophilic bacilli in milk powders from different countries. Int J Food Microbiol 96(3):263–272

    Article  CAS  PubMed  Google Scholar 

  23. Rueckert A, Ronimus RS, Morgan HW (2005) Development of a rapid detection and enumeration method for thermophilic bacilli in milk powders. J Microbiol Methods 60(2):155–167

    Article  CAS  PubMed  Google Scholar 

  24. Rueckert A, Ronimus RS, Morgan HW (2005) Rapid differentiation and enumeration of the total, viable vegetative cell and spore content of thermophilic bacilli in milk powders with reference to Anoxybacillus flavithermus. J Appl Microbiol 99(5):1246–1255

    Article  CAS  PubMed  Google Scholar 

  25. Rueckert A, Ronimus RS, Morgan HW (2006) Development of a real-time PCR assay targeting the sporulation gene, spo0A, for the enumeration of thermophilic bacilli in milk powder. Food Microbiol 23(3):220–230

    Article  CAS  PubMed  Google Scholar 

  26. Sasaki K, Shintani H, Itoh J et al (2000) Effect of calcium in assay medium on D value of Bacillus stearothermophilus ATCC 7953 spores. Appl Environ Microbiol 66(12):5509–5513

    Article  CAS  PubMed  Google Scholar 

  27. Scheldeman P, Herman L, Goris J et al (2002) Polymerase chain reaction identification of Bacillus sporothermodurans from dairy sources. J Appl Microbiol 92(5):983–991

    Article  CAS  PubMed  Google Scholar 

  28. Shaw M (1928) Thermophilic bacteria in canned foods. J Infect Dis 43(5):461–474

    Google Scholar 

  29. Weisburg WG, Barns SM, Pelletier DA et al (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173(2):697–703

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work received financial support from FranceAgriMer and French canners.

Author information

Authors and Affiliations

  1. CTCPA Centre Technique de la Conservation des Produits Agricoles, Site Agroparc, ZA de l’aéroport, BP 21 203, 84911, AVIGNON cedex 9, France

    S. Prevost, S. Andre & F. Remize

Authors
  1. S. Prevost
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Andre
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. F. Remize
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to F. Remize.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Prevost, S., Andre, S. & Remize, F. PCR Detection of Thermophilic Spore-Forming Bacteria Involved in Canned Food Spoilage. Curr Microbiol 61, 525–533 (2010). https://doi.org/10.1007/s00284-010-9648-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00284-010-9648-8

Keywords

Search

Navigation