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Current Microbiology

, Volume 61, Issue 6, pp 525–533 | Cite as

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

  • S. Prevost
  • S. Andre
  • F. RemizeEmail author
Article

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.

Keywords

Internal Transcribe Spacer Thermophilic Bacterium Geobacillus Bacillus Coagulans Geobacillus Stearothermophilus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work received financial support from FranceAgriMer and French canners.

References

  1. 1.
    Anonymous (1997) Contrôle de la stabilité des produits appertisés et assimilés—Méthode de routine, in Microbiologie des aliments. AFNOR (Association Française de Normalisation), ParisGoogle Scholar
  2. 2.
    Ashton D (1981) Thermophilic microorganisms involved in food spoilage: thermophilic anaerobes not producing hydrogen sulfide. J Food Protect 44(2):146–148Google Scholar
  3. 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 309–316Google Scholar
  4. 4.
    Byrer DE, Rainey FA, Wiegel J (2000) Novel strains of Moorella thermoacetica form unusually heat-resistant spores. Arch Microbiol 174(5):334–339CrossRefPubMedGoogle Scholar
  5. 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):581–588CrossRefPubMedGoogle Scholar
  6. 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):358–364CrossRefPubMedGoogle Scholar
  7. 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 1450Google Scholar
  8. 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 1051–1092Google Scholar
  9. 9.
    Dotzauer C, Ehrmann M, Vogel R (2002) Occurence and detection of Thermoanaerobacterium and Thermoanaerobacter in canned food. Food Technol Biotechnol 40(1):21–26Google Scholar
  10. 10.
    Esty JR, Meyer KF (1922) The heat resistance of the spores of B. botulinus and allied anaerobes XI. J Infect Dis 31:650–663Google Scholar
  11. 11.
    Feeherry FE, Munsey DT, Rowley DB (1987) Thermal inactivation and injury of Bacillus stearothermophilus spores. Appl Environ Microbiol 53(2):365–370PubMedGoogle Scholar
  12. 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):3139–3143PubMedGoogle Scholar
  13. 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):9–15CrossRefPubMedGoogle Scholar
  14. 14.
    Karnholz A, Kusel K, Gossner A et al (2002) Tolerance and metabolic response of acetogenic bacteria toward oxygen. Appl Environ Microbiol 68(2):1005–1009CrossRefPubMedGoogle Scholar
  15. 15.
    Kotzekidou P (1996) A microtitre tray procedure for a simplified identification of Bacillus spp. in spoiled canned foods. Food Microbiol 13:35–40CrossRefGoogle Scholar
  16. 16.
    Liu D (2008) Preparation of Listeria monocytogenes specimens for molecular detection and identification. Int J Food Microbiol 122(3):229–242CrossRefPubMedGoogle Scholar
  17. 17.
    Mafart P (2000) Taking injuries of surviving bacteria into account for optimising heat treatments. Int J Food Microbiol 55:175–179CrossRefPubMedGoogle Scholar
  18. 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 299–308Google Scholar
  19. 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):85–94CrossRefPubMedGoogle Scholar
  20. 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):398–412PubMedGoogle Scholar
  21. 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):53–59CrossRefPubMedGoogle Scholar
  22. 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):263–272CrossRefPubMedGoogle Scholar
  23. 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):155–167CrossRefPubMedGoogle Scholar
  24. 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):1246–1255CrossRefPubMedGoogle Scholar
  25. 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):220–230CrossRefPubMedGoogle Scholar
  26. 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):5509–5513CrossRefPubMedGoogle Scholar
  27. 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):983–991CrossRefPubMedGoogle Scholar
  28. 28.
    Shaw M (1928) Thermophilic bacteria in canned foods. J Infect Dis 43(5):461–474Google Scholar
  29. 29.
    Weisburg WG, Barns SM, Pelletier DA et al (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173(2):697–703PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.CTCPA Centre Technique de la Conservation des Produits Agricoles, Site Agroparc, ZA de l’aéroportAVIGNON cedex 9France

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