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Characterization of Clostridium tyrobutyricum Strains Using Three Different Typing Techniques.

Johanna Burtscher1, Franziska Küller1,2, Matthias Dreier3

  • 1Institute of Food Science, University of Natural Resources and Life Sciences Vienna (BOKU), 1190 Vienna, Austria.

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Clostridium tyrobutyricum causes significant cheese spoilage. This study found high strain diversity within the species, with PCR-based methods like hexaplex-PCR proving most effective for differentiation.

Keywords:
Clostridium tyrobutyricumMALDI-TOF MScheesefingerprintingspoilagetyping

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Area of Science:

  • Food microbiology
  • Bacteriology
  • Dairy science

Background:

  • Clostridium tyrobutyricum is a primary cause of severe cheese spoilage.
  • Bacterial metabolism during cheese ripening leads to defects and economic losses.
  • Strain-level variation in spoilage potential necessitates better differentiation methods.

Purpose of the Study:

  • To compare the efficacy of three typing techniques for differentiating Clostridium tyrobutyricum strains.
  • To assess the intra-species diversity of C. tyrobutyricum relevant to the dairy industry.

Main Methods:

  • Repetitive element palindromic PCR (rep-PCR) with agarose gel electrophoresis.
  • Hexaplex-PCR followed by automated capillary electrophoresis.
  • Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) typing.

Main Results:

  • MALDI-TOF MS showed moderate reproducibility and low discriminatory power.
  • Both PCR-based methods (rep-PCR and hexaplex-PCR) were highly reproducible and discriminative.
  • Hexaplex-PCR exhibited slightly higher discriminatory power than rep-PCR.

Conclusions:

  • A high degree of intra-species diversity was observed in the tested C. tyrobutyricum strains.
  • PCR-based methods, particularly hexaplex-PCR, are effective for differentiating C. tyrobutyricum strains.
  • Further strain-level investigations are warranted for practical dairy applications.