Risk assessment of the growth of Clostridium botulinum and spores germination induced by high hydrostatic pressure in seafood

Aims: In the present study, a risk assessment for growth of Clostridium botulinum in model seafood deli salads was performed along with the effect of growth of resident microbes on the intrinsic and extrinsic properties of the product. Furthermore, high hydrostatic pressure (HHP) was applied to induce the germination of the surrogate spores. Materials and Methods: Five batches of seafood deli salads were stored at 4°C for up to 42 days, with samples being withdrawn periodically for microbial and chemical analysis. The extrinsic (oxygen content of the headspace) and intrinsic (redox potential, pH, and water activity) properties were determined along with microbial counts (total viable counts [TVCs], yeast and mold, lactic acid bacteria) over the product shelf-life. The data generated were then uploaded into a predictive model, and the potential growth of C. botulinum was assessed under different storage conditions. Furthermore, product inoculated with Bacillus atrophaeus was pressure treated at 400, 500, and 600 MPa for 1, 2, and 3 min at 20°C. Results: From analysis of the deli salads during storage at 4°C, the TVC remained below 2 log colony forming unit (CFU)/g with no lactic acid bacteria being detected. The yeast and mold count progressively increased during the storage period attaining 6 log CFU/g at the end of the 45-day storage period. There was no change in the pH and oxidation-reduction potential of the deli salads during storage. Predictive model indicated that in storage at 4°C and pH 5.1 and a water activity of 0.974, the generation time was 561.37 h. Only one log spore germination induction was observed when HHP treatment was performed at 600 MPa for 3 min. Conclusions: From predictive modeling, it was determined that based on the intrinsic and extrinsic factors of stored deli salad, the growth of C. botulinum was unlikely. In addition, the application of HHP did not significantly induce germination of the surrogate spores.