Cardinal parameter growth and growth boundary model for non-proteolytic Clostridium botulinum – Effect of eight environmental factors

A new cardinal parameter growth and growth boundary model for non-proteolytic C. botulinum was developed and validated for fresh and lightly preserved seafood and poultry products. 523 growth rates in broth were used to determine cardinal parameter values and terms for temperature, pH, NaCl/water activity, acetic, benzoic, citric, lactic and sorbic acids. The new growth and growth boundary model included the inhibiting interactive effect between these factors and it was calibrated using growth curves from 10 challenge tests with unprocessed seafood. For model evaluation, 40 challenge tests with well characterized fresh and lightly preserved seafood were performed. Comparison of these observed growth curves and growth rates (μmax-values) predicted by the new model resulted in a bias factor (Bf) of 1.12 and an accuracy factor (Af) of 1.40. Furthermore, the new model was evaluated with 94 growth rates and 432 time to toxin formation data extracted from the scientific literature for seafood, poultry, meat, pasta and prepared meals. These data included responses for 36 different toxigenic strains of non-proteolytic C. botulinum. The obtained Bf-/Af-values were 0.97/2.04 for μmax-values and 0.96/1.80 for time to toxin formation. The model correctly predicted 93.8% of the growth responses with 5.6% being fail-safe and <1% fail-dangerous. A cocktail of four non-toxin producing Clostridium spp. isolates was used to develop the new model and these isolates had more than 99.8% 16S rRNA gene similarity to non-proteolytic C. botulinum (Group II). The high number of environmental factors included in the new model makes it a flexible tool to facilitate development or reformulation of seafood and poultry products that do not support the growth of non-proteolytic C. botulinum. Further, evaluation of the new model with well characterized products is desirable particularly for meat, vegetables, pasta and prepared meals as well as for dairy products that was not included in the present study.