Intra- and inter-batch variability in raw pork challenge test studies and their consequences on model predictions: An intricate interplay between L. monocytogenes, the microbiome, and packaging atmosphere

Abstract

The purpose of this study was to conduct challenge studies in raw pork by strictly following all aspects of the 2014 EURL technical guidance document for conducting shelf-life studies on Listeria monocytogenes. Growth potential was assessed on three batches of self-cut pork chops and one batch of in-house prepared pure minced pork without any additives in air and MAP (70 % O2/30% CO2) packaging. Pork chops did not support the growth of the pathogen throughout the shelf-life, given the specific conditions used in this study, with growth potential values of 0.28 and 0.46 log CFU/g, respectively, for both air and MAP. Substantial growth (>0.5 log CFU/g) was obtained in minced pork after investigating only one batch, with growth potential values of 1.69 and 0.80 log CFU/g, for air and MAP. However, both intra- and inter-batch variability for pork chops and intra-batch variability for minced pork was observed; with elevated growth being evened out by the way growth potential is calculated in the EURL 2014 document, leading to underestimations and posing a potential risk to public health. Maximum growth rate in minced pork at a constant temperature of 7 °C was estimated at μmax = 0.680 log CFU/day and μmax = 0.489 log CFU/day in air and MAP, respectively. Model predictions for the growth potential showed acceptable results for air-packed minced pork with better accuracy when the lag phase was implemented as indicated in the renewed protocol (CRL EU, 2021). In MAP, all models used, including the Combase Growth model and to a lesser extent the DMRI dynamic safety model, overestimate the growth potential probably due to a lack of integration of the changing CO2 levels in the packages. The predictive models used in this study do not adequately account for the dynamics in the raw pig matrix, which may have an inhibitory effect on the growth of L. monocytogenes, including interaction with the microbiome and CO2, and emphasize the importance of remaining critical of predictive model outcomes. In addition, the experimental intra- and inter-batch variability raise questions about the sense or nonsense of using predictive microbiology in these raw pork products.