ABSTRACT
Microbial cells respond to sub-lethal stresses with several physiological changes to increase their chance of survival. These changes are of high relevance when combined treatments (hurdle technology) are applied during food production, as the cells surviving the first hurdle may have greater resistance to subsequent treatments than untreated cells. In this study, we analyzed if Salmonella develops increased resistance to thermal treatments after the application of an acid shock. We compared the heat resistance of acid-shocked (pH 4.5 achieved with citric acid) Salmonella cells with that of cells maintained at pH 7 (control cells). Thermal treatments were performed between 57.5 and 65°C. We observed a differential response between the two strains studied. Acid-shocked cells of Salmonella Senftenberg exhibited reduced heat resistance, e.g., for a treatment at 60.0°C and pH 7.0 the time required to reduce the population by 3 log cycles was lowered from 10.75 to 1.98min with respect to control cells. Salmonella Enteritidis showed a different response, with acid-shocked cells having similar resistance than untreated cells (the time required to reduce 3 log cycles at 60.0°C and pH 7.0 was 0.30min for control and 0.31min for acid-shock cells). Based on results by differential plating (with or without adding the maximum non-inhibitory concentration of NaCl to the recovery medium), we hypothesize that the differential response between strains can be associated to sub-lethal damage to the cell membrane of S. Senftenberg caused by the acid shock. These results provide evidence that different strains of the same species can respond differently to an acid shock and highlight the relevance of cross-resistances for microbial risk assessment.
ABSTRACT
Heterogeneity in the response of microbial cells to environmental conditions is inherent to every biological system and can be very relevant for food safety, potentially being as important as intrinsic and extrinsic factors. However, previous studies analyzing variability in the microbial response to thermal treatments were limited to data obtained under isothermal conditions, whereas in the reality, environmental conditions are dynamic. In this article we analyse both empirically and through mathematical modelling the variability in the microbial response to thermal treatments under isothermal and dynamic conditions. Heat resistance was studied for four strains of Listeria monocytogenes (Scott A, CECT 4031, CECT 4032 and 12MOB052), in three different matrices (buffered peptone water, pH 7 Mcllvaine buffer and semi-skimmed milk). Under isothermal conditions, between-strain and between-media variability had no impact in the heat resistance, whereas it was very relevant for dynamic conditions. Therefore, the differences observed under dynamic conditions can be attributed to the variability in the ability for developing stress acclimation. The highest acclimation was observed in strain CECT 4031 (10-fold increase of the D-value), while the lowest acclimation was observed in strain CECT 4032 (50% increase of the D-value). Concerning the different media, acclimation was higher in buffered peptone water and semi-skimmed milk than in Mcllvaine buffer of pH 7.0. To the knowledge of the authors, this is the first research work that specifically analyses the variability of microbial adaptation processes that take place under dynamic conditions. It highlights that microbial heat resistance under dynamic conditions are sometimes determined by mechanisms that cannot be observed when cells are treated in isothermal conditions (e.g. acclimation) and can also be affected by variability. Consequently, empirical evidence on variability gathered under isothermal conditions should be extrapolated with care for dynamic conditions.
Subject(s)
Hot Temperature , Listeria monocytogenes , Milk , Animals , Colony Count, Microbial , Models, TheoreticalABSTRACT
Mathematical models developed in predictive microbiology are nowadays an essential tool for food scientists and researchers. However, advanced knowledge of scientific programming and mathematical modelling are often required in order to use them, especially in cases of modelling of dynamic and/or non-linear processes. This may be an obstacle for food scientists without such skills. Scientific software can help making these tools more accessible for scientists lacking a deep mathematical or computing background. Recently, the R package bioinactivation was published, including functions (model fitting and predictions) for modelling microbial inactivation under isothermal or dynamic conditions. It was uploaded to the Comprehensive R Archive Network (CRAN), but users need basic R programming knowledge in order to use it. Therefore, it was accompanied by Bioinactivation SE, a user-friendly web application including selected functions in the software for users without a programming background. In this work, a new web application, Bioinactivation FE, is presented. It is an extension of Bioinactivation SE which includes an interface to every function in the bioinactivation package: model fitting of isothermal and non-isothermal experiments, and generation of survivor curves and prediction intervals. Moreover, it includes several improvements in the user interface based on the users' feedback. The capabilities of the software are demonstrated through two case studies using data published in the scientific literature. In the first case study, the response of Escherichia coli to isothermal and non-isothermal treatments is compared, illustrating the presence of an induced thermal resistance. In the second, the effect of nanoemulsified d-limonene on the thermal resistance of Salmonella Senftenberg is quantified.
