Abstract

In this study we investigated how cell origin could affect the efficacy of an antimicrobial treatment (mild heating combined with terpenoids) in Listeria monocytogenes Scott A, considering cells from: 1. single colony, 2. glycerol stock, 3. cold adapted culture, and 4. fresh culture in stationary phase. After treatment, culturability on BHI medium and viability assessed by flow cytometry were evaluated. Our results showed that the cell origin significantly impacted viability and culturability of L. monocytogenes towards antimicrobial treatment. The mild heat treatment combined or not with terpenoids mainly affected culturability rather than viability, although the culturability of cells from single colony was less impacted. Therefore, to mimic the worst scenario, these latter were selected to contaminate Gorgonzola rind and roast beef slices and we evaluated the ability of L. monocytogenes cells to recover their culturability (on ALOA agar medium) and to growth on the food matrix stored at 4 °C for 7 days. Our results suggest that only Gorgonzola rind allowed a partial recovery of the culturability of cells previously heated in presence or not of terpens. In conclusion, we found a connection between the cell history and sensitivity toward an antimicrobial treatment, underlying the importance to standardize the experimental procedures (starting from the cells to be used in the assay) in the assessment of cell sensitivity to a specific treatment. Finally, our study clearly indicated that VBNC cells can resuscitate under favorable conditions on a food matrix, becoming a threat for consumer’s health.

Highlights

  • IntroductionIn this study we investigated how cell origin could affect the efficacy of an antimicrobial treatment (mild heating combined with terpenoids) in Listeria monocytogenes Scott A, considering cells from: 1. single colony, 2. glycerol stock, 3. cold adapted culture, and 4. fresh culture in stationary phase

  • In this study we investigated how cell origin could affect the efficacy of an antimicrobial treatment in Listeria monocytogenes Scott A, considering cells from: 1. single colony, 2. glycerol stock, 3. cold adapted culture, and 4. fresh culture in stationary phase

  • To examine the impact of cell origin on the efficacy of mild heat treatment combined or not with terpenoids, L. monocytogenes cells were cultivated in different conditions (Fig. 1)

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Summary

Introduction

In this study we investigated how cell origin could affect the efficacy of an antimicrobial treatment (mild heating combined with terpenoids) in Listeria monocytogenes Scott A, considering cells from: 1. single colony, 2. glycerol stock, 3. cold adapted culture, and 4. fresh culture in stationary phase. Therefor strategies based on synergistic effects of thermal treatment with antimicrobials are exploited to avoid excessive heat damages In this perspective, the use of essential oils or their constituents (added below their minimum inhibitory concentration) can be a suitable approach: the presence of thymol, carvacrol, citral, (E)-2-hexenal, vanillin is reported to increase the inactivation rate of thermal ­treatments[6,7,8,9,10,11], due to cumulative antimicrobial effects of heat and aroma compounds. Laboratory tests on stress responses of microorganisms of concern in foods, such as thermal treated L. monocytogenes, are often carried out using liquid batch cultures, which are assumed to be phenotypically homogeneous. The cells deriving from the condition able to guarantee the higher recovery after heat treatments were chosen to perform a trial in food systems, namely Gorgonzola rind and roast beef slices

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