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Mature Listeria monocytogenes biofilms exhibit reduced susceptibility to sanitizers - relevance to the (leafy green) fresh food supply chain

Mature Listeria monocytogenes biofilms exhibit reduced susceptibility to sanitizers - relevance to the (leafy green) fresh food supply chain
Mature Listeria monocytogenes biofilms exhibit reduced susceptibility to sanitizers - relevance to the (leafy green) fresh food supply chain

Salads and herbs are important for a healthy diet but during their processing and packaging, they may be exposed to environmental contamination from foodborne pathogens. Of particular concern is Listeria monocytogenes, a Gram-positive, facultative anaerobe, found ubiquitously in the environment. L. monocytogenes can survive and grow under harsh conditions such as refrigeration temperatures, low oxygen or nutrient concentrations, which is why it is a problem in the fresh food supply chain. Infection with L. monocytogenes can result in listeriosis, potentially fatal in immunocompromised patients, pregnant women, newborn babies, and the elderly. This study aims to evaluate the efficacy of common sanitizing methods used in the fresh food supply chain, where biofilm formation has raised concerns, using appropriately developed laboratory models of Listeria biofilms. L. monocytogenes Scott A, L. monocytogenes CECT 936, and L. innocua NCTC 12210 biofilms were grown at 20 °C or 4 °C, on stainless steel coupons for 7 days, and treated with high concentrations of chlorine (up to 300 ppm) or peracetic acid (up to 500 ppm) on days 1, 3, 5, and 7. Coupons were then processed for culturable cell counts on treatment days, and imaged using episcopic differential interference microscopy, coupled with epi-fluorescence microscopy (EDIC/EF) on day 7 of growth. This determined that temperature affects biofilm growth on stainless steel, as biofilm growth reached ∼8 log 10 CFU/cm 2 at 20 °C, but was significantly lower at 4 °C (∼4 log 10 CFU/cm 2) – highlighting the importance of maintaining a cold chain. Chlorine and peracetic acid were shown to be effective at treating Listeria in the planktonic form but were not effective at treating aged biofilms at both temperatures and the high concentrations of sanitizers used. This work provides important information on sanitizing efforts in the fresh food supply chain, concerning factory temperature, processing surfaces, and the age of biofilm.

Biofilm, Food industry, Foodborne pathogen chlorine, Listeria monocytogenes, Peracetic acid
0362-028X
Sutton, Lucy
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Humphreys, Megan
fe624f75-6db5-4248-8a9d-94cb1a97c15f
Highmore, Callum
04809bd8-7cad-4dcf-b67d-264697f893b9
Wilks, Sandra
86c1f41a-12b3-451c-9245-b1a21775e993
Keevil, Charles William
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Sutton, Lucy
aa82c927-bcba-4a6d-8d88-fbf23045d2ad
Humphreys, Megan
fe624f75-6db5-4248-8a9d-94cb1a97c15f
Highmore, Callum
04809bd8-7cad-4dcf-b67d-264697f893b9
Wilks, Sandra
86c1f41a-12b3-451c-9245-b1a21775e993
Keevil, Charles William
cb7de0a7-ce33-4cfa-af52-07f99e5650eb

Sutton, Lucy, Humphreys, Megan, Highmore, Callum, Wilks, Sandra and Keevil, Charles William (2025) Mature Listeria monocytogenes biofilms exhibit reduced susceptibility to sanitizers - relevance to the (leafy green) fresh food supply chain. Journal of Food Protection, 88 (12), [100652]. (doi:10.1016/j.jfp.2025.100652).

Record type: Article

Abstract

Salads and herbs are important for a healthy diet but during their processing and packaging, they may be exposed to environmental contamination from foodborne pathogens. Of particular concern is Listeria monocytogenes, a Gram-positive, facultative anaerobe, found ubiquitously in the environment. L. monocytogenes can survive and grow under harsh conditions such as refrigeration temperatures, low oxygen or nutrient concentrations, which is why it is a problem in the fresh food supply chain. Infection with L. monocytogenes can result in listeriosis, potentially fatal in immunocompromised patients, pregnant women, newborn babies, and the elderly. This study aims to evaluate the efficacy of common sanitizing methods used in the fresh food supply chain, where biofilm formation has raised concerns, using appropriately developed laboratory models of Listeria biofilms. L. monocytogenes Scott A, L. monocytogenes CECT 936, and L. innocua NCTC 12210 biofilms were grown at 20 °C or 4 °C, on stainless steel coupons for 7 days, and treated with high concentrations of chlorine (up to 300 ppm) or peracetic acid (up to 500 ppm) on days 1, 3, 5, and 7. Coupons were then processed for culturable cell counts on treatment days, and imaged using episcopic differential interference microscopy, coupled with epi-fluorescence microscopy (EDIC/EF) on day 7 of growth. This determined that temperature affects biofilm growth on stainless steel, as biofilm growth reached ∼8 log 10 CFU/cm 2 at 20 °C, but was significantly lower at 4 °C (∼4 log 10 CFU/cm 2) – highlighting the importance of maintaining a cold chain. Chlorine and peracetic acid were shown to be effective at treating Listeria in the planktonic form but were not effective at treating aged biofilms at both temperatures and the high concentrations of sanitizers used. This work provides important information on sanitizing efforts in the fresh food supply chain, concerning factory temperature, processing surfaces, and the age of biofilm.

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Accepted/In Press date: 19 October 2025
e-pub ahead of print date: 23 October 2025
Published date: 10 December 2025
Additional Information: Copyright © 2025. Published by Elsevier Inc.
Keywords: Biofilm, Food industry, Foodborne pathogen chlorine, Listeria monocytogenes, Peracetic acid

Identifiers

Local EPrints ID: 507052
URI: http://eprints.soton.ac.uk/id/eprint/507052
ISSN: 0362-028X
PURE UUID: 14d1bf16-6618-43fc-adc3-89a777a5deea
ORCID for Callum Highmore: ORCID iD orcid.org/0000-0003-0388-4422
ORCID for Sandra Wilks: ORCID iD orcid.org/0000-0002-4134-9415
ORCID for Charles William Keevil: ORCID iD orcid.org/0000-0003-1917-7706

Catalogue record

Date deposited: 25 Nov 2025 18:05
Last modified: 26 Nov 2025 02:56

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Contributors

Author: Lucy Sutton
Author: Megan Humphreys
Author: Callum Highmore ORCID iD
Author: Sandra Wilks ORCID iD

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