{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T06:13:54Z","timestamp":1764828834001,"version":"build-2065373602"},"reference-count":71,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2021,1,24]],"date-time":"2021-01-24T00:00:00Z","timestamp":1611446400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"CIISA - Centro de Investiga\u00e7\u00e3o Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterin\u00e1ria, Universidade de Lisboa","award":["Project UIDP\/CVT\/00276\/2020 (funded by FCT)"],"award-info":[{"award-number":["Project UIDP\/CVT\/00276\/2020 (funded by FCT)"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>Listeriosis is almost entirely transmitted through foods contaminated with Listeria monocytogenes. Ready-to-eat foods present a particular challenge due to their long refrigerated shelf-life, not requiring any heat treatment before consumption. In this work, a shelf-life assessment of an industrially produced ready-to-eat salad was performed using conventional culture-based and molecular methods. L. monocytogenes isolates were confirmed and serogrouped using multiplex PCR, and genetic subtyping was performed by pulsed-field gel electrophoresis (PFGE). PMAxx-qPCR was used as an alternative method for L. monocytogenes quantification in foods. Salad samples were kept at 4 \u00b0C, 12 \u00b0C, and 16 \u00b0C for eight days and analysed. At 4 \u00b0C, acceptable results were obtained considering hygiene indicators, i.e., Enterobacteriaceae (ranging from 3.55 \u00b1 0.15 log cfu\/g to 5.39 \u00b1 0.21 log cfu\/g) and aerobic mesophilic colony counts (5.91 \u00b1 0.90 log cfu\/g to 9.41 \u00b1 0.58 log cfu\/g) throughout the study, but the same did not happen at 12 \u00b0C and 16 \u00b0C. L. monocytogenes culture-based quantification exhibited low numbers (&lt;1 log cfu\/g) for all temperatures. From 30 presumptive isolates, 10 (33.3%) were confirmed as L. monocytogenes with the majority belonging to serogroup IVb. PFGE subtyping showed that 7 of the 10 L. monocytogenes isolates had 100% of pulsotype similarity, suggesting a possible common contamination source. PMAxx-qPCR revealed a statistically higher L. monocytogenes quantification (&gt;3 log cfu\/g) when compared to the conventional culture-based method, suggesting viable but non-culturable forms. Taken together, results underline the need to combine conventional methods with more sensitive, specific, and rapid ones for L. monocytogenes assessment in ready-to-eat foods shelf-life studies to reduce the potential risk for consumers.<\/jats:p>","DOI":"10.3390\/foods10020235","type":"journal-article","created":{"date-parts":[[2021,1,25]],"date-time":"2021-01-25T02:07:00Z","timestamp":1611540420000},"page":"235","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Listeria monocytogenes Assessment in a Ready-to-Eat Salad Shelf-Life Study Using Conventional Culture-Based Methods, Genetic Profiling, and Propidium Monoazide Quantitative PCR"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7703-6683","authenticated-orcid":false,"given":"Rita","family":"Bernardo","sequence":"first","affiliation":[{"name":"CIISA\u2013Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, 1300-477 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4001-0871","authenticated-orcid":false,"given":"Ana","family":"Duarte","sequence":"additional","affiliation":[{"name":"CIISA\u2013Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, 1300-477 Lisbon, Portugal"}]},{"given":"Lu\u00eds","family":"Tavares","sequence":"additional","affiliation":[{"name":"CIISA\u2013Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, 1300-477 Lisbon, Portugal"}]},{"given":"Ant\u00f3nio Salvador","family":"Barreto","sequence":"additional","affiliation":[{"name":"CIISA\u2013Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, 1300-477 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9098-4389","authenticated-orcid":false,"given":"Ana Rita","family":"Henriques","sequence":"additional","affiliation":[{"name":"CIISA\u2013Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, 1300-477 Lisbon, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,1,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2153","DOI":"10.1094\/PDIS-03-19-0472-FE","article-title":"Ready-to-eat salad crops: A plant pathogen\u2019s heaven","volume":"103","author":"Gullino","year":"2019","journal-title":"Plant Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1016\/j.foodcont.2010.07.026","article-title":"Listeria monocytogenes in ready-to-eat, sliced, cooked ham and salami products, marketed in the city of S\u00e3o Paulo, Brazil: Occurrence, quantification, and serotyping","volume":"22","author":"Martins","year":"2011","journal-title":"Food Control"},{"key":"ref_3","first-page":"1407216","article-title":"Is your lunch salad safe to eat? 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