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Fate of Listeria monocytogenes on Fresh Apples under Different Storage Temperatures.

ABSTRACT: Fresh apples are typically stored for up to 1 year commercially; different apple varieties require different storage temperatures to maintain their quality characteristics. There is sparse information available about Listeria monocytogenes survival on fresh apples under various storage temperatures. The objective of this study was to comprehensively evaluate the effect of storage temperature on apple fruit decay and L. monocytogenes survival. Unwaxed apple fruits of selected varieties (Fuji and Granny Smith) were dip inoculated in a three-strain L. monocytogenes cocktail to establish ∼3.5 and 6.0 Log10 CFU/apple. Twenty-four hours post-inoculation, apples were subjected to 1, 4, 10, or 22°C storage for up to 3 months. Apples under the different storage treatments were sampled at 1-, 4-, 7- and 14-day for short-term storage under all four tested temperatures, and 2-, 4-, 8-, and 12-week for long-term storage at 1, 4, and 10°C. A set of uninoculated and unwaxed apples were simultaneously subjected to the previously mentioned storage temperatures and sampled biweekly for their total bacterial count (TPC) and yeasts/molds (Y/M) count. During the 2-week short-term storage, L. monocytogenes population on organic Granny Smith apples stored at 1, 4, or 10°C was reduced by 0.2-0.3 Log. When apples were stored at 22°C, there was a 0.5-1.2 Log10 CFU/apple reduction 14-day post storage dependent on the initial inoculation level. During the 12-week cold storage under 1, 4, and 10°C, L. monocytogenes count on organic Granny Smith apples decreased by 0.5-1.5 Log10 CFU/apple for both inoculation levels. L. monocytogenes had similar survival pattern on conventional Granny Smith and Fuji apples with 0.8-2.0 Log10 CFU/apple reduction over a 3-month cold storage period. Interestingly, both TPC and Y/M count were stable regardless of apple variety or cultivation practice during the 12-week storage at all tested temperatures. In summary, while L. monocytogenes did not proliferate on apple surfaces during 12 weeks of refrigerated storage, only a limited reduction of L. monocytogenes was observed in this study. Therefore, the apple industry cannot rely on cold storage alone to control this pathogen. Additional interventions are needed to eradicate Listeria on fresh apples during long-term cold storage.

SUBMITTER: Sheng L

PROVIDER: S-EPMC5522875 | biostudies-other | 2017

REPOSITORIES: biostudies-other

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Project description:Listeria monocytogenes is a foodborne pathogen that can cause invasive severe human illness (listeriosis) in susceptible patients. Most human listeriosis cases appear to be caused by consumption of refrigerated ready-to-eat foods. Although initial contamination levels in foods are usually low, the ability of these bacteria to survive and multiply at low temperatures allows it to reach levels high enough to cause disease. This study explores the set of proteins that might have an association with L. monocytogenes adaptation to different temperatures. Cultures were grown in biofilm, the most widespread mode of growth in natural and industrial realms. Protein extractions were performed from three different growth temperatures (10, 25, and 37°C) and two growth phases (early stage and mature biofilm). L. monocytogenes subproteomes were targeted using three extraction methods: trypsin-enzymatic shaving, biotin-labeling and cell fractionation. The different subproteomes obtained were separated and analyzed by shotgun proteomics using high-performance liquid chromatography combined with tandem mass spectrometry (LC-OrbiTrap LTQVelos, ThermoFisher Scientific). A total of 141 (biotinylation), 98 (shaving) and 910 (fractionation) proteins were identified. Throughout the 920 unique proteins identified, many are connected to basic cell functions, but some are linked with thermoregulation. We observed some noteworthy protein abundance shifts associated with the major adaptation to cold mechanisms present in L. monocytogenes, namely: the role of ribosomes and the stressosome with a higher abundance of the general stress protein Ctc (Rl25) and the general stress transcription factor sigma B (σB), changes in cell fluidity and motility seen by higher levels of foldase protein PrsA2 and flagellin (FlaA), the uptake of osmolytes with a higher abundance of glycine betaine (GbuB) and carnitine transporters (OpucA), and the relevance of the overexpression of chaperone proteins such as cold shock proteins (CspLA and Dps). As for 37°C, we observed a significantly higher percentage of proteins associated with transcriptional or translational activity present in higher abundance upon comparison with the colder settings. These contrasts of protein expression throughout several conditions will enrich databases and help to model the regulatory circuitry that drives adaptation of L. monocytogenes to environments.

Project description:Whole and fresh-cut produce are minimally processed and susceptible to microbial contamination. This study evaluated the survival or growth of L. monocytogenes on peeled rinds, and fresh-cut produce at different storage temperatures. Fresh-cut fruits and vegetables, including cantaloupe, watermelon, pear, papaya, pineapple, broccoli, cauliflower, lettuce, bell pepper, and kale (25 g pieces) were spot inoculated with 4 log CFU/g of L. monocytogenes and stored at 4 or 13°C for 6 days. Cantaloupe and bell pepper rind disks (20 cm2), mimicking whole produce were inoculated with low inoculum level (4 log CFU/mL) and high inoculum level (6 log CFU/mL) and stored at 24°C up to 8 days and 4°C up to 14 days, respectively. L. monocytogenes counts on fresh-cut pear samples stored at 4°C increased significantly by 0.27 log CFU/g. However, Listeria levels on kale (day 4), cauliflower (day 6), and broccoli (day 2) were significantly reduced by 0.73, 1.18, and 0.80 log CFU/g, respectively, at 4°C. At 13°C, the bacterial counts increased significantly after a day of storage on fresh-cut watermelons (increasing by 1.10 log CFU/g) and cantaloupes (increasing by 1.52 log CFU/g). Similar increases were observed on pears (1.00 log CFU/g), papayas (1.65 log CFU/g), and green bell peppers (1.72 log CFU/g). Pineapple samples did not support the growth of L. monocytogenes at 13°C with a significant reduction of 1.80 log CFU/g by day 6. L. monocytogenes levels significantly increased in fresh-cut lettuce at 13°C but remained stable on kale, cauliflower, and broccoli after 6 days of storage. Stable population was observed also on cantaloupe rinds up to 8 days at 24°C. While on the outer surface of bell peppers, the population level decreased below the detectable limit of the test (10 CFU/20 cm2) after 14 days of storage at 4°C. The results demonstrated variable survival behavior of L. monocytogenes on fresh-cut produce with produce type and storage temperature.

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Fate of Listeria monocytogenes on Fresh Apples under Different Storage Temperatures.

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