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This study aimed to investigate the effects of low-dose continuous ozone gas in controlling Listeria innocua and quality attributes and disorders of Red Delicious apples during long-term commercial cold storage. Red Delicious apples were inoculated with a three-strain L. innocua cocktail at ∼6.2 log 10 CFU/apple, treated with or without 1-methylcyclopropene, and then subjected to controlled atmosphere (CA) storage with or without continuous gaseous ozone in a commercial facility for 36 weeks. Uninoculated Red Delicious apples subjected to the above storage conditions were used for yeast/mold counts and quality attributes evaluation. The 36 weeks of refrigerated air (RA) or CA storage caused ∼2.2 log 10 CFU/apple reduction of L. innocua . Ozone gas application caused an additional > 3 log 10 CFU/apple reduction of L. innocua compared to RA and CA storage alone. During the 36-week CA storage, low-dose continuous gaseous ozone application significantly retarded the growth of yeast/mold, delayed apple firmness loss, and had no negative influence on ozone burn, lenticel decay, russet, CO 2 damage, superficial scald, and soft scald of Red Delicious apples compared to CA-alone storage. In summary, the application of continuous low-dose gaseous ozone has the potential to control Listeria on Red Delicious apples without negatively influencing apple quality attributes., Competing Interests: RB was employed by Stemilt Growers LLC. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Shen, Su, Hua, Sheng, Mendoza, He, Green, Hanrahan, Blakey and Zhu.)

Apples are naturally coated with a water-repelling hydrophobic wax layer, which may limit the antimicrobial efficacies of surface sanitizer solutions. Lauric arginate (LAE) is a cationic surfactant with antimicrobial efficacy against Listeria monocytogenes . In this study, we investigated the antimicrobial and the wettability effects of LAE in enhancing anti- L. monocytogenes efficacy of peracetic acid (PAA) and further verified the optimized treatment combinations in a pilot spray-bar brush bed system. Apples after 48 h of inoculation were treated with PAA surface sanitation in combination with different concentrations of LAE at 22 or 46°C. The effectiveness of PAA with LAE solutions in decontaminating L. monocytogenes significantly increased with the increased concentration of PAA (60-80 ppm) or LAE (0.01-0.05%) or the treatment temperature (from 22 to 46°C). A 30-120-sec wash by 80 ppm PAA with 0.01 and 0.05% LAE at 22°C reduced L. monocytogenes on apples by 2.10-2.25 and 2.48-2.58 log 10 CFU/apple, respectively. Including LAE in the PAA solution decreased contact angles on apple surfaces. However, the increased wettability of the sanitizer solution may not be the main contributor to the enhanced antimicrobial efficacy of the PAA solution, given that the addition of Tween 80 or Tween 85 only slightly boosted the anti- L. monocytogenes efficacy of PAA solutions though both increased the wettability of the PAA solutions. The synergistic effects of PAA and LAE were further validated in a pilot spray-bar brush bed packing system, where a 30-sec spray wash with 80 ppm PAA and 0.05% LAE at 22 and 46°C caused 1.68 and 2.08 log reduction of Listeria on fresh apples, respectively. This study provides an improved PAA process/preventive strategy for ensuring microbial food safety of fresh apples that is applicable to commercial apple packing lines., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Shen, Cong, Mugendi, Hanrahan and Zhu.)

Recent multistate outbreaks and recalls of fresh apples due to Listeria monocytogenes contamination have increased consumer concerns regarding fresh and processed apple safety. This study aimed to evaluate the antimicrobial efficacy of two sanitizers, mineral oxychloride (JC9450) and neutral electrolyzed water (NEW), for inactivation of L. monocytogenes on fresh apples. A 2-min treatment of 0.125% (v/v) JC9450 with 100 ppm free available chlorine (FAC) or NEW with 110 ppm FAC caused 0.9-1.2 log 10 CFU/apple reduction of L. monocytogenes on both Granny Smith and Fuji apples 24 h post-inoculation. Increasing JC9450 concentration to 0.25 and 0.50% significantly improved its bactericidal effect and reduced L. monocytogenes on Granny Smith apples by ~2.0 and 3.8 log 10 CFU/apple, respectively, after a contact time of 2 min. At a shorter contact time of 30 sec, the inactivation efficacy of chlorine and 0.25-0.50% JC9450 against L. monocytogenes on apples was significantly reduced compared with the respective 2-min wash. Furthermore, no L. monocytogenes was recovered in deionized water prepared antimicrobial wash solution or on non-inoculated apples post-NEW with 110 ppm FAC or 0.125-0.5% JC9450 washes, indicating their ability to prevent cross-contamination. In addition, a 2-min exposure to NEW with 110 ppm FAC and 0.50% JC9450 reduced apple native microbiota including total plate count by 0.14 and 0.65 log 10 CFU/apple, respectively, and yeast and mold counts by 0.55 and 1.63 log 10 CFU/apple, respectively. In summary, L. monocytogenes attached on apples was difficult to eliminate. JC9450 and NEW demonstrated a dose-dependent reduction in L. monocytogenes on apples and successfully prevented cross-contamination, indicating their application potential in post-harvest washes of apples., (Copyright © 2020 Sheng, Shen, Ulloa, Suslow, Hanrahan and Zhu.)

Peroxyacetic acid (PAA) is the most commonly used antimicrobial in spray bar antimicrobial treatment during fresh apple packing and processing. However, there are limited data regarding its practical efficacy against Listeria monocytogenes on fresh apples. This study evaluated the antimicrobial activity of PAA against L. monocytogenes on fresh apples applicable to current industry practice, and further examined practical parameters impacting its efficacy to maximize the biocidal effects. Apples were inoculated with a three-strain L. monocytogenes cocktail at ~6.0 Log 10 CFU/apple and then subjected to comparative antimicrobial treatments after 48 h post-inoculation. An 80 ppm PAA treatment, at 30-s and 2-min exposure, reduced L. monocytogenes on fresh apples by ~1.3 or 1.7 Log 10 CFU/apple, respectively. The anti- Listeria efficacy of PAA was not affected by the water hardness and pH of PAA solution, while it improved dramatically when applied at elevated temperature. A 2-min exposure of 80 ppm PAA at 43 and 46°C resulted in a 2.3 and 2.6 Log 10 CFU/apple reduction, respectively. A 30-s contact time of 80 ppm PAA at 43-46°C reduced L. monocytogenes on apples by 2.2-2.4 Log 10 CFU/apple. Similarly, PAA intervention at elevated temperatures significantly strengthened its effectiveness against naturally occurring apple microbiota. PAA treatment at 43-46°C can provide a vital method to improve antimicrobial efficacy against both L. monocytogenes and indigenous microbiota on fresh apples. Our data provide valuable information and reference points for the apple industry to further validate or verify process controls.

Listeria monocytogenes contaminated processing equipment and the general packing environment have been implicated in deadly foodborne listeriosis outbreaks, highlighting the significance of proper sanitization and disinfection of food contact surfaces. This study aims to comprehensively evaluate antimicrobial efficacy of commercially available, economical sanitizers at practical concentrations against L. monocytogenes biofilm formed on polystyrene surfaces under different conditions. Ozonated water 1-min treatment at 1.0, 2.0, and 4.0 ppm resulted in ∼0.9, 3.4, and 4.1 log reduction of L. monocytogenes single strain biofilm grown on polystyrene surfaces, respectively. However, its efficacy was dramatically diminished in multi-strain L. monocytogenes biofilm and was further compromised by aged biofilm and in the presence of organic matter. Quaternary ammonium compounds (QAC) at 100/400 ppm, chlorine at 100/200 ppm, chlorine dioxide at 2.5/5.0 ppm and peroxyacetic acid (PAA) at 80/160 ppm resulted in 2.4/3.6, 2.0/3.1, 2.4/3.8, and 3.6/4.8 log reduction of L. monocytogenes single strain biofilm, respectively. Antimicrobial efficacies of all tested sanitizers against 7-day-old biofilm were much lower when compared to 2-day-old biofilm, with PAA being the least influenced by the age of the biofilm. Organic matter conditioning with diluted milk or apple juice dramatically impacted the antimicrobial efficacy of all sanitizers. PAA treatment of 1 min at 160-200 ppm resulted in a 3.2-3.5 log reduction against 7-day-old biofilm in the presence of organic matter, thus showing its effectiveness in eradicating L. monocytogenes biofilm on polystyrene surface. Collectively, data highlight the importance of timely and thoroughly cleaning food contact surfaces before disinfection and provides practical information and guidance for the food industry in selecting the most effective sanitizer in their sanitizing regimes to eliminate L. monocytogenes biofilm.

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 Log 10 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 Log 10 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 Log 10 CFU/apple for both inoculation levels. L. monocytogenes had similar survival pattern on conventional Granny Smith and Fuji apples with 0.8-2.0 Log 10 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.