Efficacy of cleaning and sanitation methods against Listeria innocua on apple packing equipment surfaces.

Previous foodborne listeriosis outbreaks and recalls of fresh produce have been linked to cross-contamination with food contact surfaces (FCS) of packing equipment. Thus, effective cleaning and sanitation practices should be implemented in the short-term to contribute to the overall food safety objective for FCS which have a suboptimal hygienic design. This research aimed to evaluate the efficacy of seven cleaning and sanitation treatment combinations against Listeria innocua populations on FCS common to produce packinghouses that have been found to have a higher prevalence of Listeria spp. harborage. Polishing brushes made of two different materials (100% nylon and nylon/horsehair mix), 100% polyethylene wash brushes, stainless steel rollers and polytetrafluorethylene (Teflon®) wrapped rollers, and interlocking conveyor belts were evaluated (n = 6 per treatment). These FCS were inoculated with L. innocua (9 log CFU/mL) and fouled with food-grade wax, with the exception of brush rollers that are encountered before waxing. Treatments included the use chlorine (200 ppm), peroxyacetic acid (PAA) (500 ppm) for 15 min, alone or in combination with an alkaline detergent (1.6%) or a degreaser, and the use of steam at 95 °C for 15 s. L. innocua was enumerated and the log reduction was calculated and compared to untreated controls. Horsehair mix polishing brushes were the surface with the lowest log reduction regardless of treatment applied (p < 0.05). Compared to 100% nylon polishing brushes, where a >3 log reduction was reached, horsehair mix brushes only reached this level of reduction when degreaser + PAA was applied. For both types of rollers and interlocking conveyor belt, an effective wax removal using a degreaser or detergent followed by sanitizer application caused the greatest L. innocua reduction (>5 log reduction). The application of steam did not show a significant log reduction on any surface (p > 0.05). This study highlights that cleaning and sanitation strategies must focus on effective wax removal if applied postharvest. In addition, 100% nylon polishing brushes could potentially offer a better hygienic design in produce packinghouses compared to the horsehair mix.

Prevalence of Listeria Species on Food Contact Surfaces in Washington State Apple Packinghouses.

The 2014 caramel apple listeriosis outbreak was traced back to cross-contamination between food contact surfaces (FCS) of equipment used for packing and fresh apples. For Washington state, the leading apple producer in the United States with 79% of its total production directed to the fresh market, managing the risk of apple contamination with Listeria monocytogenes within the packing environment is crucial. The objectives of this study were to determine the prevalence of Listeria spp. on FCS in Washington state apple packinghouses over two packing seasons and to identify those FCS types with the greatest likelihood to harbor Listeria spp. Five commercial apple packinghouses were visited quarterly over two consecutive year-long packing seasons. A range of 27 to 50 FCS were swabbed at each facility to detect Listeria spp. at two sample times, (i) postsanitation and (ii) in-process (3 h of packinghouse operation), following a modified protocol of the FDA's Bacteriological Analytical Manual method. Among 2,988 samples tested, 4.6% (n = 136) were positive for Listeria spp. Wax coating was the unit operation from which Listeria spp. were most frequently isolated. The FCS that showed the greatest prevalence of Listeria spp. were polishing brushes, stainless steel dividers and brushes under fans/blowers, and dryer rollers. The prevalence of Listeria spp. on FCS increased throughout apple storage time. The results of this study will aid apple packers in controlling for contamination and harborage of L. monocytogenes and improving cleaning and practices for sanitation of the FCS on which Listeria spp. are the most prevalent.IMPORTANCE Since 2014, fresh apples have been linked to outbreaks and recalls associated with postharvest cross-contamination with the foodborne pathogen L. monocytogenes These situations drive both public health burden and economic loss and underscore the need for continued scrutiny of packinghouse management to eliminate potential Listeria niches. This research assesses the prevalence of Listeria spp. on FCS in apple packinghouses and identifies those FCS most likely to harbor Listeria spp. Such findings are essential for the apple-packing industry striving to further understand and exhaustively mitigate the risk of contamination with L. monocytogenes to prevent future listeriosis outbreaks and recalls.

Comparison of Three Air Sampling Methods for the Quantification of Salmonella, Shiga-toxigenic Escherichia coli (STEC), Coliforms, and Generic E. coli from Bioaerosols of Cattle and Poultry Farms.

Recent fresh produce outbreaks potentially associated with bioaerosol contamination from animal operations in adjacent land highlighted the need for further study to better understand the associated risk. The purpose of this research was to evaluate three sampling methods for quantifying target bacterial bioaerosols from animal operations. A dairy cattle and poultry farm located in Georgia, U.S. were visited six times each. Air was collected for 10 min using: 2-stage Andersen impactor with and without mineral oil overlay and impingement samplers. Sampling devices were run concurrently at 0.1, 1, and 2 m heights (n = 36). Andersen samplers were loaded with CHROMagar™ Salmonella, CHROMagar™ STEC, or Brilliance™ coliforms/E. coli. The impingement sampler contained buffered peptone water (20 mL) which was vacuum filtered through a 0.45 µm filter and placed onto the respective media. Plates were incubated at 37 ℃ for 48 h. PCR confirmation followed targeting ttr for Salmonella and stx1, stx2, and eae genes for STEC. No significant differences were found among methods to quantify coliforms and E. coli. Salmonella and STEC bioaerosols were not detected by any of the methods (Limit of detection: 0.55 log CFU/m3). E. coli bioaerosols were significantly greater in the poultry (2.76-5.00 log CFU/m3) than in the cattle farm (0.55-2.82 log CFU/m3) (p < 0.05), and similarly distributed at both stages in the Andersen sampler (stage 1:>7 µm; stage 2: 0.65-7 µm particle size). Sampling day did not have a significant effect on the recovery of coliforms/E. coli bioaerosols in the poultry farm when samples were taken at the broiler house exhaust fan (p > 0.05). A greater and constant emission of coliforms and E. coli bioaerosols from the poultry farm warrants further investigation. These data will help inform bioaerosol sampling techniques which can be used for the quantification of bacterial foodborne pathogens and indicator organisms for future research.

Persistence of Listeria innocua on Fresh Apples during Long-Term Controlled Atmosphere Cold Storage with Postharvest Fungal Decay.

ABSTRACT: Recent apple-related recall and outbreak events have exposed a need for better food safety controls along the supply chain. Following harvest, apples can be stored under a controlled atmosphere for up to 1 year after harvest before packing and distribution, making the crop susceptible to many opportunities for contamination that increase the quantity of postharvest losses. Botrytis cinerea and Penicillium expansum cause significant rot-associated losses to the apple industry. These fungi can colonize and destroy apple tissue as storage duration increases, which may also impact the growth of saprophytic foodborne pathogens like Listeria monocytogenes. Thus, the objective of this study was to observe population changes of Listeria innocua as a surrogate for L. monocytogenes on apples inoculated with B. cinerea or P. expansum under long-term controlled atmosphere cold storage conditions to identify the effect of postharvest mold growth on growth patterns of a microorganism relevant to food safety. 'Gala' and 'WA 38' apples (n = 1,080) were harvested, treated with pyrimethanil, and inoculated with L. innocua only or with L. innocua and one of the mold species on wounded and unwounded portions of the apple equator. Apples were treated with 1-methylcyclopropene and stored at a controlled atmosphere (2 kPa O2, 1 kPa CO2, 1°C) for 1 week and 1, 3, 6, 9, and 11 months before enumeration. After 3 months, L. innocua consistently fell below the limit of detection (2.35 Log CFU/g), and samples were enriched following a modified Bacteriological Analytical Manual method with PCR confirmation. Listeria persistence was dependent on the storage duration and type of fungal contamination (P ≤ 0.05). Surface wounding may impact these trends, depending on the apple variety. Prevalence of L. innocua was greater in Gala apples. Future studies should more closely examine the interactions on the fruit surface that occur during the seemingly critical time frame of 3 to 6 months in storage.

Utility of rapid tests to assess the prevalence of indicator organisms (Aerobic plate count, Enterobacteriaceae, coliforms, Escherichia coli, and Listeria spp.) in apple packinghouses.

The 2014 listeriosis outbreak caused by caramel-coated apples was linked to apples cross-contaminated within an apple packing facility. This outbreak has increased the focus on effective cleaning and sanitation methods that must be validated and monitored during apple packing. Thus, rapid and reliable testing methods are necessary for assessing cleanliness in the apple packing industry. The objectives of this study were to assess the prevalence of common indicator organisms [Aerobic plate count (APC), Enterobacteriaceae, coliforms, Escherichia coli, and Listeria spp.] on food contact surfaces (zone 1) in apple packinghouses and to evaluate the utility and accuracy of currently used rapid tests (ATP and glucose/lactose residue swabs). Food contact surfaces were sampled over a 100 cm2 area in five commercial apple packinghouses to evaluate populations of indicator organisms APC, Enterobacteriaceae, coliforms, E. coli (n = 741), and rapid test readings (n = 659). Petrifilm plates were used for the quantification of APC, Enterobacteriaceae, and coliform/E. coli. Rapid tests [ATP swabs (UltraSnap) and glucose/lactose residue swabs (SpotCheck Plus)] were processed on-site. A larger area (0.93 m2) was sampled for the detection of Listeria spp. (n = 747), following a modified protocol of the FDA's Bacteriological Analytical Manual method, and confirmed with PCR and gel electrophoresis via the iap gene. No significant association was found between either rapid test and populations of APC, Enterobacteriaceae, coliforms, E. coli, and Listeria spp. detection. However, recovery of APC (log CFU/100 cm2) was higher with a failed glucose/lactose residue swab surface hygiene result (3.1) than a passed result (2.9) (p = 0.03). Populations of APC, Enterobacteriaceae, and coliforms were significantly different at each unit operation during the packing process (p ≤ 0.05). This study concluded that ATP and glucose/lactose residue rapid tests were poorly suited for determining microbial load since they were not related to populations of any common indicator organisms or the detection of Listeria spp. These findings emphasize the need to utilize a rapid test, which can be a good indicator of residual matter on a surface, along with traditional microbiological methods to assess cleaning and sanitation practices in apple packinghouses.