Irrigation method matters: Contamination and die-off rates of Escherichia coli on dry bulb onions after overhead and drip irrigation in Washington State (2022-2023).

Two U.S. outbreaks of salmonellosis in 2020 and 2021 were epidemiologically linked to red onions. The 2020 outbreak investigation implicated production agricultural water as a likely contamination source. Field trials were designed to investigate prevalence and survival of Escherichia coli (surrogate for Salmonella) on dry bulb onions after application of contaminated irrigation water at the end of the growing period. Irrigation water was inoculated at 3 log most probable number (MPN)/100 mL (2022 and 2023) or 5 log MPN/100 mL (2023, drip only) with a cocktail of rifampin-resistant E. coli and applied with the final irrigation (0.4 acre-inch/0.4 hectare-cm) to onions. Onion bulbs (40 or 80) were sampled immediately after irrigation and throughout field curing (4 weeks) and E. coli was enumerated using a MPN method. For drip irrigation, at 3 log MPN/100 mL E. coli was detected on 13% of onions at 24 h but not detected at 0 h; at 5 log MPN/100 mL for drip irrigation applied to saturated soil, E. coli was detected in 63% of onions at 0 h. Prevalence significantly (P<0.05), decreased after 7 d of curing with cell densities of 1-1,400 MPN/onion. At the end of field curing in 2023, 1/80 onions had detectable E. coli (2.04 MPN/onion). E. coli was detected in a significantly smaller percentage of onions (2022: 13%; 2023: 68%) after a contaminated drip irrigation event compared to overhead irrigation (98-100%; P<0.05). After overhead irrigation E. coli was detected in onions (1-1,000 MPN/onion) on day 0. Prevalence decreased significantly (P <0.05) after 7 d of field curing in both years (2022: 15%; 2023: 7%). E. coli was not detected on Calibra onions (80/year) at the end of field curing in either year but was detected at <12 MPN/onion in 2.5-3.75% of onions (n=80) for other cultivars. These data confirm limited contamination risk associated with drip irrigation water quality and begin to quantify contamination risks associated with overhead irrigation of dry bulb onions.

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.

Ranking Food Safety Priorities of the Fresh Produce Industry in the United States.

A broad understanding of community member food safety priorities in the fresh produce supply chain does not currently exist. This information is essential to improve food safety knowledge and practices effectively and efficiently throughout the fresh produce industry; therefore, the goal of this study was to identify and rank community produce safety priorities in the United States. Survey questions were designed and approved by food safety experts for participants to rank 24 fresh produce safety priorities. The anonymous survey was distributed online via Qualtrics™ to fresh produce community members from November 2020 to May 2021. A score was calculated for each priority by summing weighted ranking scores across responses. Descriptive statistics and logistic regression were used to determine frequencies and distribution of response and identify factors (e.g., role in produce safety, size/location of organization/operation) that influenced rankings. A total of 281 respondents represented fourteen different roles in the fresh produce industry, with most identified as growers (39.5%). Produce operations were distributed across the U.S. and annual produce sales ranged from below $25,000 to over $5,000,000. Health and hygiene, training, postharvest sanitation, traceability, and harvest sanitation were ranked as the top five food safety priorities. These findings provide insight into community member priorities in fresh produce safety and can be used to inform intervention efforts, ranging from specialized training for produce growers and packers, industry-driven research projects, and gaps in risk communication strategies.

Impact of chlorine or peracetic acid on inactivation of Salmonella, Escherichia coli, and Listeria monocytogenes in agricultural water.

Preharvest agricultural water has been recognized as one of the routes of contamination for foodborne pathogens during fruit and vegetable production. Several strategies have been proposed to reduce the risk of pathogens, including preharvest water chemigation, but literature is lacking with regards to microbiological inactivation of common bacterial foodborne pathogens associated with fresh produce contamination, Salmonella enterica, Shiga-toxigenic Escherichia coli (STEC), and Listeria monocytogenes, in surface irrigation water after exposure to chlorine and peracetic acid (PAA). Surface water supplied by a local irrigation district was collected over the summer of 2019. Water was autoclaved, divided into 100 mL samples, and inoculated with a cocktail of five Salmonella, STEC, or Listeria monocytogenes strains or a single strain non-pathogenic E. coli. Samples were then treated with 3, 5, or 7 ppm of free chlorine or PAA, and surviving populations were evaluated using a time-kill assay. A first-order kinetic model was used to fit the inactivation data and obtain the D-values. A secondary model was used to explain the changes due to the type of water, treatment, and microorganism. At a concentration of 3 ppm, the observed and predicted D-values of free chlorine treatments were higher than that of PAA treatments for ground and surface water. Results indicated that PAA was more effective inactivating bacteria than sodium hypochlorite at concentrations of 3 and 5 ppm for both water sources (surface and ground). However, at 7 ppm, the effectiveness of PAA and sodium hypochlorite showed no statistically significant difference for both surface and groundwater. Findings will provide information regarding efficacy of chemical sanitizers like chlorine and PAA for inactivation of Salmonella, Listeria, and STEC in surface water from which treatments can be derived. Ultimately benefitting growers in the selection of an appropriate method for in-field treatment of irrigation water if deemed necessary.

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.

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.

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.

Prevalence of Salmonella enterica in Flies on a Diversified Cattle and Fresh Produce Farm across Two Growing Seasons.

ABSTRACT: Flies are a vector for spreading foodborne pathogens pertinent to fresh produce, such as Shiga toxigenic Escherichia coli and Salmonella; however, most studies focus on concentrated animal feeding operations, which do not reflect low-density animal farming practices that often adjoin fruit and vegetable acreage. In this study, we determined the prevalence of Salmonella in flies collected biweekly on an integrated animal and produce operation over two growing seasons. Eleven of 889 pooled samples tested positive for Salmonella. Flies from the Calliphoridae, Muscidae, Sarcophagidae, and Tachinidae families were associated with Salmonella carriage, but fly family was not a significant factor for isolation of Salmonella (P = 0.303). Fly species were a significant factor (P = 0.026), with five Pentacricia aldrichii pools testing positive for Salmonella. With the exception of single specimen isolation, prevalence ranged from 2.2 to 15.2%. With the exception of the Tachinidae family, these results reflect a strong association of flies that are commonly associated with feces or are pests of animals. Trap location was not significantly associated with isolation of Salmonella-positive flies (P = 0.236). Overall, the population of flies was not as abundant as studies conducted with produce grown close to concentrated animal feeding operations, indicating a reduced risk of transmission; however, similar to these studies, fly families that are commonly isolated from fecal and decaying matter were most frequently associated with Salmonella isolation. Further work is warranted to elucidate the foodborne pathogen transmission rates to produce and subsequent survival over time.

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.

Optimization of a Method for the Concentration of Genetic Material in Bacterial and Fungal Communities on Fresh Apple Peel Surfaces.

Apples are the most consumed fruit in the United States and have recently been shown to exhibit some vulnerability to contamination across the supply chain. It is unclear what role a fruit microbiome analysis may serve in future food safety programs interested in understanding changes in the product and the processing environment. Ultimately, sample integrity is key if any of these approaches are to be employed; low microbial loads on apple surfaces, the inability to sample the entire surface, and inefficiency of removal may act as barriers to achieving high-quality DNA. As such, the objective of this study was to identify a reproducible method to concentrate and quantify bacterial and fungal DNA from fresh apple surfaces. Five methods were evaluated: two variations of wash solutions for bath sonication, wash filtration, epidermis excision, and surface swabbing. Epidermis excision returned the highest mean DNA quantities, followed by the sonicated washes and wash filtration. Surface swabbing was consistently below the limit of detection. Based on the quantity of host DNA contamination in surface excision, the sonicated wash solution containing a surfactant presents the greatest opportunity for consistent, high-yielding DNA recovery from the entire apple surface.

Prevalence, Persistence, and Diversity of Listeria monocytogenes and Listeria Species in Produce Packinghouses in Three U.S. States.

ABSTRACT: Listeria monocytogenes has emerged as a food safety concern for several produce commodities. Although L. monocytogenes contamination can occur throughout the supply chain, contamination from the packinghouse environment represents a particular challenge and has been linked to outbreaks and recalls. This study aimed to investigate the prevalence, persistence, and diversity of L. monocytogenes and other species of Listeria in produce packinghouses. A longitudinal study was performed in 11 packinghouses (whose commodities included microgreen, peach, apple, tomato, broccoli, cauliflower, and cucumber) in three U.S. states. In each packinghouse, 34 to 47 sites representing zones 2 to 4 were selected and swabbed. Packinghouses were visited four times over the packing season, and samples were tested for Listeria by following the U.S. Food and Drug Administration's Bacteriological Analytical Manual methods. Presumptive Listeria-positive isolates were confirmed by PCR. Species and allelic type (AT) were identified by sigB sequencing for up to eight isolates per sample. Among 1,588 samples tested, 50 (3.2%), 42 (2.7%), and 10 (0.6%) samples were positive for L. monocytogenes only, Listeria spp. (excluding L. monocytogenes) only, and both L. monocytogenes and Listeria spp., respectively. Five species of Listeria (L. monocytogenes, L. innocua, L. seeligeri, L. welshimeri, and L. marthii) were identified, and L. monocytogenes was the most prevalent species. The 102 Listeria-positive samples yielded 128 representative isolates (i.e., defined as isolates from a given sample with a different AT). Approximately 21% (21 of 102) of the Listeria-positive samples contained two or more ATs. A high AT diversity (0.95 Simpson's diversity index) was observed among Listeria isolates. There were three cases of L. monocytogenes or Listeria spp. repeated isolation (site testing positive at least twice) based on AT data. Data from this study also support the importance of drain and moisture management, because Listeria were most prevalent in samples collected from drain, cold storage, and wet nonfood contact surface sites.

Transcriptomic Behavior of Salmonella enterica Newport in Response to Oxidative Sanitizers.

ABSTRACT: Agricultural water is a known source of contamination to fresh produce and can contain foodborne pathogens including Salmonella enterica, pathogenic Escherichia coli, Listeria monocytogenes, and Campylobacter jejuni. To mitigate such risks, antimicrobial agents such as hypochlorites and peroxyacetic acid (PAA) can be applied to in-line irrigation systems as well as to water used in postharvest washing. Although these compounds are effective and widely used, some pathogenic bacteria adapt to survive exposure. RNA sequencing was used to analyze the Salmonella Newport transcriptome after exposure to sodium hypochlorite (NaOCl) and PAA in a simulated agricultural water system. Overall cellular adaptive response was determined quantitatively as a function of overall gene expression of the >4,000 genes in the Salmonella Newport genome. Differentially expressed genes ranged from 11 due to 10-ppm NaOCl treatment, 316 due to 20-ppm NaOCl treatment, 1,719 due to 10-ppm PAA treatment, and 2,010 due to 20-ppm PAA treatment compared with that of the controls (water only). Differentially expressed transcripts included cellular functions such as biosynthesis, degradation, energy generation, and nonmetabolically linked functions. Oxidative exposure upregulated genes associated with key virulence, attachment, and gene transfer. Amino acid biosynthesis was upregulated due to NaOCl exposure but primarily downregulated when Salmonella Newport was exposed to PAA. Slight upregulation occurred in nucleoside and nucleotide biosynthesis, a known DNA repair mechanism seen during exposure to sanitizers. Our results indicate that Salmonella Newport reacts differently when exposed to NaOCl versus PAA, despite oxidative activity being the primary modes of antimicrobial action of both compounds.

Potential of Cinnamon Oil Emulsions as Alternative Washing Solutions of Carrots.

The objective of this study was to evaluate the potential of cinnamon oil emulsions as alternative washing solutions to improve the microbial safety of carrots. Whey protein concentrate (WPC), gum arabic (GA), lecithin, and their combinations were used to prepare cinnamon oil emulsions. The emulsions were characterized for their hydrodynamic diameter (Dh) during 7 days of storage and their antimicrobial activity against cocktails of Salmonella enterica , Escherichia coli O157:H7, and Listeria monocytogenes . The Dh of the emulsion prepared with the GA+WPC blend did not change significantly (195.0 to 184.1 nm), whereas all other emulsions showed varying degrees of increases in Dh. Compared with free cinnamon oil dissolved in 5% ethanol, all emulsions showed similar or lower MICs and MBCs. Emulsions prepared with GA and equal masses of GA and WPC were chosen and diluted to 0.2 and 0.5% cinnamon oil to wash carrots that were surface inoculated with bacterial cocktails because of their lower MICs and MBCs than free oil. Emulsions resulted in significantly higher reductions of pathogens on carrots than free cinnamon oil, 3.0 to 3.7 versus 2.1 to 2.3 log CFU/g at 0.5% cinnamon oil and 2.0 to 3.0 versus 1.0 to 1.7 log CFU/g at 0.2% cinnamon oil. No transfer of bacteria from inoculated carrots to wash solutions and no effects of organic load on log reductions were only observed for wash treatments with 0.5% emulsified cinnamon oil. Thus, the cinnamon oil emulsions are potential alternative postharvest washing solutions for fresh produce production.

Quality attributes and microbial survival on whole cantaloupes with antimicrobial coatings containing chitosan, lauric arginate, cinnamon oil and ethylenediaminetetraacetic acid.

Cantaloupes are susceptible to microbiological contamination in pre- or postharvest environments. Novel intervention strategies, such as antimicrobial coatings, are needed to improve the microbiological safety of cantaloupes. The objective of this study was to prepare whole cantaloupes coated with mixtures containing chitosan, lauric arginate (LAE), cinnamon oil (CO), and ethylenediaminetetraacetic acid (EDTA) and determine survival characteristics of inoculated foodborne pathogens during storage as well as cantaloupe quality attributes. Chitosan coating with 0.1% LAE, 0.1% EDTA, and 1% CO was the most effective for inactivating foodborne pathogens inoculated on cantaloupes. This coating caused a >3logCFU/cm(2) reduction of Escherichia coli O157:H7 and Listeria monocytogenes immediately after coating and reduced Salmonella enterica to below the detection limit during a 14-day storage. Total molds and yeasts also were reduced to the detection limit by the coating. The redness and yellowness of uncoated cantaloupes were significantly higher than coated ones from day 6. The firmness of uncoated cantaloupes and those coated with chitosan only was significantly lower than other treatments from day 10. No significant differences were found in total soluble solids content or weight loss between coated and uncoated cantaloupes. Results showed the potential benefits of applying the coating mixtures to improve the quality and microbiological safety of cantaloupes.

Antimicrobial Efficacy of an Array of Essential Oils Against Lactic Acid Bacteria.

The essential oils of clove bud, cinnamon bark and thyme, and their individual compounds including allyl isothiocyanate (AIT), carvacrol, cinnamaldehyde, cinnamic acid, eugenol, and thymol were initially assessed for antimicrobial activity against 9 lactic acid bacteria (LAB) species. Carvacrol and thymol were the most inhibitory with MICs of 0.1% (v/v and w/v, respectively). Cinnamaldehyde, cinnamon bark oil, clove bud oil, eugenol, and thyme oil were moderately inhibitive (MICs = 0.2% v/v), while cinnamic acid required a concentration of 0.5% (w/v). AIT was not effective with MICs in excess of concentrations tested (0.75% v/v). The bactericidal capability of the oil components carvacrol, cinnamaldehyde, eugenol, and thymol were further examined against Pediococcus acidilactici, Lactobacillus buchneri, and Leuconostoc citrovorum. Thymol at 0.1% (w/v) was bactericidal against L. citrovorum (>4-log reduction), but resulted in a 2-log CFU/mL reduction against L. buchneri and P. acidilactici. Cinnamaldehyde at 0.2% to 0.25% (v/v) was effective against L. citrovorum, L. buchneri, and P. acidilactici, resulting in a >2-log reduction. All 3 organisms were susceptible to 0.2% carvacrol with >3-log reduction observed after exposure for 6 h. Eugenol was the least effective. Concentrations of 0.2% and 0.25% (v/v) were needed to achieve an initial reduction in population, >3-log CFU/mL after 6 h exposure. However, at 0.2%, P. acidilactici and L. buchneri recovered to initial populations in 48 to 72 h. Results indicate essential oils have the capacity to inactivate LAB that are commonly associated with spoilage of shelf stable low-acid foods.

Effect of alginate coatings with cinnamon bark oil and soybean oil on quality and microbiological safety of cantaloupe.

The quality and microbiological safety of cantaloupes can potentially be improved using antimicrobial coatings that are able to maintain effectiveness throughout storage. The objective of this work was to study the effect of coating mixtures containing sodium alginate and cinnamon bark oil (CBO) on the quality of cantaloupes and the survival of inoculated bacterial pathogens and naturally occurring yeasts and molds during ambient storage at 21 °C. Cantaloupes were dipped in mixtures containing 1% sodium alginate with or without 2% CBO and 0 or 0.5% soybean oil (SBO). Weight loss and total soluble solids content of the flesh were not significantly different among coating treatments. However, changes in color and firmness of cantaloupes were delayed to different extents after coating, most significantly for the CBO+SBO treatment. Cocktails of Salmonella enterica, Escherichia coli O157:H7, or Listeria monocytogenes inoculated on cantaloupes were reduced to the detection limit (1.3 log CFU/cm(2)) and completely inhibited during the 15-day storage by the CBO+SBO treatment, while L. monocytogenes and S. enterica reached populations of 2.9 log CFU/cm(2) and 2.4 log CFU/cm(2), respectively, on cantaloupes coated with CBO alone. Antimicrobial coatings, especially with SBO, also reduced yeast and mold counts on cantaloupes by 2.6 log CFU/cm(2). SBO improved the retention of CBO during storage suggesting it is related to the enhancement of quality and microbiological safety. Findings demonstrated the potential of the antimicrobial coating system studied to improve microbiological safety and quality of cantaloupes.

Antimicrobial Activity of Cinnamaldehyde, Carvacrol, and Lauric Arginate against Salmonella Tennessee in a Glycerol-Sucrose Model and Peanut Paste at Different Fat Concentrations.

The objective of this study was to investigate the antimicrobial activities of carvacrol, cinnamaldehyde, and lauric arginate (LAE) against Salmonella in a low water activity (aw ) glycerol-sucrose model and in peanut paste with different fat concentrations. Salmonella Tennessee was inoculated into the model and the low fat (<5%) and high fat (50%) peanut paste adjusted to aw 1.0, 0.7, 0.5, and 0.3 and with or without cinnamaldehyde, carvacrol, or LAE. The survival of the bacterium over 3 or 5 days at 25°C was evaluated. Reduced aw alone decreased the viable population over time, with the highest reduction at the lowest aw. In the glycerol-sucrose model, all antimicrobial agents significantly reduced the population over time (P < 0.05) compared with the controls. LAE was more lethal than the essential oil components, reducing the population to undetectable levels by day 2 for all aw. Cinnamaldehyde was more effective than carvacrol at aw 0.5 and 0.3 (2.7- to 2.9-log versus 0.39- to 1.97-log reductions on day 3). In low-fat peanut paste, none of the antimicrobial agents inhibited growth of the pathogen at aw 1.0. However, inactivation was enhanced at reduced aw. Cinnamaldehyde and LAE both reduced the pathogen population to undetectable levels on day 5 at the highest concentration tested (ca. 10 times higher than that in the glycerol-sucrose model). Inactivation efficacy of all antimicrobial agents was greatly decreased but not eliminated in 50% fat peanut paste. Results suggest that the test antimicrobial agents were effective under low aw conditions, but significantly higher concentrations are needed for potential food applications, and fat concentration can negatively impact the efficacy of these antimicrobial agents.

Formulating essential oil microemulsions as washing solutions for organic fresh produce production.

Applications of plant-derived organic essential oils (EOs) as antimicrobials for post-harvest produce operations are limited by their low water solubility. To dissolve EOs in water, microemulsions were studied using two surfactants permitted for organic production, sucrose octanoate ester (SOE) and soy lecithin that were mixed at various mass ratios before dilution with water to 40% w/w. EOs were then mixed with the surfactant solution by hand shaking. Based on visual transparency, intermediate lecithin:SOE mass ratios favoured the formation of microemulsions, e.g., up to 4.0% clove bud oil at ratios of 2:8 and 3:7, and 4.0% cinnamon bark oil and 3.0% thyme oil at ratios of 2:8 and 1:9, respectively. Microemulsions with intermediate lecithin:SOE mass ratios had a relatively low viscosity and better ability to wet fresh produce surfaces. The microemulsions established in this work may be used as washing solutions to enhance the microbial safety of organic fresh produce.

Self-emulsification of alkaline-dissolved clove bud oil by whey protein, gum arabic, lecithin, and their combinations.

Low-cost emulsification technologies using food ingredients are critical to various applications. In the present study, a novel self-emulsification technique was studied to prepare clove bud oil (CBO) emulsions, without specialized equipment or organic solvents. CBO was first dissolved in hot alkaline solutions, added at 1% v/v into neutral solutions with 1% w/v emulsifier composed of whey protein concentrate (WPC), gum arabic, lecithin, or their equal mass mixtures, and adjusted to pH 7.0. The self-emulsification process did not affect UV-vis absorption spectrum, reversed-phase HPLC chromatogram, or antimicrobial activity of CBO against Escherichia coli O157:H7, Listeria monocytogenes Scott A, and Salmonella Enteritidis. The entrapment efficiency after extraction by petroleum ether was determined to be about 80%. Most emulsions were stable during 7 days of storage. Emulsions prepared with WPC had smaller particles, whereas emulsions prepared with emulsifier mixtures had more stable particle dimensions. The studied self-emulsification technique may find numerous applications in the preparation of low-cost food emulsions.

Survival of Salmonella enterica in poultry feed is strain dependent.

Feed components have low water activity, making bacterial survival difficult. The mechanisms of Salmonella survival in feed and subsequent colonization of poultry are unknown. The purpose of this research was to compare the ability of Salmonella serovars and strains to survive in broiler feed and to evaluate molecular mechanisms associated with survival and colonization by measuring the expression of genes associated with colonization (hilA, invA) and survival via fatty acid synthesis (cfa, fabA, fabB, fabD). Feed was inoculated with 1 of 15 strains of Salmonella enterica consisting of 11 serovars (Typhimurium, Enteriditis, Kentucky, Seftenburg, Heidelberg, Mbandanka, Newport, Bairely, Javiana, Montevideo, and Infantis). To inoculate feed, cultures were suspended in PBS and survival was evaluated by plating samples onto XLT4 agar plates at specific time points (0 h, 4 h, 8 h, 24 h, 4 d, and 7 d). To evaluate gene expression, RNA was extracted from the samples at the specific time points (0, 4, 8, and 24 h) and gene expression measured with real-time PCR. The largest reduction in Salmonella occurred at the first and third sampling time points (4 h and 4 d) with the average reductions being 1.9 and 1.6 log cfu per g, respectively. For the remaining time points (8 h, 24 h, and 7 d), the average reduction was less than 1 log cfu per g (0.6, 0.4, and 0.6, respectively). Most strains upregulated cfa (cyclopropane fatty acid synthesis) within 8 h, which would modify the fluidity of the cell wall to aid in survival. There was a weak negative correlation between survival and virulence gene expression indicating downregulation to focus energy on other gene expression efforts such as survival-related genes. These data indicate the ability of strains to survive over time in poultry feed was strain dependent and that upregulation of cyclopropane fatty acid synthesis and downregulation of virulence genes were associated with a response to desiccation stress.