Hypervirulent clonal complex (CC) of Listeria monocytogenes in fresh produce from urban communities.

Introduction: This study aimed to determine the prevalence and virulome of Listeria in fresh produce distributed in urban communities. Methods: A total of 432 fresh produce samples were collected from farmer's markets in Michigan and West Virginia, USA, resulting in 109 pooled samples. Listeria spp. were isolated and L. monocytogenes was subjected to genoserogrouping by PCR and genotyping by pulsed-field gel electrophoresis (PFGE). Multi-locus sequence typing (MLST) and core-genome multi-locus sequence typing (cgMLST) were conducted for clonal identification. Results: Forty-eight of 109 samples (44.0%) were contaminated with Listeria spp. L. monocytogenes serotype 1/2a and 4b were recovered from radishes, potatoes, and romaine lettuce. Four clonal complexes (CC) were identified and included hypervirulent CC1 (ST1) and CC4 (ST219) of lineage I as well as CC7 (ST7) and CC11 (ST451) of lineage II. Clones CC4 and CC7 were present in the same romaine lettuce sample. CC1 carried Listeria pathogenicity island LIPI-1 and LIPI-3 whereas CC4 contained LIPI-1, LIPI-3, and LIPI-4. CC7 and CC11 had LIPI-1 only. Discussion: Due to previous implication in outbreaks, L. monocytogenes hypervirulent clones in fresh produce pose a public health concern in urban communities.

Microbial profile of broiler carcasses processed at a university scale mobile poultry processing unit.

Chicken and chicken products have been associated with foodborne pathogens such as Salmonella, Campylobacter, and Escherichia coli (E. coli). Poultry comprises an important segment of the agricultural economy (75 million birds processed as of 2019) in West Virginia (WV). The risk of pathogens on processed chickens has risen with the increased popularity of mobile poultry processing units (MPPUs). This study evaluated the microbial safety of broilers processed in a MPPU in WV. This study assessed aerobic plate counts (APCs), E. coli counts and the presence/absence of Salmonella and Campylobacter on 96 broiler carcasses following each MPPU step of scalding, eviscerating, and chilling. Samples were either chilled in ice water only (W) or ice water with 5 ppm chlorine (CW). The highest number of bacteria recovered from carcasses were APCs (4.21 log10CFU/mL) and E. coli (3.77 log10CFU/mL; P = 0.02). A total reduction of 0.30 (P = 0.10) and 0.63 (P = 0.01) log10CFU/mL for APCs and E. coli, respectively, occurred from chilling carcasses in CW. Overall, results show that E. coli, Salmonella, and Campylobacter were significantly (P < 0.05) reduced from the initial scalding to the chilling step. However, Salmonella frequency doubled (15.63-34.38%) after the evisceration step, indicating that washing carcasses after evisceration may be a critical control point in preventing cross-contamination by Salmonella. Proper chilling is also an important microbial mitigation step in MPPU processing. Results indicate that Campylobacter was more resistant to chilling than Salmonella. Campylobacter was not completely inactivated until carcasses were chilled in CW, whereas W was sufficient to reduce Salmonella on carcasses. The results led to the conclusion that although 5 ppm chlorine (Cl2) achieved more bacterial reductions than water alone, the reductions were not always significant (P > 0.05). Further MPPU studies are needed to verify more effective chilling and processing strategies.

Inactivation of Staphylococcus aureus by Levulinic Acid Plus Sodium Dodecyl Sulfate and their Antibacterial Mechanisms on S. aureus Biofilms by Transcriptomic Analysis.

The combination of levulinic acid (LVA) and sodium dodecyl sulfate (SDS) in recent years has shown a considerable potential to use as an antimicrobial intervention. The objectives of this study were to evaluate the antimicrobial efficacy of the combination against Staphylococcus aureus in both planktonic and biofilm states and to investigate the transcriptional changes in S. aureus biofilms coincubated with sublethal concentrations of LVA and/or SDS. The minimum inhibitory concentrations (MICs) of LVA and SDS determined by the microdilution method were 3.125 and 0.039 mg/mL, respectively. An additive bacteriostatic interaction (fractional inhibitory concentration index = 1) between the two compounds was observed by the checkerboard assay, whereas a synergistic bactericidal activity was displayed by the time-kill assay. The biomass and viable cells in the biofilms were reduced by both antimicrobials either alone or in combination in a dose-dependent manner. Transcriptomics indicated that more differentially expressed (DE) genes were observed in the biofilm treated with SDS (103 up- and 205 downregulated DE genes) and LVA + SDS (187 up and 162 down) than that coincubated with LVA (34 up and 32 down). The SDS and LVA + SDS treatments mainly affected the expression of genes responsible for cell surface proteins, virulence factors, adhesins, and capsular polysaccharides. Both the antibiofilm assay and the transcriptomics indicated that SDS, not LVA, was the major chemical contributing to the antibacterial efficacy of the combination. This study reveals the behavioral responses and protective mechanisms of S. aureus to LVA and SDS applied individually or in combination.

Enhanced inactivation of Salmonella and Pseudomonas biofilms on stainless steel by use of T-128, a fresh-produce washing aid, in chlorinated wash solutions.

The effect of the washing aid T-128 (generally recognized as safe [GRAS] formulation, composed mainly of phosphoric acid and propylene glycol) on inactivation of Salmonella and Pseudomonas populations in biofilms on stainless steel was evaluated under conditions of increasing organic matter loads in chlorinated wash solutions dominated by hypochlorous acid. Biofilms were formed statically on stainless steel coupons suspended in 2% lettuce extract after inoculation with Salmonella enterica serovar Thompson or Newport or with Pseudomonas fluorescens. Coupons with biofilms were washed in chlorine solutions (0, 0.5, 1, 2, 5, 10, or 20 mg/liter at pH 6.5, 5.0 and 2.9), with or without T-128, and with increasing loads of organic matter (0, 0.25, 0.5, 0.75, or 1.0% lettuce extract). Cell populations on coupons were dispersed using intermittent, pulsed ultrasonication and vortexing and enumerated by colony counts on XLT-4 or Pseudomonas agars. Cell responses to fluorescent viability staining of biofilm treatment washing solutions were examined using confocal laser scanning microscopy. Results showed that 0.1% T-128 (without chlorine) reduced P. fluorescens biofilm populations by 2.5 log(10) units but did not reduce Salmonella populations. For both Salmonella and Pseudomonas, the sanitizing effect of free chlorine (1.0 to 5.0 mg/liter) was enhanced (P < 0.05) when it was combined with T-128. Application of T-128 decreased the free chlorine depletion rate caused by increasing organic matter in wash waters and significantly (P < 0.05) augmented inactivation of bacteria in biofilms compared to treatments without T-128. Image analysis of surfaces stained with SYTO and propidium iodide corroborate the cultural assay results showing that T-128 can aid in reducing pathogen viability in biofilms and thus can aid in sanitizing stainless steel contact surfaces during processing of fresh-cut produce.

Antimicrobial Nonwoven Fabrics Incorporated with Levulinic Acid and Sodium Dodecyl Sulfate for Use in the Food Industry.

Safe and cost-effective antimicrobial fabrics (e.g., face masks and air filters) are conducive to preventing the spread and transmission of respiratory microorganisms in food processing plants and retail establishments. The objective of this study was to coat fabrics with two commonly used compounds in the food industry: levulinic acid (LVA) and sodium dodecyl sulfate (SDS) and determine the antimicrobial efficacy of the coated fabrics against bacterial solutions, aerosols, and influenza A virus subtype H1N1. In addition, air permeability and shelf-life of the LVA/SDS coated fabrics were also examined. Nonwoven fabrics were dip-coated with three concentrations (w/v, 0.5% LVA + 0.1% SDS, 1% LVA + 0.5% SDS, and 2% LVA + 1% SDS) of LVA and SDS and challenged with bacterial solutions (Staphylococcus aureus and Escherichia coli, ca. 7.0 log CFU/coupon) for a contact time of 3, 5, and 10 min. The coated fabrics were also challenged with S. aureus aerosol and H1N1 virus following standard operations of ASTM F2101-19 and ISO 18184:2019, respectively. The 1% LVA + 0.5% SDS coated fabrics showed potent antibacterial efficacy against both bacterial solutions (>6.0-log reduction to under the detection limit of 1.0 log CFU/coupon for S. aureus; ca. 1.0-log reduction for E. coli) and aerosols (>3.6-log reduction to under the detection limit), with greater inactivation occurring at higher concentrations and longer exposure time. Moreover, the coated fabrics inactivated >99% of the H1N1 virus. The shelf-life of the coated fabrics was stable within 12 months and the air permeability was not adversely affected with the coating concentrations less than 1% LVA + 0.5% SDS. Results reveal these low-cost and safe materials have the potential to be used to coat fabrics in the food industry to combat the spread and transmission of pathogens.

The Efficacy of Conventional Spray, Electrostatic Spray, and Dip with a Combination of Hydrogen Peroxide and Peroxyacetic Acid To Inactivate Listeria monocytogenes on Apples.

ABSTRACT: This study aimed to evaluate the efficacy of a hydrogen peroxide (H2O2) and peroxyacetic acid (PAA) mixer delivered by conventional garden spray (GS), electrostatic spray (ES), and dip methods to inactivate Listeria monocytogenes on apples. Organic Honeycrisp, Fuji, and Pink Lady apples were dip inoculated with L. monocytogenes (two strains, serotype 1/2b), which were then kept untreated (control), sprayed with water only, or treated with the H2O2-PAA mixer (0.0064, 0.1, 0.25, and 0.50%) for 20 s via GS, ES, or dip, followed by draining (for 2 min) on aluminum foil. Surviving bacteria were recovered on modified Oxford agar. Atomic force microscopy was used to detect the structural changes of inactivation of L. monocytogenes in broth medium by the H2O2-PAA mixer solution. Data (two replicates, with six samples per replicate) were analyzed using the mixed model procedure of SAS (P = 0.05). Initial counts of L. monocytogenes on untreated apples were 6.80 to 6.90 log CFU per apple. The dip method was the most effective treatment (P < 0.05) for pathogen reductions (2.31 to 2.41 log CFU per apple), followed by GS (1.44 to 1.70 log CFU per apple) and then ES (0.84 to 1.20 log CFU per apple). Reductions of L. monocytogenes were greatest (P < 0.05) when apples were treated with H2O2-PAA mixer -0.25 and -0.50%. Atomic force microscopy analyses indicated that inactivation of L. monocytogenes cells in H2O2-PAA mixer solutions resulted from disruption of the outer membrane. The H2O2-PAA mixer-treated cells had increased width and height and decreased roughness compared with the untreated cells. Results suggested that applying a H2O2-PAA mixer by dip or GS methods is better for pathogen reduction than ES on apples.

Thermal inactivation of Salmonella Typhimurium and surrogate Enterococcus faecium in mash broiler feed in a laboratory scale circulated thermal bath.

This study compares kinetic parameters of Salmonella and surrogate Enterococcus faecium in mash broiler feed during thermal inactivation. Two-gram samples of mash broiler feed were added into a filtered sample bag and inoculated with nalidixic acid (NaL, 200 ppm) resistant S. Typhimurium or Enterococcus faecium, followed by vacuum-packaging and heating in a circulated thermal water bath at 75°, 85°, and 95°C for 0 to 180 s. Counts of bacterial survival were analyzed on tryptic soy agar and bile esculin agar plus 200 ppm of NaL. Microbial data and thermal kinetic parameters (n = 8, Global-Fit and United States Department of Agriculture [USDA]-Integrated-Predictive-Modeling-Program software) were analyzed by JMP software. Heating mash broiler feed at 75°, 85°, and 95°C decreased (P < 0.05) Salmonella cell counts by >6 log10CFU/g after 180, 60, and 50 s, respectively. Heating E. faecium in feed at 75°, 85°, and 95°C for 180, 120, and 70 s achieved reductions of 3, 6, and >6.5 log10CFU/g, respectively. D-values of linear, Weibull models, and z-value of Salmonella at 75°, 85°, and 95°C were 1.8 to 11.2, 4.2 to 21.8, and 28.6 s, respectively, which were lower (P < 0.05) than those of E. faecium (3.7-18.1, 8.5-34.4, and 34.1 s). Linear with Tail, Linear with Tail and Shoulder, and Weibull with tail equations revealed that E. faecium were more resistant (P < 0.05) to heat than Salmonella as shown by longer "Shoulder-time" (26.5 vs. 16.2 s) and greater "Tail" effect (4.4-4.5 vs. 2.5-2.6 log10CFU/g). Results clearly suggested that E. faecium can be used as a surrogate for Salmonella to validate thermal inactivation during feed manufacture.

Survival of Salmonella and the surrogate Enterococcus faecium in cooking of moisture enhanced reconstructed comminuted chicken patties by double pan-broiling.

This study compares kinetic parameters of Salmonella and Enterococcus faecium in moisture enhanced, reconstructed comminuted chicken patties prepared with different pump rates during double pan-broiling with various set-up temperatures. Fresh 1.5-kg chicken breast meat was course grounded, inoculated with S. Typhimurium and Tennessee, or E. faecium, followed by adding NaCl (2.0%) + Na-tripolyphosphate (0.5%) solutions to achieve pump rates of 1%, 5%, or 11.1%. Meat samples were manually manufactured into patties with the thickness of 2.1 cm and diameter of 10.4 cm. Patties were packaged with polyvinyl chloride films in the foam-tray stored at 4°C for 42 h before double pan-broiling set at 200°, 300°, or 425°F for 0 to 420 s. Counts of pathogens were analyzed on xylose-lysine-Tergitol-4 and bile esculin agars with tryptic soy agar layers. Microbial data and kinetic parameters (n = 9, United States Department of Agriculture [USDA]-Integrated-Predictive-Modeling-Program/USDA-Global-Fit software) were analyzed by the Mixed Model Procedure (SAS). Double pan-broiling reduced >5-log10 CFU/g (P < 0.05) of Salmonella after 360 (200°F), 180 to 225 (300°F), and 150 to 165s (425°F), and of E. faecium after 270 s (300°F), and 180 s (425°F) across all samples. D-values (Mafart-Weibull model) of Salmonella and E. faecium in 1% moisture enhanced samples cooked at 200 to 425°F (102.7-248.2 and 115.5-271.0 s) were lower (P < 0.05) than 11.1% samples (119.8-263.7 and 122.5-298.3 s). Salmonella were more susceptible (P < 0.05) to heat than E. faecium. "Shoulder-time" (Buchanan-Two-Phase model) of Salmonella cooking at 200° to 425°F increased (P < 0.05) from 82.3-229.0 to 116.6-246.2 s as pump rate increased from 1 to 11.1%, whereas this phenomenon was not shown for E. faecium. Results indicate that Salmonella were resistant to heat in chicken patties with greater pump rate. E. faecium can be used as a surrogate for Salmonella to validate thermal inactivation in chicken products.

Inactivation of Escherichia coli O157:H7 in nonintact beefsteaks of different thicknesses cooked by pan broiling, double pan broiling, or roasting by using five types of cooking appliances.

This study compared thermal inactivation of Escherichia coli O157:H7 in nonintact beefsteaks of different thicknesses by different cooking methods and appliances. Coarsely ground beef was inoculated with rifampin-resistant E. coli O157:H7 (eight-strain composite, 6 to 7 log CFU/g) and then mixed with sodium chloride (0.45%) plus sodium tripolyphosphate (0.23%); the total water added was 10%. The meat was stuffed into bags (10-cm diameter), semifrozen (-20 degrees C, 6 h), and cut into 1.5-, 2.5-, and 4.0-cm-thick steaks. Samples were then individually vacuum packaged, frozen (-20 degrees C, 42 h), and tempered (4 degrees C, 2.5 h) before cooking. Partially thawed (-2 +/- 1 degrees C) steaks were pan broiled (Presto electric skillet and Sanyo grill), double pan broiled (George Foreman grill), or roasted (Oster toaster oven and Magic Chef standard kitchen oven) to a geometric center temperature of 65 degrees C. Extent of pathogen inactivation decreased in order of roasting (2.0 to 4.2 log CFU/g) > pan broiling (1.6 to 2.8 log CFU/g) >/= double pan broiling (1.1 to 2.3 log CFU/g). Cooking of 4.0-cm-thick steaks required a longer time (19.8 to 65.0 min; variation was due to different cooking appliances), and caused greater reductions in counts (2.3 to 4.2 log CFU/g) than it did in thinner samples (1.1 to 2.9 log CFU/g). The time to reach the target temperature increased in order of George Foreman grill (3.9 to 19.8 min) < Oster toaster oven (11.3 to 45.0 min) < Presto electric skillet (16.3 to 55.0 min) < Sanyo grill (14.3 to 65.0 min) < standard kitchen oven (20.0 to 63.0 min); variation was due to steak thickness. Results indicated that increased steak thickness allowed greater inactivation of E. coli O157:H7, as time to reach the target internal temperature increased. Roasting in a kitchen oven was most effective for pathogen inactivation.

Control of Listeria monocytogenes on frankfurters by dipping in hops beta acids solutions.

Hops beta acids (HBA) are parts of hops flowers used in beer brewing and have shown antilisterial activity in bacteriological broth. The U.S. Department of Agriculture, Food Safety and Inspection Service has approved HBA for use to control Listeria monocytogenes on ready-to-eat meat products. This study evaluated the effects of HBA as dipping solutions to control L. monocytogenes during storage of frankfurters. Frankfurters (two replicates and three samples each) were inoculated (1.9 +/- 0.1 log CFU/cm2) with L. monocytogenes (10-strain mixture), dipped (2 min, 25 +/- 2 degrees C) in HBA solutions (0.03, 0.06, and 0.10%) or distilled water, and then vacuum packaged and stored at 4 or 10 degrees C for up to 90 and 48 days, respectively. Samples were periodically analyzed for microbial survival and growth on tryptic soy agar plus 0.6% yeast extract and PALCAM agar. Dipping in HBA solutions caused immediate L. monocytogenes reductions (P < 0.05) of 1.3 to 1.6 log CFU/cm2, whereas distilled water reduced counts by 1.0 log CFU/cm2. Pathogen growth was completely suppressed (P < 0.05) for 30 to 50 (4 degrees C) or 20 to 28 (10 degrees C) days on frankfurters dipped in HBA solutions, with antilisterial effects increasing with higher concentrations (0.03 to 0.10%). Fitting the data with the Baranyi model confirmed that the lag-phase duration of the pathogen was extended, and the growth rate was decreased on samples dipped in HBA solutions. Therefore, HBA may be considered for use to improve the microbial safety of ready-to-eat meat products, provided that future studies show no adverse effects on sensory qualities and that their use is economically feasible.

Comparison of the Efficacy of Electrostatic versus Conventional Sprayer with Commercial Antimicrobials To Inactivate Salmonella, Listeria monocytogenes, and Campylobacter jejuni for Eggs and Economic Feasibility Analysis.

This study was conducted to compare the efficacy of antimicrobials sprayed by electrostatic versus conventional sprayer for inactivation of Salmonella, Listeria monocytogenes, and Campylobacter jejuni on eggs and to determine the economic feasibility of these treatments. Eggs were dip inoculated with overnight cultures (18 h) of Salmonella Typhimurium, Salmonella Tennessee, a two-strain mixture of L. monocytogenes, and a three-strain mixture of C. jejuni (microaerophilic condition). Inoculated eggs were then not sprayed or subjected to electrostatic and conventional spraying with peroxyacetic acid (PAA; 0.1%), lactic acid (5.0%), lactic and citric acid blend (2.5%), sodium hypochlorite (SH; 50 ppm), and SaniDate-5.0 (SD [a mixture of PAA and H2O2]; 0.25%) for 30 s (15 s each side). Surviving bacteria on eggshells were recovered on xylose lysine Tergitol 4 agar ( Salmonella), modified Oxford agar ( L. monocytogenes), or Brucella agar ( C. jejuni). Compared with conventional spraying, electrostatic spraying of PAA, SD, and SH achieved significant additional reductions ( P < 0.05) of Salmonella, L. monocytogenes, and C. jejuni of 0.96 to 3.18, 1.19 to 3.05, and 0.96 to 1.62 log CFU per egg, respectively. A simple cost comparison suggests that regardless of the antimicrobial agent used, the cost of using an electrostatic sprayer is 20 to 40% lower than that of a conventional sprayer for a small poultry farm that produces 1,500 eggs per day. Among the five antimicrobials, the total sanitizing cost was lowest for SH, followed by PAA and SD. The results indicated that electrostatic spraying of commercial antimicrobials can be considered an effective and economical approach to enhancing the microbial safety of eggs, especially for small poultry processors.

Microbiological quality assessment and validation of antimicrobials against unstressed or cold-stress adapted Salmonella and surrogate Enterococcus faecium on broiler carcasses and wings.

This study aims to evaluate the microbiological quality and efficacy of antimicrobials to inactivate unstressed or cold-stress adapted Salmonella and Enterococcus on broiler carcasses and wings processed at a small USDA-inspected slaughter facility in West Virginia. The first part of the study included 42 carcasses that were pre- and secondarily-enriched in bacterial media followed by streak-plating onto XLT-4 and HardyCHROM™-agar Salmonella and confirmation using an API20E-kit. The aerobic plate counts (APC), Escherichia coli (ECC), total coliforms (TCC), and yeast/molds were analyzed on petri-films. The second part of the study included fresh broiler carcasses and wings that were inoculated with unstressed and cold-stress-adapted (4 °C, 7-day) Salmonella Typhimurium and Tennessee, and Enterococcus faecium ATCC 8459 (5.5 to 6.0 log10CFU/mL) and later dipped into peroxyacetic acid (PAA; 1,000 ppm), lactic acid (LA; 5%), lactic and citric acid blend (LCA; 2.5%), and sodium hypochlorite (SH; 70 ppm) for 30 s without (carcasses) or with 2-min drainage (wings). The surviving bacteria were recovered onto non-selective and selective agar to analyze the total microbial population, Salmonella and Enterococcus. APC, TCC, and Yeast/Molds were 2.62, 1.08, and 2.37 log10CFU/mL on broiler carcasses, respectively. A total of 30 and 40% of the carcasses tested positive for Salmonella spp. and E. coli (0.48 to 1.70 log10CFU/mL), respectively. For carcasses, antimicrobial reductions of cold-stress-adapted cells of Salmonella and Enterococcus were greater (P < 0.05) than the unstressed cells. For wings, cold-stress-adapted Salmonella were more (P < 0.05) sensitive to antimicrobials than unstressed cells; however, unstressed and cold-stress-adapted Enterococcus behaved similarly (P > 0.05). The reduction of Salmonella and Enterococcus on carcasses and wings increased in the order of SH ≤ LCA < LA < PAA and irrespective of unstressed or cold-stress-adapted cells. Applying post-chilling antimicrobial dipping treatments could be an intervention approach to control Salmonella on locally processed broilers. In addition, Enterococcus faecium could be a Salmonella surrogate for in-plant validation studies.

A Comparison Study of Quality Attributes of Ground Beef and Veal Patties and Thermal Inactivation of Escherichia coli O157:H7 after Double Pan-Broiling Under Dynamic Conditions.

This study compared the quality variation and thermal inactivation of Escherichia coli O157:H7 in non-intact beef and veal. Coarse ground beef and veal patties (2.1 cm thick, 12.4 cm diameter, 180 g) inoculated with E. coli O157:H7, aerobically stored before double pan-broiling for 0-360 s without rest or to 55, 62.5, 71.1, and 76 °C (internal temperature) with 0.5- or 3.5-min rest. Microbial population and qualities including color, cooking losses, pH, water activity, fat, and moisture content, were tested. After cooking the beef and veal patties, the weight losses were 17.83-29%, the pH increased from 5.53-5.60 to 5.74-6.09, the moisture content decreased from 70.53-76.02% to 62.60-67.07%, and the fat content increased (p < 0.05) from 2.19-6.46% to 2.92-9.45%. Cooking beef and veal samples with increasing internal temperatures decreased a* and b* values and increased the L* value. Escherichia coli O157:H7 was more sensitive to heat in veal compared to beef with shorter D-value and "shoulder" time. Cooking to 71.1 and 76 °C reduced E. coli O157:H7 by >6 log CFU/g regardless of rest time. Cooking to 55 °C and 62.5 °C with a 3.5-min rest achieved an additional 1-3 log CFU/g reduction compared to the 0.5-min rest. Results should be useful for developing risk assessment of non-intact beef and veal products.

Impact of Built-up-Litter and Commercial Antimicrobials on Salmonella and Campylobacter Contamination of Broiler Carcasses Processed at a Pilot Mobile Poultry-Processing Unit.

The small-scale mobile poultry-processing unit (MPPU) produced raw poultry products are of particular food safety concern due to exemption of USDA poultry products inspection act. Limited studies reported the microbial quality and safety of MPPU-processed poultry carcasses. This study evaluated the Salmonella and Campylobacter prevalence in broiler ceca and on MPPU-processed carcasses and efficacy of commercial antimicrobials against Campylobacter jejuni on broilers. In study I, straight-run Hubbard × Cobb broilers (147) were reared for 38 days on clean-shavings (CS, 75) or built-up-litter (BUL, 72) and processed at an MPPU. Aerobic plate counts (APCs), coliforms, Escherichia coli, and yeast/molds (Y/M) of carcasses were analyzed on petrifilms. Ceca and carcass samples underwent microbial analyses for Salmonella and Campylobacter spp. using the modified USDA method and confirmed by API-20e test (Salmonella), latex agglutination immunoassay (Campylobacter), and Gram staining (Campylobacter). Quantitative polymerase chain reaction (CadF gene) identified the prevalence of C. jejuni and Campylobacter coli in ceca and on carcasses. In study II, fresh chilled broiler carcasses were spot inoculated with C. jejuni (4.5 log10 CFU/mL) and then undipped, or dipped into peroxyacetic acid (PAA) (1,000 ppm), lactic acid (5%), lactic and citric acid blend (2.5%), sodium hypochlorite (69 ppm), or a H2O2-PAA mix (SaniDate® 5.0, 0.25%) for 30 s. Surviving C. jejuni was recovered onto Brucella agar. APCs, coliforms, and E. coli populations were similar (P > 0.05) on CS and BUL carcasses. Carcasses of broilers raised on BUL contained a greater (P < 0.05) Y/M population (2.2 log10 CFU/mL) than those reared on CS (1.8 log10 CFU/mL). Salmonella was not detected in any ceca samples, whereas 2.8% of the carcasses from BUL were present with Salmonella. Prevalence of Campylobacter spp., C. jejuni was lower (P < 0.05), and C. coli was similar (P > 0.05) in CS-treated ceca than BUL samples. Prevalence of Campylobacter spp., C. jejuni, and C. coli was not different (P > 0.05) on CS- and BUL-treated carcasses. All antimicrobials reduced C. jejuni by 1.2-2.0 log CFU/mL on carcasses compared with controls. Hence, raising broilers on CS and applying post-chilling antimicrobial treatment can reduce Salmonella and Campylobacter on MPPU-processed broiler carcasses.

Reductions in Natural Microbial Flora, Nonpathogenic Escherichia coli , and Pathogenic Salmonella on Jalapeno Peppers Processed in a Commercial Antimicrobial Cabinet: A Pilot Plant Trial.

This experiment aimed to validate the use of antimicrobial solutions in a spray cabinet to inactivate natural microbial flora, nonpathogenic Escherichia coli , and Salmonella on jalapeno peppers. Jalapeno peppers, uninoculated or inoculated with a five-strain mixture of rifampin-resistant E. coli (3.9 log CFU/g) or novobiocin- and nalidixic acid-resistant Salmonella (4.2 log CFU/g), were passed through a commercial antimicrobial cabinet containing both a top and bottom bar spraying (1.38 bar and 2 liters/min) water, sodium hypochlorite (50 ppm), sodium hypochlorite with pH adjusted to 6.7, peroxyacetic acid (PAA; 80 ppm), PAA with pH adjusted to 6.7, lactic with citric acid (1%), lactic with citric acid with sodium lauryl sulfate (1,200 ppm), or chlorine dioxide (5 ppm). Bacteria were recovered in 0.1% buffered peptone water plus 0.1% sodium thiosulfate, which was followed by spread plating onto tryptic soy agar (TSA), TSA plus rifampin (100 µg/ml), and TSA plus novobiocin (25 µg/ml) and nalidixic acid (20 µg/ml). There were no significant differences (P ≥ 0.05) in recovered natural microbial flora, E. coli , and Salmonella populations between untreated peppers (3.5 to 4.2 log CFU/g) and peppers treated with water (3.4 to 3.8 log CFU/g). Significantly fewer (P < 0.05) natural microbial flora, E. coli , and Salmonella populations were recovered on the peppers after they were treated with a majority of the antimicrobials applied in the commercial antimicrobial cabinet. The largest population reduction was observed on peppers sprayed with PAA. Interestingly, the pH adjustment did not make a difference (P ≥ 0.05) in the recovered bacterial populations. These results validate the use of a commercial antimicrobial spray cabinet, and they are useful for developing application protocols for antimicrobials to control Salmonella during the postharvest processing of jalapeno peppers.

Generation of chlorine by-products in simulated wash water.

Free chlorine (FC) reacting with organic matter in wash water promotes the formation of chlorine by-products. This study aims to evaluate the dynamic impact of FC and organic load on the generation of haloacetic acids (HAAs) and trihalomethanes (THMs) in simulated wash water. Lettuce juice was sequentially added into FC solution with FC periodically replenished. Water samples were collected after each lettuce juice addition to measure water qualities and determine HAAs and THMs using US-Environmental-Protection-Agency (EPA) methods. Concentrations of 88-2103 µg/l of total HAAs and 20.79-859.47 µg/l of total THMs were detected during the study. Monobromoacetic, tribromoacetic, chlorodibromoacetic and trichloroacetic acid were the major HAAs components. Chloroform (trichloromethane) was the primary THMs present. A significant correlation of HAAs with chemical oxygen demand and THMs with FC was observed. Results indicated that optimizing wash water sanitizing systems to limit organic matters and maintain minimal effective FC concentration is critical.

Effect of Hops Beta Acids on the Survival of Unstressed- or Acid-Stress-Adapted-Listeria Monocytogenes and on the Quality and Sensory Attributes of Commercially Cured Ham Slices.

This study evaluated the antilisterial activity of hops beta acids (HBA) and their impact on the quality and sensory attributes of ham. Commercially cured ham slices were inoculated with unstressed- and acid-stress-adapted (ASA)-L. monocytogenes (2.2 to 2.5 log CFU/cm(2) ), followed by no dipping (control), dipping in deionized (DI) water, or dipping in a 0.11% HBA solution. This was followed by vacuum or aerobic packaging and storage (7.2 °C, 35 or 20 d). Samples were taken periodically during storage to check for pH changes and analyze the microbial populations. Color measurements were obtained by dipping noninoculated ham slices in a 0.11% HBA solution, followed by vacuum packaging and storage (4.0 °C, 42 d). Sensory evaluations were performed on ham slices treated with 0.05% to 0.23% HBA solutions, followed by vacuum packaging and storage (4.0 °C, 30 d). HBA caused immediate reductions of 1.2 to 1.5 log CFU/cm(2) (P < 0.05) in unstressed- and ASA-L. monocytogenes populations on ham slices. During storage, the unstressed-L. monocytogenes populations on HBA-treated samples were 0.5 to 2.0 log CFU/cm(2) lower (P < 0.05) than control samples and those dipped in DI water. The lag-phase of the unstressed-L. monocytogenes population was extended from 3.396 to 7.125 d (control) to 7.194 to 10.920 d in the HBA-treated samples. However, the ASA-L. monocytogenes population showed resistance to HBA because they had a higher growth rate than control samples and had similar growth variables to DI water-treated samples during storage. Dipping in HBA solution did not adversely affect the color or sensory attributes of the ham slices stored in vacuum packages. These results are useful for helping ready-to-eat meat processors develop operational procedures for applying HBA on ham slices.

Survival of Unstressed and Acid-, Cold-, and Starvation-Stress-Adapted Listeria monocytogenes in Ham Extract with Hops Beta Acids and Consumer Acceptability of HBA on Ready-to-Eat Ham.

The efficacy of hops beta acids (HBA) against unstressed and stress-adapted Listeria monocytogenes in ham extract and the consumers' acceptability of HBA on ready-to-eat (RTE) hams were investigated. Unstressed or acid-, cold-, or starvation-stress-adapted L. monocytogenes was inoculated (1.3-1.5 log CFU/mL) into 10% ham extract, without (control) or with HBA (4.44 or 10.0 µg/mL). Survival/growth of the pathogen during storage (7.2 °C, 26 days) was monitored periodically. Sensory evaluation (30 participants, 9-point hedonic scale) was performed with hams dipped into 0.05, 0.11, and 0.23% HBA solution. Ham extracts without HBA supported rapid growth of unstressed and stress-adapted cells with growth rates of 0.39-0.71 log CFU/mL/day and lag phases of 0-3.26 days. HBA inhibited growth of unstressed L. monocytogenes by slowing (P < 0.05) growth rate (0.24-0.29 log CFU/mL/day) and increasing (P < 0.05) length of the lag phase (3.49-12.98 days) compared to control. Acid-, cold-, or starvation-stress-adapted cells showed cross protection against HBA with greater (P < 0.05) growth rates (0.44-0.66 log CFU/mL/day) and similar or shorter lag phases (0-5.44 days) than unstressed cells. HBA did not (P > 0.05) affect sensory attributes of RTE ham. These results are useful for RTE meat processors to develop operational protocols using HBA to control L. monocytogenes.

Dynamic effects of free chlorine concentration, organic load, and exposure time on the inactivation of Salmonella, Escherichia coli O157:H7, and non-O157 Shiga toxin-producing E. coli.

This study evaluated the dynamic effects of free-chlorine (FC) concentration, contact time, and organic load on the inactivation of Salmonella, Escherichia coli O157:H7, and non-O157 Shiga toxin-producing E. coli (STEC) in suspension. Bacterial cells from four strains each of Salmonella, E. coli O157:H7, and non-O157 STEC were inoculated separately or as a multistrain cocktail into solutions with varying FC concentrations. Lettuce or tomato extract was used to simulate the organic matter present during commercial fresh and fresh-cut produce wash operations. After exposure to FC for various lengths of time, the bacterial survival and water-quality changes were determined. In the absence of organic matter in a wash solution, pathogen inactivation is primarily a function of initial FC concentration (P < 0.0001), exposure time (P < 0.0001), and pathogen strains (P < 0.0001). In general, an over 4.5-log CFU/ml pathogen reduction was found after exposure to >0.5 mg/liter FC for over 30 s, or to >1.0 mg/liter FC for over 5 s. When the combination of FC concentration and contact time were less than or equal to the above conditions, survival of pathogens was strain dependant and ranked as: Salmonella > E. coli O157:H7 > non-O157 STEC. When organic matter was present in the wash solution, pathogen inactivation efficacy was specifically dependent on the residual FC concentration, which directly relates to both the initial FC concentration and the organic load. Prevention of pathogen survival in chlorinated produce wash solutions can be achieved by maintaining sufficient FC concentration and reducing the accumulation of organic matter.

A pilot plant scale evaluation of a new process aid for enhancing chlorine efficacy against pathogen survival and cross-contamination during produce wash.

Developing food safety intervention technology that can be readily adopted by the industry often requires test conditions that match as closely as possible to those of commercial food processing operations; yet biosafety risks inherent in pathogen studies constrain most experiments to laboratory settings. In this study, we report the first semi-commercial pilot-scale evaluation of a new process aid, T128, for its impact on enhancing the antimicrobial efficacy of chlorinated wash water against pathogen survival and cross-contamination. A non-pathogenic, BSL-1, strain of Escherichia coli O157:H7 was inoculated onto freshly harvested baby spinach leaves and washed with large amounts of freshly cut un-inoculated iceberg lettuce shreds in wash water with free chlorine periodically replenished, in the presence or absence of T128. Changes in water quality and pathogen survival and cross-contamination were monitored at every 2 min intervals for up to 36 min for each treatment during the wash operation. Results indicated that the use of T128 did not significantly (P>0.05) influence the rate of wash water deterioration, nor the pathogen populations remaining on the inoculated spinach leaves. However, in the absence of T128 (control), survival of E. coli O157:H7 in wash water and cross-contamination of un-inoculated lettuce frequently occurred when free chlorine in solution dropped below 1mg/l during the wash process. In contrast, the use of T128 significantly reduced the occurrence of E. coli O157:H7 surviving in wash water and of cross-contamination to un-inoculated shredded iceberg lettuce under the same operational conditions, suggesting that the application of T128 in a chlorine-based fresh produce sanitization system could increase the safety margin of process control on fresh-cut operations.