Effect of Product Dimensions and Surface Browning Method on Salmonella Contamination in Frozen, Surface-Browned, Breaded Chicken Products Treated with Antimicrobials.

Not-ready-to-eat breaded chicken products formulated with antimicrobial ingredients were tested for the effect of sample dimensions, surface browning method and final internal sample temperature on inoculated Salmonella populations. Fresh chicken breast meat portions (5 × 5 × 5 cm), inoculated with Salmonella (7-strain mixture; 5 log CFU/g), were mixed with (5% v/w total moisture enhancement) (i) distilled water (control), (ii) caprylic acid (CAA; 0.0625%) and carvacrol (CAR; 0.075%), (iii) CAA (0.25%) and ε-polylysine (POL; 0.5%), (iv) CAR (0.15%) and POL (0.5%), or (v) CAA (0.0625%), CAR (0.075%) and POL (0.5%). Sodium chloride (1.2%) and sodium tripolyphosphate (0.3%) were added to all treatments. The mixtures were then ground and formed into 9 × 5 × 3 cm (150 g) or 9 × 2.5 × 2 cm (50 g) portions. The products were breaded, browned in (i) an oven (208 °C, 15 min) or (ii) deep fryer (190 °C, 15 s), packaged, and stored at -20 °C (8 d). Overall, maximum internal temperatures of 62.4 ± 4.0 °C (9 × 2.5 × 2 cm) and 46.0 ± 3.0 °C (9 × 5 × 3 cm) were reached in oven-browned samples, and 35.0 ± 1.1 °C (9 × 2.5 × 2 cm) and 31.7 ± 2.6 °C (9 × 5 × 3 cm) in fryer-browned samples. Irrespective of formulation treatment, total (after frozen storage) reductions of Salmonella were greater (P < 0.05) for 9 × 2.5 × 2 cm oven-browned samples (3.8 to at least 4.6 log CFU/g) than for 9 × 5 × 3 cm oven-browned samples (0.7 to 2.5 log CFU/g). Product dimensions did not (P ≥ 0.05) affect Salmonella reductions (0.6 to 2.8 log CFU/g) in fryer-browned samples. All antimicrobial treatments reduced Salmonella to undetectable levels (<0.3 log CFU/g) in oven-browned 9 × 2.5 × 2 cm samples. Overall, the data may be useful for the selection of antimicrobials, product dimensions, and surface browning methods for reducing Salmonella contamination.

Effects of Plant-Derived Extracts, Other Antimicrobials, and Their Combinations against Escherichia coli O157:H7 in Beef Systems.

The antimicrobial effects of thyme oil (TO), grapefruit seed extract (GSE), and basil essential oil, alone or in combination with cetylpyridinium chloride (CPC), sodium diacetate, or lactic acid, were evaluated against Escherichia coli O157:H7 in a moisture-enhanced beef model system. The model system was composed of a nonsterile beef homogenate to which NaCl (0.5%) and sodium tripolyphosphate (0.25%) were added, together with the tested antimicrobial ingredients. Beef homogenate treatments were inoculated (ca. 3 log CFU/ml) with rifampin-resistant E. coli O157:H7 (eight-strain mixture) and incubated at 15 °C (48 h). The most effective individual treatments were TO (0.25 or 0.5%) and GSE (0.5 or 1.0%), which immediately reduced (P < 0.05) pathogen levels by ≥ 3.4 log CFU/ml. Additionally, CPC (0.04%) reduced initial E. coli O157:H7 counts by 2.7 log CFU/ml. Most combinations of the tested plant-derived extracts with CPC (0.02 or 0.04%) and sodium diacetate (0.25%) had an additive effect with respect to antibacterial activity. In a second study, antimicrobial interventions were evaluated for their efficacy in reducing surface contamination of E. coli O157:H7 on beef cuts and to determine the effect of these surface treatments on subsequent internalization of the pathogen during blade tenderization. Beef cuts (10 by 8 by 3.5 cm) were inoculated (ca. 4 log CFU/g) on one side with the rifampin-resistant E. coli O157:H7 strain mixture and were then spray treated (20 lb/in(2), 10 s) with water, GSE (5 and 10%), lactic acid (5%), or CPC (5%). Untreated (control) and spray-treated surfaces were then subjected to double-pass blade tenderization. Surface contamination (4.4 log CFU/g) of E. coli O157:H7 was reduced (P < 0.05) to 3.4 (5% CPC) to 4.1 (water or 5% GSE) log CFU/g following spray treatment. The highest and lowest transfer rates of pathogen cells from the surface to deeper tissues of blade-tenderized sections were obtained in the untreated control and CPC-treated samples, respectively.

Lactic acid resistance of Shiga toxin-producing Escherichia coli and multidrug-resistant and susceptible Salmonella Typhimurium and Salmonella Newport in meat homogenate.

This study compared lactic acid resistance of individual strains of wild-type and rifampicin-resistant non-O157 Shiga toxin-producing Escherichia coli (STEC) and of susceptible and multidrug-resistant (MDR) and/or MDR with acquired ampC gene (MDR-AmpC) Salmonella against E. coli O157:H7. After inoculation of sterile 10% beef homogenate, lactic acid was added to a target concentration of 5%. Before acid addition (control), after acid addition (within 2 s, i.e. time-0), and 2, 4, 6 and 8 min after addition of acid, aliquots were removed, neutralized, and analyzed for survivors. Of wild-type and of rifampicin-resistant non-O157 STEC strains, irrespective of serogroup, 85.7% (30 out of 35 strains) and 82.9% (29 out of 35 strains), respectively, reached the detection limit within 0-6 min. Of Salmonella strains, 87.9% (29 out of 33 isolates) reached the detection limit within 0-4 min, irrespective of antibiotic resistance phenotype. Analysis of non-log-linear microbial survivor curves indicated that non-O157 STEC serogroups and MDR and susceptible Salmonella strains required less time for 4D-reduction compared to E. coli O157:H7. Overall, for nearly all strains and time intervals, individual strains of wild-type and rifampicin-resistant non-O157 STEC and Salmonella were less (P < 0.05) acid tolerant than E. coli O157:H7.

Effect of age of cook-in-bag delicatessen meats formulated with lactate-diacetate on the behavior of Listeria monocytogenes contamination introduced when opening the packages during storage.

This study evaluated the potential effect of age of cook-in-bag ham and turkey breast delicatessen meats formulated with lactate-diacetate on survival and/or growth of Listeria monocytogenes introduced after opening of packages and slicing of product. Commercially prepared cured ham and turkey breast products formulated with potassium lactate and sodium diacetate were stored at 1.7°C unsliced, in their original cook-in-bags, and without postlethality exposure. On days 5, 90, 120, and 180 of storage, product slices (10.2 by 7.6 cm) were surface inoculated (1 to 2 log CFU/cm²) with a 10-strain mixture of L. monocytogenes, vacuum packaged (seven slices per bag), and stored at 4°C for up to 13 weeks. Inoculated levels of L. monocytogenes on both products were 1.4 to 1.5 log CFU/cm². Irrespective of product age at slicing and inoculation, after 13 weeks of vacuum-packaged storage (4°C), pathogen counts on product slices were 1.5 to 2.3 (ham) and 2.3 to 2.5 (turkey) log CFU/cm². Overall, the results of the study showed that the age of the cook-in-bag products prior to slicing and inoculation with the pathogen did not (P ≥ 0.05) affect the behavior of L. monocytogenes during vacuum-packaged storage (4°C, up to 13 weeks) of ham and turkey slices. Mean counts of lactic acid bacteria and yeasts and molds, when detected, did not exceed approximately 1 and 2 log CFU/cm², respectively, among all stored samples. Findings of the study will be useful to the meat industry and risk assessors in their efforts to control L. monocytogenes in ready-to-eat meat products.

Effects of cooking methods and chemical tenderizers on survival of Escherichia coli O157:H7 in ground beef patties.

This study evaluated chemical tenderizers and cooking methods to inactivate Escherichia coli O157:H7 in ground beef patties (model system for non-intact beef). Ground beef was inoculated with E. coli O157:H7 and mixed with (i) nothing (control), (ii) calcium chloride (CC) and flavoring agents (FA), (iii) CC, FA, and acetic acid (AA), (iv) sodium chloride (SC), sodium tripolyphosphate (ST), and potassium lactate (PL), and (v) the combination of SC, ST, PL, and AA. Patties were stored in aerobic or vacuum bags at -20, 4, and 12°C. Samples were grilled, broiled, or pan-fried to 60 or 65°C. Total bacterial and E. coli O157:H7 populations remained unchanged during storage. Broiling was more effective in reducing E. coli O157:H7 than grilling and pan-frying, and acidified tenderizers reduced E. coli O157:H7 more than non-acidified tenderizers in broiling. Higher reductions were observed at 65°C than 60°C in broiled and grilled samples. These results indicate that acidified tenderizers and broiling may be useful in non-intact beef safety.

Biofilm formation of O157 and non-O157 Shiga toxin-producing Escherichia coli and multidrug-resistant and susceptible Salmonella typhimurium and newport and their inactivation by sanitizers.

This study compared biofilm formation by 7 serogroups of pathogenic Escherichia coli and 2 or 3 phenotypes of Salmonella (susceptible, multidrug-resistant [MDR], and/or multidrug resistant with ampC gene [MDR-AmpC]). One-week mature biofilms were also exposed to water, quaternary ammonium compound-based (QAC), and acid-based (AB) sanitizers. Seven groups (strain mixture) of above-mentioned pathogens were separately spot-inoculated onto stainless steel coupons surfaces for target inoculation of 2 log CFU/cm2, then stored statically, partially submerged in 10% nonsterilized meat homogenate at 4, 15, and 25 °C. Biofilm cells were enumerated on days 0, 1, 4, and 7 following submersion in 30 mL for 1 min in water, QAC, and AB. Counts on inoculation day ranged from 1.6 ± 0.4 to 2.4 ± 0.6 log CFU/cm2 and changed to 1.2 ± 0.8 to 1.9 ± 0.8 on day 7 at 4 °C with no appreciable difference among the 7 pathogen groups. After treatment with QAC and AB on day 7, counts were reduced (P < 0.05) to less than 0.7 ± 0.6 and 1.2 ± 0.5, respectively, with similar trends among pathogens. Biofilm formation at higher temperatures was more enhanced; E. coli O157:H7, as an example, increased (P < 0.05) from 1.4 ± 0.6 and 2.0 ± 0.3 on day 0 to 4.8 ± 0.6 and 6.5 ± 0.2 on day 7 at 15 and 25 °C, respectively. As compared to 4 °C, after sanitation, more survivors were observed for 15 and 25 °C treatments with no appreciable differences among pathogens. Overall, we observed similar patterns of growth and susceptibility to QAC and AB sanitizers of the 7 tested pathogen groups with enhanced biofilm formation capability and higher numbers of treatment survivors at higher temperatures.

Antilisterial properties of marinades during refrigerated storage and microwave oven reheating against post-cooking inoculated chicken breast meat.

This study evaluated growth of Listeria monocytogenes inoculated on cooked chicken meat with different marinades and survival of the pathogen as affected by microwave oven reheating. During aerobic storage at 7 °C, on days 0, 1, 2, 4, and 7, samples were reheated by microwave oven (1100 W) for 45 or 90 s and analyzed microbiologically. L. monocytogenes counts on nonmarinated (control) samples increased (P < 0.05) from 2.7 ± 0.1 (day-0) to 6.9 ± 0.1 (day-7) log CFU/g during storage. Initial (day-0) pathogen counts of marinated samples were <0.5 log CFU/g lower than those of the control, irrespective of marinating treatment. At 7 d of storage, pathogen levels on samples marinated with tomato juice were not different (P ≥ 0.05; 6.9 ± 0.1 log CFU/g) from those of the control, whereas for samples treated with the remaining marinades, pathogen counts were 0.7 (soy sauce) to 2.0 (lemon juice) log CFU/g lower (P < 0.05) than those of the control. Microwave oven reheating reduced L. monocytogenes counts by 1.9 to 4.1 (45 s) and >2.4 to 5.0 (90 s) log CFU/g. With similar trends across different marinates, the high levels of L. monocytogenes survivors found after microwave reheating, especially after storage for more than 2 d, indicate that length of storage and reheating time need to be considered for safe consumption of leftover cooked chicken.

Efficacy of chemical interventions against Escherichia coli O157:H7 and multidrug-resistant and antibiotic-susceptible Salmonella on inoculated beef trimmings.

Studies were conducted to compare the decontamination efficacy of six chemical treatments against Escherichia coli O157:H7 and multidrug-resistant and antibiotic-susceptible Salmonella inoculated on beef trimmings. The inocula, comprising four-strain mixtures of rifampin-resistant E. coli O157:H7 and antibiotic-susceptible or multidrug-resistant (MDR and/or MDR-AmpC) Salmonella Newport and Salmonella Typhimurium, were inoculated (3 log CFU/cm(2)) separately onto samples (10 by 5 by 1 cm) derived from beef chuck rolls. Samples were left untreated (control), were immersed for 30 s in acidified sodium chlorite (0.1%, pH 2.5), peroxyacetic acid (0.02%, pH 3.8), sodium metasilicate (4%, pH 12.6), Bromitize Plus (0.0225% active bromine, pH 6.6), or AFTEC 3000 (pH 1.2), or were immersed for 5 s in SYNTRx 3300 (pH 1.0). Levels of surviving Salmonella on treated trimmings were not influenced by serotype or antibiotic resistance phenotype and were generally similar (P ≥ 0.05) or lower (P < 0.05) than levels of surviving E. coli O157:H7 regardless of antimicrobial treatment. Overall, depending on chemical treatment (reductions within each chemical treatment were similar among all tested inocula), initial counts of E. coli O157:H7 (2.7 to 3.1 log CFU/cm(2)) were reduced (P < 0.05) by 0.2 to 1.4 log CFU/cm(2). Similarly, initial counts of the tested Salmonella inocula (2.8 to 3.3 log CFU/cm(2)) were reduced (P < 0.05) by 0.4 to 1.4 (Salmonella Newport, antibiotic susceptible), 0.3 to 1.4 (Salmonella Newport, MDR-AmpC), 0.2 to 1.5 (Salmonella Typhimurium, antibiotic susceptible), 0.4 to 1.3 (Salmonella Typhimurium, MDR), and 0.4 to 1.5 (Salmonella Typhimurium, MDR-AmpC) log CFU/cm(2), depending on antimicrobial treatment. Reductions obtained with sodium metasilicate were 1.3 to 1.5 log CFU/cm(2), regardless of inoculum, and reductions obtained with the five remaining antimicrobial treatments were 0.2 to 0.7 log CFU/cm(2) (depending on treatment). Findings of this study should be useful to regulatory authorities and the meat industry as they consider Salmonella contamination on beef trimmings.

Sensitivity of Shiga toxin-producing Escherichia coli, multidrug-resistant Salmonella, and antibiotic-susceptible Salmonella to lactic acid on inoculated beef trimmings.

Studies were performed to determine whether lactic acid treatments used to reduce Escherichia coli O157:H7 on beef trimmings are also effective in controlling non-O157 Shiga toxin-producing E. coli (nSTEC), and multidrug-resistant and antibiotic-susceptible Salmonella. Beef trimming pieces (10 by 5 by 1 cm) were inoculated (3 log CFU/cm(2)) separately with four-strain mixtures of rifampin-resistant E. coli O157:H7, O26, O45, O103, O111, O121, and O145. Similarly, in a second study, trimmings were separately inoculated with rifampin-resistant E. coli O157:H7, and antibiotic-susceptible or multidrug-resistant (MDR and/or MDR-AmpC) Salmonella Newport and Salmonella Typhimurium. Inoculated trimmings were left untreated (control) or were immersed for 30 s in 5% lactic acid solutions (25 or 55°C). No differences (P ≥ 0.05) were obtained among surviving counts of E. coli O157:H7 and those of the tested nSTEC serogroups on lactic acid-treated (25 or 55°C) samples. Counts (3.1 to 3.3 log CFU/cm(2)) of E. coli O157:H7 and nSTEC were reduced (P < 0.05) by 0.5 to 0.9 (25°C lactic acid) and 1.0 to 1.4 (55°C lactic acid) log CFU/cm(2). Surviving counts of Salmonella on treated trimmings were not influenced by serotype or antibiotic resistance phenotype and were similar (P ≥ 0.05) or lower (P < 0.05) than surviving counts of E. coli O157:H7. Counts (3.0 to 3.3 log CFU/cm(2)) were reduced (P < 0.05) by 0.5 to 0.8 (E. coli O157:H7) and 1.3 to 1.5 (Salmonella) log CFU/cm(2) after treatment of samples with 25°C lactic acid. Corresponding reductions following treatment with lactic acid at 55°C were 1.2 to 1.5 (E. coli O157:H7) and 1.6 to 1.9 (Salmonella) log CFU/cm(2). Overall, the results indicated that lactic acid treatments used against E. coli O157:H7 on beef trimmings should be similarly or more effective against the six nSTEC serogroups and against multidrug-resistant and antibiotic-susceptible Salmonella Newport and Salmonella Typhimurium.

Adaptive acid tolerance response of Listeria monocytogenes strains under planktonic and immobilized growth conditions.

The acid resistance of Listeria monocytogenes was evaluated: (i) after short (shock) or long-term (adaptation during growth) exposure to reduced (5.5) or neutral (7.2) pH in a liquid (broth) medium or on a solid surface (agar), and (ii) after growth on the surface of ham and turkey slices or in homogenates of these products. Three L. monocytogenes strains (serotypes 1/2a, 1/2b and 4b) were individually inoculated at: (i) 10(4)-10(5)CFU/ml in tryptic soy broth with 0.6% yeast extract (TSBYE) or on tryptic soy agar with 0.6% yeast extract (TSAYE) at pH 7.2 with 1% (+G) or without (-G) glucose of or TSBYE and TSAYE with 0.25% glucose at pH 5.5 (lactic acid) and incubated at 20°C, and (ii) 10(2)-10(3)CFU/cm(2) on ham and turkey slices (pH 6.39-6.42; formulated with potassium lactate and sodium diacetate) or in their homogenates (1:4 and 1:9; representing viscous [slurry] and liquid residues [purge], respectively), and stored at 10°C. The acid resistance of each strain was assessed in TSBYE of pH 3.5 (lactic acid) for strains growing in broth or on agar surfaces, and in TSBYE of pH 1.5 (HCl) for strains growing on ham and turkey slices or in their homogenates. Habituation at pH 5.5 for 3 or 24h at 20°C increased acid (pH 3.5) resistance of all strains compared to the control (pH 7.2). Cells grown on the surface of TSAYE-G (pH 7.2 or 5.5) showed higher resistance than cells grown in broth (TSBYE-G), whereas the opposite was observed for cells grown on TSAYE + G or in TSBYE + G. Growth of L. monocytogenes on meat product slices was markedly slower than in homogenates. Pathogen reductions following exposure to pH 1.5, after 10 and 27days of storage were strain-dependent and in the ranges of 0.5-2.5, 1.3-4.5 and 4.0-7.6 log units for cells grown on product slices in 1:4 and 1:9 homogenates, respectively. The results suggest that L. monocytogenes cells growing on food surfaces or in viscous matrices may show higher resistance to lethal acid conditions than cells growing in liquid substrates.

Comparison of decontamination efficacy of antimicrobial treatments for beef trimmings against Escherichia coli O157:H7 and 6 non-O157 Shiga toxin-producing E. coli serogroups.

UNLABELLED: The decontamination efficacy of 6 chemical treatments for beef trimmings were evaluated against Escherichia coli O157:H7 and 6 non-O157 Shiga toxin-producing E. coli (nSTEC) serogroups. Rifampicin-resistant 4-strain mixtures of E. coli O157:H7 and nSTEC serogroups O26, O45, O103, O111, O121, and O145 were separately inoculated (3 to 4 log CFU/cm(2)) onto trimmings (10 × 5 × 1 cm; approximately 100 g) fabricated from beef chuck rolls, and were immersed for 30 s in solutions of acidified sodium chlorite (0.1%, pH 2.5), peroxyacetic acid (0.02%, pH 3.8), sodium metasilicate (4%, pH 12.5), Bromitize(®) Plus (0.0225% active bromine, pH 6.6), or AFTEC 3000 (pH 1.2), or for 5 s in SYNTRx 3300 (pH 1.0). Each antimicrobial was tested independently together with an untreated control. Results showed that all tested decontamination treatments were similarly effective against the 6 nSTEC serogroups as they were against E. coli O157:H7. Irrespective of pathogen inoculum, treatment of beef trimmings with acidified sodium chlorite, peroxyacetic acid, or sodium metasilicate effectively (P < 0.05) reduced initial pathogen counts (3.4 to 3.9 log CFU/cm(2)) by 0.7 to 1.0, 0.6 to 1.0, and 1.3 to 1.5 log CFU/cm(2), respectively. Reductions of pathogen counts (3.1 to 3.2 log CFU/cm(2)) by Bromitize Plus, AFTEC 3000, and SYNTRx 3300 were 0.1 to 0.4 log CFU/cm(2), depending on treatment. Findings of this study should be useful to regulatory authorities and the meat industry as they consider nSTEC contamination in beef trimmings. PRACTICAL APPLICATIONS: Findings of this study should be useful to: (i) meat processors as they design and conduct studies to validate the efficacy of antimicrobial treatments to control pathogen contamination on fresh beef products; and (ii) regulatory agencies as they consider approaches for better control of the studied pathogens.

Activity of caprylic acid, carvacrol, ε-polylysine and their combinations against Salmonella in not-ready-to-eat surface-browned, frozen, breaded chicken products.

UNLABELLED: Caprylic acid (CAA), carvacrol (CAR), ε-polylysine (POL), and their combinations were evaluated for reduction of Salmonella contamination in not-ready-to-eat surface-browned, frozen, breaded chicken products. Fresh chicken breast meat pieces (5 × 5 × 5 cm) were inoculated with Salmonella (7-strain mixture; 4-5 log CFU/g) and mixed with distilled water (control) or with CAA, CAR, and POL as single or combination treatments of 2 or 3 ingredients. Sodium chloride (1.2%) and sodium tripolyphosphate (0.3%) were added to all formulations, followed by grinding of the mixtures and forming into 9 × 5 × 3 cm portions. Sample surfaces were brushed with egg whites, coated with breadcrumbs, surface-browned in an oven (208 °C, 15 min), packaged, and stored at -20 °C (7 d). Total reductions of inoculated Salmonella in untreated (control) surface-browned, breaded products after frozen storage were 0.8 to 1.4 log CFU/g. In comparison, single treatments of CAA (0.25% to 1.0%), CAR (0.3% to 0.5%), and POL (0.125% to 1.0%) reduced counts by 2.9 to at least 4.5, 3.4 to at least 4.4, and 1.4 to 2.3 log CFU/g, respectively, depending on concentration. Pathogen counts of products treated with 2- or 3-ingredient combination treatments (0.03125% to 0.25% CAA, 0.0375% to 0.3% CAR, and/or 0.5% POL) were 0.4 to at least 3.3 log CFU/g lower (depending on treatment) than those of the untreated controls. The antimicrobial activity of 2-ingredient combinations comprised of 0.125% CAA, 0.15% CAR, or 0.5% POL was enhanced (P < 0.05) when applied as a 3-ingredient combination (that is, 0.125% CAA + 0.15% CAR + 0.5% POL). These data may be useful for the selection of antimicrobial treatments to reduce Salmonella contamination in not-ready-to-eat processed chicken products. PRACTICAL APPLICATION: Findings from the study may be useful for the selection of suitable antimicrobials, concentrations, and combinations to reduce Salmonella contamination in not-ready-to-eat surface-browned, frozen, breaded chicken products.

Antimicrobials for reduction of Salmonella contamination in uncooked, surface-browned breaded chicken products.

Surface-browned but uncooked frozen breaded chicken products have been associated with salmonellosis outbreaks due to inadequate or no cooking of the products before consumption. This study was conducted to evaluate the effect of three antimicrobials against Salmonella during manufacture of a surface-browned, uncooked frozen breaded chicken meat product. Fresh chicken breast meat portions (5 by 5 by 5 cm) were inoculated (4 to 5 log CFU/g) with Salmonella and mixed with caprylic acid (CAA; 0.5 and 1.0%), carvacrol (CAR; 0.3 and 0.5%), ε-polylysine (POL; 0.125 and 0.25%), or distilled water (control). Sodium chloride (1.2%) and sodium tripolyphosphate (0.3%) were added to all treatments, and the mixtures were ground (5% total moisture enhancement level) and formed into portions (9 by 5 by 3 cm). The products were breaded and surface browned by baking in an oven (208°C for 15 min) or deep frying in vegetable oil (190°C for 15 s), packaged in polyethylene bags, and stored at -20°C for 7 days. Total reductions of inoculated Salmonella in untreated control oven- or fryer-browned products after frozen storage were 1.2 and 0.8 log CFU/g, respectively. In comparison, treatment with CAA, CAR, or POL reduced initial pathogen counts by 3.3 to >4.5, 4.1 to >4.7, and 1.1 to 1.6 log CFU/g, respectively, regardless of the antimicrobial concentration and browning method. Treatment with 1.0% CAA (oven browned) or 0.5% CAR (oven or fryer browned) reduced Salmonella to nondetectable levels (<0.3 log CFU/g) in stored frozen products. These data may be useful for development of suitable antimicrobial treatments to reduce the risk of Salmonella contamination in surface-browned, uncooked frozen breaded chicken products.

Transfer, attachment, and formation of biofilms by Escherichia coli O157:H7 on meat-contact surface materials.

UNLABELLED: Studies examined the effects of meat-contact material types, inoculation substrate, presence of air at the liquid-solid surface interface during incubation, and incubation substrate on the attachment/transfer and subsequent biofilm formation by Escherichia coli O157:H7 on beef carcass fabrication surface materials. Materials studied as 2 × 5 cm coupons included stainless steel, acetal, polypropylene, and high-density polyethylene. A 6-strain rifampicin-resistant E. coli O157:H7 composite was used to inoculate (6 log CFU/mL, g, or cm²) tryptic soy broth (TSB), beef fat/lean tissue homogenate (FLH), conveyor belt-runoff fluids, ground beef, or beef fat. Coupons of each material were submerged (4 °C, 30 min) in the inoculated fluids or ground beef, or placed between 2 pieces of inoculated beef fat with pressure (20 kg) applied. Attachment/transfer of the pathogen was surface material and substrate dependent, although beef fat appeared to negate differences among surface materials. Beef fat was the most effective (P < 0.05) inoculation substrate, followed by ground beef, FLH, and TSB. Incubation (15 °C, 16 d) of beef fat-inoculated coupons in a beef fat homogenate (pH 4.21) allowed the pathogen to survive and grow on coupon surfaces, with maximal biofilm formation observed between 2 and 8 d of storage and when air was present at the liquid-solid interface. The results indicated that the process of fabricating beef carcasses may be conducive to the attachment of E. coli O157:H7 onto meat-contact surfaces and subsequent biofilm formation. Furthermore, it is recommended that substrates found in beef fabrication settings, rather than laboratory culture media, be used in studies designed to investigate E. coli O157:H7 biofilm development and control in these environments. PRACTICAL APPLICATION: Findings of this study provide knowledge on the effect of type of beef carcass fabrication surface material, fabrication-floor fluids and residues, and incubation conditions on attachment/transfer and subsequent biofilm formation by E. coli O157:H7. The results highlight the importance of thoroughly cleaning soiled surfaces to remove all remnants of beef fat or other organic material that may harbor or protect microbial contaminants during otherwise lethal antimicrobial interventions.

Thermal inactivation of Escherichia coli O157:H7 inoculated at different depths of non-intact blade-tenderized beef steaks.

UNLABELLED: Studies evaluated thermal inactivation of Escherichia coli O157:H7 inoculated at different depths of simulated blade-tenderized non-intact steaks. Fresh beef slices (0.3 or 0.6 cm thick) were stacked on top of each other to form 2.4 or 1.2 cm thick steaks. Steaks were blade-tenderized and then inoculated with rifampicin-resistant Escherichia coli O157:H7 (8 strain mixture; 4 log CFU/cm(2)) on the surface or between slices, vacuum-packaged, and stored at 4 or -20 °C for 5 d before cooking. Steaks were cooked by pan-broiling or roasting to a geometric center temperature of 60 °C. Frozen samples were either cooked from the frozen state or after thawing to approximately 4 or 25 °C. In steaks inoculated on the external surface and cooked by pan-broiling, pathogen survivors recovered from thinner (1.2 cm) steaks were greater (P < 0.05) than those recovered from thicker (2.4 cm) steaks. Cooking steaks from a frozen state or after thawing (4 or 25 °C) did not (P ≥ 0.05) affect extent of pathogen inactivation. Survivors after pan-broiling of 2.4 cm thick steaks increased (P < 0.05) from 0.3 to 1.3 log CFU/cm(2) for surface-inoculated steaks to 2.5 to 3.2 log CFU/cm(2) for samples inoculated at the center (1.2 cm depth). In comparison, overall thermal destruction of the pathogen in steaks cooked by roasting was less, and survivor counts were generally not different (P ≥ 0.05) at each depth of inoculation. These data should be useful in development of lethality guidelines to ensure safe consumption of non-intact meat products. PRACTICAL APPLICATION: Results of this study should be useful for developing cooking guidelines, for foodservice establishments and consumers, to ensure safe consumption of non-intact meat products.

Attachment and biofilm formation by Escherichia coli O157:H7 at different temperatures, on various food-contact surfaces encountered in beef processing.

Escherichia coli O157:H7 attached to beef-contact surfaces found in beef fabrication facilities may serve as a source of cross-contamination. This study evaluated E. coli O157:H7 attachment, survival and growth on food-contact surfaces under simulated beef processing conditions. Stainless steel and high-density polyethylene surfaces (2×5cm) were individually suspended into each of three substrates inoculated (6log CFU/ml or g) with E. coli O157:H7 (rifampicin-resistant, six-strain composite) and then incubated (168h) statically at 4 or 15°C. The three tested soiling substrates included sterile tryptic soy broth (TSB), unsterilized beef fat-lean tissue (1:1 [wt/wt]) homogenate (10% [wt/wt] with sterile distilled water) and unsterilized ground beef. Initial adherence/attachment of E. coli O157:H7 (0.9 to 2.9log CFU/cm(2)) on stainless steel and high-density polyethylene was not affected by the type of food-contact surface but was greater (p<0.05) through ground beef. Adherent and suspended E. coli O157:H7 counts increased during storage at 15°C (168h) by 2.2 to 5.4log CFU/cm(2) and 1.0 to 2.8log CFU/ml or g, respectively. At 4°C (168h), although pathogen levels decreased slightly in the substrates, numbers of adherent cells remained constant on coupons in ground beef (2.4 to 2.5log CFU/cm(2)) and increased on coupons in TSB and fat-lean tissue homogenate by 0.9 to 1.0and 1.7 to 2.0log CFU/cm(2), respectively, suggesting further cell attachment. The results of this study indicate that E. coli O157:H7 attachment to beef-contact surfaces was influenced by the type of soiling substrate and temperature. Notably, attachment occurred not only at a temperature representative of beef fabrication areas during non-production hours (15°C), but also during cold storage (4°C) temperatures, thus, rendering the design of more effective sanitation programs necessary.

Survival of Escherichia coli O157:H7 in meat product brines containing antimicrobials.

UNLABELLED: Brine solution injection of beef contaminated with Escherichia coli O157:H7 on its surface may lead to internalization of pathogen cells and/or cross-contamination of the brine, which when recirculated, may serve as a source of new product contamination. This study evaluated survival of E. coli O157:H7 in brines formulated without or with antimicrobials. The brines were formulated in sterile distilled water (simulating the composition of freshly prepared brines) or in a nonsterile 3% meat homogenate (simulating the composition of recirculating brines) at concentrations used to moisture-enhance meat to 110% of initial weight, as follows: sodium chloride (NaCl, 5.5%) + sodium tripolyphosphate (STP, 2.75%), NaCl + sodium pyrophosphate (2.75%), or NaCl + STP combined with potassium lactate (PL, 22%), sodium diacetate (SD, 1.65%), PL + SD, lactic acid (3.3%), acetic acid (3.3%), citric acid (3.3%), nisin (0.0165%) + ethylenediamine tetraacetic acid (EDTA, 200 mM), pediocin (11000 AU/mL) + EDTA, sodium metasilicate (2.2%), cetylpyridinium chloride (CPC, 5.5%), or hops beta acids (0.0055%). The brines were inoculated (3 to 4 log CFU/mL) with rifampicin-resistant E. coli O157:H7 (8-strain composite) and stored at 4 or 15 °C (24 to 48 h). Immediate (0 h) pathogen reductions (P < 0.05) of 1.8 to ≥ 2.4 log CFU/mL were observed in brines containing CPC or sodium metasilicate. Furthermore, brines formulated with lactic acid, acetic acid, citric acid, nisin + EDTA, pediocin + EDTA, CPC, sodium metasilicate, or hops beta acids had reductions (P < 0.05) in pathogen levels during storage; however, the extent of pathogen reduction (0.4 to > 2.4 log CFU/mL) depended on the antimicrobial, brine type, and storage temperature and time. These data should be useful in development or improvement of brine formulations for control of E. coli O157:H7 in moisture-enhanced meat products. PRACTICAL APPLICATION: Results of this study should be useful to the meat industry for developing or modifying brine formulations to reduce the risk of E. coli O157:H7 in moisture-enhanced meat products.

Inactivation of Escherichia coli O157:H7 in moisture-enhanced nonintact beef by pan-broiling or roasting with various cooking appliances set at different temperatures.

This study evaluated inactivation of Escherichia coli O157:H7 in moisture-enhanced restructured nonintact beef cooked to 65 °C using different cooking appliances set at different temperatures. Batches (2 kg) of coarse-ground beef (approximately 5% fat) were mixed with an 8-strain composite (100 mL) of rifampicin-resistant E. coli O157:H7 (6.4 ± 0.1 log CFU/g) and a solution (100 mL) of sodium chloride plus sodium tripolyphosphate to yield concentrations (wt/wt) of 0.5% and 0.25%, respectively, in the final product. Beef portions of 2.54 cm thickness (15 cm dia) were prepared and were vacuum-packaged and frozen (-20 °C, 42 h). Partially thawed (-2.5 ± 1.0 °C) portions were pan-broiled (Presto electric skillet and Sanyo grill) or roasted (Oster toaster oven and Magic Chef kitchen oven) to 65 °C. The appliances were set at, and preheated before cooking to 149 or 204 °C (electric skillet), 149 or 218 °C (grill), 149 or 232 °C (toaster oven), and 149, 204, or 260 °C (kitchen oven). Temperatures of appliances and beef samples were monitored with thermocouples, and meat samples were analyzed for surviving microbial populations. In general, the higher the appliance temperature setting, the shorter the time needed to reach 65 °C, and the higher the edge and surface temperatures of the meat samples. Temperatures of 204 to 260 °C, regardless of appliance, resulted in greater (P < 0.05) pathogen reductions (3.3 to 5.5 log CFU/g) than those obtained at 149 °C (1.5 to 2.4 log CFU/g). The highest (P < 0.05) reduction (5.5 log CFU/g) was obtained in samples cooked in the kitchen oven set at 260 °C. The results should be useful to the food service industry for selection of effective nonintact beef cooking protocols, and for use in risk assessments for nonintact meat products. Practical Application: Results of this study should be useful for developing cooking recommendations to enhance the safety of nonintact beef products, and for use in risk assessments of such products.

Thermal inactivation of acid, cold, heat, starvation, and desiccation stress-adapted Escherichia coli O157:H7 in moisture-enhanced nonintact beef.

This study was conducted to compare thermal inactivation of stress-adapted and nonadapted Escherichia coli O157:H7 in nonintact beef moisture enhanced with different brine formulations and cooked to 65°C. Coarsely ground beef was mixed with acid, cold, heat, starvation, or desiccation stress-adapted or nonadapted rifampin-resistant E. coli O157:H7 (eight-strain mixture, 5 to 6 log CFU/g) and a brine solution for a total moisture enhancement level of 10%. The brine treatments included distilled water (control), sodium chloride (0.5% NaCl) plus sodium tripolyphosphate (0.25% STP), or NaCl + STP combined with cetylpyridinium chloride (0.2% CPC), lactic acid (0.3% LA), or sodium metasilicate (0.2% SM). The treated meat was extruded into bags (15 cm diameter), semifrozen (-20°C for 4.5 h), and cut into 2.54-cm (1-in.)-thick portions. Samples were individually vacuum packaged, frozen (-20°C for 42 h), and tempered at 4°C for 2.5 h before cooking. Partially thawed (-1.8 ± 0.4°C) samples were pan broiled to an internal temperature of 65°C. Pathogen counts of partially thawed (before cooking) samples moisture enhanced with brines containing CPC, LA, or SM were 0.7 to 1.1, 0.0 to 0.4, and 0.2 to 0.4 log CFU/g, respectively, lower than those of the control. Compared with microbial count reductions obtained after pan broiling of beef inoculated with nonadapted E. coli O157:H7 cells, count reductions during cooking of meat inoculated with cold and desiccation stress-adapted, acid stress-adapted, and heat and starvation stress-adapted cells indicated sensitization, cross protection, and no effect, respectively, of these stresses on the pathogen during subsequent exposure to heat. Among all stressed cultures, CPC-treated samples (0.8 to 3.6 log CFU/g) and LA-treated samples (0.8 to 3.5 log CFU/g) had the lowest numbers of E. coli O157:H7 survivors after cooking.