Thermal Inactivation of Escherichia coli O157:H7 in Cooked Turkey Products

Survival of Escherichia coli O157:H7 when heated in commercial-type turkey products was determined. Thermal death times (TDT) were determined at 52–60°C in ground turkey with no additives, 3% fat; ground turkey with no additives, 11% fat; turkey ham batter, 11% fat; turkey frank batter, 17% fat; and turkey sausage batter, 31% fat. Mean D₅₂-values ranged from 44.9 to 116 min; D₅₅-values from 6.63 to 39.4 min; D₅₇-values from 2.20 to 11.7 min; D₆₀-values from 0.68 to 5.86 min. At all temperatures, survival of E. coli O157:H7 was greater in formulated products than in turkey meat with no additives. Greatest survival occurred in the turkey frank batter. Using our z-value data, times to provide a 5 D kill of E. coli O157:H7 in turkey franks cooked at 60°C, 65.6°C, or 71°C would be 26, 3.1, or 0.37 min, respectively.     

Thermal inactivation ofClostridium perfringensvegetative cells in ground beef and turkey as affected by sodium pyrophosphate

The heat resistance (55–65°C) ofClostridium perfringensvegetative cells in ground beef and turkey that included 0, 0.15 or 0.3% (w/w) sodium pyrophosphate (SPP) was assessed in bags heated using a water bath. The surviving cell population was assayed on tryptose–sulfite– cycloserine agar. The decimal reduction (D)-values in beef that included no SPP were 21.6, 10.2, 5.3, and 1.6min at 55, 57.5, 60, 62.5°C, respectively; the values in turkey ranged from 17.5min at 55°C to 1.3min at 62.5°C. Addition of 0.15% SPP resulted in concomitant decrease in heat resistance as evidenced by reduced bacterialD-values. TheD-values in beef that included 0.15% SPP were 17.9, 9.4, 3.5, and 1.2min at 55, 57.5, 60, and 62.5°C, respectively; the values in turkey ranged from 16.2min at 55°C to 1.1min at 62.5°C. The heat resistance was further decreased when the SPP level in beef and turkey was increased to 0.3%. Heating such products to an internal temperature of 65°C for 1min killed >8log₁₀cfug^-1. The z-values in beef and turkey for all treatments were similar, ranging from 6.22 to 6.77°C. Thermal death time values from this study should assist institutional food service settings in the design of thermal processes that ensure safety againstC. perfringensin cooked beef and turkey.     

Thermal Inactivation of Clostridium perfringens After Growth at Several Constant and Linearly Rising Temperatures

Inactivation kinetics of Clostridium perfringens strains NCTC 8238 and NCTC 8798 vegetative cells were evaluated in autoclaved ground beef after growth at constant (37, 41, 45, or 49°C) or linearly rising temperatures (4.0, 6.0, or 7.5 C°/hr) representative of long-time, low-temperature (LTLT) cooking. Inactivation temperatures of 55, 57, 59, 60, and 61°C were used. D values and z values were determined. For strain NCTC 8798 cells grown at 45°C, the average D₅₉°C was 7.2 min and the z_D was 3.8 C°. Both strains exhibited greater heat resistance after growth at higher constant temperatures. Also, NCTC 8798 was more heat resistant than NCTC 8238. With linearly rising temperature, terminal growth temperatures appeared dominant in resistance to inactivation. These data will permit predictions of growth and survival of C. perfringens during LTLT cooking of beef roasts.     

Heat-Resistance of Escherichia Coli O157:H7 in Meat and Poultry as Affected by Product Composition

The effects of fat level and low fat formulation on survival of Escherichia coli O157:H7 isolate 204P heated in ground beef [7%, 10% and 20% fat], pork sausage [7%, 10%, and 30% fat], chicken (3% and 11% fat), and turkey (3% and 11% fat) were determined by D- and z-values. D-values for E. coli 0157:H7 in lowest fat products were lower than in traditional beef and pork products (P < 0.05). Overall, higher fat levels in all products resulted in higher D-values. D₆₀ values (min) ranged from 0.45–0.47 in beef, 0.37–0.55 in pork sausage, 0.38–0.55 in chicken and 0.55–0.58 in turkey. D₅₅ and D₅₀ values were respectively longer. Z-values ranged from 4.4–4.8°C. Product composition affected lethality of heat to E. coli O157:H7.     

Changes in heat tolerance of Escherichia coli O157:H7 after exposure to acidic environments

Acid-adapted bacterial cells are known to have enhanced tolerance to various secondary stresses. However, a comparison of heat tolerance of acid-adapted and acid-shocked cells of Escherichia coli O157:H7 has not been reported. D - and z -values of acid-adapted, acid-shocked, and control cells of an unusually heat-resistant strain (E0139) of E. coli O157:H7, as well as two other strains of E. coli O157:H7, were determined based upon the number of cells surviving heat treatment at 52, 54 or 56°C in tryptic soy broth (pH 7·2) for 0, 10, 20 or 30 min. The unusual heat tolerance of E. coli O157:H7 strain E0139 was confirmed. D -values for cells from 24-h cultures were 100·2, 28·3, and 6·1 min at 52, 54 and 56°C, respectively, with a z -value of 3·3°C. The highest D -values of other E. coli O157:H7 strains were 13·6 and 9·2 min at 52 and 54°C, respectively, whereas highest D -values of non-O157:H7 strains were 78·3 and 29·7 min at 52 and 54°C. D -values of acid-adapted cells were significantly higher than those of unadapted and acid-shocked cells at all temperatures tested. In a previous study, we observed that both acid-adapted cells and acid-shocked cells of strain E0139 had enhanced acid tolerance. This suggests that different mechanisms protect acid-adapted and acid-shocked cells against subsequent exposure to heat or an acidic environment. The two types of cells should be considered separately when evaluating survival and growth characteristics upon subsequent exposure to different secondary stress conditions.     

Extraordinary Heat Resistance of Talaromyces flavus and Neosartorya fischeri Ascospores in Fruit Products

Ascospores of three strains each of Talaromyces flavus, Neosartorya fischeri and Byssochlamys fulva/nivea were analyzed for resistance to thermal inactivation in five fruit-based (blueberry, cherry, peach, raspberry and strawberry) products. D_91°C values for two strains of T. flavus ranged from 2.9–5.4 min; D_88°C values ranged from 7.1–22.3 min. Ascospores of N. fischeri were somewhat less heat resistant; D_91°C values were < 2.0 min and D_88°C were 4.2–16.2 min. Ascospores of Byssochlamys spp. were considerably less heat resistant. The type of fruit product did not appear to substantially influence rates of thermal inactivation. No heat-resistant ascospores of T. flavus or N. fischeri, i.e., ascospores capable of surviving 15 min at 75°C, were formed on fruit products stored at 10°C for 137 days. However, T. flavus and N. fischeri formed ascospores on cherry substrate stored at 25°C within 65 and 137 days, respectively, that survived 15 min at 88°C.     

Thermal inactivation of polyphenoloxidase and peroxidase in green coconut (Cocos nucifera) water

Coconut water is an isotonic beverage naturally obtained from the green coconut. After extracted and exposed to air, it is rapidly degraded by enzymes peroxidase (POD) and polyphenoloxidase (PPO). To study the effect of thermal processing on coconut water enzymatic activity, batch process was conducted at three different temperatures, and at eight holding times. The residual activity values suggest the presence of two isoenzymes with different thermal resistances, at least, and a two‐component first‐order model was considered to model the enzymatic inactivation parameters. The decimal reduction time at 86.9 ^°C (D_{86.9 °C}) determined were 6.0 s and 11.3 min for PPO heat labile and heat resistant fractions, respectively, with average z‐value = 5.6 °C (temperature difference required for tenfold change in D). For POD, D_{86.9 °C} = 8.6 s (z = 3.4 °C) for the heat labile fraction was obtained and D_{86.9 °C} = 26.3 min (z = 6.7 °C) for the heat resistant one.     

Differences in thermotolerance of variousEscherichia coliO157:H7 strains in a salami matrix

Three strains ofEscherichia coliO157:H7 (ATCC 43895, Ent C9490 and 380–94) were inoculated into salami and heated in water baths at 50, 55 or 60°C. At intervals between 1 and 360 min, salami samples were removed from the water bath and examined for the presence of survivingE. coliO157:H7. Samples were directly plated onto sorbitol MacConkey (SMAC) agar, and onto tryptone soya agar (TSA) with SMAC overlay. The number of sub-lethally damaged cells in each sample was estimated from the differences between the resultant direct (uninjured cells only) and overlay (total recovery) counts. In samples heated at 50°C, the percentage of cell injury ranged from 71·8–88% for all strains. In samples heated at 55°C the percentage of sub-lethally damaged cells in strains ATCC 43895 and Ent C9490 was significantly higher (P< 0·001) at 97% than that observed in strain 380–94 (64%). Cell injury was not measured at 60°C. There were significant differences between the derived decimal reduction times (D-values) related to the different strains ofE. coliO157:H7, the heat treatment applied and the recovery/enumeration agars used. Significant interstrain differences (P< 0·05) in thermotolerance were noted. Strain Ent C9490 was significantly more heat resistant at 50°C and 60°C (D-values of 116·9 and 2·2 min, respectively), while at 55°C strain 380–94 was more thermotolerant (D-value of 21·9 min). The implications of these findings for the design of studies investigating the heat resistance ofE. coliO157:H7 in fermented meat environments are discussed.     

Heat resistance of Escherichia coli O157:H7 in cook‐in‐bag ground beef as affected by pH and acidulant†

Summary The heat resistance of a four‐strain mixture of Escherichia coli O157:H7 was tested. The temperature range was 55–62.5 °C and the substrate was beef at pH 4.5 or 5.5, adjusted with either acetic or lactic acid. Inoculated meat, packaged in bags, was completely immersed in a circulating water bath and cooked to an internal temperature of 55, 58, 60, or 62.5 °C in 1 h, and then held for pre‐determined lengths of time. The surviving cell population was enumerated by spiral plating meat samples on tryptic soy agar overlaid with Sorbitol MacConkey agar. Regardless of the acidulant used to modify the pH, the D ‐values at all temperatures were significantly lower (P < 0.05) in ground beef at pH 4.5 as compared with the beef at pH 5.5. At the same pH levels, acetic acid rendered E. coli O157:H7 more sensitive to the lethal effect of heat. The analysis of covariance showed evidence of a significant acidulant and pH interaction on the slopes of the survivor curves at 55 °C. Based on the thermal‐death–time values, contaminated ground beef (pH 5.5/lactic acid) should be heated to an internal temperature of 55 °C for at least 116.3 min and beef (pH 4.5/acetic acid) for 64.8 min to achieve a 4‐log reduction of the pathogen. The heating time at 62.5 °C, to achieve the same level of reduction, was 4.4 and 2.6 min, respectively. Thermal‐death–time values from this study will assist the retail food processors in designing acceptance limits on critical control points that ensure safety of beef originally contaminated with E. coli O157:H7.     

Determination of the effect of sodium lactate on the survival and heat resistance of Escherichia coli O157:H7 in two commercial beef patty formulations

The effect of 4% sodium lactate (NaL) in beefburger patty formulations on the survival and heat resistance of Escherichia coli O157:H7 was investigated. Fresh beef trimmings were inoculated with E. coli O157:H7 to a concentration of 6·0–7·0 log₁₀ cfu g⁻¹ and subjected to the processing stages of beefburger patty production. Two commercial beefburger patty formulations were produced: a ‘quality’ patty (100% beef) and an ‘economy’ patty (70% beef, 30% other ingredients, including onion, water, salt, seasoning, rusk and soya concentrate). Sodium lactate (4% w/v) was added to the beefburger patties during mincing and the formed patties were frozen and stored for 1 month. Beefburger patties without added NaL were used as controls. After frozen storage for 1 month, patties were examined for E. coli O157:H7 counts. There was a synergistic effect between freezing and NaL, which resulted in a small but significant reduction (P<0·05) (approximately 0·5 log₁₀ cfu g⁻¹) in E. coli O157:H7 numbers. The frozen beefburger patties were also heat-treated at 50, 55 and 60°C and the data analysed to derive D -values for E. coli O157:H7 cells. At each temperature treatment, theD -values of the quality and economy beefburger patties with 4% NaL were significantly lower (P<0·001) than the D -values of the patty formulations without NaL. The study demonstrates that the presence of 4% NaL in beefburger patty formulations can reduce the overall risks posed to consumers by the presence ofE. coli O157:H7 by, first; reducing pathogen survival during freezing and frozen storage of the uncooked product; and, second, by increasing the susceptibility of the pathogen to heat during normal cooking processes.     

Recovery of heat-stressed E. coli O157:H7 from ground beef and survival of E. coli O157:H7 in refrigerated and frozen ground beef and in fermented sausage kept at 7°C and 22°C

A study was undertaken to obtain information on survival of Escherichia coli O157:H7 in ground beef subjected to heat treatment, refrigeration and freezing and on survival of E. coli O157:H7 in fermented sausage kept at 7°C and 22°C. For the challenge test, a mixture of E. coli O157:H7 strains (EH 321, EH 385, EH 302) was used and enumeration was performed on an isolation medium suitable for recovery of stressed organisms: modified Levine's eosin methylene blue agar (mEMB). Heat resistance of E. coli O157:H7 decreased after pre-incubation at a reduced temperature.Escherichia coli O157:H7 was more susceptible to heat inactivation after storage at 7°C and die-off was even more enhanced if cultures were frozen prior to heat inactivation. The enhanced reduction of the pathogen at 56°C after prior storage under refrigeration was confirmed in a test with inoculated ground beef.Escherichia coli O157:H7 was able to survive in ground beef at 7°C for 11 days and at −18°C for 35 days showing maximal one log reduction during the storage period. Thus, ground beef contaminated with E. coli O157:H7 will remain a hazard even if the ground beef is held at low or freezing temperatures. At both 7°C and 22°C, a gradual reduction of E. coli O157:H7 was noticed in fermented sausage over the 35 days storage period resulting in a 2 log decrease of the high inoculum (10⁶cfu 25 g⁻¹). For the low inoculum (10³cfu 25 g⁻¹) a 2·5 log reduction was obtained in 7 and 28 days storage at respectively 22 and 7°C. Application of good hygienic practices and implementation of HACCP in the beef industry are important tools in the control of E. coli O157:H7.     

Thermal Resistance of Saccharomyces cerevisiae in Pilsen Beer

The thermal resistance of S. cerevisiae (CMOJ896) was determined in Pilsen beer (pH = 4.28 ± 0.05; extract of original wort (EOW %) = 11.30 ± 0.08; percentage of alcohol by volume (% at 20°C) = 4.97 ± 0.05; total nitrogen content (mg/L) = 590 ± 37; bitterness units (BU) = 20.5 ± 1.3; carbonation of the beer (Vol. CO₂) = 2.89 ± 0.09; color (SRCM) = 3.3 ± 0.4). The flask method was used for an initial population of 1 × 10⁴ cells/mL. Decimal reduction times of D_47°C = 3.16 min, D_48°C = 2.65 min, D_49°C = 1.74 min and D_50°C = 0.68 min were obtained at the temperatures studied. Values of D_60°C = 0.01 min and z = 4.6° were obtained for this microorganism.     

Enhancing the thermal destruction of Escherichia coli O157:H7 in ground beef patties by trans-cinnamaldehyde

The effect of trans-cinnamaldehyde (TC) on the inactivation of Escherichia coli O157:H7 in undercooked ground beef patties was investigated. A five-strain mixture of E. coli O157:H7 was inoculated into ground beef (7.0 log CFU/g), followed by addition of TC (0, 0.15, and 0.3%). The meat was formed into patties and stored at 4 °C for 5 days or at −18 °C for 7 days. The patties were cooked to an internal temperature of 60 or 65 °C, and E. coli O157:H7 was enumerated. The numbers of E. coli O157:H7 did not decline during storage of patties. However, cooking of patties containing TC significantly reduced (P < 0.05) E. coli O157:H7 counts, by >5.0 log CFU/g, relative to the reduction in controls cooked to the same temperatures. The D-values at 60 and 65 °C of E. coli O157:H7 in TC-treated patties (1.85 and 0.08 min, respectively) were significantly lower (P < 0.05) than the corresponding D-values for the organism in control patties (2.70 and 0.29 min, respectively). TC-treated patties were more color stable and showed significantly lower lipid oxidation (P < 0.05) than control samples. TC enhanced the heat sensitivity of E. coli O157:H7 and could potentially be used as an antimicrobial for ensuring pathogen inactivation in undercooked patties. However detailed sensory studies will be necessary to determine the acceptability to consumers of TC in ground beef patties.     

Increased thermal tolerance in Cronobacter sakazakii strains in reconstituted milk powder due to cross protection by physiological stresses

Cronobacter sakazakii (C. sakazakii) is an opportunistic, neonatal, and food borne pathogen primarily associated with the contamination of powdered infant formula (PIF). The pathogen is reported to overcome the food safety barriers such as increased acidity, heat treatment, and so on. This study evaluates the thermal tolerance of C. sakazakii strains independently at 52, 55, and 58°C in reconstituted PIF after exposure to physiological stresses: refrigeration (4°C for 24 hr), starvation (37°C for 48 hr), and desiccation (25°C for 4 days). The Log₁₀ CFU/ml and D-values indicated that survival rate of all the strains decreased significantly (p < .05) after desiccation as compared to those of the control condition (without stress exposure). However, cold stress increased the thermal tolerance of all strains at all temperatures (52, 55, and 58°C) as indicated by increased D-values. Among the tested strains, C. sakazakii strain N15 was found to be the most resistant to thermal treatment after each stress exposure as depicted by principal component analysis (PCA). No apparent correlation between thermal tolerance and starvation stress was observed. The findings indicate that prior exposure to stress conditions may induce cross protection to thermal treatment in C. sakazakii.     

Thermal inactivation of Escherichia coli O157:H7 and Escherichia coli isolated from morcilla as affected by composition of the product

Morcilla is a link sausage quite similar to black pudding, consisting of an inert casing stuffed with a mixture of beef blood, fat, and seasonings. Thirty samples of morcilla showed total microbial counts (6.3×10³–2.1×10⁸ Cfu/g ), molds and yeasts (8.9×10¹–6.3×10⁴ Cfu/g), sulfite-reducing microorganism (2.0×10¹–2.1×10²MPN/g); total coliforms (1.4×10¹–1.1×10³ MPN/g); fecal coliforms (7.0–1.5×10²MPN/g); Enterobactereaceae (1.6×10²–5.0×10⁵ Cfu/g). S. aureus and B. cereus were not detected. E. coli was detected in 76.6% of the samples analyzed. The thermal resistance (D and z-values) of Escherichia coli O157:H7 and E. coli isolated from morcilla were determined in nutrient broth and in a heating menstruun prepared with ground morcilla (discarding the casing) and added fat or starch. Higher fat and starch levels resulted in higher D-values (min) at 54, 58 and 62 °C for both strains. The z-values (°C) for isolated E. coli in nutrient broth (M1), ground morcilla (M2), M2+10% fat (M3), M2+20% fat (M4), M2+10% starch (M5), and M2+20% starch (M6) were 7.9, 7.8, 10.5, 10.4, 10.3, and 10.4, respectively, and for E. coli O157:H7 were 7.8, 7.4, 9.8, 10.2, 10.3, and 10.7. The composition of product affected heat lethality of the two strains of E. coli.     

Optimization and evaluation of heat-shock condition for spore enumeration being used in thermal-process verification: Differential responses of spores and vegetative cells of Clostridium sporogenes to heat shock

To evaluate a heat-shock condition for the enumeration of Clostridium sporogenes spores, a surrogate for C. botulinum spores, we examined the heat tolerance of C. sporogenes spores and vegetative cells exposed to a heat shock at 90°C. From the D values of the spores determined in the temperature range of 113–121°C, z value (±SD) and D_90°C value were estimated to be 10.16±0.90°C and 1,071.52 min, respectively, and the inactivation rates were predicted to be only approximately 2% at 90°C for up to 10 min. Meanwhile, the viable count of spores was significantly higher when activated under a heat-shock condition of 90°C for over 9 min than those activated for shorter time periods. The heat tolerance of vegetative cells was extremely low, showing a D_90°C value (±SD) of 0.21±0.01 min. Finally, 3 different heat-shock conditions were compared: 70°C for 30 min, 80°C for 20 min, and 90°C for 10 min, and the experimental comparative data showed no significant differences in viable spore counts. Consequently, these results support that the heat-shock treatment at 90°C for 10 min is suitable to activate spores and to inactivate vegetative cells of C. sporogenes.     

Heating and Storage Conditions Affect Survival and Recovery of Listeria monocytogenes in Ground Pork

The effect of heating rate, heating atmosphere, and meat age on survival of Listeria monocytogenes was examined in ground pork, as well as the ability of injured cells to recover during storage under air and vacuum packaging at 4 and 30°C. Significantly more survivors were observed when samples were heated at 1.3°C/min than at 8.0°C/ min. Cells inoculated into 3-month-old pork were more sensitive to heating than cells inoculated into fresh ground pork (D_62°C= 5.2 min and 7.7 min, respectively). More survivors were detected when the meat was heated aerobically than anaerobically. However, storage under vacuum at 4 or 30°C resulted in faster recovery compared with cells packaged in air. Thus, heating and packaging conditions affected ability of this pathogen to survive and recover after a heat process in pork.     

Resistance of Yeast to Dry Heat

The dry heat resistance of 10 strains of yeast was investigated to develop data useful for the evaluation of aseptic systems for packaging acid products and which sterilize containers with hot air. Although three of the strains tested showed little survival at 110°C, four other strains had Duo-c values between 1 and 4 min. Torulopsis glabrata had a D_126.7o.C of 0.78 min. Saccharomyces strains showed the highest dry heat resistance, with the most heat resistant strain tested having a D_126.7o of 5 min. The z values for these strains ranged from 9.1° to 13.3°C.     

Heat-Stability of Milk-Clotting Enzymes in Conditions Encountered in Swiss Cheese Making

Chymosin is inactivated in gelled milk at pH 6.50 at 53°C according to a biphasic kinetic. Both phases appear to follow first-order kinetics. The D-value for the first phase is 30 min. In the same medium, the Mucor miehei and Mucor pusillus proteases are much more stable (D_53°C > 100 min) while bovine pepsin, a heat-labile Mucor miehei protease and the Endothia parasitica protease are rapidly inactivated (D_53°C < 10 min). pH appears to be the most important parameter for heat stability. Protein and calcium concentrations affect the resistance to heat treatment. A residual activity of chymosin in Swiss-type curd will be very weak at the most unless the curd is cooked at pH < 6.50.     

Determination of Enterococcus faecium thermal reduction in normal and high oleic peanut products

During processing, peanut butter can become contaminated with pathogenic bacteria (e.g., Salmonella). The introduction of an additional heat treatment step after roasting can help inactivate these microorganisms. In this study, trials were conducted to determine Enterococcus faecium (Salmonella surrogate) reduction rates during the roasting of high oleic (HO) peanuts and heat-treatment of normal oleic (NO) and HO peanut butters. HO peanuts were inoculated with E. faecium and roasted in a convection oven at 190°C. There was a 2 and 6 log CFU/g reduction at 300 and 480 s, respectively. D-values for HO peanut butter at 110, 120, and 125°C were 438.9, 165.1, and 80.6 s, respectively. The z-value was calculated to be 20.8°C. There was no significant difference in D-values and z-values between NO and HO peanut butter. In a pilot scale experiment, HO peanut butter was inoculated with E. faecium and agitated in a heated mixer for 21.5 min. E. faecium was reduced by 5.1 log CFU/g after 16.5 min with no apparent change in viscosity or texture. This study demonstrated that significant reductions in E. faecium can be achieved during roasting and through an additional heat-treatment step.