JFS:

Food Microbiology and Safety

Thermal Inactivation of Salmonella spp., Salmonella typhimurium DT104, and Escherichia coli O157:H7 in Ground Beef
S.E. SMITH, J.L. MAURER, A. ORTA-RAMIREZ, E.T. RYSER, AND D.M. SMITH ABSTRACT: In 19.1% fat ground beef, Escherichia coli O157:H7 was less heat- resistant at 58 C than the Salmonella typhimurium DT104 and Salmonella senftenberg, but at 55 C the D value was similar to DT104 strains and higher than an eight-strain Salmonella cocktail. Inactivation of E. coli O157:H7 was more temperature-dependent than the cocktail and DT104 strains. E. coli and DT104 strains were more heat-resistant in beef containing 19% fat than 4.8% fat. The cocktail was more thermally stable in stationary as compared to log phase. Freezing of inoculated raw meat decreased heat resistance of the cocktail. The pathogenic strain, growth phase of the organism, state of the meat (fresh or frozen) and meat composition must be considered when designing protocols to verify thermal processes. Keywords: Salmonella, E. coli O157:H7, thermal inactivation, growth phase, freezing

Introduction
that beef patties be cooked to a certain temperature for a specified amount of time to eliminate pathogens. Proposed regulations (USDA-FSIS 1999) would supplement specific time temperature requirements with lethality performance standards. A 5-log reduction in Salmonella has been proposed as the lethality performance standard for fully cooked, uncured meat patties (USDA-FSIS 1999). The FSIS has not finalized this standard due to concerns that a process equivalent to a 5-log reduction in Salmonella may not be sufficient to protect public health. The lethality performance standard for roast beef requires the use of a combination of thermal and nonthermal processes sufficient to achieve a 6.5-log reduction in Salmonella (USDA-FSIS 1999). The USDA-FSIS (1999) recommends the use of a cocktail or a combination of Salmonella serotypes to verify compliance to the performance standard. This cocktail should consist of Salmonella strains that exhibit relatively high heat resistance and have been previously implicated in foodborne outbreaks (UDSA-FSIS 1999). D and z values for different Salmonella serotypes were determined in chicken broth and meat products in order to identify the most appropriate strains to use for defining heat resistance in meat products (Juneja and others 2001). A comparison of these strains to E. coli O157:H7 and other Salmonella strains would be useful to verify that the recommendations are appropriate. Salmonella typhimurium DT104 has emerged as a particularly dangerous Salmonella phage type that is resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline (Hollingsworth and others 1997). In 1996, 32% of the isolates of S. typhimurium tested at the Center for Disease Prevention and Control demonstrated the ACSSuT resistant pattern associated with Definitive Type 104 (Morbidity and Mortality Weekly Report 1997). D values for a clinical isolate of S. typhimurium DT104 in chicken broth ranged from 4.16 min at 55 C to 0.27 min at 62 C ( Juneja and others 2001). Several factors influence the thermal resistance of Salmo1164 JOURNAL OF FOOD SCIENCEVol. 66, No. 8, 2001

URRENT COMMAND AND CONTROL REGULATIONS REQUIRE

nella, including age of the culture, growth temperature, and initial population. According to Heddleson and others (1991) the maximum heat resistance of Salmonella spp. occurred in cells that had reached stationary phase in tryptic soy broth after 24 h of incubation. Since muscle type, pH, fat, and moisture content of the heating menstruum all influence thermal resistance of microorganisms, thermal inactivation parameters of a microorganism must be established in the food of interest for accurate process control (Murphy and others 1999). Our objectives were to determine the thermal resistance of Salmonella spp., including S. typhimurium DT104, and E. coli O157:H7 in ground beef at two fat contents and to assess the effects of microbial growth phase and freezing on bacterial heat resistance. D values were calculated using first-order kinetics.

Materials and Methods

Bacterial strains
Three human isolates of S. typhimurium DT104 (10601, 10127, 01071) were obtained from B. Swaminathan (Centers for Disease Control and Prevention, Atlanta, Ga., U.S.A.). S. senftenberg (ATCC 43854) and Escherichia coli O157:H7 (ATCC 43894) type 204P were obtained from the Microbiological Culture Lab at Michigan State University. The following eight Salmonella strains were obtained from V.K. Juneja (Agricultural Research Service, Eastern Regional Research Center, USDA-ARS, Philadelphia, Pa., U.S.A.): S. thompson FSIS 120 (chicken isolate) , S. enteriditis H3527 and H3502 (clinical isolates phage type 13A and 4, respectively), S. typhimurium H3380 (human isolate DT104), S. hadar MF60404 (turkey), S. copenhagen 8457 (pork), S. montevideo FSIS 051 (beef ), and S. heidelberg F5038BG1 (human isolate). All strains were preserved in vials containing tryptic soy broth (TSB) (Difco, Detroit, Mich., U.S.A.) with 10% glycerol at 80C.

Food Microbiology and Safety

Culture preparation
To propagate the cultures, one loop of frozen culture was
2001 Institute of Food Technologists

Inactivation of Salmonellae and E. coli . . .

transferred to 9 mL of TSB in 20-mL culture tubes. Static cultures were maintained by daily transfers in TSB (37 C, 18 to 24 h), with a minimum of 2 consecutive transfers. Each inoculum was prepared from either an 18-to-24-h (log phase) or 48-h (stationary phase) culture containing approximately 109 CFU/ml. In preliminary work, each culture was grown separately in TSB, incubated at 37 C, and plated at predetermined intervals to identify the log and stationary growth phases. The 8 Salmonella strains received from Juneja were grown in separate culture tubes and then combined in equal volumes just before centrifugation to produce a cocktail for thermal death time studies. On the day of each experiment, cultures were pelleted by centrifugation at 6000 g for 10 min at 4 C and resuspended in sterile 0.1% buffered peptone water to a concentration of about 1010 CFU/ml. The cultures were enumerated by serial dilutions in 0.1% peptone water and plated on Petrifilm aerobic count plates (3M, St. Paul, Minn., U.S.A.) in duplicate. tored using a thermocouple (Type T, length 8.6 cm, width 1.0 mm, accuracy 0.1 C, Omega Engineering, Stamford, Conn., U.S.A.) connected to a data logger system (Daqbook 100, Omega) inserted and sealed into the pouches. The heating lag time was 8 s. Time zero was defined as the time when the meat reached the target temperature. Preliminary studies were conducted to determine appropriate time-temperature combinations to achieve greater than or equal to a 5-log reduction in pathogens. Meat was removed at specific time intervals, held on ice at 4 C, and used within 4 h. Three replicate batches of ground beef were heated at each temperature.

Enumeration of Salmonella and E. coli

Cooked ground beef was transferred to sterile Whirlpak bags (18 oz, Nasco, Ft. Atkinson, Wis., U.S.A.) and manually homogenized for 1 min with 9 mL of 0.1% sterile peptone water. Bacterial counts were determined by decimal dilution of the ground beef in sterile 0.1% peptone water and plated in duplicate on Petrifilm Aerobic count plates. All plates were incubated at 37 C for 24 to 36 h.

Ground beef preparation

Low-fat beef was obtained from the Michigan State University Meat Laboratory. High-fat beef was received from a local processor. The meat was ground twice through a 4-mm dia plate in a Kitchen Aid grinder (Model K5-A, Hobart, Troy, Ohio, U.S.A.), vacuum-packaged in 60-g portions, and frozen at 12 C. The frozen meat was transported on dry ice to Iowa State University and irradiated (10 kGy) to eliminate indigenous microflora. The irradiated beef was tested for sterility by plating a 1:10 dilution onto Petrifilm Aerobic count plates. Moisture, fat, and protein contents of low- and high-fat beef were determined in triplicate by AOAC (1990) methods 950.46B, 991.36, and 981.1, respectively. For determination of pH, 10-g ground beef were homogenized with 90 mL of distilled water using a Polytron homogenizer (Model PT 10/35, Brinkman Instruments, Westbury, N.J., U.S.A.) at speed setting 3 for 30 s. The pH of the homogenized mixture was measured using a combination electrode (Model 145, Corning, Medfield, Mass., U.S.A.).

Calculation of D values and z values

Bacterial counts were converted to logarithms. D values were estimated assuming linear inactivation kinetics. D values (in minutes) were calculated by linear regression of surviving bacteria as compared with time from at least 4 data points with a correlation coefficient > 0.90 using Microsoft Excel Version 5.0a (Microsoft Corp., Redmond, Wash., U.S.A.). Thermal resistance curves were determined by plotting log D values as compared with temperature. The z value was determined as the negative reciprocal of the slope of the thermal resistance curve.

Identification of Salmonella survivors

Preliminary experiments were conducted to determine the heating times at 55 and 63 C that produced < 5-CFU/ plate. After heating at 55 C for 75 min and 63 C for 50 s, viable colonies were plated onto tryptic soy agar (TSA). Colonies were isolated and then grown at 37 C for 24 h on TSA. Survivors were tentatively identified by comparing responses on API 20 E test strips (Biomerieux Vitek, Inc., St. Louis, Mo., U.S.A.).

Inoculation of ground beef

Microorganisms were added dropwise, using aseptic procedures, to obtain a target concentration of 10 8 CFU/g ground beef. The meat was mixed in a sterile bowl using sterile gloves to ensure even distribution. Inoculated meat was transferred into a 60-mL syringe (Becton Dickinson and Co., Franklin Lakes, N.J., U.S.A.), after which 1-g aliquots were extruded into 5-cm 25.5-cm sterile polyethylene laminated nylon bags (Butcher and Packer, Detroit, Mich., U.S.A.). Sterility of the bags was confirmed by mixing 9 mL of 0.1% peptone water in random bags and plating the resultant liquid on Petrifilm Aerobic count plates. Bags containing the meat were rolled to a thickness of 1 mm using a template, and the bags were then heat sealed. For the freezing studies, the inoculated meat was held at 9 C for 7 or 14 d. Meat was tempered to 4 C by holding each bag under cold running water for 5 min prior to thermal death time studies. Freezing for 7 or 14 d, followed by thawing, did not alter initial inoculum concentrations.

Results and Discussion

Composition
Low-fat ground beef (pH 5.65 0.01) contained 4.8% 1.1% fat, 72.4% 0.2% moisture, and 15.8% 2.6% protein. High-fat ground beef (pH 5.72 0.01) contained 19.1% 0.7% fat, 63.4 % 0.6% moisture, and 14.4% 1.9% protein. No microorganisms were detected in the uninoculated irradiated meat.

Heat resistance of Salmonella senftenberg

Early studies have demonstrated the unusual heat resistance of S. senftenberg in buffer (D57 C = 31 min) (Ng and others 1969) and poultry (D66 C = 3.5 min) (Milone and Watson 1970). As expected, S. senftenberg was the most heat-resistant Thermal inactivation procedure bacteria in ground beef (19.1% fat), with D values ranging The bags containing meat were placed in a rack and com- from 21.79 min at 58 C to 0.92 min at 64 C (Table 1). Ortapletely submerged in a Polystat circulating water bath (Model Ramirez and others (1997) reported D values of 15.17 min at 1268-52, Cole-Palmer Instrument Co., Chicago, Ill., U.S.A.) 58 C and 2.08 min at 63 C in ground beef cooked in 10 by 75 set at 55, 58, 61, 63, 64, or 66 C. The temperature was moni- mm dia glass test tubes. Variations between these studies
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Table 1D values and regression parameters of Salmonella senftenberg and three strains of Salmonella typhimurium DT104 in high (19.1%) and low (4.8%) fat ground beef Fat yCon- Temp. Intertent (C) cept Slope High 66 64 61 58 64 61 58 55 64 61 58 55 64 61 58 55 64 61 58 55 6.34 6.42 6.56 6.40 6.30 6.32 6.57 6.49 7.13 6.97 7.01 6.97 7.03 6.67 7.87 6.40 6.97 7.49 7.24 7.09 2.91 1.09 0.30 0.05 6.20 1.54 0.38 0.05 6.70 1.75 0.44 0.11 14.43 2.47 0.49 0.10 7.20 2.33 0.49 0.10 D value (min) a 0.34 0.01 0.92 0.02 3.38 0.32 21.79 1.38 0.16 0.01 0.65 0.03 2.63 0.19 21.98 2.15 0.15 0.01 0.57 0.01 2.26 0.10 9.05 0.13 0.07 0.01 0.41 0.01 2.15 0.07 10.55 1.04 0.14 0.01 0.43 0.02 2.06 0.13 10.27 0.78 Table 2D values and regression parameters obtained in ground beef (19.1% fat) inoculated with an 8-strain Salmonella cocktail in log (24 h) and stationary (48 h) phase and after frozen storagea Temp. y(C) Intercept Slope Log phase 63 61 58 55 63 61 58 55 63 61 58 55 63 61 58 55 6.93 6.95 7.04 7.08 6.88 6.88 6.85 7.09 6.21 6.58 6.78 6.58 6.47 7.08 6.74 6.87 6.54 2.25 0.37 0.06 5.01 1.75 0.30 0.06 7.18 3.41 0.70 0.10 5.13 3.54 0.42 0.08 R2 0.97 0.98 0.97 0.98 0.98 0.97 0.97 0.99 0.81 0.76 0.86 0.92 0.90 0.94 0.85 0.85 D value (min) b 0.15 0.01 0.44 0.02 2.72 0.21 16.34 0.72 0.20 0.01 0.57 0.03 3.39 0.06 18.66 0.65 0.14 0.03 0.29 0.06 1.43 0.22 9.85 0.63 0.20 0.01 0.28 0.02 2.36 0.14 12.52 1.27

Bacterium S. senftenberg

R2 0.93 0.94 0.86 0.90 0.97 0.97 0.95 0.95 0.98 0.96 0.95 0.95 0.99 0.98 0.95 0.93 0.95 0.94 0.97 0.96

DT104-10127

High

Stationary phase

DT104-10127

Low

Log phase frozen

DT104-10601

Low

Stationary phase frozen

DT104-01071

Low

a Inoculated meat was frozen at 9C for 14 d for the log phase and 7 d for b The D value is expressed as the mean standard error of the mean of the

the stationary phase.

a The D value is expressed as the mean standard error of the mean of the

three replicate determinations.

three replicate determinations.

could be due to the shorter come-up time in pouches (8 s) as compared to test tubes (60 s). Hence, the longer come-up time for meat in the test tubes will decrease the initial microbial load at the target temperature, thereby decreasing the slope of the linear regression line that was fitted to the data and increasing the D value. Variations could also be attributed to differing fat contents 3.8% as compared with 19% in our study. Ahmed and others (1995) reported that D values increased with increasing fat content in ground beef.

Heat resistance of S. typhimurium DT104

Thermal death time studies were performed using 3 strains of S. typhimurium DT104 in ground beef (4.8% fat). The D values of the 3 strains were not different ( p > 0.05) from each other at 55 or 58 C. At 61 C, the D value of strain 10127 (0.57 min) was higher (p < 0.05) than those of strains 01071 (0.43 min) and 10601 (0.41 min). At 64 C, the D value for strain 10601 (0.07 min) was about half that of the other 2 strains (p < 0.05). Although reasons for these differences are not clear, the D values calculated for the 3 strains at 58 C were similar to those reported by Humphrey and others (1997) for 3 wild strains of S. typhimurium DT104 on pork muscle (D58 C = 2.61 min). The D values for strain 10127 in ground beef were similar at 58, 61, and 64 C, regardless of fat content. However, at 55 C the D value in high-fat ground beef (21.98 min) was more than twofold greater than that in the low-fat ground beef (9.05 min).

Heat resistance of the 8-strain Salmonella cocktail

In log phase, the Salmonella cocktail exhibited D values of 0.15, 0.44, 2.72, and 16.34 min at 63, 61, 58, and 55 C, respectively, in 19.1% fat ground beef (Table 2). These values were almost tenfold lower than those for S. senftenberg in ground beef at the same fat content. Using ground beef inoc- Survivor identification After heating, the inoculated meat was examined for each ulated with the same 8-strain cocktail, Juneja and others (2001) obtained D values of 8.65 min at 58 C and 1.5 min at of the 8 strains comprising the cocktail. API 20 E test strips
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62.5 C. Variations in values reported could be due to differences in sample size or meat composition. Juneja and others (2001) used a 3-g sample that might have a longer come-up time leading to a larger D value as mentioned previously. In addition, their beef had a pH of 6.0 as opposed to pH 5.7 in our study. Murphy and others (1999) reported that a 6-strain Salmonella cocktail exhibited a greater heat resistance in ground chicken breast than the cocktail used in this study (D67.5 C = 0.286 min). However, inclusion of S. senftenberg in the cocktail selected by Murphy and others (1999) could explain the higher D values. In some cases, the D values for the S. typhimurium DT104 strains were greater than those for the cocktail. For example, the D values for S . typhimurium DT104 strain 10127 were higher than those for the 8-strain cocktail (log phase) at 61 and 55 C in 19.1% fat ground beef. If beef was cooked according to the processes established with this cocktail, a 5D reduction in S. typhimurium DT104 might not be achieved. When inoculated into ground beef, the Salmonella cocktail was more thermostable during stationary phase than during log phase at all temperatures tested. Enhanced thermal resistance also has been reported for stationary phase cultures of Salmonella spp. (Heddleson and others 1991), Staphylococcus aureus (Kornacki and Marth 1989), and Pediococcus (Annous and others 1999). Thermal resistance of the Salmonella cocktail in both log and stationary phases decreased after inoculation and frozen storage of ground beef at 55, 58, and 61 C. Formation of ice crystals during freezing which disrupt the cell membrane is likely responsible for the observed decrease in viability (Doyle and Cliver 1990; Smith 1995). Given these findings, greater total lethality may be achieved when cooking previously frozen raw products as compared to products that have not been frozen.

Food Microbiology and Safety

Inactivation of Salmonellae and E. coli . . .

Table 3D values and regression parameters of Escherichia coli O157:H7 in low (4.8%) and high (19.1%) fat ground beef Fat Content Low Temp. (C) 63 61 58 55 63 61 58 55 yIntercept Slope 6.32 6.72 6.85 6.91 6.32 6.73 6.68 6.91 6.97 3.11 0.82 0.05 6.97 3.10 0.49 0.04 R2 0.91 0.91 0.86 0.95 0.91 0.95 0.92 0.96 D value (min)a 0.160.02 0.320.02 1.220.13 20.080.45 0.180.01 0.320.01 2.050.13 22.470.78 Table 4The z values and regression parameters of Salmonella senftenberg, Salmonella syphimurium DT104, Salmonella cocktail, and Escherichia coli O157:H7 in high(19.1%) and low- (4.8%) fat ground beef Bacterium Fat ycontent Intercept Slope 14.15 14.11 11.80 14.38 12.50 15.67 16.53 15.29 14.91 13.74 14.18 0.22 0.23 0.20 0.24 0.21 0.26 0.28 0.26 0.25 0.23 0.24 R2 0.99 0.99 1.00 1.00 0.99 0.96 0.99 1.00 1.00 0.99 0.97 z Value (C)a 4.510.03 4.280.05 5.070.02 4.130.07 4.770.06 3.790.10 3.600.03 3.900.03 4.080.02 4.290.18 4.200.06

High

a The D value is expressed as the mean standard error of the mean of the three replicate determinations.

could not differentiate between S. typhimurium, S. hadar, and S. copenhagen (designated as Group A) or S. enteriditis and S. heidelberg (designated as Group B). All other strains could be individually identified. S. montevideo, S. thompson, and microorganisms from Groups A and B were present after cooking at 55 C for 75 min (4.6D process), whereas S. Montevideo was the only strain not detected after cooking at 63 C for 50 s (5.6D process). Thus these 8 strains exhibited similar thermal stability, as was also reported by Juneja and others (2001).

S. senftenberg High DT104-10127 High DT104-10127 Low DT104-10601 Low DT104-01071 Low Escherichia Low coli 0157:H7 Escherichia High coli 0157:H8 Salmonella High cocktail (24h) Salmonella High cocktail (48h) Salmonella High cocktail (24h frozen) Salmonella High cocktail(48h frozen)
replicate determinations.

a The z value is expressed as the mean standard error of the mean of three

Heat resistance of Escherichia coli O157:H7

D values for E. coli O157:H7 in ground beef (19.1% fat) were 0.18, 0.32, 2.05, and 22.47 min at 63, 61, 58, and 55 C, respectively (Table 3). When ground beef (10% fat) containing E. coli O157:H7 was heated at 55C, Juneja and others (1997) reported a D of 21.13 min compared to 22.47 min in our study. However, the D values were sevenfold higher at 63 C and twofold higher at 58 C than those in our study. These variations might be attributed to differences in the strains of E. coli O157:H7 used, along with differences in recovery methods and sample size. Using high-fat ground beef (19.1%), all D values calculated for E. coli O157:H7 were lower than the Salmonella cocktail except at 55 C (16.34 min), where the D value for E. coli was 6.13 min higher. The thermal resistance of E. coli O157:H7 was more temperature-dependant than the cocktail in ground beef as shown by the lower z value and higher D value at lower temperatures. According to these results, E. coli O157:H7 would not be subjected to a 5D process at 55 C if the Salmonella cocktail was used to establish thermal processing procedures, potentially leading to the consumption of unsafe ground beef. from 4.35 to 4.78 C in ground beef as the fat content decreased from 20 to 7%. The z value of the cocktail in log phase was 3.90 C, whereas the stationary phase had a value of 4.08 C, thus indicating that Salmonella in log phase was more sensitive to temperature changes. The z value found in this study was lower than that reported by Juneja and others (2001) (6.62 C) for the same 8-strain Salmonella cocktail. Variations in fat content and pH of the ground beef could account for the differences.

Conclusions

processors to validate the efficacy of their processes to reduce microbial contamination if the procedure varies from published compliance or safe harbor guidelines. Proper calculation of D and z values is necessary to ensure safety during thermal processing of meat products. Results of this study indicate that the particular pathogen, the growth phase of the organism (log or stationary), state of the meat (fresh or frozen) and the fat content must be considered when designing protocols to verify thermal processes. Based on our z Values findings, the 8-strain Salmonella cocktail may not be able to The z value for S. senftenberg (4.51 C) in 19.1% fat ground adequately predict a 5-log reduction for some pathogens, beef was lower, and therefore D values were more tempera- such as S. typhimurium DT104 strain 10127 and E. coli ture-dependent (Table 4), than reported by Orta-Ramirez 0157:H7, especially at lower cooking temperatures. and others (1997) in 4% fat ground beef (6.25 C). This difference could be due to the fat content of the ground beef. References Thermal inactivation of all three S. typhimurium DT104 AOAC. 1990. Official Methods of Analysis. 15th ed. Arlington, Virginia: Assn. of Official Anal Chem. strains (z = 4.13 to 5.07 C) was less temperature-dependant Ahmed NM, Conner DE, Huffman DL. 1995. Heat-resistance of Escherichia coli than that of E. coli O157:H7 and the Salmonella cocktail as O157:H7 in meat and poultry as affected by product composition. J Food Sci 60(3):606-610. indicated by the higher z values. Annous BA, Kozempel MF, Kurantz MJ. 1999. Changes in membrane fatty acid The z values for S. typhimurium DT104 strain 10127 and composition of Pediococcus sp. Strain B-2354 in response to growth conditions and its effects on thermal resistance. Appl Environ Microbiol 65:2857E. coli O157:H7 were higher in low-fat as compared to high2862. fat ground beef. Ahmed and others (1995) reported similar Doyle MP, Cliver DO. 1990. Salmonella. In: Oliver DO, editor. Foodborne diseasfindings in which the z value for E. coli O157:H7 increased es. New York: Academic Press. P186-204.
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Inactivation of Salmonellae and E. coli . . .

Heddleson RA, Doores S, Anantheswaran RC, Kuhn GD, Mast MG. 1991. Survival of Salmonella species heated by microwave energy in a liquid menstruum containing food components. J Food Prot 54:637-642. Hollingsworth J, Kaplan B. 1997. Federal agencies collaborate to control dangerous new Salmonella strain. J Am Vet Med Assoc 210:1712-1716. Humphrey TJ, Wilde SJ, Rowbury RJ. 1997. Heat tolerance of Salmonella Typhimurium DT104 attached to muscle tissue. Lett Appl Microbiol. 25:265-268. Juneja VK, Eblen, BS, Ransom, GM. 2001. Thermal inactivation of Salmonella spp. in chicken broth, beef, pork, turkey, and chicken: Determination of D- and z-values. J Food Prot. 66:146-152 Juneja VK, Snyder OP, Marmer BS. 1997. Thermal destruction of Escherichia coli O157:H7 in beef and chicken: Determination of D and z values. Int J Food Microbiol 35:231-237. Kornacki JL, Marth EH. 1989. Thermal inactivation of Staphylococcus aureus in retentates from ultrafiltered milk. J Food Prot 52:631-637. Milone NA, Watson JA. 1970. Thermal inactivation of Salmonella senftenberg 775W in poultry meat. Health Lab Sci 7:199-225. Morbidity Mortality Weekly Report. 1997. Multidrug-resistance Salmonella serotype Typhimurium - United States 1996. 46:308-310. Murphy RY, Marks BP, Johnson ER, Johnson MG. 1999. Inactivation of Salmonella and Listeria in ground chicken breast meat during thermal processing. J Food Prot 62:980-985. Ng H, Bayne HG, Garibaldi JA. 1969. Heat resistance of Salmonella: The uniqueness of Salmonella senftenberg 775W. Appl Microbiol 17:78-82. Orta-Ramirez A, Price JF, Hsu YC, Veeramuthu GJ, Cherry-Merritt JS, Smith DM. 1997. Thermal inactivation of Escherichia coli O157:H7, Salmonella Senftenberg , and enzymes with potential as time-temperature indicators in ground beef. J Food Prot 60:471-475. Smith MG. 1995. Survival of E. coli and Salmonella after chilling and freezing in liquid media. J Food Sci 60(3):509-512. USDA-FSIS. 1999. Performance standards for the production of certain meat and poultry products. U.S. Department of Agriculture, Food Safety Inspection Service, Washington, D.C. Federal Register 64:732-749 ( January 6). MS 20000425
This work was supported by the Michigan State University Crop and Food Bioprocessing Center, the Michigan State University Agriculture Experiment Station, and the Cooperative State Research, Education and Extension Service, United States Department of Agriculture, under Agreement Number 96-35500-3551. Any opinions, findings, conclusions, or recommendations expressed in this document are those of the authors and do not necessarily reflect the view of the USDA.

Authors are with the Dept. of Food Science and Human Nutrition, Michigan State Univ., East Lansing MI 48824-1224. Direct inquiries to author Smith, now at the Univ. of Idaho (E-mail: dsmith@uidaho.edu).

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