Thermal Inactivation of Escherichia coli O157: H7, Salmonella senftenberg, and Enzymes with Potential as Time-Temperature Indicators in Ground Beef

1997 ◽  
Vol 60 (5) ◽  
pp. 471-475 ◽  
Author(s):  
ALICIA ORTA-RAMIREZ ◽  
JAMES F. PRICE ◽  
YIH-CHIH HSU ◽  
GIRIDARAN J. VEERAMUTHU ◽  
JAMIE S. CHERRY-MERRITT ◽  
...  

The USDA has established processing schedules for beef products based on the destruction of pathogens. Several enzymes have been suggested as potential indicators of heat processing. However, no relationship between the inactivation rates of these enzymes and those of pathogenic microorganisms has been determined. Our objective was to compare the thermal inactivation of Escherichia coli O157:H7 and Salmonella senftenberg to those of endogenous muscle proteins. Inoculated and noninoculated ground beef samples were heated at four temperatures for predetermined intervals of time in thermal-death-time studies. Bacterial counts were determined and enzymes were assayed for residual activity. The D values for E. coli O157:H7 were 46.10, 6.44, 0.43, and 0.12 min at 53, 58, 63, and 68°C, respectively, with a z value of 5.60°C. The D values for S. senftenberg were 53.00, 15.17, 2.08, and 0.22 min at 53, 58, 63, and 68°C, respectively, with a z value of 6.24°C. Apparent D values at 53, 58, 63, and 68°C were 352.93, 26.31, 5.56, and 3.33 min for acid phosphatase; 6968.64, 543.48, 19.61, and 1.40 min for lactate dehydrogenase; and 3870.97, 2678.59, 769.23, and 42.92 min for peroxidase; with z values of 7.41,3.99, and 7.80°C, respectively. Apparent D values at 53, 58, 63, and 66°C were 325.03, 60.07, 3.07, and 1.34 min for phosphoglycerate mutase; 606.72, 89.86, 4.40, and 1.28 min for glyceraldehyde-3-phosphate dehydrogenase; and 153.06, 20.13, 2.25, and 0.74 min for triose phosphate isomerase; with z values of 5.18, 4.71, and 5.56°C, respectively. The temperature dependence of triose phosphate isomerase was similar to those of both E. coli O157 :H7 and S. senftenberg, suggesting that this enzyme could be used as an endogenous time-temperature indicator in beef products.

2004 ◽  
Vol 67 (7) ◽  
pp. 1394-1402 ◽  
Author(s):  
R. Y. MURPHY ◽  
E. M. MARTIN ◽  
L. K. DUNCAN ◽  
B. L. BEARD ◽  
J. A. MARCY

At 55 to 70°C, thermal inactivation D-values for Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes were 19.05 to 0.038, 43.10 to 0.096, and 33.11 to 0.12 min, respectively, in ground turkey and 21.55 to 0.055, 37.04 to 0.066, and 36.90 to 0.063 min, respectively, in ground beef. The z-values were 5.73, 5.54, and 6.13°C, respectively, in ground turkey and 5.43, 5.74, and 6.01°C, respectively, in ground beef. In both ground turkey and beef, significant (P < 0.05) differences were found in the D-values between E. coli O157:H7 and Salmonella or between E. coli O157:H7 and L. monocytogenes. At 65 to 70°C, D-values for E. coli O157:H7, Salmonella, and L. monocytogenes were also significantly (P < 0.05) different between turkey and beef. The obtained D- and z-values were used in predicting process lethality of the pathogens in ground turkey and beef patties cooked in an air impingement oven and confirmed by inoculation studies for a 7-log (CFU/g) reduction of E. coli O157:H7, Salmonella, and L. monocytogenes.


1998 ◽  
Vol 61 (3) ◽  
pp. 285-289 ◽  
Author(s):  
M. ROCELLE S. CLAVERO ◽  
LARRY R. BEUCHAT ◽  
MICHAEL P. DOYLE

Rates of thermal inactivation of five strains of Escherichia coli O157:H7 isolated from ground beef implicated in outbreaks of hemorrhagic colitis and five strains isolated from bovine feces were determined. Ground beef (22% fat, 10 g), inoculated with individual test strains at populations ranging from 6.85 to 7.40 log10 CFU g−1 of beef, was formed into patties (0.3 cm thick and 8.0 cm in diameter) and sealed in polyethylene bags. For each strain and treatment temperature (54.4, 58.9, 62.8, 65.6, or 68.3°C), 6 bags were simultaneously immersed into a recirculating water bath. Viable cells in patties heated for various lengths of time were enumerated by plating diluted samples on sorbitol MacConkey agar supplemented with 4-methylumbelliferyl-β-d-glucuronide (MSMA) and modified eosin methylene blue (MEMB) agar. Regardless of strain or treatment temperature, higher numbers of E. coli O157:H7 cells were generally recovered on MEMB agar than on MSMA, indicating the inferiority of MSMA as a recovery medium for quantitative determination of E. coli O157:H7 cells in heat-processed ground beef. Significantly (P ≤ 0.05) higher D values when enumeration was done using MEMB agar compared with MSMA. Mean D values for combined strain data at 54.4, 58.9, 62.8, and 65.6°C from cultures on MEMB agar were 123.90, 6.47, 0.62, and 0.20 min, respectively, whereas D values of 25.5, 5.21, 0.57, and 0.18 min were obtained at the same temperatures from cultures on MSMA. Results suggest that cooking ground beef patties to an internal temperature of 68.3°C for 40 s will inactivate at least 99.99% of E. coli O157:H7 cells; z values of 4.0 and 5.1°C were calculated from mean D values obtained from MEMB agar and MSMA, respectively, as recovery media. Differences in D values and z values existed among strains but rates of thermal inactivation do not appear to be correlated with the sources of the isolates.


2008 ◽  
Vol 71 (7) ◽  
pp. 1349-1356 ◽  
Author(s):  
AVIK MUKHERJEE ◽  
YOHAN YOON ◽  
KEITH E. BELK ◽  
JOHN A. SCANGA ◽  
GARY C. SMITH ◽  
...  

Internalization of Escherichia coli O157:H7 in nonintact beef products during mechanical tenderization or during injection of marination and tenderization ingredients is of concern if such products are undercooked. This study tested organic acids (0.2% citric acid and 0.3% acetic acid), potassium and calcium salts (1.8% potassium lactate, 0.63% calcium lactate, 0.86% calcium ascorbate, and 0.23% calcium chloride), and sodium chloride (2.5%) for their influence on thermal destruction of E. coli O157:H7 in ground beef serving as a model system. Ground beef batches (700 g; 5% fat) were mixed with equal volumes (22 ml) of each treatment solution or distilled water and portions (30 g) of treated ground beef were extruded in test tubes (2.5 by 10 cm). A five-strain mixture of E. coli O157:H7 (0.3 ml; 7 log CFU/g) was introduced at the center of the sample with a pipette. After overnight storage (4°C), simulating product marination, samples were heated to 60 or 65°C internal temperature, simulating rare and medium rare doneness of beef, in a circulating water bath. At 65°C, treatments with citric and acetic acid showed greater (P < 0.05) reduction (4 to 5 log CFU/g) of E. coli O157:H7 than all the other ingredients and the control (3 to 4 log CFU/g). Sodium chloride reduced weight losses (16 to 18% compared with 20 to 27% by citric or acetic acid) and resulted in a 4-log reduction in counts during cooking to 65°C. Ingredients such as citric or acetic acid may improve thermal inactivation of E. coli O157:H7 internalized in nonintact beef products, while sodium chloride may reduce cooking losses in such products.


2003 ◽  
Vol 66 (4) ◽  
pp. 664-667 ◽  
Author(s):  
LIHAN HUANG ◽  
VIJAY K. JUNEJA

A study was conducted to investigate the antimicrobial effect of sodium lactate (NaL) (0, 1.5, 3.0, and 4.5%) on the survival of Escherichia coli O157:H7 in 93% lean ground beef. Samples inoculated with a mixture of four strains of E. coli O157:H7 (107 to 108 CFU/g) were subjected to immersion heating in a water bath stabilized at 55, 57.5, 60, 62.5, or 65°C. Results of statistical analysis indicated that the heating temperature was the only factor affecting the decimal reduction times (D-values) of E. coli O157:H7 in 93% lean ground beef. The change in temperature required to change the D-value (the z-value) was determined as 7.6°C. The thermal resistance of this organism was neither affected by the addition of NaL nor by the interactions between NaL and temperature. Adding NaL to ground beef to reduce the thermal resistance of E. coli O157: H7 is therefore not recommended.


2004 ◽  
Vol 67 (8) ◽  
pp. 1760-1764 ◽  
Author(s):  
TONG ZHAO ◽  
MICHAEL P. DOYLE ◽  
MAURICE C. KEMP ◽  
RHONDA S. HOWELL ◽  
PING ZHAO

Undercooked ground beef is a leading vehicle for acquiring Escherichia coli O157:H7 infections through consumption of foods. Studies have been performed to determine the effect of freezing and the combined effect of freezing and addition of a mixture of 20% acidic calcium sulfate (final concentration of 0.4% in ground beef) and 10% lactic acid (final concentration of 0.2% in ground beef) (ACS-LA) on the thermal sensitivity of E. coli O157:H7 in ground beef. Five strains of E. coli O157: H7 were separately inoculated into ground beef and held at 5°C for up to 10 days or −20°C for up to 3 weeks then heated at 57, 60, 62.8, 64.3, and 68.3°C to determine rates of thermal inactivation. Results revealed that D-values (decimal reduction times) at equivalent temperatures for four of five E. coli O157:H7 strains were less in the previously frozen than in the refrigerated ground beef and that strains isolated from ground beef in 1993 and 1994 were generally more sensitive to thermal inactivation than those isolated in 1999 and 2000. Only one strain of E. coli O157:H7 was used to determine the effect of ACS-LA in previously frozen or refrigerated ground beef on rates of thermal inactivation. The addition of ACS-LA to ground beef at 20 ml/kg increased the thermal sensitivity of E. coli O157:H7 in both previously frozen and refrigerated ground beef, with greatest rates of inactivation occurring in previously frozen ground beef containing ACS-LA. D-values at 57°C obtained for E. coli O157:H7 in previously refrigerated and frozen ground beef containing ACS-LA and ACS-LA diluted by half were significantly (P < 0.05) less than those obtained in ground beef with no ACS-LA added. D-values at 60 and 62.8°C were consistently less in ACS-LA treated ground beef, but for most treatments the results were not significantly (P > 0.05) different than the controls. Results revealed that the addition of ACS-LA to ground beef, whether frozen or refrigerated, can reduce the temperature or time required to kill E. coli O157:H7 during heating.


2006 ◽  
Vol 69 (5) ◽  
pp. 1080-1086 ◽  
Author(s):  
T. OSAILI ◽  
C. L. GRIFFIS ◽  
E. M. MARTIN ◽  
B. L. BEARD ◽  
A. KEENER ◽  
...  

Thermal inactivation studies were used to determine the D- and z-values of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes in ready-to-eat chicken-fried beef patties. Inoculated meat was packaged in sterile bags, which were immersed in a circulated water bath and held at 55, 57.5, 60, 62.5, 65, 67.5, and 70°C for different lengths of time. D- and z-values were determined with a linear regression model. Average D-values at temperatures 55 to 70°C were 27.62 to 0.04 min for E. coli O157:H7, 67.68 to 0.22 min for Salmonella, and 81.37 to 0.31 min for L. monocytogenes. The z-values were 5.2°C for E. coli O157:H7, 6.0°C for Salmonella, and 6.1°C for L. monocytogenes. The results of this study can be used by food processors to validate their processes and help eliminate pathogenic bacteria associated with chicken-fried beef products.


Author(s):  
Susmita Phattepuri ◽  
Prince Subba ◽  
Arjun Ghimire ◽  
Shiv Nandan Sah

Milk is an excellent medium for the growth of many bacteria. This study aimed to determine antibiotic profiling and thermal inactivation of Staphylococcus aureus and Escherichia coli isolated from raw milk of Dharan. Total viable count, total Staphylococcal count, and total coliform count were carried out by conventional microbiological methods. Identification was done on the basis of Gram staining and biochemical tests. The antibiotic susceptibility test of the isolates carried out by the modified Kirby-Baur disc diffusion method. Thermal inactivation of S. aureus and E. coli were carried out by subjecting to thermal treatment in a water bath. Total plate count ranged from 204×104 CFU/mL to 332×105 CFU/mL. Total staphylococcal count and total coliform count ranged from 14×105 CFU/mL to 8×106 CFU/mL and 11×104 CFU/mL to 3×106 CFU/mL respectively. S. aureus showed an increasing resistance patterns towards Ampicillin, Cefotixin, Carbenicillin and Cefotaxime. Ciprofloxacin, Erythromycin, Amikacin, Gentamycin, Azithromycin, and Chloramphenicol were found to be effective against S. aureus. All the E. coli isolates were resistant to Ampicillin and least resistant to Cefotixin. Chloramphenicol, Amikacin, Azithromycin, and Nalidixic acid were found highly effective to E. coli. The D-values for S. aureus at 56°C, 58°C and 60°C were 1.36 min, 1.19 min, and 1.09 min respectively. The Z-value was 14.92°C. While D-values were obtained as 0.98 min, 0.75 min, and 0.57 min for E. coli at 56° C, 58° C and 60° C respectively, and Z-value was 9.75° C. Hence, S. aureus was found to be more heat resistant than E. coli.


2007 ◽  
Vol 70 (10) ◽  
pp. 2230-2234 ◽  
Author(s):  
T. W. THOMPSON ◽  
T. P. STEPHENS ◽  
G. H. LONERAGAN ◽  
M. F. MILLER ◽  
M. M. BRASHEARS

Rapid enzyme-linked immunosorbent assays (ELISAs) are approved for detection of Escherichia coli O157 in beef products. However, these kits have also been used in the industry to detect this pathogen on hides or in feces of cattle, although this use has not been validated. The objective of this study was to compare commercially available ELISAs (E. coli Now, Reveal, and VIP) with immunomagnetic separation along with selective media to detect E. coli O157 on hides, in feces, and in medium- and low-level-inoculated ground beef and carcasses (simulated by using briskets) samples. Naturally infected hide and fecal samples were subjected to both the immunomagnetic separation method and ELISAs for the detection of E. coli O157. Additionally, E. coli O157 inoculated and noninoculated ground beef and beef briskets were used to simulate meat and carcass samples. When comparing the detection results from the ELISAs (E. coli Now, Reveal, and VIP) to the immunomagnetic separation method, poor agreement was observed for fecal samples (kappa = 0.10, 0.02, and 0.03 for E. coli Now, Reveal, and VIP, respectively), and fair-to-moderate agreement was observed for hide samples (kappa = 0.30, 0.51, and 0.29 for E. coli Now, Reveal, and VIP, respectively). However, there was near-perfect agreement between the immunomagnetic separation method and ELISAs for ground beef (kappa = 1, 1, and 0.80 for E. coli Now, Reveal, and VIP, respectively) and brisket (kappa = 1, 1, and 1 for E. coli Now, Reveal, and VIP, respectively) samples. Assuming immunomagnetic separation is the best available method, these data suggest that the ELISAs are not useful in detecting E. coli O157 from hide or fecal samples. However, when ELISAs are used on ground beef and beef brisket samples they can be used with a high degree of confidence.


1998 ◽  
Vol 61 (2) ◽  
pp. 171-175 ◽  
Author(s):  
GIRIDARAN J. VEERAMUTHU ◽  
JAMES F. PRICE ◽  
CARL E. DAVIS ◽  
ALDEN M. BOOREN ◽  
DENISE M. SMITH

The USDA Food Safety and Inspection Service has proposed to amend cooking regulations to require that any thermal process used for poultry products be sufficient to cause a 7 D reduction in salmonellae. Several enzymes have been suggested as potential indicators of heat processing in poultry, yet no relationship between the inactivation rates of these enzymes and salmonellae has been determined. The thermal inactivation kinetics of endogenous muscle proteins, Escherichia coli O157:H7 and Salmonella senftenberg were compared in ground turkey thigh meat in thermal death time studies. Bacteria counts were determined and muscle extracts were assayed for residual enzyme activity or protein concentration. D and z values were calculated using regression analysis. S. senftenberg had higher D values at all temperatures and was more heat resistant than E. coli. The z values of E. coli on Petrifilm Coliform Count plates and phenol red sorbitol agar plates were 6.0 and 5.7°C, respectively. The z values of S. senftenberg were 5.6 and 5.4°C on Petrifilm and agar, respectively. Lactate dehydrogenase (LDH) was the most heat stable protein at 64°C. LDH, glyceraldehyde-3-phosphate dehydrogenase, creatine kinase, triose phosphate isomerase (TPI), acid phosphatase, serum albumin, and immunoglobulin G had z values of 3.8, 4.3, 4.8, 5.8, 6.3, 6.7, and 8.6°C, respectively, in turkey containing 4.3% fat. The z values for TPI decreased to 5.4°C in thigh meat containing 9.8% fat. Temperature dependence of TPI was most similar to that of S. senftenberg, suggesting it might function as an endogenous time-temperature integrator to monitor adequacy of processing when a performance standard based on this pathogen is implemented.


2011 ◽  
Vol 74 (4) ◽  
pp. 622-626 ◽  
Author(s):  
CHENG-AN HWANG ◽  
VIJAY JUNEJA

Ground beef products are susceptible to contamination with Escherichia coli O157:H7. The objective of this study was to examine the effect of salt, sodium pyrophosphate (SPP), and sodium lactate on the probability of growth of E. coli O157:H7 in ground beef under a temperature abuse condition. Ground beef containing 0 to 2.25% salt, 0 to 0.5% SPP, and 0 to 3% lactate was inoculated with a four-strain mixture of E. coli O157:H7, vacuum packaged, and stored at 10°C for 15 days. A total of 25 combinations of the three additives, each with 20 samples, were tested. A logistic regression was used to model the probability of growth of E. coli O157:H7 (with a 1.0-log CFU/g increase during storage) as a function of salt, SPP, and lactate. The resultant probability model indicated that lactate at higher concentrations decreased the probability of growth of E. coli O157:H7 in ground beef, and the effect was more pronounced at higher salt concentrations. At salt concentrations below 1.3%, the increase of SPP concentration marginally increased the growth probabilities of E. coli O157:H7. The model illustrated the effect of salt, SPP, and lactate on the growth probabilities and growth or no-growth behavior of E. coli O157:H7 in ground beef and can be used to improve the microbial food safety of ground beef products.


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