Thermal Inactivation of a Single Strain Each of Serotype O26:H11, O45:H2, O103:H2, O104:H4, O111:H−, O121:H19, O145:NM, and O157:H7 Cells of Shiga Toxin–Producing Escherichia coli in Wafers of Ground Beef†

2013 ◽  
Vol 76 (8) ◽  
pp. 1434-1437 ◽  
Author(s):  
J. B. LUCHANSKY ◽  
A. C. S. PORTO-FETT ◽  
B. A. SHOYER ◽  
J. PHILLIPS ◽  
D. EBLEN ◽  
...  

For each of two trials, freshly ground beef of variable fat content (higher: 70:30 %lean:%fat; lower: 93:7 %lean:%fat) was separately inoculated with ca. 7.0 log CFU/g of a single strain of Escherichia coli serotypes O26:H11, O45:H2, O103:H2, O104:H4, O111:H−, O121:H19, O145:NM, and O157:H7. Next, ca. 3-g samples of inoculated beef were transferred into sterile filter bags and then flattened (ca. 1.0 mm thick) and vacuum sealed. For each temperature and sampling time, three bags of the inoculated wafers of beef were submerged in a thermostatically controlled water bath and heated to an internal temperature of 54.4°C (130°F) for up to 90 min, to 60°C (140°F) for up to 4 min, or to 65.6°C (150°F) for up to 0.26 min. In lower fat wafers, D-values ranged from 13.5 to 23.6 min, 0.6 to 1.2 min, and 0.05 to 0.08 min at 54.4, 60.0, and 65.6°C, respectively. Heating higher fat wafers to 54.4, 60.0, and 65.6°C generated D-values of 18.7 to 32.6, 0.7 to 1.1, and 0.05 to 0.2 min, respectively. In addition, we observed reductions of ca. 0.7 to 6.7 log CFU/g at 54.4°C after 90 min, ca. 1.1 to 6.1 log CFU/g at 60.0°C after 4 min, and 0.8 to 5.8 log CFU/g at 65.6°C after 0.26 min. Thus, cooking times and temperatures effective for inactivating a serotype O157:H7 strain of E. coli in ground beef were equally effective against the seven non-O157:H7 Shiga toxin–producing strains investigated herein.

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.


2019 ◽  
Vol 82 (8) ◽  
pp. 1308-1313 ◽  
Author(s):  
QUINCY J. SUEHR ◽  
NATHAN M. ANDERSON ◽  
SUSANNE E. KELLER

ABSTRACT Non-O157 Shiga toxin–producing Escherichia coli infections have recently been associated with wheat flour on two separate accounts in the United States and Canada. However, there is little information regarding the thermal resistance and longevity of non-O157 Shiga toxin–producing Escherichia coli during storage in low-moisture environments. The objectives of this study were to determine the thermal inactivation kinetics of E. coli O121 in wheat flour and to compare the thermal inactivation rates with those of other pathogens. Wheat flour, inoculated with E. coli O121, was equilibrated at 25°C to a water activity of 0.45 in a humidity-controlled conditioning chamber. Inoculated samples were treated isothermally at 70, 75, and 80°C, and posttreatment population survivor ratios were determined by plate counting. D- and z-values calculated with a log-linear model, were compared with those obtained in other studies. At 70, 75, and 80°C, the D-values for E. coli O121 were 18.16 ± 0.96, 6.47 ± 0.50, and 4.58 ± 0.40 min, respectively, and the z-value was 14.57 ± 2.21°C. Overall, E. coli O121 was observed to be slightly less thermally resistant than what has been previously reported for Salmonella Enteritidis PT30 in wheat flour as measured under the same conditions with the same methods.


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.


2014 ◽  
Vol 77 (7) ◽  
pp. 1201-1206 ◽  
Author(s):  
JOHN B. LUCHANSKY ◽  
ANNA C. S. PORTO-FETT ◽  
BRADLEY A. SHOYER ◽  
HARSHAVARDHAN THIPPAREDDI ◽  
JESUS R. AMAYA ◽  
...  

Preflattened veal cutlets (ca. 71.5 g, ca. 0.32 cm thick) were surface inoculated with ca. 6.8 log CFU/g of a multistrain cocktail of Escherichia coli O157:H7 (ECOH) or a cocktail made of single strains of serogroups O26, O45, O103, O104, O111, O121, and O145 of Shiga toxin–producing E. coli (STEC) cells and then were mechanically tenderized by passing once through a “Sir Steak” tenderizer. For each cooking time, in each of at least three trials, three inoculated and tenderized cutlets, with and without breading, were individually cooked in 15 or 30 ml of canola oil for 0.0, 0.75, 1.0, 1.25, 1.5, 1.75, or 2.25 min per side on an electric skillet set at 191.5°C. The temperatures of the meat and of the skillet were monitored and recorded using a type J thermocouple. Regardless of the breading or volume of oil used to cook the meat, the longer the cooking times, the higher was the internal temperature of the meat, along with a greater reduction of both ECOH and STEC. The average final internal temperature of the meat at the approximate geometric center ranged from 56.8 to 93.1°C. Microbial reductions of ca. 2.0 to 6.7 log CFU/g and ca. 2.6 to 6.2 log CFU/g were achieved for ECOH and STEC, respectively. Our data also revealed no differences in thermal inactivation of ECOH relative to the volume of oil used to cook nonbreaded cutlets. However, when cooking breaded cutlets, the use of more (30 ml) compared with less (15 ml) cooking oil resulted in greater reductions in pathogen numbers. To deliver about a 5.0-log reduction of ECOH and STEC, and to achieve the recommended internal temperature of 71.1°C, it was necessary to cook mechanically tenderized veal cutlets for at least 1.5 min per side on a preheated electric skillet set at 191.5°C and containing 15 ml of cooking oil. These data also established that cooking times and temperatures effective for inactivating serotype O157:H7 strains of E. coli in tenderized veal are equally effective against the additional six non-O157 Shiga toxin–producing strains investigated herein.


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 (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.


2018 ◽  
Vol 81 (6) ◽  
pp. 986-992 ◽  
Author(s):  
JAGPINDER S. BRAR ◽  
JOLENA N. WADDELL ◽  
MATTHEW BAILEY ◽  
SYDNEY CORKRAN ◽  
CARMEN VELASQUEZ ◽  
...  

ABSTRACT Decimal reduction time (D-value) was calculated for six non-O157 Shiga toxin–producing Escherichia coli (STEC) in a laboratory medium and ground beef. For the laboratory medium, an overnight culture of each strain of STEC was divided into 10-mL sample bags and heated in a water bath for a specific time on the basis of the temperatures. Survival curves were generated by plotting the surviving bacterial population against time, and a linear-log primary model was used to estimate the D-values from survival curves. The z-values (the temperature raised to reduce the D-value by one-tenth) were calculated by plotting the log D-values against temperature. Similarly, for ground beef, six fat contents, 5, 10, 15, 20, 25, and 30% of ground beef were formulated for this study. Inoculated meat was divided into 5-g pouches and submerged in a water bath set at specific temperatures (55, 60, 65, 68, and 71.1°C). The average D-value for these strains in a laboratory medium was 17.96 min at 55°C, which reduced significantly (P < 0.05) to 1.58 min at 60°C, and then further reduced (P < 0.05) to 0.46 min at 65°C. In ground beef, a negative correlation (P < 0.05) between fat content of ground beef and D-values was observed at 55°C. However, at temperatures greater than 60°C, there was no impact (P > 0.05) of fat content of ground beef on the thermal resistance of non-O157 STECs. Irrespective of the fat content of ground beef, the D-values ranged from 15.93 to 11.69, 1.15 to 1.12, and 0.14 to 0.09 min and 0.05 at 55, 60, 65, and 68°C, respectively. The data generated from this study can be helpful for the meat industry to develop predictive models for thermal inactivation of non-O157 STECs in ground beef with varying fat content.


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.


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.


2017 ◽  
Vol 80 (12) ◽  
pp. 2105-2111 ◽  
Author(s):  
Gavin Bailey ◽  
Long Huynh ◽  
Lachlan Govenlock ◽  
David Jordan ◽  
Ian Jenson

ABSTRACT Salmonella contamination of ground beef has been viewed as originating from the surface of carcasses. Recent studies have identified lymph nodes as a potential source of Salmonella contamination because these tissues play an active role in containment of pathogens in the live animal and because some lymph nodes are unavoidably present in manufacturing beef trimmings or primal cuts that may be incorporated into ground beef. A survey was conducted of the microbiological status of lymph nodes from Australian cattle at the time of slaughter to determine the prevalence of microbiological contamination. Sets of lymph nodes (n = 197), consisting of the superficial cervical (prescapular), prepectoral, axillary, presternal, popliteal, ischiatic, subiliac (precrural), coxalis, and iliofemoralis (deep inguinal), were collected from five geographically separated Australian abattoirs over a period of 14 months. Samples were tested for the presence of Salmonella spp. and Shiga toxin–producing Escherichia coli by BAX PCR assay. Aerobic plate count, E. coli, and coliforms were enumerated with a lower limit of detection of 80 CFU per node. The observed prevalence of Salmonella within peripheral lymph nodes was 0.48% (7 of 1,464). Two of the seven lymph nodes in which Salmonella organisms were detected came from the same animal. Grass-fed, grain-fed, and cull dairy cattle were all found to have detectable Salmonella in lymph nodes. All Salmonella detections occurred during cooler months of the year. No Shiga toxin–producing E. coli were detected. Aerobic microorganisms were detected above the limit of quantification in 3.2% of nodes (median count 2.24 log per node), and E. coli was detected in 0.8% of nodes (median count 3.05 log per node). The low prevalence of Salmonella and low concentration of aerobic microorganisms in Salmonella-positive lymph nodes of Australian cattle at the time of slaughter suggest that the likelihood of lymph nodes contributing significantly to the presence of Salmonella in ground beef is low.


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