Evaluation of Potential for Inhibition of Growth of Escherichia coli O157:H7 and Multidrug-Resistant Salmonella Serovars in Raw Beef by Addition of a Presumptive Lactobacillus sakei Ground Beef Isolate

2009 ◽  
Vol 72 (2) ◽  
pp. 251-259 ◽  
Author(s):  
JOHN R. RUBY ◽  
STEVEN C. INGHAM

The efficacy of adding presumptive Lactobacillus sakei (LS) strain 10-EGR-a, the most inhibitory from among 12 ground beef Lactobacillus isolates, to inhibit growth by Escherichia coli O157:H7 and multidrug-resistant (MDR) Salmonella (serovars Newport and Typhimurium) was evaluated in a beef-derived broth medium at 10°C and in fresh raw ground beef at 10 and 5°C. Pathogen inhibition was observed in the broth medium at both high (108:105 to 107:105) and low (106:105 to 105:105) LS:pathogen ratios. After 9 days at 10°C, in broth medium with high LS:pathogen ratios, growth of E. coli O157:H7 and MDR Salmonella was inhibited by an average of 2.6 and 3.2 log CFU/ml, respectively, whereas in broth medium with low LS:pathogen ratios, E. coli O157:H7 and MDR Salmonella growth was inhibited by an average of 2.8 and 1.8 log CFU/ml, respectively. However, in raw ground beef no significant inhibition was seen with LS:pathogen ratios of 105:102 to 105:103. Significant inhibition was seen at very high LS:pathogen ratios (106 to 107:102 to 103), but gross spoilage of the product occurred by day 6. Although presumptive LS 10-EGR-a can inhibit growth of E. coli O157:H7 and MDR Salmonella in a beef-derived broth medium, the inability to produce similar results in ground beef without deleteriously affecting the quality of the product is a limitation that needs further investigation.

2014 ◽  
Vol 77 (1) ◽  
pp. 106-111 ◽  
Author(s):  
TAREQ M. OSAILI ◽  
ANAS A. AL-NABULSI ◽  
REYAD R. SHAKER ◽  
ZIAD W. JARADAT ◽  
MOHAMMAD TAHA ◽  
...  

The presence of Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7 in ready-to-eat (RTE) meat products is considered a major concern for food control authorities worldwide. The aims of this study were to determine (i) the prevalence of Salmonella, L. monocytogenes, and E. coli O157:H7 in Mediterranean RTE chicken and beef (CB) products sold in Jordanian restaurants and (ii) the susceptibility of the isolates to antibiotics. A total of 1,028 samples of various types of RTE CB products (550 RTE chicken and 478 RTE beef products) were analyzed by methods described by the International Organization for Standardization followed by molecular confirmation of the isolates. The VITEK2 automated system was used for testing antibiotic susceptibility of the isolates. The overall prevalence of Salmonella serovars in RTE CB products was 0.5%, with 0.8 and 0.2% in RTE chicken and RTE beef, respectively. The overall prevalence of L. monocytogenes in RTE CB products was 2%, with 2.7 and 1.5% in RTE chicken and RTE beef products, respectively. E. coli O157:H7 was not isolated from any of the tested samples. Multidrug-resistant Salmonella and L. monocytogenes isolates were found. The majority of Salmonella isolates were sensitive to most of the tested antibiotics, and all of the isolates were resistant to more than one antibiotic. Similarly, more than 85% of L. monocytogenes isolates were sensitive to nine antibiotics, and the majority of L. monocytogenes isolates were resistant to fosfomycin and oxacillin.


2006 ◽  
Vol 69 (8) ◽  
pp. 1978-1982 ◽  
Author(s):  
J. E. MANN ◽  
M. M. BRASHEARS

In order to provide beef processors with valuable data to validate critical limits set for temperature during grinding, a study was conducted to determine Escherichia coli O157:H7 growth at various temperatures in raw ground beef. Fresh ground beef samples were inoculated with a cocktail mixture of streptomycin-resistant E. coli O157:H7 to facilitate recovery in the presence of background flora. Samples were held at 4.4, 7.2, and 10°C, and at room temperature (22.2 to 23.3°C) to mimic typical processing and holding temperatures observed in meat processing environments. E. coli O157:H7 counts were determined by direct plating onto tryptic soy agar with streptomycin (1,000 μg/ml), at 2-h intervals over 12 h for samples held at room temperature. Samples held under refrigeration temperatures were sampled at 4, 8, 12, 24, 48, and 72 h. Less than one log of E. coli O157:H7 growth was observed at 48 h for samples held at 10°C. Samples held at 4.4 and 7.2°C showed less than one log of E. coli O157:H7 growth at 72 h. Samples held at room temperature showed no significant increase in E. coli O157:H7 counts for the first 6 h, but increased significantly afterwards. These results illustrate that meat processors can utilize a variety of time and temperature combinations as critical limits in their hazard analysis critical control point plans to minimize E. coli O157:H7 growth during the production and storage of 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 (3) ◽  
pp. 591-595 ◽  
Author(s):  
LARRY R. BEUCHAT ◽  
ALAN J. SCOUTEN

The effects of lactic acid, acetic acid, and acidic calcium sulfate (ACS) on viability and subsequent acid tolerance of three strains of Escherichia coli O157:H7 were determined. Differences in tolerance to acidic environments were observed among strains, but the level of tolerance was not affected by the acidulant to which cells had been exposed. Cells of E. coli O157:H7 adapted to grow on tryptic soy agar acidified to pH 4.5 with ACS were compared to cells grown at pH 7.2 in the absence of ACS for their ability to survive after inoculation into ground beef treated with ACS, as well as untreated beef. The number of ACS-adapted cells recovered from ACS-treated beef was significantly (α = 0.05) higher than the number of control cells recovered from ACS-treated beef during the first 3 days of a 10-day storage period at 4°C, suggesting that ACS-adapted cells might be initially more tolerant than unadapted cells to reduced pH in ACS-treated beef. Regardless of treatment of ground beef with ACS or adaptation of E. coli O157:H7 to ACS before inoculating ground beef, the pathogen survived in high numbers.


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.


2009 ◽  
Vol 72 (10) ◽  
pp. 2038-2045 ◽  
Author(s):  
YAGUANG LUO ◽  
QIANG HE ◽  
JAMES L. McEVOY ◽  
WILLIAM S. CONWAY

This study investigated the effect of storage temperature and time on the survival and growth of Escherichia coli O157:H7, the growth of indigenous microorganisms, and the changes in product quality of packaged baby spinach. Commercial packages of spinach within 2 days of processing were cut open at one end, sprayed with fine mists of E. coli O157:H7 inoculum, resealed, and then stored at 1, 5, 8, and 12°C for 12 days until their labeled best-if-used-by dates. Microbial enumeration and product quality evaluation were conducted on day(s) 0, 3, 6, 9, and 12 postinoculation. Spinach held at 12°C supported significant (P < 0.001) E. coli O157:H7 growth, with a 1.0-log CFU/g increase within 3 days postinoculation, which was followed by additional growth during continued storage. E. coli O157:H7 grew slowly when held at 8°C, with a significant (P < 0.01) level of growth reached after 6 days of storage. However, on products held at 1 and 5°C, E. coli O157:H7 populations declined significantly (P < 0.01 and P < 0.001, respectively) within 3 days of storage. Aerobic mesophilic bacteria, psychrotrophic bacteria, and yeast and mold populations increased significantly at all storage temperatures, with more growth on products held at elevated temperatures. Product quality scores remained high within the first 6 days of storage, with a sharp decline noted on samples held at 12°C on day 9. Results suggest that E. coli O157:H7 can grow significantly on commercially packaged spinach held at 8°C or above before significant product quality deterioration occurs.


2002 ◽  
Vol 65 (12) ◽  
pp. 1894-1902 ◽  
Author(s):  
ROLANDO A. FLORES ◽  
MARK L. TAMPLIN

This study determined the distribution patterns of Escherichia coli O157:H7 in ground beef when a contaminated beef trim was introduced into a batch of uncontaminated beef trims prior to grinding in a small-scale laboratory grinder. A beef trim (15.3 ± 2 g) was inoculated with a rifampicin-resistant strain of E. coli O157:H7 (E. coli O157:H7rif) and introduced into a stream of noncontaminated beef (322 ± 33 g) prior to grinding. Seven inoculum levels (6, 5, and 4 total log CFU [high]; and 3, 2, 1, and 0 total log CFU [low]) were studied in triplicate. E. coli O157:H7rif was not detected in 3.1 to 43% of the ground beef inoculated with the high levels or in 3.4 to 96.9% of the ground beef inoculated with the low levels. For all inoculum levels studied, the five ground beef fractions (each 7.8 ± 0.6 g) with the highest pathogen levels accounted for 59 to 100% of the total pathogens detected. For all inoculum levels, there was a linear relationship between the quantity of ground beef containing E. coli O157:H7rif and the inoculum level. The quantity of E. coli O157:H7rif in the beef remaining in the grinder was proportional to the inoculum level and was related to the location in the grinder. Different components of the grinder accumulated E. coli O157:H7rif in different quantities, with the most significant accumulation being in the nut (collar) that attaches the die to the blade. This study determined specific distribution patterns of E. coli O157:H7rif after the grinding of a contaminated beef trim along with uncontaminated trims, and the results indicate that the grinding operation should be regarded as a means of distribution of microbial contamination in risk analyses of ground beef operations.


2007 ◽  
Vol 70 (7) ◽  
pp. 1663-1669 ◽  
Author(s):  
PINA M. FRATAMICO ◽  
LORI K. BAGI

A comparison was made of the relative efficiencies of three enrichment media, RapidChek Escherichia coli O157:H7 enrichment broth (REB), R&F broth (RFB), and modified E. coli broth containing novobiocin (mEC+n), and four selective plating media for detection of cold- and freeze-stressed E. coli O157:H7 in raw ground beef. Ground beef (25 g) was inoculated with E. coli O157:H7 at ≤0.5 and ≤2 CFU/g, and samples were then enriched immediately or were stored at 4°C for 72 h or at −20°C for 2 weeks and then enriched. After 8 or 20 h of enrichment, the cultures were plated onto R&F E. coli O157: H7 chromogenic plating medium, cefixime-tellurite sorbitol MacConkey agar, CHROMagar O157, and Rainbow agar O157 and tested using the RapidChek E. coli O157 lateral flow immunoassay and a multiplex PCR assay targeting the E. coli O157: H7 eae, stx1, and stx2 genes. Recovery of E. coli O157:H7 on the four agar media was 4.0 to 7.9 log CFU/ml with the REB enrichment, 1.4 to 7.4 log CFU/ml with RFB, 1.7 to 6.7 log CFU/ml with mEC+n incubated at 42°C, and 1.3 to 3.3 log CFU/ml from mEC+n incubated at 35°C. The percentages of positive ground beef samples containing nonstressed, cold-stressed, and freeze-stressed E. coli O157:H7 as obtained by plating, the immunoassay, and the PCR assay were 97, 88, and 97%, respectively, with REB, 92, 81, and 78%, respectively, with RFB, 97, 58, and 53%, respectively, with mEC+n incubated at 42°C, and 22, 31, and 25%, respectively, with mEC+n incubated at 35°C. Logistic regression analyses of the data indicated significant main effects of treatment, type of medium, enrichment time, inoculum concentration, and detection method. In particular, a positive result was 1.1 times more likely to occur after 20 h of enrichment than after 8 h, 25 times more likely with RFB and REB than with mEC+nat35°C, 3.7 times more likely with an initial inoculum of ≤2.0 CFU/g than with ≤0.5 CFU/g, 2.5 to 3 times more likely using freeze-stressed or nonstressed bacteria than with cold-stressed bacteria, and 2.5 times more likely by plating than by the immunoassay or the PCR assay. REB had better overall performance for enrichment of cold- and freeze-stressed E. coli O157:H7 present in ground beef than did the other media examined.


2001 ◽  
Vol 84 (3) ◽  
pp. 752-760 ◽  
Author(s):  
Yvette M Henry ◽  
Nandini Natrajan ◽  
Wendy F Lauer

Abstract A method for detection of Escherichia coli O157 in beef and poultry is presented. The method is antibody-based and uses a patented antibody-specific metal-plating procedure for the detection of E. coli O157 in enriched meat samples. Both raw ground beef and raw ground poultry were tested as matrixes for the organism. The sensitivity and specificity of the assay were 98 and 90%, respectively. The accuracy of the assay was 96%. Overall, the method agreement between the E. coli O157 Detex assay and the U.S. Department of Agriculture/Food Safety Inspection Service method was 96%. Sample temperature upon loading of the apparatus was critical to the observed false-positive rate of the system.


2005 ◽  
Vol 68 (3) ◽  
pp. 462-468 ◽  
Author(s):  
ALECIA LAREW NAUGLE ◽  
KRISTIN G. HOLT ◽  
PRISCILLA LEVINE ◽  
RON ECKEL

We analyzed raw ground beef testing data to determine whether a decrease in the rate of Escherichia coli O157:H7–positive raw ground beef samples has occurred since the inception of Food Safety and Inspection Service (U.S. Department of Agriculture) regulatory actions and microbiological testing concerning this commodity and pathogen. A main effects log-linear Poisson regression model was constructed to evaluate the association between fiscal year and the rate of E. coli O157: H7–positive raw ground beef samples while controlling for the effect of season for the subset of test results obtained from fiscal year (FY)2000 through FY2003. Rate ratios were used to compare the rate of E. coli O157:H7–positive raw ground beef samples between sequential years to identify year-to-year differences. Of the 26,521 raw ground beef samples tested from FY2000 through FY2003, 189 (0.71%) tested positive for E. coli O157:H7. Year-to-year comparisons identified a 50% reduction in the rate of positive ground beef samples from FY2002 to FY2003 when controlling for season (95% CI, 10 to 72% decrease; P = 0.02). This decrease was the only significant year-to-year change in the rate of E. coli O157:H7–positive raw ground beef samples but was consistent in samples obtained from both federally inspected establishments and retail outlets. We believe this decrease is attributed to specific regulatory actions by Food Safety and Inspection Service and subsequent actions implemented by the industry, with the goal of reducing E. coli O157:H7 adulteration of raw ground beef. Continued monitoring is necessary to confirm that the decrease in the rate of E. coli O157:H7 in raw ground beef samples we observed here represents the beginning of a sustained trend.


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