Validation of Pepperoni Processes for Control of Escherichia coli O157:H7‡

1996 ◽  
Vol 59 (12) ◽  
pp. 1260-1266 ◽  
Author(s):  
JAY C. HINKENS ◽  
NANCY G. FAITH ◽  
TIMOTHY D. LORANG ◽  
PHILLIP BAILEY ◽  
DENNIS BUEGE ◽  
...  

The outbreak of Escherichia coli O157:H7 linked with dry-cured salami in late 1994 prompted regulatory action that required manufacturers of fermented products to demonstrate a 5-log unit reduction in counts of this pathogen during processing. Therefore, pepperoni batter (75% pork:25% beef with a fat content of ca. 32%) was inoculated with a pediococcal starter culture and a five-strain mixture of E. coli O157:H7 (≥2 × 107 CFU/g) and stuffed into 55-mm diameter fibrous casings 47 cm in length. The viability of the pathogen was monitored before stuffing, after fermentation, after thermal processing, and/or after drying. Chubs were fermented at 96°F (36°C) and 85% relative humidity (RH) to pH ≤ 5.0 and then dried at 55°F (13°C) and 65% RH to a moisture/protein ratio of ≤1.6:1 (modified method 6 process). Counts of the pathogen decreased about 1.2 log units after fermentation and drying. In subsequent experiments, heating chubs after fermentation to internal temperatures of 145°F (63°C) instantaneous or 128°F (53°C) for 60 min resulted in a ≥5-log unit decrease in numbers of strain O157:H7 without visibly affecting the texture or appearance of the product. These data revealed that a traditional nonthermal, process for pepperoni was only sufficient to eliminate relatively low levels (ca. 2 log CFU/g) of E. coli O157:H7, whereas heating to internal temperatures of 145°F (63°C) instantaneous or 128°F (53°C) for 60 min delivered a 5 to 6 log unit reduction in counts of the pathogen in pepperoni.


1998 ◽  
Vol 61 (4) ◽  
pp. 383-389 ◽  
Author(s):  
NANCY G. FAITH ◽  
RACHEL K. WIERZBA ◽  
ANNE M. IHNOT ◽  
ANN M. ROERING ◽  
TIMOTHY D. LORANG ◽  
...  

Pepperoni batter was prepared with fat contents of about 15, 20, and 32% (wt/wt) and inoculated with a pediococcal starter culture and ≥2.0 × 107 CFU/g of a five-strain inoculum of Escherichia coli O157:H7. The batter was fermented at 96°F (ca. 36°C) and 85% relative humidity (RH) to pH ≤ 4.8 and then dried at 55°F (ca. 13°C) and 65% RH to a moisture/protein ratio of ≤1.6:1. For storage, slices were packaged under air or vacuum and stored at 39°F (ca. 4°C) and 70°F (ca. 21°C). For baking, frozen slices were placed on retail frozen cheese pizzas that were subsequently baked at 275°F (ca. 135°C), 375°F (ca. 191°C), or 475°F (ca. 246°C) for 0 to 20 min. Appreciable differences related to fat levels were observed after drying; pathogen numbers decreased by 1.04, 1.31 and 1.62 log10 units in sticks prepared from batter at initial fat levels of 15, 20, and 32%, respectively. During storage, the temperature rather than the atmosphere had the greater effect on pathogen numbers, with similar viability observed among the three fat levels tested. At 70°F (ca. 21°C), compared to original levels, pathogen numbers decreased by ≥5.56 and ≥4.53 log10 units within 14 days in slices stored under air and vacuum, respectively, whereas at 39°F (ca. 4°C) numbers decreased by ≤2.43 log10 CFU/g after 60 days of storage under either atmosphere. Baking, as expected, resulted in greater reductions in pathogen numbers as the temperature and/or time of baking increased. However, it was still possible to recover the pathogen by enrichment after baking frozen slices on frozen pizza at 475°F (ca. 246°C) for 10 min or at 375°F (ca. 191°C) for 15 min. The calculated D values for all three temperatures tested increased as the fat content of the batter increased from 15 to 20 to 32%. The present study confirmed that fermentation and drying were sufficient to reduce levels of E. coli O157:H7 in pepperoni sticks by <2.0 log10 CFU/g. Storage of slices for at least 14 days at ambient temperature under air resulted in a >5.5-log10-unit total reduction of the pathogen. Baking slices on frozen pizza for at least 15 min at 475°F (ca. 246°C) or 20 min at 375°F (ca. 191°C) was necessary to reduce pathogen numbers to below detection by both direct plating and enrichment.



1998 ◽  
Vol 61 (4) ◽  
pp. 377-382 ◽  
Author(s):  
NANCY G. FAITH ◽  
NELLY PARNIERE ◽  
TRINA LARSON ◽  
TIMOTHY D. LORANG ◽  
CHARLES W. KASPAR ◽  
...  

The fate of Escherichia coli O157:H7 was monitored in salami during conditioning of batter, fermentation and drying of sticks, and storage of slices. The raw batter (75% pork:25% beef, wt/wt, fat content about 20%) was inoculated with a pediococcal starter culture (about 108 CFU/g) and a five-strain cocktail of E. coli O157:H7 (≥2 × 107 CFU/g) and stuffed into 104-mm diameter fibrous casings. After being refrigerated at 4°C or being tempered at 13°C, frozen at −20°C, and thawed at 4°C, or being frozen at −20°C, and thawed at 4°C, the inoculated batter was fermented at 24°C and 90% relative humidity (RH) to pH ≤4.8, dried at 13°C and 65% RH to a moisture/protein ratio of ≤1.9:1, and then stored at 4 or 21°C under air or vacuum. For salami sticks sampled immediately after drying, appreciable differences were evident among the various batter-conditioning treatments; pathogen numbers were reduced from original levels by 2.1, 1.6, or 1.1 log10 units when batter was tempered, frozen, and thawed, frozen and thawed, or refrigerated, respectively. Similarly, regardless of storage temperature or atmosphere, within 7 days salami slices cut from sticks prepared from batter that was tempered, frozen, and thawed (2.7- to 4.9-log10-unit reduction) or frozen and thawed (2.3- to 4.8-log10-unit reduction) displayed a greater impact on pathogen numbers than slices cut from sticks prepared from batter that was refrigerated (1.6- to 3.1-log10-unit reduction). The effects of batter conditioning notwithstanding, a greater reduction in levels of E. coli O157:H7 was observed when slices were stored at 21°C compared to otherwise similar slices stored at 4°C. After storage for 60 days the pathogen was only detected by enrichment in slices stored at 21°C, whereas pathogen levels ranged from 1.4 to 4.5 log10 CFU/g in slices stored at 4°C. Differences related to storage atmosphere were first observed after slices were stored for 21 days. Such differences were more readily demonstrable after 60 and 90 days, with pathogen numbers for treatments that were statistically different ranging from 0.6- to 1.5-log10 units higher on slices stored under vacuum than in air. These data emphasize the need to implement multiple barriers to appreciably reduce numbers of E. coli O157:H7 in salami.



2009 ◽  
Vol 72 (12) ◽  
pp. 2476-2482 ◽  
Author(s):  
NIGEL M. HARPER ◽  
MICHELLE N. ROBERTS ◽  
KELLY J. K. GETTY ◽  
ELIZABETH A. E. BOYLE ◽  
DANIEL Y. C. FUNG ◽  
...  

Foodborne outbreaks have been linked to jerky produced under insufficient thermal processing schedules. Reduction of Escherichia coli O157:H7 and Salmonella serovars during thermal processing of chopped and formed beef jerky was evaluated under two processing schedules representative of those used by large-scale (LS) and small-scale (SS) jerky production facilities. Fresh chopped and formed all-beef jerky batter was inoculated with 5.8 to 7.3 log CFU of E. coli O157:H7 or Salmonella per g, extruded into strips, and thermally processed by LS or SS schedules. A $5.0-log CFU/g reduction of both pathogens occurred with <10% relative humidity and a cumulative process of 44 min at 55.6°C followed by 46 min at 77.8°C into the LS schedule. Additional drying at 77.8°C for 3.5 h was needed to achieve a water activity of 0.67 and a moisture-to-protein ratio (MPR) of 0.77. For the SS process, a $5.0-log CFU/g reduction of both pathogens occurred with 15 to 20% relative humidity and a cumulative process of 45 min at 52°C, 60 min at 57°C, 45 min at 60°C, 45 min at 63°C, 90 min at 68°C, and finishing with 30 min at 77°C. After processing for an additional 90 min at 77°C, water activity was 0.60 while the MPR was 0.82. The LS and SS processes for producing chopped and formed jerky provided $5.0 log lethality to control E. coli O157:H7 and Salmonella. However, both processes would require additional drying to achieve an MPR of 0.75 to be labeled as jerky.



2004 ◽  
Vol 67 (12) ◽  
pp. 2651-2656 ◽  
Author(s):  
P. McGEE ◽  
L. SCOTT ◽  
J. J. SHERIDAN ◽  
B. EARLEY ◽  
N. LEONARD

Ruminant livestock, particularly cattle, is considered the primary reservoir of Escherichia coli O157:H7. This study examines the transmission of E. coli O157:H7 within groups of cattle during winter housing. Holstein Friesian steers were grouped in six pens of five animals. An animal inoculated with and proven to be shedding a marked strain of E. coli O157: H7 was introduced into each pen. Fecal (rectal swabs) and hide samples (900 cm2 from the right rump) were taken from the 36 animals throughout the study. Water, feed, and gate or partition samples from each pen were also examined. Within 24 h of introducing the inoculated animals into the pens, samples collected from the drinking water, pen barriers, and animal hides were positive for the pathogen. Within 48 h, the hides of 20 (66%) of 30 cohort animals from the six pens were contaminated with E. coli O157:H7. The first positive fecal samples from the noninoculated cohort animals were detected 3 days after the introduction of the inoculated steers. During the 23 days of the study, 15 of 30 cohort animals shed the marked E. coli O157: H7 strain in their feces on at least one occasion. Animal behavior in the pens was monitored during a 12-h period using closed circuit television cameras. The camera footage showed an average of 13 instances of animal grooming in each pen per hour. The study suggests that transmission of E. coli O157:H7 between animals may occur following ingestion of the pathogen at low levels and that animal hide may be an important source of transmission.



2008 ◽  
Vol 71 (3) ◽  
pp. 486-493 ◽  
Author(s):  
GARY H. GRAUMANN ◽  
RICHARD A. HOLLEY

Compounds generated by the enzymatic hydrolysis of glucosinolates naturally present in mustard powder are potently bactericidal against Escherichia coli O157:H7. Because E. coli O157:H7 can survive the dry fermented sausage manufacturing process, 2, 4, and 6% (wt/wt) nondeheated (hot) mustard powder or 6% (wt/wt) deheated (cold) mustard powder were added to dry sausage batter inoculated with E. coli O157:H7 at about 7 log CFU/g to evaluate the antimicrobial effectiveness of the powders. Reductions in E. coli O157:H7 populations, changes in pH and water activity (aw), effects on starter culture (Pediococcus pentosaceus and Staphylococcus carnosus) populations, and effects of mustard powder on sausage texture (shear) were monitored during ripening. Nondeheated mustard powder at 2, 4, and 6% in dry sausage (0.90 aw) resulted in significant reductions in E. coli O157:H7 (P < 0.05) of 3.4, 4.4, and 6.9 log CFU/g, respectively, within 30 days of drying. During fermentation and drying, mustard powder did not affect P. pentosaceus and S. carnosus activity in any of the treatments. Extension of drying to 36 and 48 days reduced E. coli O157:H7 by >5 log CFU/g in the 4 and 2% mustard powder treatments, respectively. The 6% deheated mustard powder treatment provided the most rapid reductions of E. coli O157:H7 (yielding <0.20 log CFU/g after 24 days) by an unknown mechanism and was the least detrimental (P < 0.05) to sausage texture.



2019 ◽  
Vol 82 (5) ◽  
pp. 815-825 ◽  
Author(s):  
MAHTA MOUSSAVI ◽  
VANESSA LIEBERMAN ◽  
CHRIS THEOFEL ◽  
JAVAD BAROUEI ◽  
LINDA J. HARRIS

ABSTRACT During harvest, pistachios are hulled, separated in water into floater and sinker streams (in large part on the basis of nut density), and then dried before storage. Higher prevalence and levels of Salmonella were previously observed in floater pistachios, but contributing factors are unclear. To examine the behavior of pathogens on hulled pistachios during simulated drying delays, floater and sinker pistachios collected from commercial processors were inoculated at 1 or 3 log CFU/g with cocktails of Salmonella and in some cases Escherichia coli O157:H7 or Listeria monocytogenes and incubated for up to 30 h at 37°C and 90% relative humidity. Populations were measured by plating onto tryptic soy agar and appropriate selective agars. In most cases, no significant growth (P > 0.05) of Salmonella was observed in the first 3 h after inoculation in hulled floaters and sinkers. Growth of Salmonella was greater on floater pistachios than on corresponding sinkers and on floater pistachios with ≥25% hull adhering to the shell surface than on corresponding floaters with <25% adhering hull. Maximum Salmonella populations (2 to 7 log CFU/g) were ∼2-log higher on floaters than on corresponding sinkers. The growth of E. coli O157:H7 and Salmonella on hulled pistachios was similar, but a longer lag time (approximately 11 h) and significantly lower maximum populations (4 versus 5 to 6 log CFU/g; P < 0.05) were predicted for L. monocytogenes. Significant growth of pathogens on hulled pistachios is possible when delays between hulling and drying are longer than 3 h, and pathogen growth is enhanced in the presence of adhering hull material.



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.



2010 ◽  
Vol 73 (12) ◽  
pp. 2217-2224 ◽  
Author(s):  
DENNIS J. D'AMICO ◽  
MARC J. DRUART ◽  
CATHERINE W. DONNELLY

This study was conducted to examine the fate of Escherichia coli O157:H7 during the manufacture and aging of Gouda and stirred-curd Cheddar cheeses made from raw milk. Cheeses were manufactured from unpasteurized milk experimentally contaminated with one of three strains of E. coli O157:H7 at an approximate population level of 20 CFU/ml. Samples of milk, whey, curd, and cheese were collected for enumeration of bacteria throughout the manufacturing and aging process. Overall, bacterial counts in both cheese types increased almost 10-fold from initial inoculation levels in milk to approximately 145 CFU/g found in cheeses on day 1. From this point, counts dropped significantly over 60 days to mean levels of 25 and 5 CFU/g in Cheddar and Gouda, respectively. Levels of E. coli O157:H7 fell and stayed below 5 CFU/g after an average of 94 and 108 days in Gouda and Cheddar, respectively, yet remained detectable after selective enrichment for more than 270 days in both cheese types. Changes in pathogen levels observed throughout manufacture and aging did not significantly differ by cheese type. In agreement with results of previous studies, our results suggest that the 60-day aging requirement alone is insufficient to completely eliminate levels of viable E. coli O157:H7 in Gouda or stirred-curd Cheddar cheese manufactured from raw milk contaminated with low levels of this pathogen.



2021 ◽  
Vol 72 (1) ◽  
pp. 2689
Author(s):  
G CELIK ◽  
A DIKICI ◽  
A KOLUMAN

In this study, the survival of Escherichia coli O157:H7 and non-O157 STEC serogroups of O26, O111, O103, and O145 were investigated during production and storage of yogurt. For this purpose, pathogens were individually inoculated into milk after pasteurization along with the starter culture (approximately 7.00±1.00 log10 cfu/g). After incubation at 44oC (about 180 min), yogurt samples were capped and stored at 4oC for 20 days. Pathogens were enumerated at 0, 5, 10, 15, and 20th days of storage. Lactic acid content (%) and pH of the samples were also screened. Moreover, mesophilic Lactococcus spp. and mesophilic Lactobacillus spp. were enumerated during production of yogurt.After incubation, the number of E. coli O157, O26, O103, O145, O111were 6.76±0.45, 6.64±0.53, 7.12±0.43, 6.00±1.39, 5.89±1.37 log10 cfu/g, respectively. A significant decrease was determined in all groups during the storage of yogurt samples at 4oC (p<0.05). It was detected on the 20th day of storage that the number of E. coli O157:H7 and non-O157 STEC serogroups of O103 and O145 were under the detection limit. However, STEC O26 and O111 were viable around 1.51±0.98 and 1.18±0.62 log10 cfu/g respectively. Results of the study showed that Escherichia coli O157:H7 and non-O157 STEC serogroups might pose a potential health risk during production and storage of yogurt.



2007 ◽  
Vol 70 (12) ◽  
pp. 2717-2724 ◽  
Author(s):  
SUNEE HIMATHONGKHAM ◽  
MARY LEE DODD ◽  
JENNY K. YEE ◽  
DAVID K. LAU ◽  
RAYMOND G. BRYANT ◽  
...  

The objective of this study was to develop a rapid, simple method for enhanced detection and isolation of low levels of Escherichia coli O157:H7 from leafy produce and surface water using recirculating immunomagnetic separation (RIMS) coupled with real-time PCR and a standard culture method. The optimal enrichment conditions for the method also were determined. Analysis of real-time PCR data (CT values) suggested that incubation of lettuce and spinach leaves rather than rinsates provides better enrichment of E. coli O157:H7. Enrichment of lettuce or spinach leaves at 42°C for 5 h provided better detection than enrichment at 37°C. Extended incubation of surface water for 20 h at 42°C did not improve the detection. The optimized enrichment conditions were also employed with modified Moore swabs, which were used to sample flowing water sites. Positive isolation rates and real-time PCR results indicated an increased recovery of E. coli O157:H7 from all samples following the application of RIMS. Under these conditions, the method provided detection and/or isolation of E. coli O157:H7 at levels as low as 0.07 CFU/g of lettuce, 0.1 CFU/g of spinach, 6 CFU/100 ml of surface water, and 9 CFU per modified Moore swab. During a 6-month field study, modified Moore swabs yielded high isolation rates when deployed in natural watershed sites. The method used in this study was effective for monitoring E. coli O157:H7 in the farm environment, during postharvest processing, and in foodborne outbreak investigations.



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