Effectiveness of Salt, pH, and Diacetyl as Inhibitors for Escherichia coli O157:H7 in Dairy Foods Stored at Refrigeration Temperatures

1998 ◽  
Vol 61 (9) ◽  
pp. 1098-1102 ◽  
Author(s):  
R. GURAYA ◽  
J. F. FRANK ◽  
A. N. HASSAN

The behavior of Escherichia coli O157:H7 inoculated in 10% rehydrated nonfat dry milk adjusted to pH levels between 3.8 and 5.4 with lactic acid, salt levels of 0 to 6%, and diacetyl levels of 0, 5, and 10 μg/g was determined at 4 and 12°C. Cell populations were determined by surface plating on tryptic soy agar after 7 and 35 days of incubation. Survival was also determined using retad cultured dairy products. E. coli O157:H7 did not survive in skim milk at pH 3.8 and was reduced by 3 log cycles at pH 4.1, regardless of salt, diacetyl, and temperature levels. At pH levels above 4.4, survival was observed at lower salt concentrations for up to 35 days at both 12 and 4°C. The organism grew (up to a 2.2-log increase) at pH 5.0 at 2% salt levels after 35 days of storage at 12°C but not at 4°C. Diacetyl at a concentration of 10 ppm had no effect on survival and growth. In all but one case, E. coli O157:H7 was inactivated in yogurt, sour cream, and buttermilk at a rate similar to or greater than what was consistent with the acidified skim milk data. Also consistent with the skim milk data, growth occurred in two of the three cottage cheese samples at 12°C after 7 days but not after 35 days or at 4°C, when a 1- to 2-log decline was observed.

1992 ◽  
Vol 55 (5) ◽  
pp. 379-381 ◽  
Author(s):  
MARLENE M. AROCHA ◽  
MELINDA MCVEY ◽  
SUSAN D. LODER ◽  
JOHN H. RUPNOW ◽  
LLOYD BULLERMAN

The ability of enterohemorrhagic Escherichia coli O157:H7 to grow and survive during the manufacture of Cottage cheese was determined. Pasteurized skim milk artificially contaminated with E. coli O157:H7 was used to make Cottage cheese by the washed curd method. E. coli O157:H7 was enumerated by surface plating samples on MacConkey sorbitol agar with 5-bromo-4-chloro-3-indoxyl-β-D-glucuronic acid cyclohexylammonium salt (MSA-BCIG) and incubating at 42°C for 24 h. The heat treated samples were previously inoculated into a modified EC broth with novobiocin and incubated static at 35°C for 24 h. Sorbitol and β-glucuronidase negative colonies were picked from MSA-BCIG, spread on Levine eosin methylene blue agar plates and phenol red sorbitol agar plates with 4-methylumbelliferyl-β-D-glucuronide (PRS-MUG) added for confirmation. E. coli O157:H7 increased 100-fold in numbers during the manufacturing process, but death occurred during cooking of the curd and whey. The pH and acidity did not halt the growth of this pathogen during the manufacture of the cheese; furthermore, the values of these parameters were the same between the contaminated and control samples.


1997 ◽  
Vol 60 (8) ◽  
pp. 891-897 ◽  
Author(s):  
L. M. HUDSON ◽  
J. CHEN ◽  
A. R. HILL ◽  
M. W. GRIFFITHS

Outbreaks of enterohemorrhagic Escherichia coli O157:H7 have been commonly associated with products derived from ground beef, but recently the organism has been implicated as the causative agent in outbreaks involving yogurt and cheese. This finding has raised concern about the potential for its growth and survival in fermented dairy products. A bioluminescent strain of E. coli O157:H7 was used to determine postprocessing survival in yogurt with live cultures at pH 4.17, 4.39, and 4.47 stored at 4 and 10°C. In addition, survival of E. coli O157:H7 was monitored during the manufacture of Cottage, Colby, Romano, and Feta cheeses. Results indicated survival for 8 and 5 days at 4 and 10°C respectively in yogurt at pH 4.17, 17 and 15 days at 4 and 10°C respectively in yogurt at pH 4.39, and 17days at both 4 and 10°C in yogurt at pH 4.47. E. coli O157:H7 did not survive cooking procedures at 56°C in Cottage cheese. However, the pathogen survived for 27, 30, and 27 days in Colby, Romano, and Feta cheeses respectively. A high correlation of r2 > 0.89 was obtained between counts of bioluminescenct colonies and standard plate count for all yogurt and cheese varieties, indicating that bioluminescence was a sensitive and rapid indicator of cellular viability for E. coli O157:H7. Survival of the pathogen, as indicated by this method, is possible in highly acidic environments even at refrigeration temperatures. This poses a potential hazard should postprocessing contamination occur.


1998 ◽  
Vol 81 (2) ◽  
pp. 403-418 ◽  
Author(s):  
Phyllis Entis ◽  
◽  
D Bryant ◽  
J Bryant ◽  
R G Bryant ◽  
...  

abstract Fifteen laboratories took part in a collaborative study to validate a method for enumerating Escherichia coli 0157:H7. The method is based on use of a hydrophobic grid membrane filter and consists of 24 h presumptive enumeration on SD-39 Agar and serological confirmation to yield a confirmed E. coli 0157:H7 count. Six food products were analyzed: pasteurized apple cider, pasteurized 2% milk, cottage cheese, cooked ground pork, raw ground beef, and frozen whole egg. The test method produced significantly higher confirmed count results than did the reference method for milk, pork, and beef. Test method results were numerically higher than but statistically equivalent to reference method results for cheese, cider, and egg. The test method produced lower repeatability and reproducibility values than did the reference method for most food/inoculation level combinations and values very similar to those of the reference method for the remaining combinations. Overall, 94% of presumptive positive isolates from the test method were confirmed serologically as E. coli 0157:H7, and 98% of these were also biochemically typical of E. coli 0157:H7 (completed test). Corresponding rates for the reference method were 69 and 98%, respectively. On the basis of the results of this collaborative study and the precollaborative study that preceded it, it is recommended that this method be adopted official first action for enumeration of E. coli 0157:H7 in meats, poultry, dairy foods, infant formula, liquid eggs, mayonnaise, and apple cider


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.


1997 ◽  
Vol 60 (12) ◽  
pp. 1478-1482 ◽  
Author(s):  
DONNA MAE GARREN ◽  
MARK A. HARRISON ◽  
SCOTT M. RUSSELL

Escherichia coli O157:H7 and non-O157:H7 survival due to induced acid tolerance or shock responses when exposed to lactic acid over time was studied. Induced acid tolerance or shock responses could allow pathogens, like E. coli O157:H7, to survive acidic conditions in foods during storage. Escherichia coli O157:H7 isolates 932 and E009 and a non-Ol57:H7 strain, 23716, were grown to stationary phase at 32°C and exposed to one of two treatments: acid shock or acid adaption. Acid-shocked cells were exposed to lactic acid at pH 3.5 or 4.0. Acid-adapted cells were exposed to pH 5.5 for an adaptation period and then exposed to an acid challenge of pH 3.5 or 4.0. Samples were incubated at either 25 or 32°C and survival of the isolates at 0, 3, 24, 48, 72, and 168 h (7 days), 336 h (14 days), and 504 h (21 days) was determined. All three isolates survived longer with larger populations at pH 4.0 and 25°C compared to the other treatments. In cases where a difference was observed in the two responses, acid-shocked cells had a higher survival rate (typically less than 2 logs) than acid-adapted cells in most cases. Isolate differences were observed at the two pH and temperature levels. Isolate 932 was the most resistant to the acidic conditions during the incubation period, E009 intermediate, and strain 23716 was the most sensitive.


1997 ◽  
Vol 60 (6) ◽  
pp. 610-613 ◽  
Author(s):  
GUODONG WANG ◽  
TONG ZHAO ◽  
MICHAEL P. DOYLE

Escherichia coli O157:H7, which causes hemorrhagic colitis and hemolytic uremic syndrome, has been responsible for several outbreaks associated with consumption of unpasteurized and improperly processed pasteurized milk, and yogurt. Studies were conducted to determine the survival and growth characteristics of this pathogen in unpasteurized milk and pasteurized milk (3.5% fat, 2% fat, skim) at 5,8, 15, and 22°C for up to 28 days. Two levels of inocula (103 and 105 CFU/ml) of a mixture of five nalidixic acid-resistant E. coli O157:H7 strains were used. E. coli O157:H7 did not grow at 5°C and decreased by 1.6 to 2.0 log CFU/ml in 28 days. Growth occurred at 8°C, with an approximately 1- to 2-log CFU/ml increase within the first 4 days. About a 3- to 5-log CFU/ml increase in E. coli O157:H7 populations was observed at 15°C within the first 3 days. In 3 pasteurized milk samples, E. coli O157:H7 continued to grow to populations of greater than 1.0 × 108 CFU/ml at day 7 and remained constant during the remainder of the incubation period. At 22°C, the pH decreased rapidly to less than 4.0 within 4 days and E. coli O157:H7 decreased to undetectable populations within 14 days. E. coli O157:H7 grew more slowly (P < 0.01) in unpasteurized milk, which had a higher initial microbial population, than in pasteurized milks at 8, 15, or 22°C, likely because of antagonistic activity from preexisting bacteria. No significant differences (P> 0.05) in survival or growth of E. coli O157:H7 were observed among the pasteurized milk samples, regardless of fat concentration, at all temperatures throughout the study. The data indicate that temperature abuse during shipping and handling can result in significant growth of E. coli O157:H7. Holding milk at ≤5°C is recommended to prevent growth of this pathogen.


2004 ◽  
Vol 67 (10) ◽  
pp. 2107-2116 ◽  
Author(s):  
MANAN SHARMA ◽  
GLENNER M. RICHARDS ◽  
LARRY R. BEUCHAT

Survival and growth of wild-type (EDL 933) and rpoS-deficient (FRIK 816-3) strains of Escherichia coli O157:H7 after exposure to an alkaline cleaner for 2 min and inoculating into roast beef (pH 6.3) and hard salami (pH 4.9) at low (0.003 to 0.52 CFU/g) and high (0.69 to 31.5 CFU/g) populations were determined. Roast beef was stored at 4 and 12°C; salami was stored at 4, 12, and 20°C. At 4°C, untreated cells of both strains showed greater reductions in populations in salami than in roast beef during a 21-day storage period. Populations of treated and untreated cells recovered from roast beef and salami stored at 4°C on tryptic soy agar were significantly (P ≤ 0.05) higher than on sorbitol MacConkey agar, indicating that a portion of the cells was injured. Treated and untreated cells grew in roast beef at 12°C. Growth of treated cells of the FRIK 816-3 strain in roast beef at 12°C was significantly slower than that of the EDL 933 strain. Populations of both strains decreased at different rates in salami stored at different temperatures (20°C > 12°C > 4°C). E. coli O157:H7 strain EDL 933 grew more rapidly at 20°C in a slurry (pH 5.97) prepared from stored salami (17 days at 20°C) on which Penicillium chrysogenum had grown than in a slurry (5.23) prepared from salami showing no mold growth. Within 2 to 3 days, populations were ca. 3 log CFU/ml higher in slurry made from infected salami than in control salami. Results indicate that treatment of E. coli O157: H7 with an alkaline cleaner for 2 min does not impair resuscitation and growth of surviving cells in roast beef at 12°C. Cross protection of cells exposed to an alkaline cleaner against subsequent stress conditions imposed by roast beef and salami stored at 4°C was not evident in either of the test strains.


2007 ◽  
Vol 70 (11) ◽  
pp. 2631-2637 ◽  
Author(s):  
SYLVIA GAYSINSKY ◽  
T. MATTHEW TAYLOR ◽  
P. MICHAEL DAVIDSON ◽  
BARRY D. BRUCE ◽  
JOCHEN WEISS

The antimicrobial activity of eugenol microemulsions (eugenol encapsulated in surfactant micelles) in ultrahigh-temperature pasteurized milk containing different percentages of milk fat (0, 2, and 4%) was investigated. Antimicrobial micro-emulsions were prepared from a 5% (wt) aqueous surfactant solution (Surfynol 485W) with 0.5% (wt) eugenol. Two strains each of Listeria monocytogenes and Escherichia coli O157:H7 previously shown to be the least and most resistant to the microemulsion in microbiological media were used to inoculate sterile milk (104 CFU/ml). Samples were withdrawn and plated at 0, 1, 3, 6, 12, and 24 h for enumeration. Microemulsions completely prevented growth of L. monocytogenes for up to 48 h in skim milk and reduced both strains of E. coli O157:H7 to less than detectable levels in less than 1 h. Similarly, in 2% fat milk, eugenol-Surfynol combinations reduced both strains of E. coli O157:H7 to less than detectable levels in less than 1 h but only increased the lag phase of both strains of L. monocytogenes. In full-fat milk (4% fat), microemulsions inhibited growth of the least resistant strains of L. monocytogenes and E. coli but were ineffective against the two resistant strains. Unencapsulated eugenol was slightly more or as inhibitory as microemulsions against target pathogens. Results were attributed to diffusional mass transport of antimicrobials from microemulsions to the macroemulsion (milk). Results suggest that food composition, especially fat level, may affect the efficiency of targeting of foodborne pathogens with surfactant-encapsulated antimicrobials.


2015 ◽  
Vol 78 (7) ◽  
pp. 1288-1295 ◽  
Author(s):  
DIKE O. UKUKU ◽  
LIHAN HUANG ◽  
CHRISTOPHER SOMMERS

For health reasons, people are consuming fresh-cut fruits with or without minimal processing and, thereby, exposing themselves to the risk of foodborne illness if such fruits are contaminated with bacterial pathogens. This study investigated survival and growth parameters of Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and aerobic mesophilic bacteria transferred from cantaloupe rind surfaces to fresh-cut pieces during fresh-cut preparation. All human bacterial pathogens inoculated on cantaloupe rind surfaces averaged ~4.8 log CFU/cm2, and the populations transferred to fresh-cut pieces before washing treatments ranged from 3 to 3.5 log CFU/g for all pathogens. A nisin-based sanitizer developed in our laboratory and chlorinated water at 1,000 mg/liter were evaluated for effectiveness in minimizing transfer of bacterial populations from cantaloupe rind surface to fresh-cut pieces. Inoculated and uninoculated cantaloupes were washed for 5 min before fresh-cut preparation and storage of fresh-cut pieces at 5 and 10°C for 15 days and at 22°C for 24 h. In fresh-cut pieces from cantaloupe washed with chlorinated water, only Salmonella was found (0.9 log CFU/g), whereas E. coli O157:H7 and L. monocytogenes were positive only by enrichment. The nisin-based sanitizer prevented transfer of human bacteria from melon rind surfaces to fresh-cut pieces, and the populations in fresh-cut pieces were below detection even by enrichment. Storage temperature affected survival and the growth rate for each type of bacteria on fresh-cut cantaloupe. Specific growth rates of E. coli O157:H7, Salmonella, and L. monocytogenes in fresh-cut pieces were similar, whereas the aerobic mesophilic bacteria grew 60 to 80% faster and had shorter lag phases.


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