Survival of Escherichia coli O157:H7 in buttermilk as affected by contamination point and storage temperature

The effects of contamination point (during fermentation versus postfermentation) and storage temperature (5 and 12 degrees C) were determined for survival of Escherichia coli O157:H7 in fermented buttermilk. E. coli O157:H7 was recovered from buttermilk inoculated during fermentation for 22 days and in buttermilk inoculated postfermentation for 32 days. For storage temperatures of 5 and 12 degrees C, D-values were lower for E. coli O157:H7 inoculated during fermentation (2.5, 2.2 days) than postfermentation (5.6, 4.8 days) (P < 0.05). Developed acidity in inoculated buttermilks was not different from controls (P > 0.05). The extended recovery of viable enterohemorrhagic E. coli O157:H7 from both processing scenarios indicates that the presence of E. coli O157:H7 in buttermilk is not limited to postprocessing contamination.     

Survival of Escherichia coli O157:H7 and Salmonella in apple cider and orange juice as affected by ozone and treatment temperature

Inactivation of Escherichia coli O157:H7 and Salmonella in apple cider and orange juice treated with ozone was evaluated. A five-strain mixture of E. coli O157:H7 or a five-serovar mixture of Salmonella was inoculated (7 log CFU/ml) into apple cider and orange juice. Ozone (0.9 g/h) was pumped into juices maintained at 4 degrees C, ambient temperature (approximately 20 degrees C), and 50 degrees C for up to 240 min, depending on organism, juice, and treatment temperature. Samples were withdrawn, diluted in 0.1% peptone water, and surface plated onto recovery media. Recovery of E. coli O157:H7 was compared on tryptic soy agar (TSA), sorbitol MacConkey agar, hemorrhagic coli agar, and modified eosin methylene blue agar; recovery of Salmonella was compared on TSA, bismuth sulfite agar, and xylose lysine tergitol 4 (XLT4) agar. After treatment at 50 degrees C, E. coli O157:H7 populations were undetectable (limit of 1.0 log CFU/ml; a minimum 6.0-log CFU/ml reduction) after 45 min in apple cider and 75 min in orange juice. At 50 degrees C, Salmonella was reduced by 4.8 log CFU/ml (apple cider) and was undetectable in orange juice after 15 min. E. coli O157:H7 at 4 degrees C was reduced by 4.8 log CFU/ml in apple cider and by 5.4 log CFU/ml in orange juice. Salmonella was reduced by 4.5 log CFU/ml (apple cider) and 4.2 log CFU/ml (orange juice) at 4 degrees C. Treatment at ambient temperature resulted in population reductions of less than 5.0 log CFU/ml. Recovery of E. coli O157:H7 and Salmonella on selective media was substantially lower than recovery on TSA, indicating development of sublethal injury. Ozone treatment of apple cider and orange juice at 4 degrees C or in combination with mild heating (50 degrees C) may provide an alternative to thermal pasteurization for reduction of E. coli O157:H7 and Salmonella in apple cider and orange juice.     

Water, sodium chloride and acidified sodium chlorite effects on Escherichia coli O157:H7 and Staphylococcus aureus on beef briskets

Effectiveness of spray application of potable water wash (WW), 25% (w/v) sodium chloride (NaCl), and 0.1% (v/v) acidified sodium chlorite (ASC) was evaluated against Escherichia coli O157:H7 and Staphylococcus aureus inoculated onto beef briskets. The purpose was to identify antimicrobial treatments which may be applied to beef carcasses and more specifically in kosher meat facilities. Treatments were applied for 10-60 s at pressure of 419 kPa. Water wash, NaCl, and ASC significantly reduced E. coli O157:H7 as compared with the control, although, only ASC resulted in improved removal with increased exposure time. Water wash did not significantly reduce S. aureus counts throughout exposure and NaCl was only effective after 60 s of exposure, while ASC reduced counts throughout exposure. E. coli O157:H7 was twice as sensitive to WW and NaCl as S. aureus in terms of percent reduction in cell count.     

Death of Salmonella,Escherichia coli O157:H7, and Listeria monocytogenes in garlic butter as affected by storage temperature

Garlic is known to have antimicrobial activity against several spoilage and pathogenic bacteria. However, the fate of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes in garlic butter has not been reported. This study was undertaken to determine the viability of these organisms in garlic butter as affected by the type of raw minced garlic added to the butter, storage temperature, and storage time. Unsalted butter at 40 degrees C was combined with raw minced jumbo, elephant, or small-cloved garlic at a 4:1 butter/garlic ratio (wt/wt), inoculated with mixed-strain suspensions of Salmonella, E. coli O157:H7, or L monocytogenes, and stored at 4.4, 21, or 37 degrees C for up to 48 h. All pathogens retained their viability at 4.4 degrees C, regardless of the presence of garlic. The addition of garlic to butter enhanced the rates of inactivation of all three pathogens at 21 and 37 degrees C. The most rapid decline in pathogen populations was observed at 37 degrees C. The inactivation of L. monocytogenes occurred more slowly than did that of Salmonella or E. coli O157:H7. The inactivation of Salmonella and L. monocytogenes was more rapid in jumbo garlic butter than in elephant or small-cloved garlic butter. It is concluded that Salmonella, E. coli O157:H7, and L. monocytogenes did not grow in unsalted butter, with or without garlic added (20%, wt/wt), when inoculated products were stored at 4.4, 21, and 37 degrees C for up to 48 h.     

Thermal inactivation of Escherichia coli O157:H7 in ground beef supplemented with sodium lactate

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 (10(7) to 10(8) CFU/g) were subjected to immersion heating in a water bath stabilized at 55, 57.5, 60, 62.5, or 65 degrees 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 degrees 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.     

Modelling the effect of pH,sodium chloride and sodium pyrophosphate on the thermal resistance of Escherichia coli O157:H7 in ground beef

The objective of this study was to assess the combined effects of temperature, pH, sodium chloride (NaCl), and sodium pyrophosphate (SPP) on the heat resistance of Escherichia coli O157:H7 in minced beef meat. A fractional factorial design consisted of four internal temperatures (55.0, 57.5, 60.0 and 62.5 °C), five concentrations of NaCl (0.0, 1.5, 3.0, 4.5 and 6.0 wt/wt.%) and SPP (0.0, 0.1, 0.15, 0.2 and 0.3 wt/wt.%), and five levels of pH (4.0, 5.0, 6.0, 7.0 and 8.0). The 38 variable combinations were replicated twice to provide a total of 76 survivor curves, which were modelled by a modified three-parameter Weibull function as primary model. The polynomial secondary models, developed to estimate the time to achieve a 3-log and a 5-log reduction, enabled the estimation of critical pH, NaCl and SPP concentrations, which are values at which the thermo-tolerance of E. coli O157:H7 reaches it maximum. The addition up to a certain critical concentration of NaCl (~ 2.7–4.7%) or SPP (~ 0.16%) acts independently to increase the heat resistance of E. coli O157:H7. Beyond such critical concentrations, the thermo-resistance of E. coli O157:H7 will progressively diminish. A similar pattern was found for pH with a critical value between 6.0 and 6.7, depending upon temperature and NaCl concentration. A mixed-effects omnibus regression model further revealed that the acidity of the matrix and NaCl concentration had a greater impact on the inactivation kinetics of E. coli O157:H7 in minced beef than SPP, and both are responsible for the concavity/convexity of the curves. When pH, SPP or NaCl concentration is far above or below from its critical value, the temperatures needed to reduce E. coli O157:H7 up to a certain log level are much lower than those required when any other environmental condition is at its critical value. Meat processors can use the model to design lethality treatments in order to achieve specific log reductions of E. coli O157:H7 in ready-to-eat beef products.     

Survival of Salmonella and Escherichia coli O157:H7 in chorizos

Mexican-style raw meat sausages (chorizos) are not regulated in California when they are produced in small ethnic food markets. These sausages are sold uncooked, but their formulation imparts a color that may lead the consumer to assume that they are already cooked, and thus the chorizos may sometimes be eaten without proper cooking. If pathogens are present in such cases, illness may result. Survival of Salmonella and Escherichia coli O157:H7 in chorizos was evaluated under different storage conditions selected based on an initial survey of uninspected chorizos in California. Chorizos were formulated with five different initial water activity (aw) values (0.85, 0.90, 0.93, 0.95, and 0.97), stored under four conditions (refrigeration at 6 to 8 degrees C, room temperature at 24 to 26 degrees C, under a hood at 24 to 26 degrees C with forced air circulation, and incubation at 30 to 31 degrees C with convective air circulation), and sampled after 1, 2, 4, and 7 days. The initial pH was 4.8 and remained near 5.0 from day 1 of the sampling period. Two separate studies of packs inoculated with five-strain cocktails of Salmonella and of E. coli O157:H7 were performed twice for each initial aw. The three lowest aw values (0.85, 0.90, and 0.93) and the incubation and hood storage conditions were more effective (P < or = 0.05) at reducing the target pathogen levels in chorizos than were the two highest aw values (0.95 and 0.97) and the refrigeration storage condition, regardless of storage time. These results provide a scientific basis for guidelines given to producers of uninspected chorizo and should reduce the probability of foodborne illness associated with these products.     

Survival of Escherichia coli O157:H7 and Salmonella Muenchen on apples as affected by application of commercial fruit waxes

Survival of Escherichia coli O157:H7, Salmonella Muenchen, and yeasts and molds on apples as affected by application of five commercial apple waxes was investigated. Red Delicious cv. apples at 21 degrees C were spot inoculated with E. coli O157:H7 and S. Muenchen and spray coated with waxes. Apples sprayed with water served as controls. Apples were dried at either 21 or 55 degrees C for 2 min before subjecting to microbiological analysis after storage for 0, 1, 3, 6, and 12 weeks at 2 or 21 degrees C. Drying temperature did not significantly influence populations of E. coli O157:H7 and S. Muenchen. Waxing reduced populations E. coli O157:H7 and S. Muenchen by up to 1.48 log10 cfu/apple. Compared to untreated apples, treatment of apples with water or waxes resulted in significant (P < or = 0.05) reductions in populations of E. coli O157:H7 and S. Muenchen during storage at 2 degrees C. Reductions on waxed apples stored at 21 degrees C were not as marked compared to reductions on waxed apples stored at 2 degrees C. With the exception of one wax, drying temperature did not significantly influence populations of yeasts and molds. Mold populations were less affected by wax applications than were yeasts, and were detected in higher numbers on apples treated with three of the five waxes compared to populations recovered from untreated control apples. None of the waxes evaluated can be relied upon to kill or remove E. coli O157:H7 and Salmonella on apples.     

大肠杆菌O157:H7的冷冻损伤及解冻存活

为探讨冷冻后残存的大肠杆菌O157:H7（Escherichia coli O157:H7）在解冻后的存活情况,本研究首先比较4 株E. coli O157:H7冷冻后的死亡和损伤情况,进而采用无营养的磷酸盐缓冲液作为基质研究冷冻后不同解冻方式对E. coli O157:H7存活的影响。结果表明：4 株E. coli O157:H7 －20 ℃冷冻24、48、72 h后均发生了一定程度的死亡和损伤,冷冻时间越长细菌致死和致伤程度越明显,且存在菌株差异,冷冻72?h时菌株CICC21530的损伤率最高,为87.70%。采用混合菌株进行解冻实验,4?株E.?coli?O157:H7磷酸盐缓冲液菌液冷冻后立即置于20、30、37?℃解冻,细菌发生了进一步的死亡,解冻温度越高死亡越明显,3?个温度组在解冻48?h时菌落数均显著低于冷冻72?h时菌落数（P＜0.05）。进一步探讨缓慢解冻方式对菌体存活的影响,菌液冷冻后先置于4?℃一定时间（0、2、6、12?h）,再置于37?℃不同时间（5、10、30?min）观察菌株存活情况,结果表明4?℃缓慢解冻时间越长,越有利于细菌的存活,4?℃、12?h/37?℃、5～30?min解冻方式下改良山梨醇麦康凯琼脂上菌落数仍显著低于胰蛋白胨大豆琼脂上的菌落数（P＜0.05）,表明仍有损伤菌的存在。本实验提示采用缓慢解冻反而有利于残存菌的存活,冷冻食品风险评估时应重视残存菌尤其是损伤菌的检测和控制。     

Viability of Salmonella,Escherichia coli O157:H7, and Listeria monocytogenes in yellow fat spreads as affected by storage temperature

A study was conducted to characterize the survival and inactivation kinetics of a five-serotype mixture of Salmonella (6.23 to 6.55 log10 CFU per 3.5-ml or 4-g sample), a five-strain mixture of Escherichia coli O157:H7 (5.36 to 6.14 log10 CFU per 3.5-ml or 4-g sample), and a six-strain mixture of Listeria monocytogenes (5.91 to 6.18 log10 CFU per 3.5-ml or 4-g sample) inoculated into seven yellow fat spreads (one margarine, one butter-margarine blend, and five dairy and nondairy spreads and toppings) after formulation and processing and stored at 4.4, 10, and 21 degrees C for up to 94 days. Neither Salmonella nor E. coil O157:H7 grew in any of the test products. The time required for the elimination of each pathogen depended on the product and the storage temperature. Death was more rapid at 21 degrees C than at 4.4 or 10 degrees C. Depending on the product, the time required for the elimination of viable cells at 21 degrees C ranged from 5 to 7 days to >94 days for Salmonella, from 3 to 5 days to 28 to 42 days for E. coli O157:H7, and from 10 to 14 days to >94 days for L. monocytogenes. Death was most rapid in a water-continuous spray product (pH 3.66, 4.12% salt) and least rapid in a butter-margarine blend (pH 6.66, 1.88% salt). E. coli O157:H7 died more rapidly than did Salmonella or L. monocytogenes regardless of storage temperature. Salmonella survived longer in high-fat (> or = 61%) products than in products with lower fat contents. The inhibition of growth is attributed to factors such as acidic pH, salt content, the presence of preservatives, emulsion characteristics, and nutrient deprivation. L. monocytogenes did not grow in six of the test products, but its population increased between 42 and 63 days in a butter-margarine blend stored at 10 degrees C and between 3 and 7 days when the blend was stored at 21 degrees C. On the basis of the experimental parameters examined in this study, traditional margarine and spreads not containing butter are not "potentially hazardous foods" in that they do not support the growth of Salmonella, E. coli O157:H7, or L. monocytogenes.     

Survival of enterohemorrhagic Escherichia coli O157:H7 in retail mustard

Escherichia coli O157:H7 survival in acid foods such as unpasteurized apple cider and fermented sausage is well documented. Researchers have determined that E. coli O157:H7 can survive in refrigerated acid foods for weeks. The potential of acid foods to serve as a vector of E. coli O157:H7 foodborne illness prompted this study to determine the fate of this organism in retail mustard containing acetic acid when stored at room and refrigerated temperatures. Various retail brands of dijon, yellow, and deli style mustard, pH ranging from 3.17 to 3.63, were inoculated individually with three test strains of E. coli O157:H7. Samples were inoculated with approximately 1.0 x 10(6) CFU/g, incubated at room (25+/-2.5 degrees C) and refrigerated (5+/-3 degrees C) temperatures, and assayed for surviving test strains at predetermined time intervals. An aliquot was appropriately diluted and plated using sorbitol MacConkey agar (SMAC). When the test strain was not recoverable by direct plating, the sample was assayed by enrichment in modified tryptic soy broth and recovered using SMAC. Growth of E. coli O157:H7 test strains was inhibited in all retail mustard styles. E. coli O157:H7 was not detected in dijon style mustard beyond 3 h at room and 2 days at refrigerated temperatures. Survival in yellow and deli style mustard was not detected beyond 1 h. Overall, test strain survival was greater at refrigerated than room temperature. Retail mustard demonstrated the ability to eliminate effectively any chance contamination by this organism within hours to days, suggesting that these products are not a likely factor in E. coli O157:H7 foodborne illness.     

Inhibition of Escherichia coli O157:H7, Listeria monocytogenes and Staphylococcus aureus on sliced roast beef by cetylpyridinium chloride and acidified sodium chlorite

The effects of 0.5% cetylpyridinium chloride (CPC), 0.12% acidified sodium chlorite (ASC) and a mix of equal volume of the two (0.25% CPC-0.06% ASC) on Escherichia coli O157:H7, Listeria monocytogenes and Staphylococcus aureus were evaluated on inoculated sliced roast beef. The antimicrobial agents were, respectively, sprayed on the beef surfaces and tray absorbent pads, and samples were stored at 4 degrees C for 10 days (d). At 0 d, L. monocytogenes and S. aureus were reduced to undetectable levels in 2 h after spraying with CPC. CPC-ASC treatment reduced E. coli O157:H7, L. monocytogenes and S. aureus by 4.07, 6.37 and 4.32 log cfu/cm2, respectively, at 0 d. ASC treatment reduced the population of E. coli O157:H7 by 6.09 log cfu/cm2 at 10 d. CPC treatment caused a slight discoloration and ASC-treated beef surfaces demonstrated the lowest redness and highest lightness. The grey colour and off-odour were significant in the ASC-treated beef samples, while CPC-treated samples demonstrated less off-odor and brown colour from 0 to 4 d. Based on our results, it appears that the application of CPC on sliced roast beef can extend the shelf-life of the product without impairing its quality.     

Survival of Escherichia coli O157:H7 in cucumber fermentation brines

Abstract: Bacterial pathogens have been reported on fresh cucumbers and other vegetables used for commercial fermentation. The Food and Drug Administration currently has a 5‐log reduction standard for E. coli O157:H7 and other vegetative pathogens in acidified pickle products. For fermented vegetables, which are acid foods, there is little data documenting the conditions needed to kill acid resistant pathogens. To address this knowledge gap, we obtained 10 different cucumber fermentation brines at different stages of fermentation from 5 domestic commercial plants. Cucumber brines were used to represent vegetable fermentations because cabbage and other vegetables may have inhibitory compounds that may affect survival. The 5‐log reduction times for E. coli O157:H7 strains in the commercial brines were found to be positively correlated with brine pH, and ranged from 3 to 24 d for pH values of 3.2 to 4.6, respectively. In a laboratory cucumber juice medium that had been previously fermented with Lactobacillus plantarum or Leuconostoc mesenteroides (pH 3.9), a 5‐log reduction was achieved within 1 to 16 d depending on pH, acid concentration, and temperature. During competitive growth at 30 °C in the presence of L. plantarum or L. mesenteroides in cucumber juice, E. coli O157:H7 cell numbers were reduced to below the level of detection within 2 to 3 d. These data may be used to aid manufacturers of fermented vegetable products determine safe production practices based on fermentation pH and temperature. Practical Application: Disease causing strains of the bacterium E. coli may be present on fresh vegetables. Our investigation determined the time needed to kill E. coli in cucumber fermentation brines and how E. coli strains are killed in competition with naturally present lactic acid bacteria. Our results showed how brine pH and other brine conditions affected the killing of E. coli strains. These data can be used by producers of fermented vegetable products to help assure consumer safety.     

Escherichia coli O157:H7 shedding in vaccinated beef calves born to cows vaccinated prepartum with Escherichia coli O157:H7 SRP vaccine

Extensive research, intervention equipment, money, and media coverage have been directed at controlling Escherichia coli O157:H7 in beef cattle. However, much of the focus has been on controlling this pathogen postcolonization. This study was conducted to examine the performance, health, and shedding characteristics of beef calves that were vaccinated with an E. coli O157:H7 SRP bacterial extract. These calves had been born to cows vaccinated prepartum with the same vaccine. Cows and calves were assigned randomly to one of four treatments: (i) neither cows nor calves vaccinated with E. coli O157:H7 SRP (CON), (ii) cows vaccinated with E. coli O157:H7 SRP prepartum but calves not vaccinated (COWVAC), (iii) calves vaccinated with E. coli O157:H7 SRP but born to cows not vaccinated (CALFVAC), (iv) cows vaccinated with E. coli O157:H7 SRP prepartum and calves also vaccinated (BOTH). Calves born to vaccinated cows had significantly higher titers of anti-E. coli O157:H7 SRP antibodies (SRPAb) in circulation at branding time (P < 0.001). Upon entry to the feedlot, overall fecal E. coli O157:H7 prevalence was 23 % among calves, with 25 % in the CON treatment group, 19 % in the CALFVAC group, 32 % in the COWVAC group, and 15 % in the BOTH group (P > 0.05). Fecal shedding of E. coli O157 on arrival to the feedlot was not correlated with fecal shedding at slaughter (Spearman's rho = -0.02; P = 0.91). No significant effects of cow or calf E. coli O157:H7 SRP vaccination treatment were found on feedlot calf health or performance (P > 0.05), prevalence of lung lesions or liver abscess (P > 0.05), or morbidity, retreatment, or mortality numbers (P > 0.05). The findings of this study indicate that the timing of vaccination of calves against E. coli O157:H7 may be an important consideration for maximizing the field efficacy of this vaccine.     

Control of Escherichia coli O157:H7 with sodium metasilicate

Three intervention strategies-trisodium phosphate, lactic acid, and sodium metasilicate--were examined for their in vitro antimicrobial activities in water at room temperature against a three-strain cocktail of Escherichia coli O157:H7 and a three-strain cocktail of "generic" E. coli. Both initial inhibition and recovery of injured cells were monitored. When 3.0% (wt/wt) lactic acid, pH 2.4, was inoculated with E. coli O157:H7 (approximately 6 log CFU/ml), viable microorganisms were recovered after a 20-min exposure to the acid. After 20 min in 1.0% (wt/wt) trisodium phosphate, pH 12.0, no viable E. coli O157:H7 microorganisms were detected. Exposure of E. coli O157:H7 to sodium metasilicate (5 to 10 s) at concentrations as low as 0.6%, pH 12.1, resulted in 100% inhibition with no recoverable E. coli O157:H7. No difference in inhibition profiles was detected between the E. coli O157:H7 and generic strains, suggesting that nonpathogenic strains may be used for in-plant sodium metasilicate studies.     

Seasonal prevalence of Escherichia coli O157:H7 in beef cattle feces

Cattle are an asymptomatic reservoir of Escherichia coli O157:H7, but the bacterial colonization and shedding patterns are poorly understood. The prevalence and shedding of this human pathogen have been reported to be seasonal with rates typically increasing during warm months. The objectives of this study were (i) to assess the prevalence of E. coli O157:H7 in feces of feedlot cattle in Kansas during summer, fall, and winter months, and (ii) to characterize E. coli O157:H7 by screening for virulence factors. Of 891 fecal samples collected, 82 (9.2%) were positive for E. coli O157:H7. No significant differences in prevalence were detected among summer, fall, and winter months. The highest monthly prevalence (18.1%) was detected in February. All tested isolates were positive for stx2 (Shiga toxin 2) and eaeA (intimin) genes; 14 isolates (12.8%) also carried stx1. Our results indicate the prevalence of E. coli O157:H7 in beef cattle feces is not necessarily season dependent.     

Chlorine inactivation of Escherichia coli O157:H7 in water

Six human isolates of Escherichia coli O157:H7 and E. coli (ATCC 11229) were used to determine the concentrations of free chlorine and exposure times required for inactivation. Free chlorine concentrations of 0.25, 0.5, 1.0, and 2.0 ppm at 23 degrees C were evaluated, with sampling times at 0, 0.5, 1.0, and 2.0 min. Results revealed that five of six E. coli O157:H7 isolates and the E. coli control strain were highly susceptible to chlorine, with >7 log10 CFU/ml reduction of each of these strains by 0.25 ppm free chlorine within 1 min. However, comparatively, one of the seven strains was unusually tolerant to chlorine at 23 degrees C for 1 min, with a 4-, 5.5-, 5.8-, and >5.8-log CFU/ml reduction at free chlorine concentrations (ppm) of 0.25, 0.5, 1.0, and 2.0. respectively. Based on these studies most isolates of E. coli O157:H7 have no unusual tolerance to chlorine; however, one strain was exceptional in being recovered after 1-min of exposure of 10(7) CFU/ml to 2.0 ppm of free chlorine. This isolate may be a useful reference strain for future studies on chlorine tolerance of E. coli O157:H7.     

Presence of Escherichia coli O157:H7 in ground beef and ground baby beef meat

A total of 114 beef and baby beef samples were examined. The samples included ground baby beef, mixed ground baby beef and pork, and chopped and shaped meat. The samples were analyzed from 30 different grocery stores in Zagreb, Croatia. The object of this study was to evaluate the prevalence of Escherichia coli O157:H7 in the samples that can enhance the potential risk of outbreaks of hemorrhagic colitis and hemolytic uremic syndrome. The results in all tested samples of E. coli O157:H7 were negative. A single sample was positive in a latex agglutination test using antiserum to O157:H7. It was identified as Proteus vulgaris at the Pasteur Institute, Paris, France. This result correlates positively with cross-contamination with Yersinia enterocolitica 09, Brucella abortus, Salmonella type N, and Pseudomonas maltophila.     

Inactivation of Escherichia coli O157:H7 in packaged ground beef by allyl isothiocyanate

Commercial allyl isothiocyanate (AIT) was examined for its ability to reduce numbers of Escherichia coli O157:H7 inoculated in fresh ground beef packaged under nitrogen and stored refrigerated or frozen. A five-strain cocktail of E. coli O157:H7 containing 3 or 6 log10 cfu/g was inoculated into 100 g ground beef and formed into 10x1-cm patties. A 10-cm diameter filter paper disk treated with AIT suspended in sterile corn oil was placed on top of a single patty. One patty and paper disk were placed in a bag of Nylon/EVOH/PE with O2 permeability of 2.3 cm3 m(-2) 24 h atm at 23 degrees C. The bags were back-flushed with 100% nitrogen, heat-sealed and stored at 10, 4 and -18 degrees C for 8, 21 or 35 days, respectively. During storage, the AIT levels in the package headspaces were determined by gas liquid chromatography, and mesophilic bacteria and E. coli O157:H7 were counted. The mesophilic aerobic bacteria in ground beef patties were largely unaffected by the addition of AIT. At an initial population of 3 log10 cfu/g, E. coli O157:H7 was reduced by AIT to undetectable levels after 18 days at 4 degrees C or 10 days at -18 degrees C. In samples inoculated with 6 log10 cfu/g, a >3 log10 reduction of E. coli O157:H7 was observed after 21 days at 4 degrees C, while a 1 log10 reduction was observed after 8 and 35 days at 10 and -18 degrees C, respectively. The final AIT concentrations in the headspaces after storage at 10, 4, and -18 degrees C were 444, 456, and 112 microg/ml at 8, 21, and 35 days, respectively. Results showed that AIT can substantially reduce numbers of E. coli O157:H7 in fresh ground beef during refrigerated or frozen storage.