Survival of Escherichia coli O157:H7 in Galotyri cheese stored at 4 and 12 degrees C

Post-process contamination of fresh acid-curd cheeses with Escherichia coli O157:H7 may pose a risk considering the low infectious dose and the ability of the pathogen to survive in acidic foods. To evaluate its survival in Galotyri, a traditional Greek acid-curd cheese, portions (0.5 kg) of two commercial fresh products, one artisan (pH 3.9+/-0.1) and the other industrial (pH 3.7+/-0.1), were inoculated with approximately 3.0 or 6.5 log cfu g(-1) of a five-strain cocktail of E. coli O157:H7, including rifampicin-resistant derivatives of the strains ATCC 43895 and ATCC 51657, and stored aerobically at 4 and 12 degrees C. Survival was monitored for 28 days by plating cheese samples on tryptic soy agar with 100 mg l(-1) rifampicin (TSA+Rif), SMAC and Fluorocult E. coli O157:H7 agar media. The pathogen declined much faster (P<0.05) in the industrial as compared to the artisan cheeses at both temperatures. Thus, while E. coli O157:H7 became undetectable by culture enrichment after 14 days at 4 degrees C in industrial samples, irrespective of the inoculation level, populations of 1.4-1.9 and 4.2-5.1 log cfu g(-1) survived after 28 days in the corresponding artisan cheeses with the low and high inocula, respectively. Survival was longer and greater (P<0.05) on TSA+Rif than on SMAC and Fluorocult, indicating the presence of acid-injured cells. Interestingly, survival of E. coli O157:H7 after 14-28 days in cheeses was better at 12 degrees C than at 4 degrees C, probably due to yeasts which grew on the surface of temperature-abused cheeses. The large difference in the pathogen's inactivation between the industrial and artisan cheeses at 4 degrees C could not be associated with major differences in pH or type/concentration of organic acids, suggesting another anti-E. coli O157:H7 activity by the industrial starter. The high survival of the pathogen in artisan Galotyri under conditions simulating commercial storage should be of concern.     

Growth and survival of Escherichia coli O157:H7 on fresh-cut apples in modified atmospheres at abusive temperatures

The effects of reduced-O2 and elevated-CO2 modified atmospheres (MAs) and abusive temperatures on the growth and survival of E. coli O157:H7, yeast, and molds and on changes in the visual quality of fresh-cut apples were evaluated. High-CO1 and low-O2 (> or = 15% and < 1%, respectively) atmospheres inhibited the growth of the pathogen on apple slices at 15 and 20 degrees C. However, the population of the pathogen increased by 1 log cycle after 2 weeks of storage in air. The high-CO2 MA resulted in the inhibition of yeast and mold growth, less browning, and better visual quality than did air and ambient-CO2 atmospheres. The results of this study confirm that E. coli O157:H7 can grow on apple slices in air. These results also show that these organisms survive but are inhibited in MAs with high CO2 levels at abusive temperatures. An MA can increase the shelf life of fresh-cut apples by improving retention of visual quality and inhibiting yeast and molds. Thus, contamination of minimally processed apples with E. coli O157:H7 can be a safety issue for both air- and MA-packaged cut 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。     

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.     

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.     

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 in bovine feces over time under various temperature conditions

Although Escherichia coli O157:H7 prevalence estimates in cattle have increased over time due to improvements in detection methods, fecal sample transport conditions from farm to microbiological laboratories for further analysis may be a factor for prevalence underestimation. The objective of this study was to compare and determine the survival characteristics of E. coli O157:H7 in bovine feces under various storage conditions that could be encountered during transport. Fecal pats were inoculated with a four-strain cocktail of antibiotic-resistant E. coli O157:H7 to contain approximately 1 x 10(5) CFU/g. Inoculated and control samples were taken after 0, 24, 48, 120, and 168 h at each storage temperature and examined for presence and numbers of E. coli O157:H7. Each sample was subdivided and placed at each of the four following temperatures: 37, 23, 4.4 degrees C, and in a plastic cooler with refrigerant packs (0, 4, 4, 21, and 23 degrees C at five sampling times, respectively) to simulate transportation conditions. A statistically significant decrease in the population of the pathogen was observed after 48 h in samples held at 37 degrees C (P < 0.01) and after 168 h at 4.4 degrees C (P = 0.02). At 37 degrees C, E. coli O157:H7 was not detected after 48 h, either by direct plating (P < 0.01) or by immunomagnetic separation. Overall, the results of this study showed that E. coli O157:H7 survived without significant detriment in bovine fecal material inside the cooler for up to 168 h. These results indicate that shipment and storage under these conditions before microbiological analysis would be acceptable and should not affect pathogen detection.     

Survival of Escherichia coli O157:H7 in traditional African yoghurt fermentation

Growth and survival of a nontoxigenic strain of Escherichia coli O157:H7 (ATCC 43888) was determined in traditionally fermented pasteurized milk. Preheated milk was inoculated with 1% (v/v) of a mixed culture of Lactobacillus delbrueckii ssp. bulgaricus (NCIMB 11778) and Streptococcus salivarius ssp. thermophilus (NCIMB 110368) and incubated at 25, 30, 37 or 43 degrees C for 24 h. E. coli O157:H7 (10(5) CFU/ml) were introduced into the milk pre- and post-fermentation. Fermented milk samples were subsequently stored at either 4 degrees C (refrigerator temperature) or 25 degrees C (to mimic African ambient temperature) for 5 days. After 24 h of fermentation, the pH of the samples fermented at the higher temperatures of 37-43 degrees C decreased from 6.8 to 4.4-4.0 ( +/- 0.2) whereas at the lower temperature of 25 degrees C, the pH decreased to pH 5.0 +/- 0.1. During this period, viable counts for E. coli O157:H7 increased from 10(5) to 10(8) - 10(9) CFU/ml except in milk fermented at 43 degrees C wherein viability declined to 10(4) CFU/ml. In fermented (25-30 degrees C) milk stored at 4 degrees C for 5 days, E. coli O157:H7 viability decreased from 10(8-9) to 10(6-7) CFU/ml whereas milk fermented at 43 degrees C resulted in loss of detectable cells. In contrast, storage of fermented milk samples at 25 degrees C for 5 days eventually resulted in complete loss of viability irrespective of fermentation temperature. Stationary phase E. coli O157:H7 inoculated post-fermentation (25 and 43 degrees C) survived during 4 degrees C storage, but not 25 degrees C storage. Fermentation temperature and subsequent storage temperature are critical to the growth and survival of E. coli O157:H7 in traditional fermented products involving yoghurt starter cultures.     

Survival characteristics of Salmonella Typhimurium and Escherichia coli O157:H7 in minimally processed lettuce during storage at different temperatures

Salmonella Typhimurium and Escherichia coli O157:H7 strains were inoculated in minimally processed lettuce samples and survival characteristics of these pathogens during storage at two different temperatures. One group of samples was kept at refrigerator at +4?°C, another group was kept at ambient temperature (+25?°C). According to data obtained, the factor ??temperature?? significantly affected the pathogen bacteria counts of the samples (P?<?0.05). Counts of these two pathogen bacteria were decreased in minimally processed lettuce during storage at +4?°C.     

Selection of recently isolated colicinogenic Escherichia coli strains inhibitory to Escherichia coli O157:H7

Escherichia coli strains were screened for their ability to inhibit E. coli O157:H7. An initial evaluation of 18 strains carrying previously characterized colicins determined that only colicin E7 inhibited all of the E. coli O157:H7 strains tested. A total of 540 strains that had recently been isolated from humans and nine different animal species (cats, cattle, chickens, deer, dogs, ducks, horses, pigs, and sheep) were tested by a flip-plating technique. Approximately 38% of these strains were found to inhibit noncolicinogenic E. coli K12 strains. The percentage of potentially colicinogenic E. coli per animal species ranged from 14% for horse isolates to 64% for sheep strains. Those isolates that inhibited E. coli K12 were screened against E. coli O157:H7, and 42 strains were found to be capable of inhibiting all 22 pathogenic strains tested. None of these 42 strains produced bacteriophages, and only 24 isolates inhibited serotype O157:H7 in liquid culture. The inhibitory activity of these strains was completely eliminated by treatment with proteinase K. When mixtures of these 24 colicinogenic strains were grown in anaerobic continuous culture, the four-strain E. coli O157:H7 population was reduced at a rate of 0.25 log10 cells per ml per h, which was fivefold faster than the washout rate. Two strains originally isolated from cat feces (F16) and human feces (H30) were identified by repetitive sequences polymerase chain reaction as the predominant isolates in continuous cultures. The results of this work indicate that animal species other than cattle can be sources of anti-O157 colicinogenic strains, and these results also lead to the identification of at least two isolates that could potentially be used in preharvest control strategies.     

不同贮藏温度下鸡肉中大肠杆菌O157:H7生长预测模型的建立

目的 建立大肠杆菌在鸡胸肉的生长预测模型,为其加工、贮藏保鲜等过程的温度控制提供参考。方法 以大肠杆菌（Escherichia coli O157:H7）为研究对象,监测其在4、8、10、12、16、20、28、37℃贮藏条件下熟鸡胸肉上的生长情况。采用Gompert模型建立熟鸡胸肉上大肠杆菌的一级生长模型,并将一级生长模型拟合得到的数据代入Ratkowsky方程建立二级模型,综合考虑准确因子Af、偏差因子Bf和均方根误差R2并检验模型的可行性。结果 10、12、16、20、28和37℃条件下拟合的一级生长模型相关系数均在0.94以上,说明能够用来预测该温度范围内大肠杆菌在熟鸡胸肉上的生长动态。在4℃和8℃条件下,大肠杆菌在熟鸡胸肉上呈现先上升后下降再上升的趋势,而在10、12、16、20、28和37℃下,大肠杆菌先进入延滞期,后进入快速增长的指数期,且温度越高,指数期越短。结论 在10～37℃下可以用Gompert模型拟合鸡胸肉中E. coli O157:H7的生长预测模型,且效果较好。     

Survival of Salmonella typhimurium and Escherichia coli O157:H7 in cheese brines

Survival of Salmonella typhimurium and Escherichia coli O157:H7 was studied in model brines and brine from three cheese plants. Three strain mixtures of S. typhimurium and E. coli O157:H7 (10(6) CFU/ml) were inoculated separately into 23% model brine with or without added pasteurized whey (2%) and as a combined inoculum into the commercial brines. The model brines were incubated at 8 and 15 degrees C for 28 days, and the commercial brines at 4 and 13 degrees C for 35 days. Populations of both pathogens in the model brine + whey decreased slowly over 28 days (1.0-2.0 log CFU/ml) with greater survival at 8 degrees C than at 15 degrees C. Corresponding decreases in model brine without whey were 1.9-3.0 log CFU/ml, with greater survival at 8 degrees C than at 15 degrees C. Both S. typhimurium and E. coli O157:H7 survived significantly better (P < 0.05) at 4 degrees C than at 13 degrees C in two of the commercial brines. The survival of each pathogen in the commercial brines at 13 degrees C was significantly influenced by brine pH. Both pathogen populations decreased most rapidly in commercial brines during the first week of storage (2.5-4.0 and 2.3-2.8 log CFU/ml for S. typhimurium and E. coli O157:H7, respectively) with significant recovery (ca. 0.5 log CFU/ml increase) often occurring in the second week of storage. Counts changed little thereafter. Overall, E. coli O157:H7 survived better than S. typhimurium, with differences of 0.1-1.2 log CFU/ml between the two pathogens. Results of this study show that cheese brine could support the survival of contaminating S. typhimurium and E. coli O157:H7 for several weeks under typical brining conditions.     

Survival of Escherichia coli O157:H7 in meat product brines containing antimicrobials

Brine solution injection of beef contaminated with Escherichia coli O157:H7 on its surface may lead to internalization of pathogen cells and/or cross-contamination of the brine, which when recirculated, may serve as a source of new product contamination. This study evaluated survival of E. coli O157:H7 in brines formulated without or with antimicrobials. The brines were formulated in sterile distilled water (simulating the composition of freshly prepared brines) or in a nonsterile 3% meat homogenate (simulating the composition of recirculating brines) at concentrations used to moisture-enhance meat to 110% of initial weight, as follows: sodium chloride (NaCl, 5.5%) + sodium tripolyphosphate (STP, 2.75%), NaCl + sodium pyrophosphate (2.75%), or NaCl + STP combined with potassium lactate (PL, 22%), sodium diacetate (SD, 1.65%), PL + SD, lactic acid (3.3%), acetic acid (3.3%), citric acid (3.3%), nisin (0.0165%) + ethylenediamine tetraacetic acid (EDTA, 200 mM), pediocin (11000 AU/mL) + EDTA, sodium metasilicate (2.2%), cetylpyridinium chloride (CPC, 5.5%), or hops beta acids (0.0055%). The brines were inoculated (3 to 4 log CFU/mL) with rifampicin-resistant E. coli O157:H7 (8-strain composite) and stored at 4 or 15 °C (24 to 48 h). Immediate (0 h) pathogen reductions (P < 0.05) of 1.8 to ≥ 2.4 log CFU/mL were observed in brines containing CPC or sodium metasilicate. Furthermore, brines formulated with lactic acid, acetic acid, citric acid, nisin + EDTA, pediocin + EDTA, CPC, sodium metasilicate, or hops beta acids had reductions (P < 0.05) in pathogen levels during storage; however, the extent of pathogen reduction (0.4 to > 2.4 log CFU/mL) depended on the antimicrobial, brine type, and storage temperature and time. These data should be useful in development or improvement of brine formulations for control of E. coli O157:H7 in moisture-enhanced meat products. PRACTICAL APPLICATION: Results of this study should be useful to the meat industry for developing or modifying brine formulations to reduce the risk of E. coli O157:H7 in moisture-enhanced meat products.     

Studies on the growth of Escherichia coli O157:H7 strains at 45.5 degrees C

The objectives of the present report were to examine the ability of 18 strains of Escherichia coli O157:H7 to grow in EC broth at 42.4, 43.5, 44.5, and 45.5 degrees C, and to document the incidence of phenotypic variants present in low numbers that are capable of growth at 45.5 degrees C in EC broth. Among the 18 strains of E. coli O157:H7 studied, only 3 were capable of producing turbid growth with gas formation in EC broth at 45.5 degrees C with 1 x 10(2) initial CFU/ml. Higher initial densities of CFU resulted in turbid growth and gas formation in EC broth at 45.5 degrees C with all strains. The presence of bile salts #3 in EC broth was found to be inhibitory at 45.5 degrees C. All 18 strains were found to be capable of growth at 45.5 degrees C in nonselective media. The ability of at least one sensitive strain to grow in EC broth at 45.5 degrees C was found to be dependent on the initial number of CFU/ml. Prior growth of cells of a sensitive strain in EC broth at 45.5 degrees C from a cell density of 2.0 x 10(7) to 8.0 x 10(7) CFU/ml followed by removal of cells and reinoculation at a cell density of 2.0 x 10(6) CFU/ml resulted in growth at 45.5 degrees C that did not occur without such conditioning of the inhibitory medium. These results indicate that the ability of most strains of E. coli O157:H7 to grow in EC broth at 45.5 degrees C is dependent on the initial density of CFU and that at low densities of CFU the ability to initiate growth is dependent on either low numbers of phenotypic variants tolerant to the presence of bile salts #3 in EC broth at 45.5 degrees C or to conditioning of the medium with prior elevated numbers of cells.     

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.     

Survival of Listeria monocytogenes and Escherichia coli O157:H7 during sauerkraut fermentation

Sauerkraut was produced from shredded cabbage, as is typical in the United States, and from whole head cabbages, which is a traditional process in parts of Eastern Europe. The sauerkraut was inoculated with five strain mixtures of Escherichia coli O157:H7 and Listeria monocytogenes, and the populations of these bacteria, as well as lactic acid bacteria, pH, and titratable acidity, were monitored over the course of fermentation. Fermentation variables were temperature (18 and 22 degrees C) and salt concentration (1.8, 2.25, and 3%). For most of the analyses, the type of cabbage processing was a significant factor, although within cabbage type, neither salt nor fermentation temperature had significant effects. The final pH of the whole-head sauerkraut was lower than the shredded sauerkraut, but the titratable acidity was significantly higher in the shredded sauerkraut. E. coli O157:H7 and L. monocytogenes persisted in the brines for most of the fermentation, although at the end of the fermentations (15 days for shredded, 28 days for whole head), neither pathogen had detectable populations. E. coli populations decreased more rapidly in the shredded sauerkraut even though the pH was higher because of the higher total acidity in the shredded sauerkraut. Acid-tolerant strains of E. coli and L. monocytogenes were isolated from both shredded and whole-head sauerkraut at different salt concentrations and temperatures after 15 days of fermentation and could be detected at 35 days in the wholehead sauerkraut.     

Survival of Escherichia coli O157:H7 during storage in pressure-treated orange juice.

The effect of a high-pressure treatment on the survival of a pressure-resistant strain of Escherichia coli O157:H7 (NCTC 12079) in orange juice during storage at 3 degrees C was investigated over the pH range of 3.4 to 5.0. The pH of shelf-stable orange juice was adjusted to 3.4, 3.6, 3.9, 4.5, and 5.0 and inoculated with 10(8) CFU ml(-1) of E. coli O157:H7. The orange juice was then pressure treated at 400 MPa for 1 min at 10 degrees C or was held at ambient pressure (as a control). Surviving E. coli O157:H7 cells were enumerated at 1-day intervals during a storage period of 25 days at 3 degrees C. Survival of E. coli O157:H7 during storage was dependent on the pH of the orange juice. The application of high pressure prior to storage significantly increased the susceptibility of E. coli O157:H7 to high acidity. For example, after pressure treatment, the time required for a 5-log decrease in cell numbers was reduced from 13 to 3 days at pH 3.4, from 16 to 6 days at pH 3.6, and from >25 to 8 days at pH 3.9. It is evident that the use of high-pressure processing of orange juice in order to increase the juice's shelf-life and to inactivate pathogens has the added advantage that it sensitizes E. coli O157:H7 to the high acid conditions found in orange juice, which results in the survival of significantly fewer E. coli O157:H7 during subsequent refrigerated storage.     

Inhibition of beef isolates of E. coli O157:H7 by orange oil at various temperatures

Plant essential oils have previously been shown to exhibit antimicrobial activities against various microorganisms. In this study, cold pressed terpeneless Valencia orange oil (CPTVO) was examined at various temperatures (37, 10, and 4 °C) to determine its antimicrobial activity against 3 strains of E. coli O157:H7 recovered from beef products. A micro broth dilution method using 96 well microtiter plates was used with trypticase soy broth with 0.15% agar and 2,3,5 tetrazolium chloride as a growth indicator as the medium. Serial dilutions of CPTVO were made, resulting in final concentrations of oil ranging from 0.2% to 25% or 0.1% to 10%. Plates were incubated statically at 4, 10, or 37 °C, and sampled hourly. After 6 h at 37 °C, all strains were inhibited at concentrations ranging from 0.2% to 0.6%, with a mean of 0.4 ± 0.01%. At 10 °C, all strains were inhibited at concentrations ranging from 0.8% to 6.3%, with a mean of 1.1% ± 0.2%, after 6 h. At 4 °C, all strains were inhibited after 6 h at concentrations ranging from 2.3% to 4.6%, with a mean of 3.5% ± 2.1%. After 24 h at 4 °C the strains were inhibited at concentrations ranging from 0.7% to 1% with a mean of 0.8% ± 0.3%. The ranges appear to be the result of effects from the variable nature of a complex media and an antimicrobial that presents potential multiple mechanisms for inhibition. It appears CPTVO is a viable option to inhibit E. coli O157:H7 growth at refrigeration temperatures. PRACTICAL APPLICATION: Beef products are often the source of foodborne illness from the organism E. coli O157:H7. Orange essential oils have been in the human diet for centuries, and the research reported here indicates that some of these oils may be used as surface applications during cold temperatures to inhibit the foodborne pathogen E. coli O157:H7.