食品标本处理方法对大肠埃希菌检出限影响

目的 研究快速、敏感、低成本的食品标本处理方法。方法 在牛肉馅中接种不同浓度的大肠埃希菌O157：H7，经离心、过滤，去除PCR抑制剂。浓缩菌株，用煮沸法和酶／冻融法裂解菌株，制备模板DNA。通过PCR检测大肠埃希菌O157：H7志贺毒素基因以评价该处理方法的可行性。结果 在未增菌条件下，PCR最低能检测到103CFU／g牛肉馅，增菌6h，最低能检测1CFU／g牛肉馅，整个检测过程只需要12h。结论 所采用的标本处理方法是一种灵敏、省时、低成本的方法，能显著地提高PCR检测食品标本中的大肠埃希菌O157：H7的灵敏度。     

大肠杆菌O157∶H7双重荧光PCR方法的建立与应用

摘要:目的　建立快速检测食品中大肠杆菌O157︰H7的双重荧光PCR 方法。方法　针对大肠杆菌O157︰H7菌体抗原O和鞭毛抗原H的特异性基因rfbE（O157）和fliC（H7）筛选设计引物和探针,优化反应条件,建立可特异性检测大肠杆菌O157︰H7的双重荧光PCR方法,并对该法的特异性、敏感性和重复性进行评价,对模拟样品和实际样品进行初步验证。结果　利用建立的方法对2株大肠杆菌O157︰H7及12株非大肠杆菌O157︰H7进行检测,O157︰H7菌株检测结果均为rfbE和fliC阳性,非O157︰H7菌株检测结果均为阴性,rfbE和fliC基因的特异性均为100%;2基因的检测灵敏度可达1.16×102 CFU/ml;批内、批间变异系数均小于3.5％;模拟样品立即检测,2个基因的最低检出限（2.7×102 CFU/ml）和细菌纯培养时接近;实际样品检测结果与我国检验检疫行业标准（SN/T 0973-2010）的检测结果一致。结论　建立的荧光定量PCR 方法特异性及重复性好,灵敏度高,可快速准确鉴定大肠杆菌O157︰H7菌株。     

Meningococcal disease at the University of Southampton: outbreak investigation

Between November 1992 and February 1993, a large outbreak of Escherichia coli O157:H7 infections occurred in the western USA and was associated with eating ground beef patties at restaurants of one fast-food chain. Restaurants that were epidemiologically linked with cases served patties produced on two consecutive dates; cultures of recalled ground beef patties produced on those dates yielded E. coli O157:H7 strains indistinguishable from those isolated from patients, confirming the vehicle of illness. Seventy-six ground beef patty samples were cultured quantitatively for E. coli O157:H7. The median most probable number of organisms was 1.5 per gram (range, < 0.3-15) or 67.5 organisms per patty (range, < 13.5-675). Correlation of the presence of E. coli O157:H7 with other bacterial indicators yielded a significant association between coliform count and the presence of E. coli O157:H7 (P = 0.04). A meat traceback to investigate possible sources of contamination revealed cattle were probably initially colonized with E. coli O157:H7, and that their slaughter caused surface contamination of meat, which once combined with meat from other sources, resulted in a large number of contaminated ground beef patties. Microbiological testing of meat from lots consumed by persons who became ill was suggestive of an infectious dose for E. coli O157:H7 of fewer than 700 organisms. These findings present a strong argument for enforcing zero tolerance for this organism in processed food and for markedly decreasing contamination of raw ground beef. Process controls that incorporate microbiological testing of meat may assist these efforts.     

大肠杆菌O157多克隆抗体及食品中双抗ELISA测定方法的研究

本研究获得了抗肠出血性大肠杆菌O157:H7的多克隆抗体 ,建立了一种适宜食品样品检测的双抗ELISA检测方法。该方法对纯培养菌液检出限为 10 3 ～ 10 4 cfu ml;只对O157菌株有特异性反应 ,对非O157菌株无交叉反应 ;经过增菌 ,鸡肉与牛奶染菌样品中的大肠杆菌O157的检出限均为 0 1cfu g(cfu ml)。     

Detection by multiplex real-time polymerase chain reaction assays and isolation of Shiga toxin-producing Escherichia coli serogroups O26, O45, O103, O111, O121, and O145 in ground beef

Six Shiga toxin-producing Escherichia coli (STEC) serogroups, which include O26, O45, O103, O111, O121, and O145, are responsible for the majority of non-O157 STEC infections in the United States, representing a growing public health concern. Cattle and other ruminants are reservoirs for these pathogens; thus, food of bovine origin may be a vehicle for infection with non-O157 STEC. Methods for detection of these pathogens in animal reservoirs and in food are needed to determine their prevalence and to develop intervention strategies. This study describes a method for detection of non-O157 STEC in ground beef, consisting of enrichment in modified tryptic soy broth at 42°C, followed by real-time multiplex polymerase chain reaction (PCR) assays targeting stx(1), stx(2), and genes in the O-antigen gene clusters of the six serogroups, [corrected] and then immunomagnetic separation (IMS) followed by plating onto Rainbow? Agar O157 and PCR assays for confirmation of isolates. All ground beef samples artificially inoculated with 1-2 and 10-20 CFU/25?g of ground beef consistently gave positive results for all of the target genes, including the internal amplification control using the multiplex real-time PCR assays after enrichment in modified tryptic soy broth for a total of 24?h (6?h at 37°C and 18?h at 42°C). The detection limit of the real-time multiplex PCR assays was ～50 CFU per PCR. IMS for O26, O103, O111, and O145 was performed with commercially available magnetic beads, and the IMS beads for O45 and O121 were prepared using polyclonal antiserum against these serogroups. A large percentage of the presumptive colonies of each serogroup picked from Rainbow Agar O157 were confirmed as the respective serogroups; however, the percent recovery of STEC O111 was somewhat lower than that of the other serogroups. This work provides a method for detection and isolation in ground beef and potentially other foods of non-O157 STEC of major public health concern.     

食品样品中大肠杆菌O157:H7复合PCR检测方法的研究

根据大肠杆菌O15 7:H7特异性基因fliC ,rfbE ,SLTI与SLTII设计四对引物 ,建立了复合PCR方法 ,扩增产物长度分别为为 5 6 0bp ,6 78bp ,2 10bp与 4 84bp。该方法可以一步检测出O15 7与H7特异性抗原基因 ,并可同时检测该菌产生SLT毒素的类型 ,与 71株试验菌株的基因型完全配合 ,是一种特异、高效的检测方法。该方法适用于牛奶样品中大肠杆菌O15 7:H7的检测 ,经过增菌步骤检测限可达 0 1cfu ml     

用最低检出限评价大肠埃希菌O157:H7的检测方法及应用研究

目的用最低检出限评价大肠埃希菌O157:H7的三种检测方法,在突发公共卫生事件及食源性致病菌流行病学调查中,将大肠埃希菌O157:H7的检测方法有机组合,提高检测的速度、灵敏度。方法将标准菌株10倍稀释,制备模拟样品,用大肠埃希菌O157:H7的三种方法进行检测,比较不同检测方法的最低检出限。结果标准菌株10倍稀释改良CHROMagar O157显色琼脂平板分离法、免疫磁珠分离法、实时荧光PCR法检测法的最低检出限分别为103、100、102cfu/ml;模拟样品中增菌前改良CHROMagar O157显色琼脂平板分离法、免疫磁珠分离法、实时荧光PCR法检测法的最低检出限分别为105、101、104cfu/ml,增菌培养后改良CHROMagar O157显色琼脂平板分离法、免疫磁珠分离法、实时荧光PCR法检测法的最低检出限分别为102、100、100cfu/ml。结论免疫磁珠分离法在增菌前和增菌培养后的检出限为最低,增菌培养后实时荧光PCR法的检出限也达到了最低,适用于食物中毒及食源性疾病的快速诊断。用改良CHROMagar O157显色琼脂平板分离法进行检测,易造成漏检。将免疫磁珠分离法、实时荧光PCR法有机组合,可以大大提高检测的速度,灵敏度,并且可获得菌株。     

Evaluation of a multiplex PCR system for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in foods and in food subjected to freezing

Conventional culture methods were compared to a multiplex polymerase chain reaction (PCR) assay for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 from enrichment cultures of various types of artificially inoculated and naturally contaminated foods. The multiplex PCR assay was evaluated in 44 types of spiked food samples, including meat, produce, fish, and dairy products targeting genes specific for each pathogen for simultaneous detection. The sensitivity of the assay was 相似文献   

Evaluation of universal pre-enrichment broth for isolation of Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes from dairy farm environmental samples

Use of universal pre-enrichment broth (UPB) as a primary enrichment medium for detection of Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes from dairy farm environmental samples was evaluated. There were no differences in bacterial growth between UPB and selective primary enrichment broths for each pathogen inoculated individually or in combination at 10(1) and 10(2) colony forming units/mL. In addition, no differences were observed when UPB and selective primary enrichment broths were compared for detection efficiency of pathogens in artificially contaminated raw milk and fecal samples. Listeria enrichment broth (LEB) was compared with UPB to support growth of L. monocytogenes from naturally contaminated environmental samples. Listeria monocytogenes was isolated from seven of 30 samples enriched in UPB and six of 30 samples enriched in LEB. Dairy farm environmental samples were examined for recovery of the three pathogens using UPB. Subsequent isolation was achieved using selective secondary enrichment of each pathogen. Listeria monocytogenes, Salmonella spp., and E. coli O157:H7 were isolated in 13.4% (30 of 224), 8.9% (20 of 224), and 2.2% (five of 224) of samples, respectively. Isolation rates of the three pathogens were somewhat higher than in previous reports. Overall, UPB supported growth of test pathogens to detectable levels within 24 h. Our results demonstrate that UPB has potential for routine use in isolation of foodborne pathogens from diverse environmental samples.     

12株疑似O157:H7大肠菌PCR鉴定研究

目的:对12株疑似O157:H7大肠菌采用PCR法进行鉴定。方法:利用单一PCR和多重聚合酶链反应(mPCR)检测不同来源菌株志贺样毒素(stx1和stx2)、溶血素(hly)、粘附抹平因子(eaeA)、β-葡糖醛酸糖苷酶(u idA)、O157抗原编码(rfbE)、H7鞭毛抗原编码(fliC)基因。结果:4株大肠菌rfbE和fliC基因检测为阳性,确认为EHEC O157:H7,其中1株菌株扩增出全部毒力基因,另3株菌株扩增出除stx1外其它全部毒力基因;2株大肠菌rfbE基因检测阳性,确认为O157:H7-大肠菌;其它均为非O157:H7其它大肠菌。结论:PCR技术的应用能对可疑O157:H7大肠菌进行有效鉴定与分析,应成为今后病原学鉴定的主要技术手段。     

Polymerase chain reaction screening for Salmonella and enterohemorrhagic Escherichia coli on beef products in processing establishments

Pathogenic Escherichia coli strains on raw or insufficiently cooked foods are of public health concern as serious disease may result from their ingestion. Therefore, many commercial producers of beef products screen for E. coli O157:H7 before shipment. While Salmonella is not considered an adulterant on raw beef products, it is used as an indication of process control. To detect these microorganisms, rapid screening methods are often used to provide results within 8-24 hours after sampling. During 2005-2008, about 971,389 samples from several commercial beef production plants were tested using a rapid screening method based on the polymerase chain reaction to determine if they were presumptively positive for bacterial cells carrying Salmonella or Shiga toxin-producing E. coli-specific genes. Of the product lots sampled (trim, ground beef, and variety meats), 15% were positive for the stx(1) and/or stx(2) (Shiga toxin genes), 9.1% for the eae gene (the attaching and effacing gene [eae] encoding intimin), 3.0% for an rfb gene region (encoding the O157-specific O side chain polysaccharide), and 1.67% for Salmonella by the polymerase chain reaction assay. In general, lots of ground beef showed the lowest frequency of contamination, and variety meats (by-products of carcass evisceration), the highest. Overall, 4.6%, 4.6%, and 0.8% samples were screen-positive for enteropathogenic E. coli, enterohemorrhagic E. coli, and E. coli O157, respectively. Of the E. coli O157-positive samples, 14% were also Salmonella positive. The frequency of screen-positive samples increases during the summer months, probably because of the prevalence of climatic conditions more conducive to microbial growth. The presence of fecal organisms in beef products suggests a failure of sanitary controls during processing and the more prevalent relatives of E. coli O157, Shiga toxin-producing Escherichia coli, enteropathogenic E. coli, and enterohemorrhagic E. coli, serve as more sensitive indicators of contamination than O157 strains alone.     

Effects of gamma ray and electron-beam irradiations on survival of anaerobic and facultatively anaerobic bacteria

An extension of the approval for food irradiation is desired due to the increase in the incidence of food poisoning in the world. One anaerobic (Clostridium perfringens) and four facultatively anaerobic (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Enteritidis) bacteria irradiated with gamma ray or electron beam (E-beam) were tested in terms of survival on agar under packaging atmosphere. Using pouch pack, effects of two irradiations on survival of anaerobic and facultatively anaerobic bacteria were evaluated comparatively. E-beam irradiation was more effective than gamma ray irradiation in decreasing the D10 value of B. cereus at 4 degrees C, slightly more effective in that of E. coli O157, and similarly effective in that of the other three bacteria at 4 degrees C. The gamma irradiation of the bacteria without incubation at 4 degrees C before irradiation was more effective than that of the bacteria with incubation overnight at 4 degrees C before irradiation in decreasing the D10 values of these bacteria (B. cereus, E. coli O157, and L. monocytogenes). Furthermore, ground beef patties inoculated with bacteria were irradiated with 1 kGy by E-beam (5 MeV) at 4 degrees C. The inoculated bacteria in the 1-9 mm beef patties were killed by 1 kGy E-beam irradiation and some bacteria in more than 9 mm beef patties were not killed by the irradiation.     

大肠埃希菌O157:H7的实时荧光PCR检测方法研究

目的建立一种敏感、特异、快速的大肠埃希菌O157:H7的检测方法,应用于突发公共卫生事件及食源性致病菌流行病学调查的检测。方法根据GenBank大肠埃希菌O157:H7rfbE基因序列,设计引物和TaqMan探针,对实时荧光PCR反应条件进行优化,建立实时荧光PCR检测大肠埃希菌O157:H7的反应体系,并对该法的特异性、敏感性和重复性进行评价。结果大肠埃希菌O157:H7菌株的检测结果均为阳性,而所有其它菌株检测结果均为阴性;该方法检测的灵敏度可达1×102cfu/ml。模拟污染的猪肉、羊肉、鸡肉、生食蔬菜样品,均可检出1×104cfu/ml的细菌。从细菌核酸提取至完成检测约需3 h。结论建立的实时荧光PCR检测方法具有灵敏度高、特异性强、快速等优点,可用于大肠埃希菌O157:H7食物中毒的快速诊断和食品微生物检测,为食源性疾病的分子流行病学调查提供新的检测手段。     

A comparison of the antimicrobial activity of garlic, ginger, carrot, and turmeric pastes against Escherichia coli O157:H7 in laboratory buffer and ground beef

The antimicrobial effects of garlic, ginger, carrot and turmeric pastes against Escherichia coli O157:H7 in laboratory buffer and model food system were investigated. Turmeric paste, fresh carrot, ginger and garlic pastes from roots, and commercial ginger and garlic paste were heated alone or with buffered peptone water (BPW) or ground beef at 70 degrees C for 7 min. All samples were inoculated with a three strain cocktail of overnight cultures of E. coli O157: H7 and stored at 4 degrees C and 8 degrees C for 2 weeks. Each paste exhibited different antimicrobial effects alone and in ground beef or BPW at 4 degrees C and 8 degrees C for 2 weeks. Commercial ginger paste and fresh garlic paste showed the strongest antimicrobial activity with complete inactivation of E. coli O157:H7 in the paste at 3 days at 4 degrees C and 8 degrees C. Carrot and turmeric pastes did not show any antimicrobial activity both at 4 degrees C and 8 degrees C. Commercial garlic showed antimicrobial activity at both 4 degrees C and 8 degrees C (about 1 log CFU/g reduction) in the paste. However, fresh ginger paste showed antimicrobial activity only at 8 degrees C. Only commercial ginger paste had antimicrobial activity in BPW at 4 degrees C for 2 weeks. However, commercial ginger paste showed antimicrobial activity in ground beef at 3 days and after (about 1-2 log CFU/g) compared to control samples at 8 degrees C for 2 weeks. Fresh garlic paste showed antimicrobial activity only in BPW (1.3 log CFU/g) at 8 degrees C. These results indicate that the antimicrobial activity of these pastes is decreased in ground beef and laboratory buffer.     

Effects of the ergot alkaloids dihydroergotamine, ergonovine, and ergotamine on growth of Escherichia coli O157:H7 in vitro

A series of experiments were conducted to evaluate the effects of ergot alkaloids (dihydroergotamine, ergonovine, and ergotamine) on E. coli O157:H7 in both pure and mixed ruminal fluid culture. Alkaloids were added to solutions of E. coli O157:H7 strains 933 (pure and ruminal cultures) and 6058 (ruminal culture only), and growth rates and colony-forming units (CFU) of E. coli O157:H7 were measured. Two mixtures of all three alkaloids at either 2 or 500 microM for each alkaloid decreased (p < 0.001) the growth rate of E. coli O157:H7 in pure culture compared to the individual alkaloids. Dihydroergotamine tended (p = 0.07) to reduce growth rate of E. coli O157:H7 in pure culture compared with ergonovine or ergotamine alone. Increased concentrations of dihydroergotamine and ergotamine decreased (p < 0.003) growth rate of E. coli O157:H7 but increasing concentrations of ergonovine did not influence (p > 0.10) E. coli O157:H7 growth rate. Similar to results in pure culture, a mixture of all three alkaloids at various concentrations for each alkaloid decreased (p < 0.001) the CFU of E. coli O157:H7 strain 6058 in mixed ruminal culture compared to the individual ergot alkaloids. Dihydroergotamine decreased (p = 0.04) CFU of E. coli O157:H7 strain 6058 when compared to ergonovine but CFU were similar (p > 0.10) between dihydroergotamine and ergotamine. Ruminal and (or) intestinal populations of E. coli O157:H7 may be influenced in livestock consuming endophyte-infected tall fescue, and these alterations could be due to the presence of ergot alkaloids in fescue plants.     

Evaluation of a multiplex real-time polymerase chain reaction for the quantification of Escherichia coli O157 in cattle feces

Cattle are asymptomatic reservoirs for Escherichia coli O157, a major foodborne pathogen. The organism generally colonizes the hindgut of cattle and is shed in the feces at low concentrations. The objective of this research was to evaluate a multiplex, real-time polymerase chain reaction (mqPCR) assay for quantification of E. coli O157 in cattle feces using stx1, stx2, and rfbE gene targets. Primer efficiency and analytical sensitivity of the assay were evaluated with a single or pooled (five strain) culture of E. coli O157. In pure culture, the minimum detection limit of the assay was 1.4×10(3) CFU/mL and 3.6×10(3) CFU/mL for the single and five-strain mixture of E. coli O157, respectively. Diagnostic sensitivity was analyzed using DNA extracted from cattle feces spiked with E. coli O157. In feces spiked with the pooled mixture of five E. coli O157 strains, the minimum detection limit was 3.6×10(4) CFU/g. We also evaluated the assay with feces from cattle experimentally inoculated with E. coli O157 by comparing the results to a culture-based method. For the majority of samples tested, the concentration of E. coli O157 detected by the real-time and culture methods was within one log difference. However, the assay could only be evaluated for cattle shedding high concentrations of E. coli O157. In conclusion, the mqPCR quantifying E. coli O157 in cattle feces using stx1, stx2, and rfbE gene targets may have use in detecting and quantifying super shedders, but is not applicable for quantification in animals shedding low concentrations (10(2) to 10(3) CFU/g feces).     

Development and assessment of a rapid method to detect Escherichia coli O26, O111 and O157 in retail minced beef

A molecular-based detection method was developed to detect Escherichia coli O26, O111 and O157 in minced (ground) beef samples. This method consists of an initial overnight enrichment in modified tryptone soya broth (mTSB) and novobiocin prior to DNA extraction and subsequent serogrouping using a triplex PCR. This method has a low limit of detection and results are available within 24 hours of receipt of samples. Once optimized, this rapid method was utilized to determine the prevalence of these E. coli serogroups in six hundred minced beef samples all of which were previously examined by immunomagnetic separation (IMS) and selective plating for E. coli O26 and O111. Using IMS, two E. coli O26 isolates were detected. No E. coli O111 were recovered. The multiplex PCR technique described here did not detect E. coli O111 nor O157 in any of the samples, however six minced beef samples were positive for E. coli O26 using our method, only two of these were previously detected by IMS and culture. Application of molecular methods are useful to support culture-based approaches thereby further contributing to risk reduction along the food chain.     

Growth and survival of Escherichia coli O157:H7 during manufacture and storage of Indian cheese (paneer)

The growth and survival of Escherichia coli O157:H7 during manufacture and storage of Indian cheese or paneer was investigated. Pasteurized and raw milk that had been inoculated with 10(4) CFU/mL of E. coli O157:H7 cultures were used for manufacturing paneer samples which were vacuum-packaged and stored at 4 degrees C, 8 degrees C, and 28 degrees C. Survival and growth of E. coli O157:H7 in paneer samples was determined after every 4 h for up to 48 h. Escherichia coli O157:H7 could survive the manufacturing process of paneer and were present at the end of the storage period (at 28 degrees C) in significantly (p < 0.05) greater numbers than the initial inoculum. No significant difference in survival and growth were noticeable in paneer samples manufactured from raw milk. Though refrigeration (4 degrees C or 8 degrees C) effectively inhibited the growth of E. coli O157:H7, this pathogen could survive over a period of 48 h. Our observations suggest that unpasteurized or improperly pasteurized milk could be an important source for transmission of E. coli O157:H7 through paneer, and appropriate steps must be taken by government agencies to ensure consumer safety for this dairy product.     

Disinfection of radish and alfalfa seeds inoculated with Escherichia coli O157:H7 and Salmonella by a gaseous acetic acid treatment

Abstract The majority of seed sprout-related outbreaks have been associated with Escherichia coli O157:H7 and Salmonella. Therefore, we aimed to find an effective method to inactivate these organisms on seeds before sprouting. Treatment with 8.7% (v/v) acetic acid at 55°C for 2-3?h reduced the population of E. coli O157:H7 and Salmonella inoculated on alfalfa (Medicago sativa L.) and radish seeds (Raphanus sativus L.) by more than 5.0?log CFU/g, and a longer treatment time completely eliminated the E. coli O157:H7 population. The E. coli O157:H7 populations were reduced to an undetectable level with a gaseous acetic acid treatment for 48?h. After enrichment, no E. coli O157:H7 were found in the alfalfa and radish seeds (25?g). However, these treatments were unable to eliminate Salmonella in both seed types. No significant difference between the germination rates of treated alfalfa seeds and control seeds was found, and germination rates greater than 95% were obtained for the radish seeds. Although chlorine washing is commonly used for seed decontamination, chlorine washing at 200 and 20,000?ppm resulted in a reduction of pathogens by less than or equal to 3?log CFU/g. Therefore, these results suggested that gaseous acetic acid is more effective than chlorine washing in controlling pathogenic bacteria on sprout seeds.     

Thermal tolerance of acid-adapted and non-adapted Escherichia coli O157:H7 and Salmonella in ground beef during storage

Thermal tolerance of acid-adapted Escherichia coli O157:H7 or Salmonella in ground beef was evaluated during storage at 4 degrees C or -20 degrees C. Both pathogens were adapted to acidic conditions (pH approximately 4.6) by growing in tryptic soy broth supplemented with 1% glucose. A five-strain cocktail of E. coli O157:H7 or Salmonella was grown separately in TSB (pH approximately 6.6) and TSB + 1% glucose for 24 h at 37 degrees C to provide cells with or without acid adaptation. Irradiated ground beef was inoculated with either acid-adapted or non-adapted E. coli O157:H7 or Salmonella; the samples stored at 4 degrees C were subjected to heat treatment at 62 degrees C or 65 degrees C on days 1, 7, 14, 21, and 28, and the samples stored at -20 degrees C were subjected to heat treatment at 62 degrees C or 65 degrees C on days 1, 30, 60, 90, and 120. Decimal reduction time (D values) of the pathogens was determined as an indicator of thermal tolerance. Significantly higher D(62) values were observed on days 21 and 28 for non-adapted E. coli O157:H7 stored at 4 degrees C and on days 90 and 120 for non-adapted E. coli O157:H7 stored at -20 degrees C (P < 0.05). Higher D(62) values were observed on days 21 and 28 among non-adapted Salmonella strains stored at 4 degrees C and on day 28 for acid-adapted strains of Salmonella stored at 4 degrees C (P < 0.05). Higher D(62) values for acid-adapted strains of Salmonella stored at -20 degrees C were observed on days 30, 60, and 90 (P < 0.05), when while no differences were observed in the D(65) values of acid-adapted and non-adapted strains of E. coli O157:H7 and Salmonella throughout storage at both temperatures (P > 0.05). This suggests that acid adaptation of foodborne pathogens provides a certain level of protection against heat treatment at lower cooking temperatures, while at higher temperatures there were no observed differences between the sensitivity of acid-adapted and non-adapted strains in an actual food system over an extended period of refrigerated and frozen storage.