Prevalence and characterization of Escherichia coli O157:H7 isolates from meat and meat products sold in Amathole District,Eastern Cape Province of South Africa

Meat and meat products have been implicated in outbreaks of Escherichia coli O157:H7 in most parts of the world. In the Amathole District Municipality of the Eastern Cape Province of South Africa, a large number of households consume meat and meat products daily, although the microbiological quality of these types of food is questionable. The present study investigated the prevalence of E. coli O157:H7 isolated from selected meat and meat products (45 samples each of biltong, cold meat, mincemeat, and polony) sold in this area. Strains of E. coli O157:H7 were isolated by enrichment culture and confirmed by polymerase chain reaction (PCR). Also investigated were the antibiogram profiles of the E. coli O157:H7 isolates. Five (2.8%) out of 180 meat and meat products examined were positive for E. coli O157:H7 that carried the fliC_H7, rfbE_O157, and eaeA genes. Two of the E. coli O157:H7 isolates were resistant against all the eight antibiotics tested. To prevent E. coli O157:H7 infections, meat and meat products such as biltong, cold meat, mincemeat and polony should be properly handled, and packed in sterile polyvinyl wrappers.     

A new 7-plex PCR assay for simultaneous detection of shiga toxin-producing Escherichia coli O157 and Salmonella Enteritidis in meat products

A 7-plex PCR assay was developed to achieve an effective detection and identification of serotype Enteritidis of Salmonella spp. and shiga toxin-producing type of Escherichia coli O157 in meat products. Six DNA sequences in the invA and sdfI genes of Salmonella Enteritidis as well as rfbE, eae, stx1, and stx2 genes of E. coli O157:H7 were employed to design primers. The multiplex PCR assay could specifically and sensitively detect and identify target pathogens. Applying the assay to meat samples, the multiplex PCR assay was able to simultaneously detect and identify the two pathogens at a sensitivity of three CFU/10?g raw meats after simple 16?h enrichment in buffered peptone water. Further applying in 21 retail meat samples revealed that two samples were positive for non-shiga toxin producing E. coli O157, one sample was positive for Stx2 producing E. coli O157 and five samples were positive for Salmonella enterica Enteritidis. Taken together, the 7-plex PCR assay is a rapid and reliable method for effectively screening single or multiple pathogens occurrences in various meat products, and could also identify the Salmonella enterica Enteritidis from all Salmonella spp. and shiga toxin producing type from all E. coli strains. Considering as a non expensive screening tool, the multiplex PCR assay has a great potential in complement for food stuff analysis by conventional microbiological tests.     

An evaluation of rapid methods for detecting Escherichia coli O157 on beef carcasses

Numbers of Escherichia coli O157 in food may be low and sensitive techniques are therefore needed for its detection. The objectives of this study were to use carcass meat samples artificially inoculated with various strains of E. coli O157 to compare the sensitivity of enrichment in three different media and to compare immunomagnetic separation followed by culture of magnetic beads to cefixime tellurite sorbitol MacConkey agar with three immunoassays for the detection of E. coli O157 in the enrichment cultures. Duplicate 250, 25 and 2-3 CFU of each of 16 strains of E. coli O157 added to 25-g samples of beef carcass meat were used to compare the sensitivity of (1) enrichment in supplemented tryptone soya broth (sTSB), Reveal 8-h and Reveal 20-h media, and (2) immunomagnetic separation and culture to cefixime tellurite sorbitol MacConkey agar (IMS/CT-SMAC) with Reveal, VIP and STAT immunoassays for detecting the organism. An initial inoculum of 250 CFU/25 g meat was detected in all 32 samples by IMS/CT-SMAC performed on all enrichment media and by Reveal performed on Reveal 8-h and Reveal 20-h media, but in only 30, 19 and 9 samples by Reveal, VIP and STAT, respectively, performed on sTSB medium. An initial inoculum of 25 CFU/25 g meat was detected in 28, 32 and 30 of 32 samples by IMS/CT-SMAC performed on sTSB, Reveal 8-h and Reveal 20-h media, respectively, and in 32 and 30 samples by Reveal performed on Reveal 8-h and Reveal 20-h media, but in only 22, 11 and 2 samples by Reveal, VIP and STAT, respectively, performed on sTSB medium. An initial inoculum of 2-3 CFU/25 g meat was detected in 25, 31 and 28 of 32 samples by IMS/CT-SMAC performed on sTSB, Reveal 8-h and Reveal 20-h media, respectively, and in 25 and 23 samples by Reveal performed on Reveal 8-h and Reveal 20-h media, but in only 14, 1 and 0 samples by Reveal, VIP and STAT, respectively, performed on sTSB medium.     

Differences in thermotolerance of variousEscherichia coliO157:H7 strains in a salami matrix

Three strains ofEscherichia coliO157:H7 (ATCC 43895, Ent C9490 and 380–94) were inoculated into salami and heated in water baths at 50, 55 or 60°C. At intervals between 1 and 360 min, salami samples were removed from the water bath and examined for the presence of survivingE. coliO157:H7. Samples were directly plated onto sorbitol MacConkey (SMAC) agar, and onto tryptone soya agar (TSA) with SMAC overlay. The number of sub-lethally damaged cells in each sample was estimated from the differences between the resultant direct (uninjured cells only) and overlay (total recovery) counts. In samples heated at 50°C, the percentage of cell injury ranged from 71·8–88% for all strains. In samples heated at 55°C the percentage of sub-lethally damaged cells in strains ATCC 43895 and Ent C9490 was significantly higher (P< 0·001) at 97% than that observed in strain 380–94 (64%). Cell injury was not measured at 60°C. There were significant differences between the derived decimal reduction times (D-values) related to the different strains ofE. coliO157:H7, the heat treatment applied and the recovery/enumeration agars used. Significant interstrain differences (P< 0·05) in thermotolerance were noted. Strain Ent C9490 was significantly more heat resistant at 50°C and 60°C (D-values of 116·9 and 2·2 min, respectively), while at 55°C strain 380–94 was more thermotolerant (D-value of 21·9 min). The implications of these findings for the design of studies investigating the heat resistance ofE. coliO157:H7 in fermented meat environments are discussed.     

Lethality of heat to Escherichia coli O157:H7: D- and z-value determinations in turkey,lamb and pork

Thermal inactivation of a four-strain mixture of E. coli O157:H7 was determined in lean ground turkey, lamb and pork. Inoculated meat was packaged in bags completely immersed in a circulating water bath and held at 55, 57.5, 60, 62.5, and 65°C for predetermined lengths of time. The surviving cell population was enumerated by spiral plating meat samples on tryptic soy agar overlaid with Sorbitol MacConkey agar. D-values, determined by linear regression, in turkey were 11.51, 3.59, 1.89, 0.81 and 0.29 min at 55, 57.5, 60, 62.5 and 65°C, respectively (z=6.5°C). When a survival model was fitted to the non-linear survival curves, D-values in turkey ranged from 11.26 min at 55°C to 0.23 min at 65°C (z=6°C). When the E. coli O157:H7 four-strain cocktail was heated in ground pork or lamb, D-values calculated by both approaches were similar at all temperatures. Thermal-death-times from this study will assist the retail food industry to design cooking regimes that ensure safety of ground muscle foods contaminated with E. coli O157:H7.     

Salmonella, Escherichia coli O157:H7 and E. coli biotype 1 in a pilot survey of imported and New Zealand pig meats

A pilot survey for the pathogens Salmonella and Escherichia coli O157:H7, and E. coli biotype 1 was conducted on 100 New Zealand-produced (domestic) pig carcasses and 110 imported pig meat samples over an 8-month period to assess the likelihood of introduction of novel pathogen strains into New Zealand (NZ), and as a guide for development of a domestic pork National Microbiological Database programme. Salmonella was not isolated from domestic pig carcasses or from pig meat imported from Canada and the USA. The prevalence of Salmonella in imported pig meat was 3.6% (95% CI 1.0–9.0) with positive samples detected from Australian pig meat. The prevalence of E. coli O157:H7 on domestic pig carcasses was 1% (95% CI 0.03–5.4) while the overall prevalence of E. coli O157:H7 in imported pig meat was 1.8% (95% CI 0.2–6.4), detected mainly from Australian but not from Canadian or US pork. All except three samples have an E. coli biotype 1 count of <100 CFU cm⁻² or g⁻¹, indicating good hygiene quality of domestic and imported pig meat. The results demonstrated that importation of uncooked pig meat is a potential route for the introduction of new clones of Salmonella and E. coli O157:H7 into New Zealand.     

Occurrence of Escherichia coli O157, O111 and O26 in raw ewe's milk and performance of two enrichment broths and two plating media used for its assessment

The occurrence of Escherichia coli O157, O111 and O26 in 159 raw ewe's milk samples was examined. Sample-aliquots were incubated simultaneously in TSB added with yeast extract (YETSB) and mTSB with novobiocin (N-mTSB). Serogroup-specific immunomagnetic separation (IMS) was then used and IMS beads were plated in a cefixime tellurite (CT)-containing media (CT-SMAC, CT-SBMAC and CT-RMAC for E. coli O157, O111 and O26, respectively) and E. coli O157:H7 chromogenic ID agar. A sweep of confluent growth from each medium was examined for the presence of E. coli O157 and O111 using PCR, and for E. coli O26 using a latex agglutination test. Enumeration of E. coli O157 and O111 was performed in the samples tested positive for the correspondent serogroup using the most probable number (MPN) method combined with PCR. Percentage occurrences of E. coli O157, O111 and O26 were 18.2, 8.2 and 5.7, respectively. Mean E. coli O157 and O111 levels were 0.22 and < 0.04 MPN/mL, respectively. Enrichment in YETSB resulted in higher detection rates of E. coli O157 and O26 than in N-mTSB. When YETSB was used as enrichment broth and for these last two serogroups, the analysis of the confluent growth from the CT-media gave more positive results than that from E. coli O157:H7-ID medium.     

Development of PCR assays for detection of Escherichia coli O157:H7 in meat products

A multiplex polymerase chain reaction (PCR) procedure based on fliC_h7 and rfbE genes was developed for the detection of Escherichia coli O157:H7 in raw pork meat and ready-to-eat (RTE) meat products. Two different DNA extraction procedures were evaluated for application on meat products. MasterPure™ DNA Purification kit in combination with immunomagnetic separation was found to be the best method in a meat system. The optimized PCR included an enrichment step in brilliant green bile 2% broth at 37 °C. This method was applied to artificially inoculated meat and RTE meat products with different concentrations of E. coli O157:H7. The results indicate that the PCR assay developed could sensitively and specifically detect E. coli O157:H7 in raw pork meat and RTE meat products in approximately 10 h, including a 6 h enrichment step. Thus, this method could be proposed for screening E. coli O157:H7 in raw pork and RTE meat products.     

Molecular detection of nine clinically important non-O157 Escherichia coli serogroups from raw sheep meat in Chaharmahal-va-Bakhtiari province,Iran

STEC isolates and also stx-negative Escherichia coli isolates from sheep meat from the Chaharmahal-va-Bakhtiari province, Iran were analyzed for nine clinically important non-O157 serotypes by PCR. A total of 90 E. coli isolates were tested. Stx-positive and eae-positive E. coli isolates did not belong to the nine most clinically relevant non-O157 STECs. Of the 80 non-STEC isolates, two belonged to the O103 and two belonged to the O128 groups. Stx-negative E. coli O103 and O128 strains isolated have potential in acquiring stx genes and continuing into the digestive system of consumers. Further studies are needed to analyze virulence characteristics of these E. coli strains to determine their potential role in causing disease in humans. For the sake of public health, it is important to monitor and investigate non-O157 diarrheagenic E. coli strains in meat in order to control and prevent them.     

The growth and survival of Escherichia coli O157:H7 on minced bison and pieces of bison meat stored at 5 and 10 °C

This study investigated the growth and survival of Escherichia coli O157:H7 on minced and whole pieces of bison meat. Growth curves of native microflora, including Pseudomonas spp. and Enterobacteriaceae were also generated. A marked E. coli O157:H7 strain was inoculated onto minced and whole pieces of bison meat at an initial level of 1.5 log₁₀ cfu g⁻¹. The inoculated meat was stored at either 5 °C for 28 days or 10 °C for 21 days. Survival, but no growth, of E. coli O157:H7 was observed on both forms of bison meat stored at 5 °C, while significant growth of the organism was observed at 10 °C. E. coli O157:H7 counts on whole pieces were generally higher than counts observed on minced bison meat, and reached their highest population by 14 days, with a total increase of 3.36 log₁₀ cfu g⁻¹ on whole pieces and 2.12 log₁₀ cfu g⁻¹on minced bison meat stored at 10 °C. Under the same storage temperature, Pseudomonas spp. and total counts displayed similar growth patterns on both pieces and minced bison meat, while the Enterobacteriaceae showed a slower growth rate. This study showed that the growth of E. coli O157:H7 on bison meat is similar to that observed in studies of beef.     

Comparison of a real-time PCR and an IMS/culture method to detect Escherichia coli O26 and O111 in minced beef in the Republic of Ireland

This study compared an immuno-magnetic separation (IMS)/culture method and a real-time PCR method to detect Verocytotoxigenic Escherichia coli (VTEC) serovar O26 and/or O111 in minced beef. A total of 65 samples were examined, 40 of which were frozen beef samples previously established as containing E. coli O157, and 25 were samples of fresh minced beef, purchased from butcher shops in the Dublin area. After selective enrichment, all samples were (a) subjected to IMS, plated on differential media and identified as E. coli O26 or O111 using biochemical and immuno-logical methods; and (b) subjected to DNA extraction and real-time PCR analysis using primers and probes against E. coli O111 and O26 serovar specific genes, and verotoxin genes. Overall, from the 65 minced beef samples collected, three were positive for E. coli O26 by real-time PCR, with only one of these samples positive for E. coli O26 by the culture method. One sample was positive for E. coli O111 by both real-time PCR and the culture method. The two samples found positive for E. coli O26 by real-time PCR method but not by culture method belonged to the group of frozen beef samples, indicating that the previously developed culture method for the detection of E. coli O26 may not be suitable for the detection of freeze injured cells. In conclusion, this study highlights the role of beef meat in the transmission of non-O157 VTEC. The results of the study emphasize that the analyses for emergent pathogens should be included in food safety surveillance systems and that the development of standard methods for the detection of E. coli O26 and O111 in routine food testing is needed in order to reduce the consumer exposure to contaminated food.     

A new 7-plex PCR assay for simultaneous detection of shiga toxin-producing Escherichia coli O157 and Salmonella Enteritidis in meat products

A 7-plex PCR assay was developed to achieve an effective detection and identification of serotype Enteritidis of Salmonella spp. and shiga toxin-producing type of Escherichia coli O157 in meat products. Six DNA sequences in the invA and sdfI genes of Salmonella Enteritidis as well as rfbE, eae, stx1, and stx2 genes of E. coli O157:H7 were employed to design primers. The multiplex PCR assay could specifically and sensitively detect and identify target pathogens. Applying the assay to meat samples, the multiplex PCR assay was able to simultaneously detect and identify the two pathogens at a sensitivity of three CFU/10 g raw meats after simple 16 h enrichment in buffered peptone water. Further applying in 21 retail meat samples revealed that two samples were positive for non-shiga toxin producing E. coli O157, one sample was positive for Stx2 producing E. coli O157 and five samples were positive for Salmonella enterica Enteritidis. Taken together, the 7-plex PCR assay is a rapid and reliable method for effectively screening single or multiple pathogens occurrences in various meat products, and could also identify the Salmonella enterica Enteritidis from all Salmonella spp. and shiga toxin producing type from all E. coli strains. Considering as a non expensive screening tool, the multiplex PCR assay has a great potential in complement for food stuff analysis by conventional microbiological tests.     

Detection and Isolation of Low Levels of E. coli O157:H7 in Cilantro by Real-Time PCR, Immunomagnetic Separation, and Cultural Methods with and without an Acid Treatment

Abstract: Leafy greens such as cilantro, contaminated with Escherichia coli O157:H7, have been implicated in cases of human illnesses. High levels of microflora in fresh cilantro make recovery of low numbers of E. coli O157:H7 difficult. To improve upon current methods, immunomagnetic separation (IMS) techniques in combination with real‐time PCR (RTiPCR) and selective enrichment protocols were examined. Rinsates were prepared from cilantro samples inoculated with low (～0.02 CFU/g) and slightly higher (～0.05 CFU/g) levels of E. coli O157:H7. Rinsate portions were enriched in modified buffered peptone water with pyruvate (mBPWp) for 5 h at 37 °C. After 5 h, selective agents were added to samples and further incubated at 42 °C overnight. Detection and recovery were attempted at 5 and 24 h with and without IMS. IMS beads were screened by RTiPCR for simultaneous detection of stx1, stx2, and uidA SNP. Additionally, broth cultures and IMS beads were streaked onto selective agar plates (Rainbow^®agar, R&F^®E. coli O157 Chromogenic medium, TC‐SMAC and CHROMagar? 0157) for isolation of E. coli O157:H7. Both broth cultures and IMS beads were also acid treated in Trypticase Soy Broth pH 2 prior to plating to selective media to improve upon cultural recovery. Although E. coli O157 strains were detected in most samples by PCR after 5 h enrichment, cultural recovery was poor. However, after 24 h enrichment, both PCR and cultural recovery were improved. Acidification of the broths and the IMS beads prior to plating greatly improved recovery from 24 h enrichment broths by suppressing the growth of competing microorganisms. Practical Application: Detection and recovery of Escherichia coli O157:H7 in fresh produce matrices (e.g., cilantro) can be complicated by high background microflora present in these foods. Rapid detection by molecular methods combined with effective enrichment and isolation procedures such as using immunomagnetic separation (IMS) techniques can quickly identify potential hazards to public health. Additional techniques such as acidification of enrichment broths can exploit acid resistance characteristics of pathogens such as E. coli O157:H7, facilitating their isolation in complex food matrices.     

Changes in heat tolerance of Escherichia coli O157:H7 after exposure to acidic environments

Acid-adapted bacterial cells are known to have enhanced tolerance to various secondary stresses. However, a comparison of heat tolerance of acid-adapted and acid-shocked cells of Escherichia coli O157:H7 has not been reported. D - and z -values of acid-adapted, acid-shocked, and control cells of an unusually heat-resistant strain (E0139) of E. coli O157:H7, as well as two other strains of E. coli O157:H7, were determined based upon the number of cells surviving heat treatment at 52, 54 or 56°C in tryptic soy broth (pH 7·2) for 0, 10, 20 or 30 min. The unusual heat tolerance of E. coli O157:H7 strain E0139 was confirmed. D -values for cells from 24-h cultures were 100·2, 28·3, and 6·1 min at 52, 54 and 56°C, respectively, with a z -value of 3·3°C. The highest D -values of other E. coli O157:H7 strains were 13·6 and 9·2 min at 52 and 54°C, respectively, whereas highest D -values of non-O157:H7 strains were 78·3 and 29·7 min at 52 and 54°C. D -values of acid-adapted cells were significantly higher than those of unadapted and acid-shocked cells at all temperatures tested. In a previous study, we observed that both acid-adapted cells and acid-shocked cells of strain E0139 had enhanced acid tolerance. This suggests that different mechanisms protect acid-adapted and acid-shocked cells against subsequent exposure to heat or an acidic environment. The two types of cells should be considered separately when evaluating survival and growth characteristics upon subsequent exposure to different secondary stress conditions.     

Growth patterns of Escherichia coli O157:H7 in ground beef treated with nisin,chelators, organic acids and their combinations immobilized in calcium alginate gels

The effects of antimicrobial substances including nisin, acetic acid, lactic acid, potassium sorbate and chelators (disodium ethylenediamine tetraacetic acid [EDTA] and sodium hexametaphosphate [HMP]), alone or in combination and, with or without immobilization in calcium alginate gels, on the growth of Escherichia coli O157:H7 in ground beef were investigated. Results showed that acetic acid and potassium sorbate could inhibit the growth of E. coli O157:H7 effectively at 10°C and at 30°C. Both EDTA and HMP did not halt the growth of E. coli O157:H7. In an antimicrobial system immobilized with calcium alginate, most of the antimicrobials could not inhibit the growth of E. coli O157:H7 in ground beef at 10°C and at 30°C, with the exception of acetic acid and lactic acid. Immobilization did not enhance the effectiveness of acetic acid against E. coli O157:H7 in ground beef at 10°C and at 30°C (P>0.05) but it did enhance the effectiveness of lactic acid at 10°C. In a system combining different antimicrobials, treatment with nisin /EDTA or nisin/potassium sorbate at 10°C revealed a significantly lower population change of E. coli O157:H7 compared to samples treated with nisin, EDTA or potassium sorbate alone. The use of calcium alginate immobilization further enhanced the effectiveness of the combination system of nisin/EDTA, nisin/acetic acid and nisin/potassium sorbate on the growth of E. coli O157:H7 in ground beef at 10°C but it was not effective at 30°C.     

The prevalence of Escherichia coli O157 and O157:H7 in ground beef and raw meatball by immunomagnetic separation and the detection of virulence genes using multiplex PCR

The present study was conducted to investigate the presence of Escherichia coli O157 and O157:H7 strains and to detect the presence of the stx1, stx2, and eaeA genes in isolates derived from 200 samples (100 samples from fresh ground beef and 100 samples from raw meatball). The samples were purchased from the Samsun Province in Turkey, over a period of 1 year. Enrichment-based immunomagnetic separation and multiplex polymerase chain reaction were applied for these analyses. E. coli O157 was detected in five of the 200 (2.5%) samples tested (one isolated from ground beef and four from meatball samples), whereas E. coli O157: H7 was not detected in any sample. During the analysis, eight strains of E. coli O157 were obtained. The genes stx1, stx2, and eaeA were detected in two E. coli O157 isolates obtained from two meatball samples, whereas only the eaeA and the stx2 genes were detected in four E. coli O157 strains that were isolated from one meatball sample. None of the stx1, stx2, and eaeA was detected in the E. coli O157 isolates obtained from the ground beef and the one meatball samples.     

The physiologic state of Escherichia coli O157:H7 does not affect its detection in two commercial real-time PCR-based tests

Multiplex real-time PCR detection of Escherichia coli O157:H7 is an efficient molecular tool with high sensitivity and specificity for meat safety assurance. The Biocontrol GDS^® and DuPont Qualicon BAX^®-RT rapid detection systems are two commercial tests based on real-time PCR amplification with potential applications for quantification of specific E. coli O157:H7 gene targets in enriched meat samples. However, there are arguments surrounding the use of these tests to predict pre-enrichment concentrations of E. coli O157:H7, as well as arguments pertaining to the influence of non-viable cells causing false positive results. The present study attempts to illustrate the effects of different bacterial physiologic states and the presence of non-viable cells on the ability of these systems to accurately measure contamination levels of E. coli O157:H7 in ground beef. While the PCR threshold cycle (C_T) values of these assays showed a direct correlation with the number of bacteria present in pure cultures, this was not the case for ground beef samples spiked with various levels of injured or healthy cells. Furthermore, comparison of post-enrichment cell densities of bacteria did not correlate with injured or healthy cell numbers inoculated before enrichment process. Ground beef samples spiked with injured or healthy cells at different doses could not be distinguished by C_T values from either assay. In addition, the contribution of nonviable cells in generating positive real-time PCR signals was investigated using both assays on pre-enriched and post-enriched beef samples, but only if inoculated at levels of 10⁶ cells/sample or higher, which are levels not typically seen in ground beef.     

Enteropathogenicity markers in Escherichia coli strains isolated from soft white cheese and poultry in Rio de Janeiro,Brazil

Diarrhoeagenic Escherichia coli is the main cause of infantile diarrhoea in Brazil. Unseasoned soft white cheese and poultry are cheap foods widely consumed by adults and children in Brazilian cities. To assess the risk involved in consumption of these foods, virulence markers were investigated in 575 E. coli colonies isolated from 44 soft white cheese and 39 chicken carcass samples. Escherichia coli colonies were tested with enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC) and O157 antisera and tested for adherence to HEp-2 cells, FAS test, enterohemolysin production, Dean assay for STa and DNA hybridization with eae, EAF, stx and LT-I probes. Shiga toxin (Stx) production by probe-positive strains was tested on Vero cells. Escherichia coli strains of EPEC serogroups O127 and O128 were found in 11·3% of cheese samples and O114, O119 and O158 were found in 10·2% of chicken samples examined. None of these produced localized adherence on HEp-2 cells or hybridized with eae and EAF probes. EIEC, enterotoxigenic E. coli (ETEC) and enterohemolysin-producing E. coli was not detected. Two non-O157 E. coli strains (serotypes O?:H28 and R:H?) isolated from chicken carcasses hybridized with stx2 probe and one of them also hybridized with eae probe. However, none of these strains were positive for enterohaemolysin, FAS, adhered to HEp-2 cells or produced cytotoxic effects on Vero cells.     

Recovery of heat-stressed E. coli O157:H7 from ground beef and survival of E. coli O157:H7 in refrigerated and frozen ground beef and in fermented sausage kept at 7°C and 22°C

A study was undertaken to obtain information on survival of Escherichia coli O157:H7 in ground beef subjected to heat treatment, refrigeration and freezing and on survival of E. coli O157:H7 in fermented sausage kept at 7°C and 22°C. For the challenge test, a mixture of E. coli O157:H7 strains (EH 321, EH 385, EH 302) was used and enumeration was performed on an isolation medium suitable for recovery of stressed organisms: modified Levine's eosin methylene blue agar (mEMB). Heat resistance of E. coli O157:H7 decreased after pre-incubation at a reduced temperature.Escherichia coli O157:H7 was more susceptible to heat inactivation after storage at 7°C and die-off was even more enhanced if cultures were frozen prior to heat inactivation. The enhanced reduction of the pathogen at 56°C after prior storage under refrigeration was confirmed in a test with inoculated ground beef.Escherichia coli O157:H7 was able to survive in ground beef at 7°C for 11 days and at −18°C for 35 days showing maximal one log reduction during the storage period. Thus, ground beef contaminated with E. coli O157:H7 will remain a hazard even if the ground beef is held at low or freezing temperatures. At both 7°C and 22°C, a gradual reduction of E. coli O157:H7 was noticed in fermented sausage over the 35 days storage period resulting in a 2 log decrease of the high inoculum (10⁶cfu 25 g⁻¹). For the low inoculum (10³cfu 25 g⁻¹) a 2·5 log reduction was obtained in 7 and 28 days storage at respectively 22 and 7°C. Application of good hygienic practices and implementation of HACCP in the beef industry are important tools in the control of E. coli O157:H7.