Use of mild irradiation doses to control pathogenic bacteria on meat trimmings for production of patties aiming at provoking minimal changes in quality attributes

The objectives of the present work were to assess the use of moderate doses of gamma irradiation (2 to 5 kGy) and to reduce the risk of pathogen presence without altering the quality attributes of bovine trimmings and of patties made of irradiated trimmings. Microbiological indicators (coliforms, Pseudomonas spp and mesophilic aerobic counts), physicochemical indicators (pH, color and tiobarbituric acid) and sensory changes were evaluated during storage. 5 kGy irradiation doses slightly increased off flavors in patties. Two pathogenic markers (Listeria monocytogenes and Escherichia coli O157:H7) were inoculated at high or low loads to trimming samples which were subsequently irradiated and lethality curves were obtained. Provided that using irradiation doses ≤ 2.5 kGy are used, reductions of 2 log CFU/g of L. monocytogenes and 5 log CFU/g of E. coli O157:H7 are expected. It seems reasonable to suppose that irradiation can be successfully employed to improve the safety of frozen trimmings when initial pathogenic bacteria burdens are not extremely high.     

Effects of packaging type and storage temperature on the growth of foodborne pathogens on shredded ‘Romaine’ lettuce

Fresh produce can be a vehicle for the transmission of pathogens capable of causing human illnesses and some of them can grow on fresh-cut vegetables. The survival and growth of Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes inoculated onto shredded lettuce was determined under modified atmosphere packaging conditions, at various storage temperatures. We also monitored changes in pH and gas atmospheres within the packages and the growth of psychrotrophic and mesophilic microorganisms. After pathogen inoculation, shredded lettuce was packaged in films of different permeability and stored at 5 and 25 °C. After 10 days at 5 °C populations of E. coli O157:H7 and Salmonella decreased approximately 1.00 log unit while L. monocytogenes increased about 1.00 log unit, in all package films. Moreover, the pathogens level increased between 2.44 and 4.19 log units after 3 days at 25 °C. Psychrotrophic and mesophilic bacteria had similar growth at both temperatures with higher populations in air than in the other atmospheres. The composition of the storage atmosphere within the packaging of lettuce had no significant effect on the survival and growth of the pathogens used in this study at refrigeration temperatures. The results obtained can be considered as a warning indicator, which reinforces the necessity for corrective measures to avoid contamination of vegetables.     

Antagonistic effect of Pseudomonas graminis CPA-7 against foodborne pathogens in fresh-cut apples under simulated commercial conditions

Recently, we reported that the application of the strain CPA-7 of Pseudomonas graminis, previously isolated from apple, could reduce the population of foodborne pathogens on minimally processed (MP) apples and peaches under laboratory conditions. Therefore, the objective of the present work was to find an antioxidant treatment and a packaging atmosphere condition to improve CPA-7 efficacy in reducing a cocktail of four Salmonella and five Listeria monocytogenes strains on MP apples under simulated commercial processing. The effect of CPA-7 application on apple quality and its survival to simulated gastric stress were also evaluated. Ascorbic acid (2%, w/v) and N-acetyl-l-cysteine (1%, w/v) as antioxidant treatments reduced Salmonella, L. monocytogenes and CPA-7 recovery, meanwhile no reduction was observed with NatureSeal^® AS1 (NS, 6%, w/v). The antagonistic strain was effective on NS-treated apple wedges stored at 10 °C with or without modified atmosphere packaging (MAP). Then, in a semi-commercial assay, efficacy of CPA-7 inoculated at 10⁵ and 10⁷ cfu mL⁻¹ against Salmonella and L. monocytogenes strains on MP apples with NS and MAP and stored at 5 and 10 °C was evaluated. Although high CPA-7 concentrations/populations avoided Salmonella growth at 10 °C and lowered L. monocytogenes population increases were observed at both temperatures, the effect was not instantaneous. No effect on apple quality was detected and CPA-7 did not survived to simulated gastric stress throughout storage. Therefore, CPA-7 could avoid pathogens growth on MP apples during storage when use as part of a hurdle technology in combination with disinfection techniques, low storage temperature and MAP.     

Development and validation of a probabilistic second-order exposure assessment model for Escherichia coli O157:H7 contamination of beef trimmings from Irish meat plants

A second-order quantitative Monte Carlo simulation model was developed for Escherichia coli O157:H7 contamination of beef trimmings in Irish abattoirs. The assessment considers initial contamination levels, cross-contamination and decontamination events during the cattle slaughter process. The mean simulated prevalence of E. coli O157:H7 on trimmings was 2.36% and the mean simulated counts of E. coli O157:H7 on contaminated trimmings was -2.69log(10)CFU/g. A parallel validation survey provided some confidence in the model predictions. An uncertainty analysis indicated that microbial test sensitivity is a significant factor contributing to model uncertainty and requires further investigation while also indicating that risk reduction measures should be directed towards reducing the hide to carcass transfer (correlation coefficient 0.25) during dehiding and reducing the initial prevalence and counts on bovine hides (correlation coefficients 0.19 and 0.16, respectively). A characterisation of uncertainty and variability indicating that further research is required to reduce parameter uncertainty and to achieve better understanding of microbial transfer in meat plants. The model developed in this study highlights the need for further development of quantitative risk assessments in the food industry.     

Pulsed UV light as an intervention strategy against Listeria monocytogenes and Escherichia coli O157:H7 on the surface of a meat slicing knife

The main objective of this work was to explore the applicability of the Intense Light Pulses (ILP) for decontamination of a stainless steel meat contact surface, exemplified by a slicing knife, as a function of time between contamination and decontamination, number of light pulses applied, and the prior contact with different meat matrices. Listeria monocytogenes and Escherichia coli O157:H7 were chosen as the challenge microorganisms. The ILP system was a laboratory-scale four-lamp batch system generating 3 J/cm² with an input voltage of 3000 V. The results obtained demonstrate successful application of ILP treatment for reduction of L. monocytogenes and E. coli O157:H7 on a surface of stainless steel slicing knife. The inactivation effectiveness depended on the type of meat product that was in the contact with the treated surface and on the time between the contamination and the ILP treatment. Statistical analysis showed the significant interaction between the time and type of meat product on the effectiveness of ILP treatment. The highest effectiveness of the ILP (the complete inactivation of 6.5 log CFU/side of knife) was obtained when the knife surface was in contact with the products containing lower fat and protein content and when it was treated with pulsed light as fast as possible after the contamination (within 60 s). The decontamination efficacy of ILP treatment could not be improved by multiple light pulses if lost due to the extended time between the moment of contamination and ILP treatment. Results showed that the suggested approach can be very effective as an intervention strategy along meat processing lines preventing cross-contamination between the equipment and the final product.     

Control of foodborne pathogens on fresh-cut fruit by a novel strain of Pseudomonas graminis

The consumption of fresh-cut fruit has substantially risen over the last few years, leading to an increase in the number of outbreaks associated with fruit. Moreover, consumers are currently demanding wholesome, fresh-like, safe foods without added chemicals. As a response, the aim of this study was to determine if the naturally occurring microorganisms on fruit are “competitive with” or “antagonistic to” potentially encountered pathogens. Of the 97 and 107 isolates tested by co-inoculation with Escherichia coli O157:H7, Salmonella and Listeria innocua on fresh-cut apple and peach, respectively, and stored at 20 °C, seven showed a strong antagonistic capacity (more than 1-log unit reduction). One of the isolates, CPA-7, achieved the best reduction values (from 2.8 to 5.9-log units) and was the only isolate able to inhibit E. coli O157:H7 at refrigeration temperatures on both fruits. Therefore, CPA-7 was selected for further assays. Dose-response assays showed that CPA-7 should be present in at least the same amount as the pathogen to adequately reduce the numbers of the pathogen. From the results obtained in in vitro assays, competition seemed to be CPA-7's mode of action against E. coli O157:H7. The CPA-7 strain was identified as Pseudomonas graminis. Thus, the results support the potential use of CPA-7 as a bioprotective agent against foodborne pathogens in minimally processed fruit.     

Efficacy of trimming chilled beef during fabrication to control Escherichia coli O157:H7 surrogates on subsequent subprimals

Effectiveness of trimming external carcass surfaces from subprimals during fabrication to reduce Escherichia coli O157:H7 surrogates was evaluated. Carcass sides (n = 10 sides) were inoculated along the hide pattern opening before entering the blast chill cooler with a gelatin slurry containing a bacterial cocktail of three rifampicin-resistant, nonpathogenic E. coli biotype I strains. Following a 48 h chill, sides were fabricated to produce eight subprimals. Microbiological samples were taken from the original carcass fat surface area, initial lean surface area, trimmed fat surface area (where applicable), and trimmed lean surface area (where applicable). Newly exposed lean surfaces had lower (P < 0.05) counts of rifampicin-resistant E. coli than did the external fat surfaces. However, fat and lean surfaces that were not inoculated became contaminated during the fabrication process. Trimming external surfaces reduced levels of pathogens, but under normal fabrication processes, pathogens were still spread to newly exposed surfaces.     

Microbiological and organoleptic characteristics of beef trim and ground beef treated with acetic acid, lactic acid, acidified sodium chlorite, or sterile water in a simulated commercial processing environment to reduce Escherichia coli O157:H7 and Salmonella

The objective of this study was to validate the effectiveness of acetic and lactic acids (2% and 5%), acidified sodium chlorite (1000 ppm), and sterile water in reducing Escherichia coli O157:H7 and Salmonella Typhimurium in inoculated beef trim in a simulated processing environment. Samples were collected to assess microbial characteristics at three processing points. Results from this study indicate that all treatments, including sterile water, reduced pathogen concentrations (P < 0.05) of both E. coli O157:H7 and Salmonella Typhimurium in ground beef up to 0.5 and 0.6 log by 24 h, respectively. In some cases, there were no significant differences between the antimicrobial treatments and the sterile water using this application method. Triangle sensory test results of non-inoculated beef indicated there were no differences (P < 0.05) in the means of correct responses between controls or antimicrobial treatments at 6 or 24 h. While interventions are important for beef trim, use of the interventions must be validated under industry conditions to ensure proper effectiveness.     

Fate of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella on fresh-cut celery

Illnesses from Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella have been associated with the consumption of numerous produce items. Little is known about the effect of consumer handling practices on the fate of these pathogens on celery. The objective of this study was to determine pathogen behavior at different temperatures under different storage conditions. Commercial fresh-cut celery was inoculated at ca. 3 log CFU/g onto either freshly cut or outer uncut surfaces and stored in either sealed polyethylene bags or closed containers. Samples were enumerated following storage for 0, 1, 3, 5, and 7 days when held at 4 °C or 12 °C, and after 0, 8, and 17 h, and 1, and 2 days when held at 22 °C. At 4 °C, all populations declined by 0.5–1.0 log CFU/g over 7 days. At 12 °C, E. coli O157:H7 and Salmonella populations did not change, while L. monocytogenes populations increased by ca. 0.5 log CFU/g over 7 days. At 22 °C, E. coli O157:H7, Salmonella, and L. monocytogenes populations increased by ca. 1, 2, or 0.3 log CFU/g, respectively, with the majority of growth occurring during the first 17 h. On occasion, populations on cut surfaces were significantly higher than those on uncut surfaces. Results indicate that populations are reduced under refrigeration, but survive and may grow at elevated temperatures.     

Antibacterial effects of natural tenderizing enzymes on different strains of Escherichia coli O157:H7 and Listeria monocytogenes on beef

This study determined the efficacy of actinidin and papain on reducing Listeria monocytogenes and three mixed strains of Escherichia coli O157:H7 populations on beef. The average reduction of E. coli O157:H7 was greater than that of L. monocytogenes and higher concentrations of either protease yielded greater reduction in bacterial populations. For instance, actinidin at 700 mg/ml significantly (p ≤ 0.05) reduced the population of L. monocytogenes by 1.49 log cfu/ml meat rinse after 3 h at 25 & 35 °C, and by 1.45 log cfu/ml rinse after 24 h at 5 °C, while the same actinidin concentration significantly reduced the populations of three mixed strains of E. coli O157:H7 by 1.81 log cfu/ml rinse after 3 h at 25 & 35 °C, and 1.94 log cfu/ml rinse after 24 h at 5 °C. These findings suggest that, in addition to improving the sensory attributes of beef, proteolytic enzymes can enhance meat safety when stored at suitable temperatures.     

ß-phenylethylamine as a novel nutrient treatment to reduce bacterial contamination due to Escherichia coli O157:H7 on beef meat

Bacterial infection by Escherichia coli O157:H7 through the consumption of beef meat or meat products is an ongoing problem, in part because bacteria develop resistances towards chemicals aimed at killing them. In an approach that uses bacterial nutrients to manipulate bacteria into behaviors or cellular phenotypes less harmful to humans, we screened a library of 95 carbon and 95 nitrogen sources for their effect on E. coli growth, cell division, and biofilm formation. In the initial screening experiment using the Phenotype MicroArray^TM technology from BioLog (Hayward, CA), we narrowed the 190 starting nutrients down to eight which were consecutively tested as supplements in liquid beef broth medium. Acetoacetic acid (AAA) and ß-phenylethylamine (PEA) performed best in this experiment. On beef meat pieces, PEA reduced the bacterial cell count by 90% after incubation of the PEA treated and E. coli contaminated meat pieces at 10 °C for one week.     

Antibacterial activity of oregano and thyme essential oils against Listeria monocytogenes and Escherichia coli O157:H7 in feta cheese packaged under modified atmosphere

The antibacterial activity of the essential oils (EO) of oregano and thyme added at doses of 0.1 or 0.2 and 0.1 ml/100 g, respectively, to feta cheese inoculated with Escherichia coli O157:H7 or Listeria monocytogenes was investigated during cheese storage under modified atmosphere packaging (MAP) of 50% CO₂ and 50% N₂ at 4 °C. Compositional analysis showed that the predominant phenols were carvacrol and thymol for both EO. In control feta inoculated with the pathogens and stored under MAP, results showed that E. coli O157:H7 and L. monocytogenes strains survived up to 32 and 28 days of storage. However, in feta cheese treated with oregano EO at the dose of 0.1 ml/100 g, E. coli O157:H7 or L. monocytogenes survived up to 22 and 18 days, respectively, whereas at the dose of 0.2 ml/100 g up to16 or 14 days, respectively. Feta cheese treated with thyme EO at 0.1 ml/100 g showed populations of E. coli O157:H7 or L. monocytogenes not significantly different (P > 0.05) than those of feta cheese treated with oregano at 0.1 ml/100 g. Although both essential oils exhibited equal antibacterial activity against both pathogens, the populations of L. monocytogenes decreased faster (P < 0.05) than those of E. coli O157:H7 during the refrigerated storage, indicating a stronger antibacterial activity of both essential oils against the former pathogen.     

A review of quantitative microbial risk assessment in the management of Escherichia coli O157:H7 on beef

Since Escherichia coli O157:H7 first emerged as a food borne pathogen in the mid 1980s, it has been linked to many cases of food poisoning across the world. While multiple sources and routes of transmission for this pathogen are now recognised, beef and beef products remain an important vehicle of the pathogen and continue to be linked to outbreaks across the developed world. Much research has been directed at E. coli O157:H7 transmission, survival and control in the beef chain and this paper presents an overview of current knowledge on this pathogen in the beef chain from primary production through slaughter, processing, distribution, final preparation and cooking. In order to strategically manage E. coli O157:H7 and to devise approaches to reduce the public health risk posed, many national and international groups have applied quantitative risk assessment techniques to model the risk posed by E. coli O157:H7 in beef, particularly in ground/minced beef which is most often linked with infection. This paper reviews these quantitative risk assessments and their application in managing the risk posed by E. coli O157:H7 in beef.     

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.     

Multiplex fiber optic biosensor for detection of Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella enterica from ready-to-eat meat samples

Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella enterica are the most common foodborne bacterial pathogens and are responsible for many outbreaks. Therefore, multiplex detection of these three using a single assay platform is highly desirable. The objective was to develop and optimize a fiber optic sensor for simultaneous detection of these three from food. The streptavidin coated optical waveguides were immobilized with biotinylated polyclonal antibodies and exposed to the bacterial suspensions or enriched food samples for 2 h. Pathogens were detected after reacting with Alexa-Fluor 647-labeled monoclonal antibodies. Ready-to-eat beef, chicken and turkey meats were inoculated with each pathogen (∼100 cfu/25 g), enriched in SEL (Salmonella, E. coli, Listeria), a multipathogen selective enrichment broth for 18 h and tested with the biosensor. The biosensor was able to detect each pathogen, individually or in a mixture with very little cross-reactivity. The limit of detection for the sensor was ∼10³ cfu/ml for all three pathogens. Furthermore, the biosensor successfully detected each pathogen, grown in a mixture from enriched meat samples under 24 h. The pathogen presence was further verified by PCR and immunofluorescence assay. The multiplex fiber optic sensor shows promise for detection of the three pathogens if present in the same sample eliminating the use of multiple single pathogen detection platforms.     

Quantification of contamination of lettuce by GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium

The primary objective of this study was to determine the possibility of internalization of GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium (S. Typhimurium) strains MAE 110 (multi-cellular morphology) and 119 (wild type morphology) into lettuce seedlings (Lactuca sativa cv. Tamburo) grown in an inoculated hydroponic and soil system. The second aim was to quantify the level of contamination with the use of a proper surface sterilization method. Silver nitrate was superior in reducing the number of viable bacteria on leave surfaces compared to sodium hypochlorite and ethanol. With the hydroponic system internal colonization of lettuce only occurred at high densities with S. Typhimurium MAE 119. With the soil system E. coli O157:H7, S. Typhimurium 110 and S. Typhimurium 119 were found at considerable densities in sterilized leaf samples (respectively, 3.95, 2.57 and 2.37 log cfu/g on average) with prevalences of 0.29, 0.23 and 0.15, respectively. No statistical differences were observed between the Salmonella strains. A negative correlation was observed between shoot weight and leaf contamination. The observed presence of the pathogens in lettuce, after thorough surface sterilization, demonstrates the possible presence of human pathogens in locations were they are unlikely to be removed by the actions of consumer washing and therefore pose a serious threat when occurring in field situations.     

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.     

A note on the incidences of Salmonella spp., Listeria spp. and Escherichia coli O157:H7 serotypes in Turkish sausage (Soudjouck)

The incidence of Salmonella spp., Listeria monocytogenes and Escherichia coli O157:H7 was determined in 100 Turkish sausage (soudjouck) samples collected from shops and markets in the Afyon province, Turkey. Salmonella spp. were detected in 7% of the samples. All of the isolates were S. enterica Paratyphi B. In addition, Listeria spp. were detected in 9% of the samples. Its distribution was 7% L. monocytogenes and 1% each of L. ivanovii and L. innocua. Serological study of the seven L. monocytogenes isolates showed that three of these were 1/2 ab, three were 5/6 ab and one was 1 ab. E. coli O157:H7 was not detected in any of the samples. The pH values of the samples ranged from 4.8 to 6.5. In conclusion, increasing number of listeriosis and salmonellosis cases in Turkey and the contamination levels found indicate that risk assessment and improved preventive measures are required for these sausages.     

Immunosensors for rapid detection of Escherichia coli O157:H7 - perspectives for use in the meat processing industry

This review critically evaluates different types of immunosensors proposed for rapid identification of Escherichia coli O157:H7. The methods are compared with approved USDA-FSIS standard procedures for determination of this pathogen in raw or ready-to-eat meat products. Major advantages and disadvantages for each method are highlighted. Our analysis suggests that application of immunosensors in the meat-processing industry may be limited to identification of uncontaminated samples after conventional selective enrichment in broth. Use for detection appears limited at the present time.     

Behavior of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium in teewurst, a raw spreadable sausage

The fate of Listeria monocytogenes, Salmonella Typhimurium, or Escherichia coli O157:H7 were separately monitored both in and on teewurst, a traditional raw and spreadable sausage of Germanic origin. Multi-strain cocktails of each pathogen (ca. 5.0 log CFU/g) were used to separately inoculate teewurst that was subsequently stored at 1.5, 4, 10, and 21 °C. When inoculated into commercially-prepared batter just prior to stuffing, in general, the higher the storage temperature, the greater the lethality. Depending on the storage temperature, pathogen levels in the batter decreased by 2.3 to 3.4, ca. 3.8, and 2.2 to 3.6 log CFU/g for E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, during storage for 30 days. When inoculated onto both the top and bottom faces of sliced commercially-prepared finished product, the results for all four temperatures showed a decrease of 0.9 to 1.4, 1.4 to 1.8, and 2.2 to 3.0 log CFU/g for E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, over the course of 21 days. With the possible exceptions for salt and carbohydrate levels, chemical analyses of teewurst purchased from five commercial manufacturers revealed only subtle differences in proximate composition for this product type. Our data establish that teewurst does not provide a favourable environment for the survival of E. coli O157:H7, S. Typhimurium, or L. monocytogenes inoculated either into or onto the product.