Growth of Escherichia coli O157:H7 and Listeria monocytogenes with prior resistance to intense pulsed light and lactic acid

Previous study showed that repetitive mild decontamination treatments with intense light pulses (ILP) and lactic acid (LA) can induce increased resistance in surviving pathogenic cells. Research has evaluated the potential of increased resistance to enhance the persistence of resistant variants of Listeria monocytogenes and Escherichia coli O157:H7 under suboptimal growth conditions. Growth of resistant variants and parental strains was determined by optical density (OD) measurements in nutrient broths with different pH values and NaCl concentration, at low temperature. The real lag phase was calculated, and results indicated that intense light pulses (ILP) resistant variants needed longer time to initiate growth compared to their parental strains, for both L. monocytogenes and E. coli O157:H7 when incubated at 7 °C and 10 °C, respectively. These selected variants were of the similar resistance towards heat and low pH (no cross-tolerance). Nevertheless, lactic acid (LA) resistant variant of L. monocytogenes was cross-protected when exposed to low pH, but not when treated with heat.     

Treatment of Escherichia coli O157:H7 with lactic acid, neutralized electrolyzed oxidizing water and chlorine dioxide followed by growth under sub-optimal conditions of temperature, pH and modified atmosphere

The utilization of sub-lethal decontamination treatments gains more and more interest due to the increased consumers' demand for fresh, minimally processed and convenient food products. These products rely on cold chain and hurdle (combination) technology to provide microbiological safety and quality during their shelf life. To investigate the ability of surviving cells to resuscitate and grow in a food simulating environment, sub-lethal decontamination treatments were coupled with subsequent storage under sub-optimal growth conditions. For this purpose chlorine dioxide (ClO₂) and neutralized electrolyzed oxidizing water (NEW)-treated cultures of Escherichia coli O157:H7 were inoculated in TSB-YE of pH 5.8 and a_w 0.99, and stored at 10 °C, 12.5 °C and 15 °C, under four different atmospheres (0%, 30% and 60% CO₂ balanced with N₂, and air). Due to the severity of injury, lactic acid-treated cells were inoculated in TSB-YE pH 7.0. Data obtained reveal that the fraction of sub-lethally injured E. coli O157:H7 undergoes an additional inhibitory effect during the storage period under of sub-optimal conditions. Observed extension in the lag growth phase was a direct consequence prior sub-lethal injury. The effects of liquid ClO₂ and NEW were less pronounced in comparison to lactic acid. The current study signifies the potential utilization of appropriate combination of different extrinsic and intrinsic factors in the elimination or growth inhibition of food-borne pathogens.     

Bacterial resistance after pulsed electric fields depending on the treatment medium pH

The objective was to evaluate and compare the pulsed electric field (PEF) resistance of four Gram-positive (Bacillus subtilis, Listeria monocytogenes, Lactobacillus plantarum, Staphylococcus aureus) and four Gram-negative (Escherichia coli, E. coli O157:H7, Salmonella serotype Senftenberg 775W, Yersinia enterocolitica) bacterial strains under the same treatment conditions. Microbial characteristics such as cell size, shape or type of the cell envelopes did not exert the expected influence on microbial PEF resistance. The most PEF resistant bacteria depended on the treatment medium pH. For instance, L. monocytogenes, which showed the highest PEF resistance at pH 7.0, was one of the most sensitive at pH 4.0. The most PEF resistant strains at pH 4.0 were the Gram-negatives E. coli O157:H7 and S. Senftenberg. A subsequent holding of PEF-treated cells in pH 4.0 for 2 h increased the degree of inactivation up to 4 extra Log₁₀ cycles depending on the bacterial strain investigated. Under these treatment conditions, the most PEF resistant bacterial strains were still the pathogens S. Senftenberg and E. coli O157:H7.

Industrial relevance

The design of appropriate food preservation processes by PEF requires the selection of an adequate target bacterial strain, which should correspond to the most PEF resistant microorganism contaminating food. This study indicates that the pH of the treatment medium plays an important role in determining this target bacterial strain. On the other hand, the combination of PEF and subsequent holding under acidic conditions has been proven to be an effective method in order to achieve a higher level of microbial inactivation.     

Combined treatment of fumaric acid with aqueous chlorine dioxide or UV-C irradiation to inactivate Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes inoculated on alfalfa and clover sprouts

Effect of fumaric acid, chlorine dioxide (ClO₂), and UV-C treatment was examined on the inactivation of microorganisms in alfalfa and clover sprouts. Clover sprouts were irradiated with UV-C light (1–10 kJ/m²), and the treatment decreased the population of total aerobic bacteria by 1.03–1.45 log CFU/g. Clover sprouts inoculated with pathogenic bacteria were treated with various concentration of fumaric acid, and 0.5 g/100 ml fumaric acid treatment was the most effective. In addition, the combined treatment of fumaric acid (0.5 g/100 ml)/UV-C (1 kJ/m²) reduced the populations of Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes inoculated on clover sprouts by 3.02, 2.88, and 2.35 log CFU/g. Alfalfa sprouts were treated with ClO₂, fumaric acid, and the combination of fumaric acid/ClO₂. The combined treatment was the most effective, and it reduced the total aerobic bacteria by 3.18 log CFU/g as well as the initial populations of E. coli O157:H7, S. typhimurium, and L. monocytogenes inoculated on alfalfa sprouts by 4.06, 3.57, and 3.69 log CFU/g. These results suggest that the combined treatment of fumaric acid with UV-C or ClO₂ can be useful for improving the microbial safety of alfalfa and clover sprouts.     

UV-C inactivation of Escherichia coli at different temperatures

The influence of treatment parameters (dose and temperature), treatment medium characteristics (absorption coefficient, pH and water activity) and microbiological factors (strain, growth phase and UV damage and repair capacity) on Escherichia coli UV-C resistance has been investigated. UV-C doses to inactivate at 25 °C 99.99% of the initial population (4D) of five strains of E. coli in McIlvaine buffer of pH 7.0 with tartrazine added (absorption coefficient of 10.77 cm⁻¹) were 16.60, 14.36, 14.36, 13.22, 11.18 J/mL for strains E. coli STCC 4201, STCC 471, STCC 27325, O157:H7 and ATCC 25922, respectively. The entrance in the stationary growth phase increased the 4D value of the most resistant strain, E. coli STCC 4201, from 13.09 to 17.23 J/mL. Survivors to UV treatments showed neither oxidative damages nor injuries in cell envelopes. On the contrary, the photoreactivation by the incubation of plates for 60 min below visible light (11.15 klx) increased the dose to 18.97 J/mL. The pH and the water activity of the treatment medium did not affect the UV tolerance of E. coli STCC 4201, but the lethal effect of the treatments decreased exponentially (Log₁₀4D = − 0.0628α + 0.624) by increasing the absorption coefficient (α). A treatment of 16.94 J/mL reached 6.35, 4.35, 2.64, 1.93, 1.63, 1.20, 1.02 and 0.74 Log₁₀ cycles of inactivation with absorption coefficients of 8.56, 10.77, 12.88, 14.80, 17.12, 18.51, 20.81 and 22.28 cm⁻¹. The temperature barely changed the UV resistance up to 50.0 °C. Above this threshold, inactivation rates due to the combined process synergistically increased with the temperature. The magnitude of the synergism decreased over 57.5 °C. An UV treatment of 16.94 J/mL in media with an absorption coefficient of 22.28 cm⁻¹ reached 1.23, 1.64, 2.36, 4.01 and 6.22 Log₁₀ cycles of inactivation of E. coli STCC 4201 at 50.0, 52.5, 55.5, 57.5 and 60.0 °C, respectively.

Industrial relevance

Results obtained in this investigation show that UV light applied at mild temperatures (57.5 to 60 °C) could be an alternative to heat treatments for 5-Log₁₀ reductions of E. coli in liquid foods. Since microbial resistance to UV-C light did not depend on the pH and water activity (a_w) of the treatment media, eventual advantages of UV light for pasteurization purposes will be higher in low a_w foods. E. coli STCC 4201 could be considered as a target when UV light processing of foods.     

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.     

Influence of partial inactivation on growth of Listeria monocytogenes under sub-optimal conditions of increased NaCl concentration or increased acidity

The effect of partial inactivation with lactic acid (LA), liquid chlorine dioxide (ClO₂) and intense light pulses (ILP) on injury and post-treatment growth under increased NaCl concentration and reduced pH values of Listeria monocytogenes strains was investigated. Inactivation levels and the percentage of sub-lethal were dependent upon strain and type of inactivation technique used. Comparison of the mean time-to-detection (TTD) values under suboptimal conditions (increased NaCl concentration or reduced pH) showed that the longest TTD was at every pH observed for the cultures treated with ClO₂, followed by LA and ILP. Under increased NaCl concentration LA treated cells required the longest TTD, followed by ClO₂ and ILP, respectively. Significant difference in TTD between untreated and cultures treated with ClO₂ and LA was observed. Recovery of ILP treated cultures was not always different from untreated cultures. The extended post-treatment effect based on the growth retardation or inhibition of injured cells under sub-optimal conditions is suggested as an important tool in conditioning of microbial food safety.Industrial relevanceSmall and medium sized enterprises (SMEs) are the backbone of the European economy. They are a key source of jobs and a breeding ground for implementation of research results. They are however the most sensitive of all to changes in the production practices coming as a consequence of increased consumers' awareness regarding fresh and natural-like foods. In order to respond to the changed consumption pattern, non-heat and so-called mild technologies have emerged. This research as a part of the EU Pathogen Combat project is tailor made to answer some of the burning issues when it comes to “mild” decontamination. Among the more established technologies used here is decontamination with lactic acid, followed by chlorine dioxide. Both are now under the eye of EU policy makers who are seeking relevant facts to adjust or not to adjust EU policy regarding decontamination of foods. Latest EFSA opinions have already pushed these agents forward, but relevant scientific facts were declared missing. Moreover, intense light pulses, as non-chemical alternative to pasteurization have a considerable potential not only in food surface decontamination, but also in the decontamination of packaging materials, industrial surfaces etc. The present research brought new facts into the light that might have a credible influence on understanding pros and cons of lactic acid, chlorine dioxide and intense light pulses as decontamination agents. The post-treatment, based on sublethal injury, effect is suggested as an important extension tool in shelf life extension and safety barrier.     

Viability of multi-strain mixtures of Listeria monocytogenes, Salmonella typhimurium, or Escherichia coli O157:H7 inoculated into the batter or onto the surface of a soudjouk-style fermented semi-dry sausage

The fate of Listeria monocytogenes, Salmonella typhimurium, or Escherichia coli O157:H7 were separately monitored both in and on soudjouk. Fermentation and drying alone reduced numbers of L. monocytogenes by 0.07 and 0.74 log₁₀ CFU/g for sausages fermented to pH 5.3 and 4.8, respectively, whereas numbers of S. typhimurium and E. coli O157:H7 were reduced by 1.52 and 3.51 log₁₀ CFU/g and 0.03 and 1.11 log₁₀ CFU/g, respectively. When sausages fermented to pH 5.3 or 4.8 were stored at 4, 10, or 21 °C, numbers of L. monocytogenes, S. typhimurium, and E. coli O157:H7 decreased by an additional 0.08–1.80, 0.88–3.74, and 0.68–3.17 log₁₀ CFU/g, respectively, within 30 days. Storage for 90 days of commercially manufactured soudjouk that was sliced and then surface inoculated with L. monocytogenes, S. typhimurium, and E. coli O157:H7 generated average D-values of ca. 10.1, 7.6, and 5.9 days at 4 °C; 6.4, 4.3, and 2.9 days at 10 °C; 1.4, 0.9, and 1.6 days at 21 °C; and 0.9, 1.4, and 0.25 days at 30 °C. Overall, fermentation to pH 4.8 and storage at 21 °C was the most effective treatment for reducing numbers of L. monocytogenes (2.54 log₁₀ CFU/g reduction), S. typhimurium (5.23 log₁₀ CFU/g reduction), and E. coli O157:H7 (3.48 log₁₀ CFU/g reduction). In summary, soudjouk-style sausage does not provide a favorable environment for outgrowth/survival of these three pathogens.     

Inhibition of Listeria monocytogenes and Escherichia coli O157:H7 in liquid broth medium and during processing of fermented sausage using autochthonous starter cultures

The antimicrobial effect of two autochthonous starter cultures of Lactobacillus sakei was evaluated in vitro (in liquid broth medium) and in situ assays. The inactivation of foodborne pathogens Listeria monocytogenes (serotype 4ab No 10) and Escherichia coli O157:H7 ATCC 43888 was investigated during the production of fermented sausage according to a typical Greek recipe using L. sakei strains as starter cultures. The inactivation kinetics were modeled using GInaFiT, a freeware tool to assess microbial survival curves. By the end of the ripening period, the inhibition of L. monocytogenes was significant in treatments with L. sakei 8416 and L. sakei 4413 compared to the control treatment. A 2.2-log reduction of the population of E. coli O157:H7 resulted from the autochthonous starter culture L. sakei 4413 during sausage processing. The use of the autochthonous starter cultures constitutes an additional improvement to the microbial safety by reducing foodborne pathogens.     

Effect of X-ray treatments on inoculated Escherichia coli O157: H7, Salmonella enterica, Shigella flexneri and Vibrio parahaemolyticus in ready-to-eat shrimp

This study was conducted to evaluate the inactivation effect of X-ray treatments on Escherichia coli O157: H7, Salmonella enteric (S. enterica), Shigella flexneri (S. flexneri) and Vibrio parahaemolyticus (V. parahaemolyticus) artificially inoculated in ready-to-eat (RTE) shrimp. A mixed culture of three strains of each tested pathogen was used to inoculate RTE shrimp. The shrimp samples were inoculated individually with selected pathogenic bacteria then aseptically placed in sterile plastic cups and air-dried at 22 °C for 30 min (to allow bacterial attachment) in the biosafety cabinet prior to X-ray treatments. The inoculated shrimp samples were then placed in sterilized bags and treated with 0.1, 0.2, 0.3, 0.5, 0.75, 1.0, 2.0, 3.0 and 4.0 kGy X-ray at ambient temperature (22 °C and 60% relative humidity). Surviving bacterial populations were evaluated using a non-selective medium (TSA) with the appropriate selective medium overlay for each bacterium; CT-SMAC agar for E. coli O157: H7, XLD for S. enterica and S. flexneri and TCBS for V. parahaemolyticus. More than a 6 log CFU reduction of E. coli O157: H7, S. enterica, S. flexneri and V. parahaemolyticus was achieved with 2.0, 4.0, 3.0 and 3.0 kGy X-ray, respectively. Furthermore, treatment with 0.75 kGy X-ray significantly reduced the initial microflora on RTE shrimp samples from 3.8 ± 0.2 log CFU g⁻¹ to less than detectable limit (<1.0 log CFU g⁻¹).     

Inactivation of Escherichia coli O157:H7, Salmonella typhimurium,and Listeria monocytogenes on Stored Iceberg Lettuce by Aqueous Chlorine Dioxide Treatment

ABSTRACT: Inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes in iceberg lettuce by aqueous chlorine dioxide (ClO₂) treatment was evaluated. Iceberg lettuce samples were inoculated with approximately 7 log CFU/g of E. coli O157:H7, S. typhimurium, and L. monocytogenes. Iceberg lettuce samples were then treated with 0, 5, 10, or 50 ppm ClO₂ solution and stored at 4 °C. Aqueous ClO₂ treatment significantly decreased the populations of pathogenic bacteria on shredded lettuce (P < 0.05). In particular, 50 ppm ClO₂ treatment reduced E. coli O157:H7, S. typhimurium, and L. monocytogenes by 1.44, 1.95, and 1.20 log CFU/g, respectively. The D₁₀‐values of E. coli O157:H7, S. typhimurium, and L. monocytogenes in shredded lettuce were 11, 26, and 42 ppm, respectively. The effect of aqueous ClO₂ treatment on the growth of pathogenic bacteria during storage was evaluated, and a decrease in the population size of these pathogenic bacteria was observed. Additionally, aqueous ClO₂ treatment did not affect the color of lettuce during storage. These results suggest that aqueous ClO₂ treatment can be used to improve the microbial safety of shredded lettuce during storage.     

Inactivation of Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes by underwater shock waves

Underwater shock waves are pressure discontinuities generated by means of high voltage electric discharges in water. Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes were exposed to shock waves. The influence of the pulse of light produced by the electric discharge, acoustic cavitation produced by the shock waves, the phase of growth and shock wave dosage on bacteria inactivation was evaluated. Inactivation of bacteria by shock waves ranged from 0 to 3.18 log₁₀ CFU/ml. L. monocytogenes was the most susceptible microorganism, 3.17 log₁₀ CFU/ml inactivation was achieved with 350 shock waves. At the same shock wave dosage, 1.68 log₁₀ CFU/ml and 0.56 log₁₀ CFU/ml inactivation was achieved for S. Typhimurium and E. coli O157:H7, respectively. Even if the achieved inactivation still is of little practical value, higher shock wave energy may enhance bactericidal activity. The analysis of variance revealed that synergy between light and the other factors contributed to bacterial inactivation. Differences in the response of bacteria, along with data analysis, suggest different mechanisms of inactivation by shock waves.Industrial relevance: Underwater shock waves generated via high voltage electric discharge or the use of explosives have been reported for microbial inactivation and for softening of animal tissue. The current paper deals with the interaction of underwater shock waves, the associated cavitation and pulse of light generated in the UV and visible spectra. Although the data generated don't show spectacular microbial inactivation, systematic studies like this can lead to the development of new combination processes with specific applications (e.g., microbial inactivation within containers).     

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.     

Inactivation of Escherichia coli O157:H7 on surface-uninjured and -injured green pepper (Capsicum annuum L.) by chlorine dioxide gas as demonstrated by confocal laser scanning microscopy

Cells of Escherichia coli O157:H7 on uninjured and injured surfaces of green pepper were inactivated by 0·15–1·2 mg l⁻¹ClO₂gas treatments. A membrane-surface-plating method was used for resuscitation and enumeration of E. coli O157:H7 treated with ClO₂. The location and viability ofE. coli O157:H7 on uninjured and injured green pepper surfaces after ClO₂gas treatments were visualized using confocal laser scanning microscopy (CLSM). Live and dead cells of E. coli O157:H7 on pepper surfaces were labeled with a fluorescein isothiocyanate-labeled antibody and propidium iodide, respectively. A 7·27 log reduction of E. coli O157:H7 on uninjured green pepper surfaces was obtained with a 0·60 mg l⁻¹ClO₂gas treatment for 30 min at 20°C under 90–95% relative humidity. For injured surfaces, a 6·45 log reduction was achieved with a 1·2 mg l⁻¹ClO₂gas treatment. Each ClO₂gas treatment (0·15–1·2 mg l⁻¹ClO₂) for inoculated bacteria on uninjured surfaces showed significantly more reductions (1·23–4·24 log) than for those on injured surfaces (P<0·05). The microphotographs of CLSM showed that bacteria preferentially attached to injured surfaces and those bacteria could be protected from bacterial reduction by the injuries. This study indicates that ClO₂gas treatment can be a potential effective method of pathogen reduction for fresh fruits and vegetables.     

Inactivation kinetics of Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and Campylobacter jejuni in ready-to-eat sliced ham using UV-C irradiation

To investigate the applicability of UV-C irradiation on the inactivation of foodborne pathogenic bacteria in ready-to-eat sliced ham, UV-C treatment was evaluated. Irradiation dose required for 90% reduction of the populations of Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and Campylobacter jejuni were determined to be 2.48, 2.39, and 2.18 J/m². Ready-to-eat sliced hams were inoculated with the pathogens and irradiated with UV-C light of 1000, 2000, 4000, 6000, and 8000 J/m². Microbiological data indicated that foodborne pathogen populations significantly (p < 0.05) decreased with increasing UV-C irradiation. In particular, UV-C irradiation at 8000 J/m² reduced the populations of L. monocytogenes, S. Typhimurium, and C. jejuni in the ham by 2.74, 2.02, and 1.72 log CFU/g. The results indicate that UV-C irradiation can be used as a microbial inactivation method for ready-to-eat sliced ham, and inactivation kinetics of the foodborne pathogens fit the Weibull model better than the first-order kinetics model.     

Inactivation of Yersinia enterocolitica by pulsed electric fields

The influence of growth conditions, treatment medium characteristics and PEF process parameters on the lethal effect on Yersinia enterocolitica of pulsed electric fields (PEF) treatments in batch has been investigated. Growth phase, temperature of growth, pH, conductivity of the treatment medium, pulse width and frequency of pulses did not influence the sensitivity of Y. enterocolitica to PEF. However, an A_w decrease from >0.99 to 0.93 of the treatment medium increased the PEF resistance of Y. enterocolitica with 3.5 log₁₀ cycles after a treatment of 22 kV/cm, 800 μs and 880 kJ/kg. Inactivation of Y. enterocolitica increased with the field strength, treatment time and total specific energy up to a maximum of 6 log₁₀ cycles after 28 kV/cm, 2000 μs and 3559 kJ/kg. A nonlinear relationship was found among the survival fraction and the treatment time or the specific energy that was accurately described by a mathematical model based on the Weibull distribution. The inactivation of Y. enterocolitica by PEF was characterized by maximum field strength thresholds. Above these thresholds, specific energy necessary to obtain a given level of inactivation scarcely decreased by increasing the electric field strength, and inactivation of Y. enterocolitica only depended on the specific energy applied.     

The prevalence of Listeria, Salmonella, Escherichia coli and E. coli O157:H7 on bison carcasses during processing

Bison meat is a relatively new, emerging meat species gaining increased popularity in the US and European meat markets, but little is known of its microflora or pathogens that may be present. This study was carried out to determine the incidence of the foodborne pathogens Listeria, Salmonella, Escherichia coli/E. coli O157:H7 on slaughtered bison and to evaluate the bison slaughter process. Bison carcass sampling was carried out at monthly intervals over a period of 1 year at a Bison processing facility in the Midwestern United States. A total of 355 Bison carcasses were sampled by surface swabbing the carcasses at five points on the production line: pre-dehiding, post-evisceration, post-USDA inspection, post-washing and 24 h chilled carcass. Overall, the prevalence of Listeria spp., Salmonella spp., E. coli and E. coli O157:H7 was 18.3%, 3.94%, 38.3% and 1.13%, respectively. The prevalence of Listeria spp. at each sampling point tested was 42.24%, 18.1%, 6.03%, 1.72% and 3.77% while the prevalence of E. coli at each sampling point was: 88.79%, 73.28%, 52.59%, 56.89% and 11.3%, respectively. The data obtained suggests that current antimicrobial intervention strategies used at the plant are relatively effective in reducing Listeria and E. coli contamination on bison carcasses to some extent, however further study is required to determine the influence of current slaughter practices on carcass contamination. The data reported in this study to the authors’ knowledge is some of the first information reporting on the bacteriological status of Bison, and provides some useful baseline information for future research.     

Characterization of Escherichia coli from raw poultry in Belgium and impact on the detection of Campylobacter jejuni using Bolton broth

A comparative study examining Bolton broth and Preston broth for enrichment and reliable detection of Campylobacter jejuni (both healthy and freeze stressed cells) was performed. Tested as pure cultures, Bolton broth enabled faster resuscitation and growth of C. jejuni compared to Preston broth. When C. jejuni was co-incubated with extended-spectrum-beta-lactamase (ESBL) producing Escherichia coli isolated from Belgian poultry meat preparations, the latter dominated in the Bolton enrichment broth and crowded the mCCDA plates. This resulted in the inability to recover C. jejuni by ISO 10272-1:2006 standard method. Preston broth did not support the growth of the ESBL E. coli isolates, but showed longer detection time of C. jejuni compared to Bolton broth. The use of the same antibiotic (sodium cefoperazone) in Bolton broth and in mCCDA plates may explain the problems encountered for detection of C. jejuni, as high numbers of ESBL E. coli present after enrichment in Bolton broth, also caused overgrowth and masked the few C. jejuni colonies present on the mCCDA plates. The use of Campylobacter spp. specific real-time PCR circumvented these problems and enabled rapid detection of the pathogen after 24 h enrichment in both Bolton and Preston broth, for both healthy and freeze stressed cells.     

Comparing predicting models for heat inactivation of Listeria monocytogenes and Pseudomonas aeruginosa at different pH

Under the same experimental conditions it has been demonstrated that whereas survival curves of Listeria monocytogenes in the range of temperatures from 54 to 62 °C followed a first-order kinetic, those of Pseudomonas aeruginosa in the range of temperatures from 50 to 56 °C were not linear showing a shoulder followed by a linear region. The first order kinetic model did not describe survival curves of P. aeruginosa. A model based on the Weibull distribution (Log₁₀(N_t/N₀)=(1/−2.303)*(t/b)ⁿ)) accurately described the inactivation kinetics of both microorganisms at the three pHs of 4, 5.5, 7.4 investigated. For both microorganisms, the b value depended on the treatment temperature and the pH of the treatment medium. Whereas for L. monocytogenes the n value was independent of the treatment conditions, for P. aeruginosa the n value depended on the pH of the treatment medium.The model based on the Weibull distribution was capable of accurately predicting the treatment time to inactivate five Log₁₀ cycles of both microorganisms at the three pHs investigated.     

Efficacy of Chlorine Dioxide, Ozone, and Thyme Essential Oil or a Sequential Washing in Killing Escherichia coli O157:H7 on Lettuce and Baby Carrots

Chlorine dioxide (ClO₂), ozone, and thyme essential oil has been found to be effective in reducing pathogens, including Escherichia coli O157:H7, on selected produce. The efficacy of these sanitizers was evaluated, alone or through their sequential washing to achieve a 3 or more log reduction of mixed strains of E. coli O157:H7 on shredded lettuce and baby carrots. Samples sprinkle inoculated with mixed strains of E. coli O157:H7 were air-dried for 1 h at 22±2°C in a biosafety cabinet, stored at 4°C for 24 h, and then treated with different concentrations of disinfectants and exposure time. Sterile deionized water washing resulted in approximately 1log reduction ofE. coli O157:H7 after 10 min washing of lettuce and baby carrots. Gaseous treatments resulted in higher log reductions in comparison to aqueous washing. However, decolorization of lettuce leaves was observed during long exposure time. A logarithmic reduction of 1.48-1.97log₁₀ cfu/g was obtained using aqueous ClO₂ (10.0 mg/L for 10 min) ozonated water (9.7 mg/L for 10 min) or thyme oil suspension (1.0 mL/L for 5 min) on lettuce and baby carrots. Of the three sequential washing treatments used in this study, thyme oil followed by aqueous ClO₂/ozonated water, or ozonated water/aqueous ClO₂ were significantly (P<0.05) more effective in reducing E. coli O157:H7 (3.75 and 3.99log, and 3.83 and 4.34 log reduction) on lettuce and baby carrots, respectively. The results obtained from this study indicate that sequential washing treatments could achieve 3-4log reduction of E. coli O157:H7 on shredded lettuce and baby carrots.