Viability Of Listeria Monocytogenes Research Articles

Viability of Listeria monocytogenes and Salmonella spp. on Slices of a German-Style Bologna Containing Blends of Organic Acid Salts During Storage at 4 or 12°C

Viability of cells of Listeria monocytogenes or Salmonella spp. was quantified on slices of a German-style bologna manufactured by a local butcher to contain no added antimicrobials or to include 0.9% or 1.3% of a blend of potassium acetate and sodium diacetate (K-Ace) or 2.5% of a blend of potassium lactate and sodium diacetate (K-Lac) as ingredients. After slicing (ca. 7.1 cm L by 6.7 cm W, ca. 0.5 cm thick, ca. 22.4 g each), a single slice of bologna was placed into a nylon–polyethylene bag and surface inoculated with 250 µL per side of a five-strain mixture of either cells of L. monocytogenes or Salmonella spp. to achieve an initial level of ca. 3.5–4.0 log CFU/slice. The packages were vacuum-sealed and then stored at 4 or 12°C for 90 and 30 days, respectively. Without antimicrobials added to the formulation, L. monocytogenes numbers increased by ca. 5.4 and 6.0 log CFU/slice at both 4 and 12°C during the entire 90- and 30-day storage period, respectively. Likewise, levels of Salmonella also increased by ca. 6.0 log CFU/slice at 12°C in the absence of added antimicrobials; however, levels of this pathogen decreased by ca. 1.7 log CFU/slice after 90 days at 4°C. With the inclusion of 0.9% or 1.3% K-Ace or 2.5% K-Lac in the bologna formulation, levels of L. monocytogenes decreased by ca. ≤0.7 log CFU/slice after 90 days at 4°C, whereas levels of Salmonella decreased by ca. 1.6–2.3 log CFU/slice. After 30 days at 12°C, levels of L. monocytogenes increased by ca. ≤3.4 log CFU/slice on product containing 0.9% K-Ace or 2.5% K-Lac but remained relatively unchanged on slices formulated with 1.3% K-Ace. For Salmonella, in the presence of 0.9% or 1.3% K-Ace or 2.5% K-Lac, pathogen levels decreased by ca. ≤0.7 log CFU/slice at 12°C after 30 days. Our data validate that the inclusion of K-Ace (0.9% or 1.3%) or K-Lac (2.5%) as ingredients is effective for controlling L. monocytogenes and Salmonella on slices of bologna during refrigerated storage.

Effects of Rubus fruticosus and Juniperus oxycedrus derivatives on culturability and viability of Listeria monocytogenes

The consumers’ demand for safe foods without chemical additives increased the research for green solutions, based on natural antimicrobials. Plants can be an important source of bioactive compounds able to prevent the development of foodborne pathogens and spoilage microflora. This paper aimed to characterize phenolic extracts (PEs) and essential oils (EOs) obtained from Mediterranean Rubus fruticosus leaves and Juniperus oxycedrus needles and to evaluate their antimicrobial effects against Listeria monocytogenes Scott A. The growth dynamics with sub-lethal concentrations of plant derivatives were modeled and flow cytometry was used to better evidence the effect on cell viability and culturability. The results showed that these plant derivatives affected the growth of L. monocytogenes, increasing lag phase (about 40 h in the presence of PEs vs. 8 h in the control) and decreasing the final cell load of at least 1 log cycle with respect to the control. R. fruticosus EO was the most effective, determining an initial decrease of cell counts of about 6 log cycles, followed by a restart of growth after 10 h, with rate similar to the control (0.08 with R. fruticosus EO vs. 0.09 ((log CFU/ml)/h in the control) but significantly lower final cell load (7.33 vs. 8.92 log CFU/ml). According to flow cytometry, only R. fruticosus EO induced a relevant increase of dead cells, while the other plant derivatives determined different extent of sub-lethal cell injury. The discrepancy observed in some cases between viability and culturability could indicate the presence of cells not able to grow in culture media, whose fate needs to be further investigated to assess their potential recovery, thus bringing to an overestimation of the antimicrobial effect of these substances. This research contributed to increase the knowledge of these underused raw materials such as blackberry leaves and juniper needles that can be exploited in food and other industries.

Inactivation of Listeria monocytogenes and Shiga Toxin-Producing Escherichia coli in “Soupie,” a Homemade Sopressata

Viability of Listeria monocytogenes and Shiga toxin-producing Escherichia coli (STEC) was monitored in “soupie,” a homemade soppressata. Coarse-ground fresh ham was mixed with nonmeat ingredients, a starter culture (ca. 6.0 log CFU/g), and one pathogen cocktail (ca. 6.5 log CFU/g). The batter was then fine ground, stuffed into fibrous casings, and fermented at 26.7°C and ca. 90 ± 5% relative humidity (RH) for ≤ 48 h to achieve a pH of ≤ 5.3. Chubs were dried at 15.6°C and ca. 87 ± 5% RH for 5 days, flattened under weights for 3 days, and then dried for an additional 21 days at 4°C and ca. 73 ± 5% RH. Half of the chubs were vacuum sealed individually in bags with 8 mL of sunflower oil, and the other half were submerged in sunflower oil (ca. 1.5 L) within covered plastic containers; all chubs were stored for 6 months at 20°C. Fermentation and drying delivered a ≤ 1.2-log reduction in levels of both pathogens. Regardless of storage conditions, a ≥ 5.0-log reduction was observed within 1 and 4 months of storage at 20°C for STEC and L. monocytogenes, respectively. These data establish that artisanal soupie, prepared and stored as described here, does not provide a favorable environment for pathogen persistence or proliferation.

Ohmic heating processing of milk for probiotic fermented milk production: Survival kinetics of Listeria monocytogenes as contaminant post-fermentation, bioactive compounds retention and sensory acceptance

The survival kinetics of Listeria monocytogenes (9 log CFU/mL) as a post-fermentation contaminant in probiotic fermented milk (Lactobacillus acidophilus La-5, 8–9 log CFU/mL) processed with milk subjected to ohmic heating (0, 4, 6, and 8 V/cm; CONV, OH4, OH6, OH8, 90–95 °C/5 min) was investigated using Weibull predictive model. Additionally, the presence of bioactive compounds (antioxidant activity, inhibition of the enzymes α-glucosidase, α-amylase, and angiotensin-converting) and sensory analysis (consumer test) of probiotic fermented milks were evaluated. Overall, OH provided a decrease in the viability of Listeria monocytogenes, suitable Lactobacillus acidophilus counts, and satisfactory results in the gastrointestinal tract survival. The Weibull model presented an excellent fit to the data of all conditions. Furthermore, lower δ values (217–298 against 665 h, CONV), and increased R2 values (0.99 against 0.98, CONV) were obtained for the OH-treated samples, emphasizing the best performance of OH data. In addition, OH improved the generation of bioactive compounds as well as the sensory acceptance. Indeed, considering functional and safety purposes, OH presented as an interesting technology to be used in milk for manufacturing probiotic fermented milk.

The effect of produce washing using electrolyzed water on the induction of the viable but non-culturable (VBNC) state in Listeria monocytogenes and Escherichia coli O157:H7

The effects of pathogen inoculum concentration, pH and organic load of electrolyzed oxidizing (EO) water treatment on the culturability and viability of Listeria monocytogenes and Escherichia coli O157:H7 was examined. Inoculum concentrations (8, 6, and 4 log CFU/mL), EO water at pH between 2.7 and 8.5 and two lettuce-to-EO water ratios (1:20 and 1:10 w/v) were tested by exposing pathogens to EO water for 5 min, followed by culturing on nonselective media and viability assessment through flow cytometry and resuscitation attempts. Treating pathogens at high initial populations with EO water led to maintenance of culturability, while lower populations lost culturability at free chlorine concentrations (FCC) above 1 mg/L. The pH of EO water significantly affected the reduction of viable populations of both pathogens (p < 0.05). The ultraviolet absorbance at 254 nm (UV254) was used as the organic load indicator and formation of VBNC cells was observed at 6 and 7.5 mg/L FCC of EO water samples with UV254 readings of 0.059 and 0.102, respectively. Increased organic load of EO water raised the VBNC-inducing FCC levels. However, keeping residual chlorine of EO water above 9 mg/L prevented the formation of VBNC cells and hence helped prevent cross-contamination during fresh produce washing treatment.

Application of EvaGreen for the assessment of Listeria monocytogenes АТСС 13932 cell viability using flow cytometry.

Determination of eukaryotic cell viability using flow cytometry is widespread and based on the use of fluorescent dyes such as SYTO, DAPI, SYBR, PI, and SYTOX. For many years, traditional microbiological methods have been used to successfully analyze prokaryotic cells, but the application of flow cytometry should be considered because it provides an opportunity for quantitative assessment. A combination of SYTO 9 or SYBR green and PI has been used successfully. DNA-binding dyes such as SYTO 9, SYBR green, and EvaGreen are used in qPCR. The aim of this study was to assess the feasibility of EvaGreen to determine the viability of Listeria monocytogenes АТСС 13932 cells using flow cytometry. RNA from Escherichia coli ATCC 25922 was isolated using the MagNA Pure LC RNA Isolation Kit-High Performance (Roche, Germany) according to the kit instructions on MagNA Pure LC® 2.0 (Roche, Switzerland). Chicken DNA was isolated using the Sorb-GMO-B kit (Syntol CJSC, Russia) according to the kit instructions. RNA from E. coli ATCC 25922, chicken DNA, a positive control, and a negative control of L. monocytogenes АТСС 13932 were stained with EvaGreen and analyzed on the Guava EasyCyte flow cytometer (Merck Millipore, Germany). Chicken DNA demonstrated both green and red fluorescence, while E. coli RNA displayed only red fluorescence. While the positive L. monocytogenes АТСС 13932 control and chicken DNA demonstrated similar fluorescence properties, the negative control showed a localization similar to that observed with E. coli RNA. Degraded ssDNA and RNA stained with EvaGreen demonstrated red fluorescence. Although EvaGreen is a class III dye, we observed fluorescence of live L. monocytogenes АТСС 13932 cells in the positive control stained with EvaGreen. The observed phenomenon was linked to the solution composition. It is necessary to repeat this analysis with various solution compositions as well as a wide range of both Gram-positive and Gram-negative bacteria to determine the effects on cell envelope permeability of EvaGreen.

Impact of environmental factors on the culturability and viability of Listeria monocytogenes under conditions encountered in food processing plants

The ability of Listeria monocytogenes to adhere to and persist on surfaces for months or even years may be responsible for its transmission from contaminated surfaces to food products. Hence the necessity to find effective means to prevent the establishment of L. monocytogenes in food processing environments. The aim of this study was to assess, through a fractional experimental design, the environmental factors that could affect the survival of L. monocytogenes cells on surfaces to thereby prevent the persistence of this pathogen in conditions mimicking those encountered in food processing plants: culture with smoked salmon juice or meat exudate, use of two materials with different hygiene status, biofilm of L. monocytogenes in pure-culture or dual-culture with a Pseudomonas fluorescens strain, application of a drying step after cleaning and disinfection (C&D) and comparison of two strains of L. monocytogenes. Bacterial survival was assessed by culture, qPCR to quantify total cells, and propidium monoazide coupled with qPCR to quantify viable cells and highlight viable but non-culturable (VBNC) cells. Our results showed that failure to apply C&D causes cell persistence on surfaces. Moreover, the sanitation procedure leads only to a loss of culturability and appearance of VBNC populations. However, an additional daily drying step after C&D optimises the effectiveness of these procedures to reduce culturable populations. Our results reinforce the importance to use molecular tools to monitor viable pathogens in food processing plants to avoid underestimating the amounts of cells using only methods based on cell culture.

Viability of Listeria monocytogenes on Boneless, Water-Added Hams, Commercially Prepared with and without Food-Grade Chemicals, during Extended Storage at 4 and/or −2.2°C

Viability of Listeria monocytogenes was monitored during refrigerated (4°C) and/or frozen (i.e., deep chilling at -2.2°C) storage on casing-cooked hams that were commercially prepared with and without potassium lactate and sodium diacetate (1.6%), buffered vinegar (2.2%), buffered vinegar and potassium lactate (1.7%), or a blend of potassium lactate, potassium acetate, and sodium diacetate (1.7%). A portion of these hams were subsequently surface treated with lauric arginate ester (LAE; 44 ppm). In phase I, hams (ca. 3.5 kg each) were sliced (ca. 0.7 cm thick, ca. 100 g), inoculated (ca. 4.0 log CFU per slice), surface treated with LAE, and stored at either 4°C for 120 days or at -2.2°C for 90 days and then at 4°C for an additional 120 days. In phase I, without antimicrobials, the population of L. monocytogenes increased by ca. 5.9 log CFU per slice within 120 days at 4°C; however, pathogen levels increased only slightly (ca. 0.45 log CFU per slice) for hams formulated with potassium lactate and sodium diacetate and decreased by ca. 1.2 log CFU per slice when formulated with the other antimicrobials. For slices held at -2.2°C and then stored at 4°C, but not treated with LAE, L. monocytogenes increased by ca. 4.5 log CFU per slice for controls, whereas when formulated with antimicrobials, pathogen levels decreased by ca. 1.4 to 1.8 log CFU per slice. For product treated with LAE, L. monocytogenes increased by ca. 4.0 log CFU per slice for controls, whereas when formulated with antimicrobials, pathogen levels decreased by ca. 0.9 to 1.9 log CFU per slice. In phase II, whole hams (ca. 1.0 kg each) containing antimicrobials were inoculated (6.8 log CFU per ham) and then stored at -2.2°C for 6 months. Pathogen levels decreased by ca. 2.0 to 3.5 log CFU per ham (without LAE treatment) and by ca. 4.2 to 5.2 log CFU per ham (with application of LAE via Sprayed Lethality in Container) when product was held at -2.2°C. In general, deep chilling hams was listericidal, and inclusion of antimicrobials in the formulation suppressed outgrowth of L. monocytogenes during extended cold storage.

A study on the effect of temperature,PH and Capsicum annuum extract on the viability of Listeria monocytogenes in kariesh cheese

A study on the effect of temperature,PH and Capsicum annuum extract on the viability of Listeria monocytogenes in kariesh cheese

Viability of Listeria monocytogenes in an experimental model of nigiri sushi of halibut (Hippoglossus hippoglossus) and salmon (Salmo salar)

Sushi is a traditional Japanese food, mostly consisting of raw seafood in combination with rice. However, eating sushi has become popular among consumers in many countries outside Japan. Sushi is not free from health risks and foodborne illness linked to this product has been caused by Listeria monocytogenes. The aim of our work was to study viability of L. monocytogenes in emulated nigiri sushi comprised of halibut and salmon. Raw samples of halibut and salmon were inoculated with the pathogen and subsequently put on top of vinegar marinated sushi rice followed by storage for 7 days at 4 and 8 °C. As controls, inoculated seafood without rice was used. The experiments demonstrated that the level of L. monocytogenes in nigiri sushi was significant lower during storage over 7 days at 4 and 8 °C compared to controls, in case of inoculated seafood only. The pH drop in the fish muscle, caused by the sushi rice, is believed to be the reason for the decrease in viability of L. monocytogenes in nigiri sushi.

Presence of Listeria spp. in fish samples, fish products and sea products

Listeria monocytogenes is a bacterium that is pathogenic for man and for most animal species. Listeriosis is a generalized infection that starts after ingestion of the causative agent L. monocytogenes (Finlay, 2001). Food that is not properly thermically processed, long storage of such food, food that is produced in inadequate hygienic food plants, as well as cooked-cooled ready to eat food (RTE) is the ideal medium for listeria multiplication. High risk food originated from fish, fish products and sea products comprised of: molluscs (fresh of frozen shellfish, crustaceans shelled or not), fresh fish (ready to eat without cooking), fish products packed with brine (NaCl<6%), (salted, marinated, fermented, cold smoked and fish in brine), thermically treated fish and crustacean products (pasteurization, cooking, hot smoking, including pre-cooking and panning). In this research, fish samples, fish products and sea products from Serbian markets were examinated such as: fresh fish (cooled), frozen food (fish and sea products - cuttlefish, squid, octopus, shellfish, crustaceans and shrimps), panned products; smoked fish, salted fish, thermally treated fish and fish products, semi-canned fish and canned fish. Microbiological testing has been performed according to internationally prescribed standards ISO 11290-1 (1996) "Microbiology of food and animal feed stuffs - Horizontal method for the detection and enumeration of Listeria monocytogenes". There were 470 samples from fish products and sea products. Presence of Listeria spp was registered in 58 samples (12.34%). Listeria monocytogenes was found in 1.92% samples (9 isolates), which makes 15.52% of the total Listeria that were isolated from the tested food. Other isolated Listeria belong to the following species: L. innocua (8.51%), L.welshimeri (1.28%), L.welshimeri/innocua (0.21%), L. grayi (0.21%) and L. seeligeri (0.21%). Exceptional viability of Listeria monocytogenes in the food tested was documented, also (freezing temperature - 18oC, five months).

Control of Listeria monocytogenes on commercially-produced frankfurters prepared with and without potassium lactate and sodium diacetate and surface treated with lauric arginate using the Sprayed Lethality in Container (SLIC®) delivery method

Viability of Listeriamonocytogenes was monitored on frankfurters formulated with or without potassium lactate and sodium diacetate at a ratio of ca. 7:1 and treated with lauric arginate (LAE; 22 or 44ppm) using the Sprayed Lethality in Container (SLIC(R)) delivery method. Without antimicrobials, pathogen numbers remained relatively constant at ca. 3.3logCFU/package for ca. 30d, but then increased to ca. 8.4logCFU/package over 120d. Regardless of whether or not lactate and diacetate were included, when treated with LAE, pathogen numbers decreased from ca. 3.3logCFU/package to ca. 1.5logCFU/package within 2h, but then increased to 7.3 and 6.7logCFU/package, respectively, after 120d. When frankfurters were formulated with lactate and diacetate and treated with LAE, pathogen numbers decreased by ca. 2.0logCFU/package within 2h and remained relatively unchanged over the 120d. These data confirm that LAE provides an initial lethality towards L. monocytogenes and when used in combination with reduced levels/ratio of lactate and diacetate as an ingredient for frankfurters provides inhibition throughout shelf life.

Effect of storage and subsequent reheating on viability of Listeria monocytogenes on pork scrapple.

We evaluated the fate of Listeria monocytogenes on commercial pork scrapple, a regionally popular, ready-to-eat (RTE) meat. We also conducted an informal survey to address consumer practices for storing and reheating scrapple. Of the 129 consumers who responded to at least one of the eight questions posed in the survey, about half (46.4%; 52 of 112) considered scrapple RTE, the majority (69.7%; 76 of 109) stored it in the refrigerator, and all (100%; 112 of 112) preferred to reheat it prior to consumption. Most respondents (83.9%; 94 of 112) reheated the scrapple by pan frying for 1 to 10 min at medium to high temperature. To study pathogen behavior, slices of pork scrapple were surface inoculated with a five-strain cocktail of L. monocytogenes (ca. 2.0 log CFU/g), vacuum sealed, and stored for up to 60 days. Pathogen levels increased to 8.9, 9.5, and 9.9 log CFU/g after 44 (4 degrees C), 21 (10 degrees C), and 5 (21 degrees C) days, respectively. When slices 1.3 cm (ca. 55 g) and 1.9 cm (ca. 85 g) thick were surface inoculated with L. monocytogenes (ca. 7.0 log CFU/g) and then reheated in a skillet (191 degrees C) for 0.5 to 4 min per side or to target instantaneous internal temperatures of 48.9 to 71.1 degrees C, it was possible to achieve pathogen reductions ranging from ca. 2.2 to 6.5 log CFU/g. These data confirm that in the unlikely event of postprocessing contamination of pork scrapple by L. monocytogenes, proper reheating can appreciably reduce levels of the pathogen before consumption.

Growth-Death Kinetics of Listeria Monocytogenes in Strained Yogurt

In this research, viability of Listeria monocytogenes was studied in strained yogurt (Turkish name: Torba yogurt) samples prepared in laboratory conditions and contaminated with various levels of L. monocytogenes before and after incubation. When strained yogurt samples (pre-incubation inoculation, A1 and post-incubation inoculation, B1) were contaminated with a 106 cfu/ml level of L. monocytogenes, their pH value reached to 4.08, and the presence of L. monocytogenes was defined only by using the enrichment method at the 30th day of the storage at the pH of 3.55, since the resistance of the bacteria had been lost in those yogurts. However, L. monocytogenes was completely inhibited at the 7th day of the storage for the strained yogurt samples (pre-incubation inoculation, A2 and post-incubation inoculation, B2) contaminated with the level of 104 cfu/ml when the pH value was 3.58. As a result, it can be stated that viability of Listeria monocytogenes may not be possible if the pH of the yogurt is around the ≤3.59 after the 7th day of the storage.

Adhesion to and Viability of Listeria monocytogenes on Food Contact Surfaces

Listeria monocytogenes is an important pathogen responsible for major outbreaks associated with food products. Adhesion to surfaces leads to significant modifications in cell physiology. The aim of this work was to determine the adhesion ability of 10 isolates of L. monocytogenes to eight materials commonly used in kitchens and to evaluate the viability of the adhered cells. The materials assayed were stainless steel 304, marble, granite, glass, polypropylene from a bowl and from a cutting board, and two kinds of silestone. All L. monocytogenes strains attached to all surfaces, although to different extents. L. monocytogenes adhered most tightly to granite and marble, followed by stainless steel 304, glass, silestones, and finally polypropylene surfaces. Surfaces at the threshold between hydrophobicity and hydrophilicity, with high electron acceptor capability and a regular pattern of roughness, were more prone to attachment. Polypropylene surfaces displayed the highest percentage of viable bacteria (nearly 100%), whereas marble and granite had a lower percentage of cultivable cells, 69.5 and 78.7%, respectively. The lowest percentage of culturable bacteria was found on white silestone (18.5%). These results indicate that there are differences in adhered cell viability on different materials. Cell viability assays are important to better understand the cross-contamination process because only adhered bacteria that remain viable are responsible for postprocess contamination.

Determination of the effect of plant essential oils obtained by supercritical fluid extraction on the growth and viability of Listeria monocytogenes in broth and food systems using flow cytometry

The antimicrobial properties of oregano, rosemary and laurel extracts obtained by supercritical fluid extraction were investigated by examining their influence on the growth and viability of Listeria monocytogenes in laboratory medium and broccoli juice at 30 and 8 °C. Important decreases in the L. monocytogenes population were shown in presence of all the extracts obtained from rosemary and one oregano extract. The counts were reduced below the level of detection after 4 h of exposure at 30 °C in laboratory medium. A bactericidal effect was observed also when L. monocytogenes was exposed to rosemary at 30 °C and 8 °C in broccoli juice. Significant reductions in growth rate and an increase in lag phase of L. monocytogenes were observed in presence of some of the laurel and oregano extracts at both temperatures. Flow cytometry was used as a rapid method to determine the antibacterial effect of supercritical extracts and the physiological state of L. monocytogenes. Bacterial viability performed by dual staining of L. monocytogenes with SYTO 9 and propidium iodide revealed three different cell populations, specifically, living, dead and compromised cells. Live cell percentage decreased with the time of exposure, whereas the percentage of compromised cells remained constant and the dead cells increased in the same period.

THE INHIBITORY EFFECT OF HONEY BEE VENOM AGAINST SOME DIFFERENT MICROOGANISMS

Venom collected from honey bee (Apis meilifera L.) workers of Egyptian, Carniolian and Carnionlian hybrid races were microbiologically examined for their inhibitory effect against different microorganisms by using disc diffusion technique. There was a clear inhibitory effect (IE) for all bee venom types and this (1E) was strongly dependent on venom type and the type of test microorganism. Bee venom from the Egyptian race showed the strongest inhibitory effect (133-23.? mm) against microorganisms while, the venom obtained from Carniolian race gave the lowest effect (10.3-15.3 mm). Bee venom from Camiolian hybrid race revealed a moderate inhibitory action (11.7490 mm). Gram—positive bacten'a as well as yeast and moulds demonstrated the most susceptible cells for venom type while, in contrary Gram- negative bacteria manifested quite tolerance for the antimicrobial action of bee venom. Antimicrobial activity of bee venom proved to be concentration—dependent and it had a bacteriostatic effect at low concentrations while. at higher concentrations it considered as bactericidal. The incorporation of bee venom in growth medium negatively affected the progress in growth behavior of microorganisms and this effect increased with increasing venom concentration. Viability of Listeria monocytogenes and E.coii cells in their growth media had been lost and completely died-off after 24 and 48 hrs of incubation in the presence of high venom concentration (1000 pgfml) respectively. The presence of a moderate venom concentration (400 pgiml) in the medium delayed the bacterial growth and finally cells disappeared after five days of exposure. Providing growth media with low venom concentrations considerably suppressed the growth of the two bacterial strains. This phenomenon can be used in medical purposes and drug preparations or by some modifications in food preservation.

Viability of Listeria monocytogenes on commercially-prepared hams surface treated with acidic calcium sulfate and lauric arginate and stored at 4 °C

We demonstrated the effectiveness of delivering an antimicrobial purge/fluid into shrink-wrap bags immediately prior to introducing the product and vacuum sealing, namely the “Sprayed Lethality In Container” (SLIC™) intervention delivery method. The pathogen was Listeria monocytogenes, the antimicrobials were acidic calcium sulfate (ACS; calcium sulfate plus lactic acid; 1:1 or 1:2 in dH 2O) and lauric arginate (LAE; Ethyl- N-dodecanoyl- l-arginate hydrochloride; 5% or 10% in dH 2O), and the product was commercially prepared “table brown” ham (ca. 3 pounds each). Hams were surface inoculated with a five-strain cocktail of L. monocytogenes (ca. 7.0 log 10 CFU per ham), added to shrink-wrap bags that already contained ACS or LAE, vacuum-sealed, and stored at 4 °C for 24 h. Pathogen levels decreased by 1.2, 1.6, 2.4, and 3.1 log 10 CFU/ham and 0.7, 1.6, 2.2, and 2.6 log 10 CFU/ham in samples treated with 2, 4, 6, and 8 mL of a 1:1 and 1:2 solution of ACS, respectively. In samples treated with 2, 4, 6, and 8 mL of a 5% solution of LAE, pathogen levels decreased by 3.3, 6.5, 5.6, and 6.5 log 10 CFU/ham, whereas when treated with a 10% solution of LAE pathogen levels decreased ca. 6.5 log 10 CFU/ham for all application volumes tested. The efficacy of ACS and LAE were further evaluated in shelf-life studies wherein hams were surface inoculated with either ca. 3.0 or 7.0 log 10 CFU of L. monocytogenes, added to shrink-wrap bags that contained 0, 4, 6, or 8 mL of either a 1:2 solution of ACS or a 5% solution of LAE, vacuum-sealed, and stored at 4 °C for 60 days. For hams inoculated with 7.0 log 10 CFU, L. monocytogenes levels decreased by ca.1.2, 1.5, and 2.0 log 10 CFU/ham and 5.1, 5.4, and 5.5 log 10 CFU/ham within 24 h at 4 °C in samples treated with 4, 6, and 8 mL of a 1:2 solution of ACS and a 5% solution of LAE, respectively, compared to control hams that were not treated with either antimicrobial. Thereafter, pathogen levels remained relatively unchanged (±1.0 log 10 CFU/ham ) after 60 days at 4 °C in hams treated with 4, 6, and 8 mL of a 1:2 solution of ACS and increased by ca. 2.0–5.0 log 10 CFU/ham in samples treated with 4, 6, and 8 mL of a 5% solution of LAE. For hams inoculated with 3.0 log 10 CFU, L. monocytogenes levels decreased by 1.3, 1.9, and 1.8 log 10 CFU/ham within 24 h at 4 °C in samples treated with 4, 6, and 8 mL of a 1:2 solution of ACS, respectively, compared to control hams that were not treated. Likewise, levels of the pathogen were reduced to below the limit of detection (i.e., 1.48 log 10 CFU/ham) in the presence of 4, 6, and 8 mL of a 5% solution of LAE within 24 h at 4 °C. After 60 days at 4 °C, pathogen levels remained relatively unchanged (±0.3 log 10 CFU/ham) in hams treated with 4, 6, and 8 mL of a 1:2 solution of ACS. However, levels of L. monocytogenes increased by ca. 2.0 log 10 CFU/ham in samples treated with 4 and 6 mL of a 5% LAE solution within 60 days but remained below the detection limit on samples treated with 8 mL of this antimicrobial. These data confirmed that application via SLIC™ of both ACS and LAE, at the concentrations and volumes used in this study, appreciably reduced levels of L. monocytogenes on the surface of hams within 24 h at 4 °C and showed potential for controlling outgrowth of the pathogen over 60 days of refrigerated storage.

Effect of Reheating on Viability of a Five-Strain Mixture ofListeria monocytogenes in Vacuum-Sealed Packages of Frankfurters following Refrigerated or Frozen Storage

The purpose of this study was to assess consumer preferences for storing and reheating frankfurters and to use this information to assess the effect of product formulation and storage times and temperatures on the viability ofListeria monocytogenes after reheating of frankfurters. Individual links were inoculated with about 8.0 log CFU per package of a five-strain mixture of the pathogen, vacuum sealed, and stored at 4°C for 3 and 15 days and at −18°C for 30 days. Frankfurters formulated with and without 2% added potassium lactate were heated to a surface temperature of 60, 70, 80, or 90°C for up to 8 min by submersing the packages in a thermostatically controlled circulating water bath. Surviving bacteria were recovered and counted by rinsing the contents of each package with sterile peptone water and plating this solution directly onto modified Oxford selective agar plates. In general, the results revealed that about a 5-log unit reduction was achieved by reheating to a surface temperature of 70°C for about 2 min or 80 or 90°C for about 0.6 min regardless of storage conditions or formulation. Product formulation did not appreciably affect the viability of the pathogen after heating; there was no appreciable difference in the number of cells surviving the heat treatment in product prepared with or without potassium lactate. These findings can be used to establish reheating guidelines for consumers to ensure that frankfurters, which may become contaminated with low levels ofL. monocytogenes prior to packaging and after unpackaging, are adequately reheated prior to consumption.

Antioxidant Power, Lipid Oxidation, Color, and Viability of Listeria monocytogenes in Beef Bologna Treated with Gamma Radiation and Containing Various Levels of Glucose

Ionizing radiation can be used to pasteurize ready-to-eat (RTE) meat products. Thermal processing of RTE meats that contain dextrose results in the production of antioxidants that may interfere with ionizing radiation pasteurization of RTE meat products. Beef bologna was manufactured with dextrose concentrations of 0, 2, 4, 6, and 8%. Antioxidant activity, as measured by the Ferric Reducing Antioxidant Power assay, increased with dextrose concentration but was unaffected by ionizing radiation. Lipid oxidation increased significantly in irradiated bologna (4 kGy) that contained dextrose. Hunter color analysis indicated that the addition of dextrose reduced the ionizing radiation-induced loss of redness (a-value) but promoted the loss of brightness (L-value). The radiation resistance, D10-value, of Listeria monocytogenes that was surface-inoculated onto bologna slices was not affected by dextrose concentration. L. monocytogenes strains isolated from RTE meats after listeriosis outbreaks were utilized. Increased antioxidant activity generated by thermal processing of dextrose in fine emulsion sausages does not present a barrier to radiation pasteurization of RTE meats. However, a high dextrose concentration in combination with gamma irradiation increases lipid oxidation significantly.