Listeria monocytogenes infection of HD11, chicken macrophage-like cells.

Listeria monocytogenes can be carried by and infect poultry, although the clinical disease in birds is rare. Escape from macrophage phagocytosis is a key step in pathogenesis for L. monocytogenes. Therefore, we investigated the infection of the chicken macrophage-like cell line HD11 with 2 strains of L. monocytogenes EGD-e and Scott A. After infection, L. monocytogenes was quantified by spread plating and HD11 was quantified with trypan blue exclusion stain before enumeration. The standard macrophage killing protocols require washing the cell monolayers 3 times with PBS, which was found to negatively influence HD11 monolayers. Maximum bacterial densities within macrophages were not different between the 2 Listeria strains. HD11 required more than 11 h to effectively reduce intracellular L. monocytogenes Scott A, and Scott A was more susceptible to HD11 killing than EGD-e. It appears that Listeria infection initially causes attenuation of HD11 growth, and infected HD11 cells do not begin to lyse until at least 11 h post infection. These results suggest that there are subtle strain to strain differences in response to HD11 macrophage phagocytosis. The long lead-time required for HD11 to kill L. monocytogenes cells means that there is sufficient time available for chicken macrophages to circulate in the blood and transfer the intracellular Listeria to multiple tissues.

Listeria monocytogenes batch culture growth response to metabolic inhibitors.

In certain environments nutrient and energy sources available to microorganisms can be limited. Foodborne pathogens must efficiently adapt in order to be successfully transmitted through the food chain to their hosts. For the intracellular foodborne pathogen Listeria monocytogenes, little is known regarding its response to nutrient/energy-limiting conditions. The alternative stress responsive sigma factor σ(B) has been reported to contribute to survival under specific stresses. Therefore, the effects of several metabolic inhibitors on growth of L. monocytogenes wild-type and a ΔsigB mutant were examined. In the absence of inhibitors, both strains reached stationary phase after 18 h at 23°C and 10 h at 37°C. All of the metabolic inhibitors slowed growth of either strain, with few differences observed among the different inhibitors.

Dietary fiber content influences soluble carbohydrate levels in ruminal fluids.

The soluble carbohydrate concentration of ruminal fluid, as affected by dietary forage content (DFC) and/or ruminally undegradable intake protein content (UIPC), was determined. Four ruminally cannulated steers, in a 4 × 4 Latin square design, were offered diets containing high (75 % of DM) or low (25 % of DM) DFC and high (6 % of DM) or low (5 % of DM) UIPC, in a 2 × 2 factorial arrangement. Zinc-treated SBM was the primary UIP source. Soluble hexose concentration (145.1 µM) in ruminal fluid (RF) of steers fed low DFC diets exhibited a higher trend (P = 0.08) than that (124.5 µM) of steers fed high DFC diets. UIPC did not modulate (P = 0.54) ruminal soluble hexose concentrations. Regardless of diet, soluble hexose concentration declined immediately after feeding and did not rise until 3 h after feeding (P < 0.0001). Cellobiose (≈90 %) and glucose (≈10 %) were the major soluble hexoses present in RF. Maltose was not detected. Soluble glucose concentration (13.0 µM) was not modified by either UIPC (P = 0.40) nor DFC (P = 0.61). However, a DFC by post-prandial time interaction was detected (P = 0.02). Pentose concentrations were greater (P = 0.02) in RF of steers fed high DFC (100.2 µM) than steers fed low DFC (177.0 µM). UIPC did not influence (P = 0.35) soluble pentose concentration. The identity of soluble pentoses in ruminal fluid could not be determined. However, unsubstituted xylose and arabinose were excluded. These data indicate that: (i) soluble carbohydrate concentrations remain in ruminal fluid during digestion and fermentation; (ii) slight diurnal changes began after feeding; (iii) DFC influences the soluble carbohydrate concentration in RF; and (iv) UIPC of these diets does not affect the soluble carbohydrate concentration of RF.

Cleaning and decontamination efficacy of wiping cloths and silver dihydrogen citrate on food contact surfaces.

AIMS: To test the efficacy of four wipe cloth types (cotton bar towel, nonwoven, microfibre and blended cellulose/cotton) with either quaternary ammonia cleaning solution or silver dihydrogen citrate (SDC) in cleaning food contact surfaces. METHODS: Swab samples collected from untreated, cloth-treated and cloth disinfectant-treated surfaces were subjected to hygiene monitoring using adenosine triphosphate (ATP) bioluminescence and aerobic total plate counting (TPC) assays. RESULTS: Adenosine triphosphate measurements taken after wiping the surfaces showed poor cleaning by nonwoven cloths (2·89 RLU 100 cm(-2) ) than the microfibre (2·30 RLU 100 cm(-2) ), cotton terry bar (2·26 RLU 100 cm(-2) ) and blended cellulose/cotton cloth types (2·20 RLU 100 cm(-2) ). The cellulose/cotton cloth showed highest log reduction in ATP-B RLU values (95%) and CFU values (98·03%) when used in combination with SDC disinfectant. CONCLUSIONS: Cleaning effect of wiping cloths on food contact surfaces can be enhanced by dipping them in SDC disinfectant. ATP-B measurements can be used for real-time hygiene monitoring in public sector, and testing microbial contamination provides more reliable measure of cleanliness. SIGNIFICANCE AND IMPACT OF THE STUDY: Contaminated food contact surfaces need regular hygiene monitoring. This study could help to estimate and establish contamination thresholds for surfaces at public sector facilities and to base the effectiveness of cleaning methods.

Assessment of a food microbiology senior undergraduate course as a potential food safety distance education course for poultry science majors.

Distance education courses have become popular due to the increased number of commuter students as well as people already in the workforce who need further education for advancement within their careers. A graduate-level Web-based course entitled Special Topics-Poultry Food Safety Microbiology was developed from an existing senior undergraduate advanced food microbiology course in the Poultry Science Department at Texas A&M University. Conversion of standard lecture material into a distance education course can provide unique challenges to maintain comparable course content in an asynchronous manner. The overall objective for this course was to examine bacterial activities including ecology in food, animals, raw and processed meat, eggs, and human pathogenesis. Students were surveyed at the end of the class and the majority agreed that they would be willing to take the course as an online course, although they were not willing to pay an extra fee for an online course. The majority of students used the online version of the course as a supplement to the classroom rather than as a substitute.

Control of Listeria monocytogenes by lauric arginate on frankfurters formulated with or without lactate/diacetate.

Listeria monocytogenes (Lm) is a food safety concern that can be associated with ready-to-eat (RTE) meat and poultry products because of its persistence in the processing environment. Listeriosis has a fatality rate of 28% in immuno-compromised individuals. RTE meats receive a lethal heat treatment but may become contaminated by Lm after this treatment. Federal regulators and manufacturers of RTE meats are working to find additional ways to control postprocess contamination by Lm in RTE meats. This research was initiated to validate combinations of antimicrobials that would produce an immediate lethality of at least 1 log of Lm on artificially contaminated frankfurters, and also suppress Lm growth to less than 2 logs throughout the extended shelf life at refrigerated temperatures (4 degrees C). Based on our studies, 22-ppm lauric arginate (LAE, ethyl-N-dodecanoyl-L-arginate hydrochloride) gave more than a 1-log reduction of Lm surface inoculated onto frankfurters within 12 h. The combination of either 1.8%/0.13% or 2.1%/0.15% potassium lactate/sodium diacetate (L/D) in combination with 22 ppm LAE caused more than a 2-log reduction at 12 h. Storage studies revealed that complementary interactions of L/D and LAE also met the 2nd requirement. This combination initially reduced Lm by 2 logs and suppressed growth to less than 2 logs even at the end of the 156-d storage life for frankfurters. These results confirmed that the combination of L/D with LAE as a postprocessing-prepackaging application could be useful in complying with the USDA's Alternative 1 that requires validation for the control of Lm on RTE frankfurters.

Identification of Listeria innocua surrogates for Listeria monocytogenes in hamburger patties.

Listeria innocua M1 has been used by many researchers as a nonpathogenic thermal processing surrogate for Listeria monocytogenes. However, L. innocua M1 has been criticized because its thermal survivability characteristics are not as closely parallel to L. monocytogenes as some would like in a variety of foods and processing conditions. The present study was conducted to compare multiple L. innocua and L. monocytogenes strains to validate L. innocua M1 as the ideal surrogate under high-temperature thermal processing conditions for L. monocytogenes. The D- and z-values of L. innocua M1, L. innocua strain SLCC 5639 serotype (6a), SLCC 5640 (6b), SLCC 2745 (4ab), and L. monocytogenes F4243 (4b) were calculated for raw hamburger patties. Hamburger patties were inoculated with 10(7-8) CFU/g of L. monocytogenes or L. innocua. Samples were heat treated at 4 temperatures (62.5 to 70 degrees C). At each temperature, the decimal reduction time (D-value) was obtained by linear regression of survival curves. The D- and z-values were determined for each bacterium. The D-values of L. innocua and L. monocytogenes serotypes ranged from 3.17 to 0.13 min at 62.5 to 70 degrees C, and the z-values of L. innocua and L. monocytogenes were 7.44 to 7.73 degrees C. Two of the 4 L. innocua serotypes used in this experiment have the potential for use as surrogates in hamburger meat with varying margins of safety. L. innocua M1 should serve as the primary nonpathogenic surrogate with the greatest margin of safety in verifying a new thermal process to destroy L. monocytogenes.

Orange essential oils antimicrobial activities against Salmonella spp.

Seven citrus essential oils (EOs) were screened by disc diffusion assay for their antibacterial act against 11 serotypes/strains of Salmonella. The 3 most active oils were selected to determine the minimal inhibitory concentration (MIC) against the same Salmonella. Orange terpenes, single-folded d-limonene, and orange essence terpenes all exhibited inhibitory activity against the Salmonella spp. on the disc diffusion assay. EOs were stabilized in broth by the addition of 0.15% (w/v) agar for performance of the MIC tests. Orange terpenes and d-limonene both had MICs of 1%. The most active compound, terpenes from orange essence, produced an MIC that ranged from 0.125% to 0.5% against the 11 Salmonella tested. Gas chromatography-mass spectrometry (GC/MS) analysis revealed that this orange essence oil was composed principally of d-limonene, 94%, and myrcene at about 3%. EOs from citrus offer the potential for all natural antimicrobials for use in improving the safety of organic or all natural foods.

Improving ground beef safety and stabilizing color during irradiation using antioxidants, reductants or TSP.

The objective of this research was to extend ground beef retail display life using antioxidants, reductants, and/or TSP treatments combined with electron beam irradiation. Ground beef was produced with added butylated hydroxyanisole (BHA) plus butylated hydroxytoluene (BHT) with the following combinations; (1) ascorbate; (2) trisodium phosphate(buffer); (3) erythorbate; (1) and (2); (1) and (3); (1), (2), and (3); and an untreated control, C. Half of the treated samples were irradiated (I) at 2.0kGy-absorbed dose under a nitrogen atmosphere, half remained non-irradiated (N). Samples were displayed under atmospheric oxygen and evaluated for total aerobic plate count (TPC), thiobarbituric acid reactive substances (TBARS) and instrumental color during 9d of simulated retail display (SRD). Controls had the highest (P<0.05) TBARS value and the lowest (P<0.05) redness (CIE a∗), proportion of oxymyoglobin and vividness. Treated irradiated samples were just as red and vivid on SRD day 9 as the non-irradiated untreated control at day 0. Treatments stabilized color and lipids of ground beef after irradiation and during SRD.

A 2-step cooking method of searing and hot water pasteurization to maximize the safety of refrigerated, vacuum packaged, chicken breast meat.

Americans consume almost 40 kg per capita of chicken each year. Increasing consumption of chicken surpassed pork in 1982 and beef in 1992. The objectives of this study were to examine the effectiveness of a novel, 2-step cooking method of grilling, slicing, vacuum packaging, and hot water pasteurization to inhibit the growth of Listeria monocytogenes in chicken breast meat. Because this study required the use of pilot plant scale pasteurization equipment, Listeria innocua M1, a nonpathogen with slightly greater heat resistance than L. monocytogenes, was used as a surrogate. We first examined the lethal effects of grilling on a boneless skinless chicken breast to mimic cross-contaminated, surface-inoculated Listeria. Searing produced a mean reduction of 2.5 log CFU/g of Listeria and a moisture loss of only 7% (w/w). A 2nd experiment studied the lethal effect of pasteurization of the sliced seared chicken breast. L. innocua M1 inoculated between the slices mimicked contamination in deep muscle. Pasteurization in a 71 degrees C bath (final internal temperature of 66 degrees C) gave an additional 2.3 log CFU/g reduction. L. innocua M1 did not show significant regrowth during a wk of refrigerated storage. The combined 2-step cooking method of searing and pasteurization gave a combined 4.8 log reduction in LI M1. In parallel tests a non-Listeria indicator, Corynebacterium glutamicum, inoculated between sliced, seared chicken, showed a 3 log reduction after pasteurization for 10 min in a 71 degrees C bath compared to 2.3 log reduction of Listeria. Corynebacterium regrowth occurred much faster than did L. innocua M1.

The pre- and post-grinding application of rosemary and its effects on lipid oxidation and color during storage of ground beef.

The timing of the application of rosemary extract was evaluated as one-way of minimizing myoglobin and lipid oxidation in ground beef. In experiment 1, rosemary extract was added to beef at four different stages namely trim, cube, coarse, and fine ground beef. The beef was evaluated for color and TBARS values during 144h of storage (4°C). Results showed that when rosemary was added to the pre-grinding treatments of trim and cube, ground beef had the highest a(∗) values (redness), oxymyoglobin content, and lowest TBARS values at 144h. In experiment 2, the effect of rosemary extract was evaluated on the color quality of case ready ground beef inoculated with 10(7)CFU/g Escherichia coli. Microbial counts, color, and TBARS values were measured during 144h of simulated storage. The results showed that both the rosemary treated samples that were inoculated and uninoculated remained redder longer and had lower TBARS values than the untreated inoculated and uninoculated controls. There was no significant inhibition of E. coli by the rosemary extract.