Aubrey F. Mendonca

Radiation Resistance and Virulence of Listeria monocytogenes Scott A following Starvation in Physiological Saline

The influence of starvation on the resistance of Listeria monocytogenes Scott A to electron beam irradiation in 0.85% (wt/vol) NaCl (saline) and in ground pork was investigated. Exponential- or stationary-phase cells (control) were grown at 35 degrees C in tryptic soy broth supplemented with 0.6% yeast extract. Washed cells were starved for 12 days in saline, and virulence of the pathogen was evaluated at 0, 8, and 12 days during starvation. Samples of saline and irradiation-sterilized ground pork, inoculated with control or starved cells, were irradiated at doses ranging from 0.0 to 2.5 kGy. L. monocytogenes survivors were determined by plating diluted samples of saline or pork on tryptic soy agar supplemented with 0.6% yeast extract and counting bacterial colonies following incubation (35 degrees C, 48 h). Virulence of starved cells and control was not significantly different (P > 0.05). Cells exhibited the highest radiation resistance at 8 days of starvation. Irradiation (0.5 kGy) in saline resulted in approximately 7.14, 5.55, and 2.38 log reduction in exponential, stationary, and starved cells, respectively. Irradiation of ground pork at 2.5 kGy reduced controls by approximately 6.0 log, whereas starved cells were reduced by only 3.8 log. Starved cells consistently exhibited higher irradiation D10-values than controls (P < 0.05). D10-values for exponential, stationary, and starved cells were 0.07, 0.09, and 0.21 kGy and 0.35, 0.42, and 0.66 kGy in saline and ground pork, respectively. These results indicate that starvation cross-protects L. monocytogenes Scott A against radiation inactivation and should be considered when determining this pathogens irradiation D-value.

Combining pediocin with postpackaging irradiation for control of Listeria monocytogenes on frankfurters

Frankfurters, in 1-link, 5-link, or 10-link packages, were surface inoculated with a five-strain mixture of Listeria monocytogenes (3.40 or 5.20 log CFU/g) after treatment with 3,000 arbitrary units (AU) or 6,000 AU of pediocin (in ALTA 2341) per link. The frankfurters were vacuum packaged, after which the 1-link and 5-link packages were irradiated at 1.2 or 2.3 kGy and the 10-link packages were irradiated at 1.4 or 3.5 kGy. L. monocytogenes was enumerated following the treatments. Selected treatments were subsequently evaluated during storage at 4, 10, and 25 degrees C for up to 12 weeks. Combination of pediocin with postpackaging irradiation at 1.2 kGy or more was necessary to achieve a 50% reduction of L. monocytogenes on frankfurters in 1-link or 5-link packages. The combination of 6,000 AU of pediocin and irradiation at 2.3 kGy or more was effective in all package sizes for inhibition of the pathogen for 12 weeks at 4 or 10 degrees C. There was a synergistic effect between pediocin and irradiation for inhibition of L. monocytogenes. Storage at 4 degrees C enhanced the antilisterial effects of the treatment combinations, with little or no growth of the pathogen in 1-link or 5-link packages during 12 weeks of storage. In general, these treatments did not affect the sensory quality of frankfurters.

Effect of dietary vitamin E and irradiation on lipid oxidation, color, and volatiles of fresh and previously frozen turkey breast patties §

Turkey breast meat patties, prepared from the turkeys fed diets containing 0, 50, 100, or 200 IU of dl-α-tocopheryl acetate (TA) per kg diet from 84 to 112 days of age, were aerobically packaged and irradiated at 0, 1.5, or 2.5 kGy. When dietary TA was increased from 0 to 200 IU/kg diet, plasma and muscle vitamin E levels increased by 5- and 4-fold, respectively. Dietary TA at 100 IU/kg diet significantly improved the storage stability of turkey breast, and it was more distinct in irradiated than nonirradiated meats. Both irradiation and dietary TA increased a*-values of turkey breast meat, but irradiation had a stronger impact. The redness of meat decreased during the 7-day storage, but irradiated meat maintained redder color than nonirradiated. Irradiated meat produced more sulfur volatiles and aldehydes than nonirradiated meats, and dietary TA effectively reduced these compounds during storage. The effects of dietary TA on the reduction of off-odor volatiles were more distinct in previously frozen-stored meats than in fresh meats.

Inhibitory Effects of Organic Acid Salts for Control of Listeria monocytogenes on Frankfurters

Sodium diacetate (SD), sodium diacetate plus potassium benzoate (SD-PB), and sodium lactate plus sodium diacetate plus potassium benzoate (SL-SD-PB) were selected for initial effectiveness against Listeria monocytogenes on frankfurters. Treatments were evaluated at -2.2, 1.1, 4.4, 10.0, and 12.8 degrees C for up to 90 days. The compounds were applied as 3 or 6% (total concentration) dipping solutions for surface treatment of the frankfurters. The treated frankfurters were inoculated with a five-strain cocktail of L. monocytogenes (Scott A 4b, H7764 1/2a, H7962 4b, H7762 4b, and H7969 4b) using 1 ml of 10(4) cells for each 90.8-g package of two frankfurters. The maximum population of L. monocytogenes was decreased and generation time and lag phase were increased after surface treatments with 6% SD, 6% SL-SD-PB, 3% SD-PB, and 6% SD-PB solutions at 1.1 degrees C. Surface treatment of frankfurters with SD at 6% was more effective for inhibiting L. monocytogenes growth than were the other treatments. Under the conditions of this study, L. monocytogenes survived in refrigerated storage even in the presence of the additives tested.

Combining Pediocin (ALTA 2341) with Postpackaging Thermal Pasteurization for Control of Listeria monocytogenes on Frankfurters

Frankfurters packaged in 1-link, 5-link, or 10-link packages were surface-inoculated with a five-strain mixture of Listeria monocytogenes (3.40 or 5.20 log CFU/g) after treatments with 3,000 arbitrary units (AU) or 6,000 AU pediocin (in ALTA 2341) per link. The frankfurters were vacuum packaged, after which the packages were heated in hot water at 71, 81, or 96 degrees C for 30, 60, or 120 s. L. monocytogenes was enumerated following the treatments. Selected treatments were subsequently evaluated during storage at 4, 10, and 25 degrees C for up to 12 weeks. L. monocytogenes was reduced by all treatments, but 81 degrees C or more for at least 60 s in combination with pediocin (Pdn-6000) was necessary to achieve a 50% reduction of initial inoculations. Heat treatments were most effective for 1-link packages and least effective for 10-link packages. Little or no growth of L. monocytogenes occurred on frankfurters for 12 weeks at 4 or 10 degrees C, and for 12 days at 25 degrees C. Generally, the treatments mentioned above did not significantly (P > 0.05) affect the sensory qualities of frankfurters. Therefore, pediocin (in ALTA 2341) in combination with postpackaging thermal treatment offers an effective treatment combination for improved control of L. monocytogenes on frankfurters.

Antilisterial effects of gravinol-s grape seed extract at low levels in aqueous media and its potential application as a produce wash.

Grape seed extract (GSE) is a rich source of proanthocyanidins, a class of natural antioxidants reported to have wide-ranging bioactivity as anti-inflammatory, anticarcinogenic, and antimicrobial agents. The ability of GSE to rapidly inactivate Listeria monocytogenes in vitro and the generally recognized as safe status of GSE make this extract an attractive candidate for control of Listeria in or on foods. Previously, GSE has been used at relatively high concentrations (1%) in complex food matrices and in combination with other antimicrobials. We sought to characterize the antilisterial effects of a commercial GSE preparation (Gravinol-S) alone at much lower concentrations (0.00015 to 0.125%) in aqueous solution and to test its possible use as an antimicrobial wash for fresh produce surfaces. Based on broth microdilution tests, the MICs of GSE against L. monocytogenes Scott A and Listeria innocua ATCC 33090 were as low as 50 and 78 mug ml(-1), respectively. GSE was evaluated in 0.85% saline against live cells of L. innocua via flow cytometry, using propidium iodide as a probe for membrane integrity. At sub-MICs and after only 2 min of exposure, treatment with GSE caused rapid permeabilization and clumping of L. innocua, results that we confirmed for L. monocytogenes using fluorescence microscopy and Live/Dead staining. At higher concentrations (0.125%), GSE reduced viable cell counts for L. monocytogenes by approximately 2 log units within 2 min on tomato surfaces. These results suggest the potential for GSE as a natural control of Listeria spp. on low-complexity foods such as tomatoes.

Effect of Ethylenediaminetetraacetate and Lysozyme on the Antimicrobial Activity of Ovotransferrin Against Listeria monocytogenes

This study evaluated the effect of EDTA and lysozyme on the antibacterial activities of activated ovotransferrin against 5 strains of Listeria monocytogenes. First, a disc test was performed to screen the concentrations of EDTA or lysozyme that showed antibacterial activities in ovotransferrin (O) or ovotransferrin in 100 mM NaHCO3 (OS) solution. Turbidity and viability tests were conducted using O or OS solution combined with either lysozyme (OL and OSL) or EDTA (OE and OSE). Also, OS combined with 2 mg/mL of lysozyme (OSL) or 1 mg/mL of EDTA (OSLE), or both, was applied on commercial hams to determine if the solutions show antibacterial activities on meat products. The effect of initial cell population on the antibacterial activities of ovotransferrin combined with either EDTA or lysozyme was also determined. The L. monocytogenes started to grow after 1 d of incubation in the presence of >2.0 mg/mL of lysozyme. The OL groups showed weak antibacterial activities against L. monocytogenes in brain heart infusion broth culture, and their activities were bacteriostatic. The OSL groups were bactericidal against L. monocytogenes, resulting in 1 log reduction from initial cell population. Even though OSL showed stronger antibacterial activity than OS, lysozyme had no significant effect on antibacterial activity of OS against L. monocytogenes. Also, EDTA itself at 1.0 and 2.0 mg/mL was bacteriostatic against 5 strains of L. monocytogenes. They were more susceptible to EDTA than lysozyme, and OSE1 and OSE2 had bactericidal activity against L. monocytogenes. There was a significant difference in the survivor cell populations between OS and OSE groups (P < 0.05). Therefore, EDTA enhanced the antibacterial activity of OS against L. monocytogenes. However, ovotransferrin plus either lysozyme or EDTA, or both, did not show any antibacterial effect in commercial hams during storage at 10 degrees C. In addition, the initial population of L. monocytogenes cells influenced the antibacterial activity of OSL or OSE.

Incubation of curing brines for the production of ready-to-eat, uncured, no-nitrite-or-nitrate-added, ground, cooked and sliced ham

Salt concentration, vegetable juice powder (VJP) concentration and temperature were investigated to determine necessary conditions for incubation of curing brines including VJP and a starter culture containing Staphylococcus carnosus prior to production of naturally cured, no-nitrate/nitrite-added meat products. Subsequently, incubated brines were utilized to produce no-nitrate/nitrite-added sliced ham in which quality characteristics and residual nitrite concentrations were measured to determine feasibility of brine incubation for nitrate conversion prior to injection. Two ham treatments (one with VJP and starter culture; one with pre-converted VJP) and a nitrite-added control were used. No differences (P>0.05) were found for color in the VJP treatments. Control sliced ham was redder after 42 days of storage, retaining significantly (P<0.05) greater a* (redness) than either of the VJP treatments. Residual nitrite concentration was greater (P<0.05) in the control hams during the first week of storage. While the nitrite-added control retained greater red color and initially had more residual nitrite than the VJP treatments, the two VJP treatments did not differ from each other.

Fate of Listeria monocytogenes in ready-to-eat turkey breast rolls formulated with antimicrobials following electron-beam irradiation

The objective of this study was to determine the effect of antimicrobials on the survival and proliferation of Listeria monocytogenes in turkey breast rolls following electron-beam irradiation. Six antimicrobial additive treatments that include no preservatives (control), 0.1% potassium benzoate (PB), 2% sodium lactate (SL), 0.1% potassium benzoate plus 2% sodium lactate (PB + SL), 2% sodium lactate plus 0.1% sodium diacetate (SL + SDA), and 0.1% potassium benzoate, 2% sodium lactate, and 0.1% sodium diacetate (PB + SL + SDA) were used. Sliced turkey breast rolls were artificially inoculated with approximately 10(6) cfu/cm(2) of 5-strain L. monocytogenes cocktails, then vacuum-packaged and irradiated at 0, 1.0, 1.5, 2.0, or 2.5 kGy. The radiation dose (kGy) that results in 90% reduction of viable cells for breast rolls, D(10) value, with various additive treatments ranged from 0.56 to 0.58 kGy. Adding PB (0.1%) or SL (2%) in turkey rolls failed to prevent L. monocytogenes from growing during refrigerated storage. In turkey rolls added with 2 (PB + SL or SL + SDA) or 3 (PB + SL + SDA) antimicrobial combinations had 2 or 3 wk of lag phases before L. monocytogenes growth, respectively. Irradiating turkey rolls, which were added with PB + SL or SL + SDA, at 1.0 kGy was effective in suppressing the growth of L. monocytogenes for about 6 wk when stored at 4 degrees C. No growth of L. monocytogenes after irradiation occurred during 42 d of storage for 2.0 kGy irradiated breast rolls formulated with 0.1% PB + 2% SL, 2% SL + 0.1% SDA or 0.1% PB + 2% SL + 0.1% SDA, and 1.0 kGy irradiated turkey breast with 0.1% PB + 2% SL + 0.1% SDA. Sensory panelists found that low-dose irradiation (1.0 kGy) had no effect on the sensory characteristics of ready-to-eat turkey breast rolls. Including SL + SDA had slightly negative effect for nonirradiated turkey breast rolls, but the sensory characteristics of 1.0 kGy irradiated turkey roll containing SL + SDA was not significantly different from the others receiving 1.0 kGy irradiation. For microbial safety, PB + SL and SL + SDA antimicrobial treatments combined with 1.0 kGy or 2.0 kGy irradiation are a promising technology.

Temperature abuse affects the quality of irradiated pork loins

The influence of temperature abuse on the quality of irradiated pork loins was investigated. Pork loins were obtained directly from a local packing plant, sliced and vacuum-packaged. Pork loins were randomly separated into 3 groups, sliced, and assigned to receive 0, 1.5, or 2.5 kGy electron-beam irradiation. Then, each chop was further cut into three equal pieces and assigned to three temperature treatments: Trt I was placed in a refrigerator directly after irradiation; Trt II was left at room temperature for 3 h before refrigeration; and Trt III was exposed at room temperature for 1 h three consecutive days with intermittent storage at 4 °C between exposures. Before irradiation, each loin pieces were vacuum-packaged. Color, 2-thiobarbituric acid reactive substances (TBARS), and volatiles were measured after 0, 14, 28 and 42 days of storage, and water-holding capacity and sensory characteristics of the loins were measured after 0, 14 and 28 days of storage. Temperature abuse had no significant effect on color, oxidation, and volatiles of irradiated pork loins. However, temperature abuse improved water-holding capacity of meat, which could be caused by the accelerated hydrolysis of muscle proteins at higher temperature. Irradiation increased redness, sulfur contents in volatiles and off-odor of pork loin. Off-odor and redness induced by irradiation sustained during storage. Among sulfur compounds, the content of dimethyl disulfide decreased gradually while the level of thiourea remained relatively constant. Irradiation also increased water loss, which might be related to the structural damage in membrane during irradiation. This study shows that temperature abuse has little effect on the quality of irradiated pork.