Leakage of intracellular UV materials of high hydrostatic pressure-injured Escherichia coli O157:H7 strains in tomato juice.

The behavior of high hydrostatic pressure-injured Escherichia coli O157:H7 cells (strain SEA13B88 and a strain from the June-July 1999 Oklahoma juice outbreak) in tomato juice (pH 4.1) and phosphate-buffered saline (PBS; pH 7.2) at final concentrations of 8.4 to 8.8 log CFU/ml, respectively, and treated at 400, 500, and 600 MPa for 40 min at 25 and 35 degrees C with storage at 5 and 23 degrees C for 1,800 min was investigated. Immediately after treatment and every 3 h for 24 h of storage, an aliquot (0.1 ml) was plated on Trypticase soy agar and sorbitol MacConkey agar to determine the percentage of injured population. Leakage of UV materials and possible recovery from injury were investigated. Pressure (600-MPa) treatment at 35 degrees C for 40 min caused a higher percentage of bacterial injury than for 10 min of treatment. A higher percentage of injured population was found among the Oklahoma strain cells than among strain SEA13B88 cells, and differences in viability loss for bacterial strains were determined. The viability loss determined in PBS was 4.8 log for SEA13B88 cells and 5.2 log for Oklahoma cells, while losses of 5.4 and 5.7 log were determined in tomato juice for SEA13B88 and Oklahoma cells, respectively. The leakage of intracellular materials of injured Oklahoma cells was higher than that observed for SEA13B88 cells, but injured Oklahoma cells recovered faster in PBS. However, injured and healthy populations for both strains were below detection in tomato juice stored at 5 degrees C for 1,440 min.

Effect of hydrostatic pressure pulsing on the inactivation of Salmonella enteritidis in liquid whole egg.

Eggs and egg-containing foods contaminated with bacterial human pathogens have been implicated in numerous foodborne outbreaks leading to costly recalls. Research was undertaken to investigate the use of high pressure-pulse treatment to inactivate Salmonella Enteritidis inoculated in liquid egg. Liquid egg was inoculated with Salmonella Enteritidis (8.0 log colony-forming units [CFU]/mL) and exposed to hydrostatic pressures (300-400 MPa) and pressure (350 MPa) pulsing at 25 degrees C, 40 degrees C, and 50 degrees C for up to 40 minutes to determine the maximum allowable pressure that can inactivate the Salmonella with minimal injury. Pressure treatments (350 and 400 MPa) at 25 degrees C for up to 40 minutes reduced the population of Salmonella Enteritidis by approximately 4.8 and 6.0 log(10) CFU/mL, respectively. High pressure (350 MPa) treatment at 50 degrees C and 2-minute pulses at four cycles for a total of 11.4 minutes, including the come-up and come-down times, led to a significant (p < 0.05) inactivation of Salmonella Enteritidis in liquid egg without causing coagulation. However coagulation occurred in the liquid egg at 400 MPa pressure treatment for 10 minutes at 50 degrees C. No Salmonella population was recovered in this liquid egg stored at 4 degrees C, 25 degrees C, and 37 degrees C for 24 hours suggesting that 350 MPa hydrostatic pressure and pulsing treatment is a better alternative for inactivation of Salmonella in liquid egg than continuous pressure treatment.

Combined efficacy of nisin and pediocin with sodium lactate, citric acid, phytic acid, and potassium sorbate and EDTA in reducing the Listeria monocytogenes population of inoculated fresh-cut produce.

The inability of chlorine to completely inactivate human bacterial pathogens on whole and fresh-cut produce suggests a need for other antimicrobial washing treatments. Nisin (50 microg/ml) and pediocin (100 AU/ml) individually or in combination with sodium lactate (2%), potassium sorbate (0.02%), phytic acid (0.02%), and citric acid (10 mM) were tested as possible sanitizer treatments for reducing the population of Listeria monocytogenes on cabbage, broccoli, and mung bean sprouts. Cabbage, broccoli, and mung bean sprouts were inoculated with a five-strain cocktail of L. monocytogenes at 4.61, 4.34, and 4.67 log CFU/g, respectively. Inoculated produce was left at room temperature (25 degrees C) for up to 4 h before antimicrobial treatment. Washing treatments were applied to inoculated produce for 1 min, and surviving bacterial populations were determined. When tested alone, all compounds resulted in 2.20- to 4.35-log reductions of L. monocytogenes on mung bean, cabbage, and broccoli, respectively. The combination treatments nisin-phytic acid and nisin-pediocin-phytic acid caused significant (P < 0.05) reductions of L. monocytogenes on cabbage and broccoli but not on mung bean sprouts. Pediocin treatment alone or in combination with any of the organic acid tested was more effective in reducing L. monocytogenes populations than the nisin treatment alone. Although none of the combination treatments completely eliminated the pathogen on the produce, the results suggest that some of the treatments evaluated in this study can be used to improve the microbial safety of fresh-cut cabbage, broccoli, and mung bean sprouts.

Effect of sanitizer treatments on Salmonella Stanley attached to the surface of cantaloupe and cell transfer to fresh-cut tissues during cutting practices.

The ability of Salmonella Stanley to attach and survive on cantaloupe surfaces, its in vivo response to chlorine or hydrogen peroxide treatments, and subsequent transfer to the interior tissue during cutting was investigated. Cantaloupes were immersed in an inoculum containing Salmonella Stanley (10(8) CFU/ml) for 10 min and then stored at 4 or 20 degrees C for up to 5 days. Periodically, the inoculated melons were washed with chlorine (1,000 ppm) or hydrogen peroxide (5%), and fresh-cut tissues were prepared. The incidence of Salmonella Stanley transfer from the rinds to the fresh-cut tissues during cutting practices was determined. A population of 3.8 log10 CFU/cm2 of Salmonella Stanley was recovered from the inoculated rinds. No significant (P < 0.05) reduction of the attached Salmonella population was observed on cantaloupe surfaces stored at 4 or 20 degrees C for up to 5 days, and the population was not reduced after washing with water. Salmonella Stanley was recovered in fresh-cut pieces prepared from inoculated whole cantaloupes with no sanitizer treatment. Washing with chlorine or hydrogen peroxide solutions was most effective immediately after inoculation, resulting in an approximate 3.0-log10 CFU/cm2 reduction, and the level of recovered Salmonella population transferred to fresh-cut samples was reduced to below detection. The effectiveness of both treatments diminished when inoculated cantaloupes stored at 4 or 20 degrees C for more than 3 days were analyzed, and the fresh-cut pieces prepared from such melons were Salmonella positive. Salmonella outgrowth occurred on inoculated fresh-cut cubes stored above 4 degrees C.

Bioluminescence ATP assay for estimating total plate counts of surface microflora of whole cantaloupe and determining efficacy of washing treatments.

The surface microflora of cantaloupes were estimated using a bioluminescence ATP assay, and results were compared to plate count data. Cantaloupes were treated as follows: (i) water washed, or (ii) washed in solutions of sodium hypochlorite (1,000 mg/liter) or hydrogen peroxide (5%) for 5 min. Bioluminescence ATP assay results showed differences in ATP level/cm2 of cantaloupes dipped in chlorine or hydrogen peroxide solution; ATP levels in these washed samples were lower than in controls due to antimicrobial action of the treatments on the cantaloupe surface. Linear correlations were found between the bioluminescence ATP assay and aerobic plate counts of unwashed cantaloupe (r2 = 0.995) and those washed with water (r2 = 0.990) determined before storage. Lower correlations between the bioluminescence ATP assay and the aerobic plate counts were observed on cantaloupes stored for 120 h at 20 degrees C (r2 = 0.751) than at 4 degrees C (r2 = 0.980) without washing treatment. Lower correlation at 20 degrees C may be the result of clusters or growth that occurred in chains. ATP levels of washed cantaloupes correlated well with bacterial plate counts (r2 = 0.999). A reliable minimum detectable threshold using the bioluminescence ATP assay was established at 3 log10 fg/cm2 corresponding to 4 log10 CFU/cm2. Bioluminescence ATP assay is not recommended for washed samples where the microbial load is near or below the threshold. Therefore, the bioluminescence ATP assay will be recommended for quick estimation of total microbial load on cantaloupe surfaces where the population is expected to exceed this threshold. The assay can save the industry time by eliminating the required incubation required by the conventional methods.

Bioluminescence measurements of the antilisterial activity of nisin: comparison with ampicillin and streptomycin.

The influence of nisin on intracellular ATP and cell numbers of Listeria monocytogenes strain Scott A was determined and compared with the effect of amplicillin and streptomycin under similar conditions. In the presence of nisin (3-12 micrograms/ml), intracellular ATP and cell numbers decreased rapidly during the first hour at 35 degrees C and extracellular ATP increased. Cell numbers and intracellular ATP increased after 3 h of incubation. No effect was observed in cells treated with ampicillin (3-12 micrograms/ml) and streptomycin (15-60 micrograms/ml) during the first hour. However, concentrations of > or = 3 micrograms/ml ampicillin and > or = 30 micrograms/ml streptomycin were listeriostatic after 3 h of incubation. Progressive loss of viability and reduction of intracellular ATP were observed in resting cells in PBS (pH 7.2) containing increasing concentrations of the antimicrobials. Rapid accumulation of extracellular ATP, observed immediately after treatment with nisin but not with the antibiotics, supports the reported collapse of proton motive force in L. monocytogenes by nisin.