Evaluation of SDS and GRAS liquid disinfectants for mitigation of hepatitis A virus contamination of berries.

AIM: To evaluate generally recognized as safe (GRAS) liquid wash formulations against hepatitis A virus-contaminated strawberries and blackberries in order to identify a formulation suitable for reducing virus contamination. METHODS AND RESULTS: Formulations included the surfactant sodium dodecyl sulfate (SDS; 0·5% w/v) by itself, and in combination, with lactic acid (LA; 0·5% v/v), levulinic acid (LVA; 0·5% v/v) and 3 ppm aqueous chlorine dioxide (ClO2 ). After contamination and drying overnight, the average total extracted contamination for both untreated strawberries and blackberries was 4·4 log PFU. Three successive distilled H2 O only treatments reduced total contamination by up to 1·8 log PFU for both strawberries and blackberries, while wash formulations showed significant (P ≤ 0·05) total reductions ranging from 2·1 to 2·9 log PFU. CONCLUSIONS: Considering results for both berry types, the combination of ClO2 and SDS was the most effective. Overall results indicate that adding surfactant and several types of sanitizers to berry wash can enhance HAV reduction on berries. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that industry could enhance the virologic safety of ready-to-eat berries by the combined use of surfactant and sanitizer.

Effect of hot water surface pasteurization of whole fruit on shelf life and quality of fresh-cut cantaloupe.

Cantaloupes are associated with recent outbreaks of foodborne illnesses and recalls. Therefore, new approaches are needed for sanitization of whole and cut fruit. In the present study, whole cantaloupes were submerged into water in the following 3 conditions: 10 degrees C water for 20 min (control), 20 ppm chlorine at 10 degrees C for 20 min, and 76 degrees C water for 3 min. Populations of microflora were measured on the rinds of the whole cantaloupes. Quality and microbial populations of fresh-cut cantaloupes prepared from whole fruit were analyzed after 1, 6, 8, 10, 13, 16, and 20 d of storage at 4 degrees C. The hot water significantly reduced both total plate count (TPC) and yeast and mold count on rind of whole fruits while chlorine or cold water wash did not result in a significant reduction of microbial population. Fresh-cut pieces prepared from hot water-treated cantaloupes had lower TPC than the other 2 treatments in the later storage periods (days 13 to 20) in 2 of 3 trials. The hot water treatment of whole fruits was inconsistent in reducing yeast and mold count of fresh-cut pieces. Soluble solids content, ascorbic acid content, fluid loss, and aroma and appearance scores were not consistently affected by either hot water or chlorine treatment. Our results suggested that hot water pasteurization of whole cantaloupes frequently resulted in lower TPCs of fresh-cut fruit during storage and did not negatively affect quality of fresh-cut cantaloupes.

A study of U.S. orchards to identify potential sources of Escherichia coli O157:H7.

The association of unpasteurized apple cider with Escherichia coli O157:H7 foodborne illness has led to increased interest in potential reservoirs of this pathogen in the orchard. Fourteen U.S. orchards were surveyed in autumn 1999 to determine the incidence and prevalence of E. coli O157:H7, E. coli, total aerobic microflora, and yeasts and molds. Fruit samples (n = 63) (eight apple and two pear varieties) and soil, water, and fecal samples were collected. Samples were plated on (i) tryptic soy agar for total mesophilic aerobic count, (ii) E. coli and coliform Petrifilm for total coliforms and E. coli, and (iii) yeast and mold Petrifilm. Samples positive for coliforms and E. coli were enriched and tested for E. coli O157:H7. Fruit was also tested for internalization of microflora by aseptically removing the core, stem, and calyx areas, and the individual sections were assessed for the categories of microflora listed above. E. coli was detected in soil and water and in 6% of fruit samples (three pear samples and one apple sample), generally collected from areas previously designated as high risk in this study. However, no E. coli O157:H7 was found. Coliforms were found in 74% of fruit samples and were internalized in the cores of 40% of fruit tested. Yeasts and molds were internalized in 96.7% of samples and aerobic bacteria in 89.6%. E. coli was not found to be internalized. Total aerobic counts and total coliforms were higher in dropped and damaged fruit (P < 0.05). Findings suggest that dropped or damaged fruit should not be included in fruit designated for the production of unpasteurized juice or for the fresh or fresh-cut market. In addition, orchards should be located away from potential sources of contamination, such as pastures.

Efficacy of washing with a commercial flatbed brush washer, using conventional and experimental washing agents, in reducing populations of Escherichia coli on artificially inoculated apples.

Conventional and experimental washing formulations were applied with a commercial flatbed brush washer under conditions representative of commercial practice to determine their efficacy in decontaminating apples inoculated with a nonpathogenic Escherichia coli strain. Golden Delicious apples (18 kg) inoculated with E. coli were mixed with approximately 109 kg of uninoculated Fuji apples (distinctly different in appearance) in a wet dump tank containing 1,325 liters of water at 20 degrees C for 15 min. The combined apples were washed in a flatbed brush washer with the following washing solutions: water at 20 degrees C, water at 50 degrees C, 200 ppm of chlorine (pH 6.4) at 20 degrees C, 8% trisodium phosphate at 20 degrees C, 8% trisodium phosphate at 50 degrees C, 5% hydrogen peroxide at 20 degrees C, 5% hydrogen peroxide at 50 degrees C, 1% APL Kleen 245 at 50 degrees C, and two-stage washing treatments using the combination of 1% APL Kleen 245 at 20 or 50 degrees C followed by 5% hydrogen peroxide at 35 or 50 degrees C. None of the washing treatments tested under the conditions of this experiment significantly reduced the E. coli populations on the inoculated apples or in cider made from these apples, probably as a consequence of the inability of this washing system to inactivate or remove the bacterial cells in inaccessible calyx and stem areas of apples. These results are important because they demonstrate the need for new fruit washing technology that can overcome this limitation. Also, there was no significant cross-contamination of the Fuji apples in the dump tank. Significant cross-contamination of cider, made with uninoculated apples, occurred in the hammer mill and/or the press cloth when these units were not sanitized following a trial with inoculated apples.

The survival of Escherichia coli O157:H7 in the presence of Penicillium expansum and Glomerella cingulata in wounds on apple surfaces.

The survival of Escherichia coli O157:H7 in the presence of one of two plant pathogens, Penicillium expansum and Glomerella cingulata, in wounds on apples was observed during 14 days storage at room temperature (RT) and at 4 degrees C. The aim of this work was to determine if changes in apple physiology caused by the proliferation of fungal decay organisms would foster the survival of E. coli O157:H7. Trials were performed where (A) plant pathogens (4 log10 spores) were added to apple wounds 4 days before the wounds were inoculated with E. coli O157:H7 (3 log10 CFU g(-1) apple) (both RT and 4 degrees C storage), (B) plant pathogens and E. coli O157:H7 were added on the same day (both RT and 4 degrees C storage), and (C) E. coli O157:H7 was added 2 days (RT storage) and 4 days (4 degrees C storage) before plant pathogens. In all trials E. coli O157:H7 levels generally declined to <1 log10 at 4 degrees C storage, and in the presence of P. expansum at 4 degrees C or RT. However, in the presence of G. cingulata at RT E. coli O157:H7 numbers increased from 3.18 to 4.03 log10 CFU g(-1) in the apple wound during trial A, from 3.26 to 6.31 log10 CFU g(-1) during trial B, and from 3.22 to 6.81 log10 CFU g(-1) during trial C. This effect is probably a consequence of the attendant rise in pH from 4.1 to approximately 6.8, observed with the proliferation of G. cingulata rot. Control apples (inoculated with E. coli O157:H7 only) were contaminated with opportunistic decay organisms at RT during trials A and B, leading to E. coli O157:H7 death. However, E. coli O157:H7 in control apples in trial C, where no contamination occurred, increased from 3.22 to 5.97 log10 CFU g(-1). The fact that E. coli O157:H7 can proliferate in areas of decay and/or injury on fruit highlights the hazards associated with the use of such fruit in the production of unpasteurized juice.

Changes in membrane fatty acid composition of Pediococcus sp. strain NRRL B-2354 in response to growth conditions and its effect on thermal resistance.

Membrane fatty acid composition and thermal resistance (D value) of Pediococcus sp. were determined for mid-exponential-phase (ME) and stationary-phase (ST) cells grown in tryptic soy broth (TSB) and tryptone-glucose-yeast extract (TGY) at 28 and 37 degrees C. As the cells entered the stationary phase of growth, the unsaturated fatty acid, C18:1 n11c, produced during the exponential phase of growth was converted to its cyclic form, C19:0 Delta9c. This shift in membrane fatty acid composition was accompanied by an increase in the D values of this bacterium. Data from this study suggest that the membrane fatty acid composition of Pediococcus sp. is dependent on the growth conditions and that membrane fatty acid composition plays a critical role in thermal resistance. Thermal inactivation curves of Pediococcus sp. cells grown in TGY at 28 degrees C indicated the presence of a cell population that is heterogeneous in thermal resistance. The growth of this bacterium in TGY at 37 degrees C and in TSB at 28 and 37 degrees C resulted in cell populations that were uniform in thermal resistance with a lag time for thermal inactivation. Thermal inactivation curves of ME and ST cultures were similar. The data presented here suggest that the cell population's uniformity of thermal inactivation is independent of the growth phase of the culture.

Inactivation of microorganisms with microwaves at reduced temperatures.

We developed a pilot-plant nonthermal flow process using microwave energy to inactivate microorganisms. The process consists of multiple passes through the microwave generator. Each passed material goes to a receiving tank for subsequent passes. The flow rate was 0.96 to 1.26 kg/min and the dwell time per pass was 1.1 to 1.5 min. Five passes were used. The microwave energy is instantaneously and simultaneously applied to the system, and thermal energy is removed by a cooling tube within the process line in the microwave generator. The cooling tube maintains the temperature below 40 degrees C. There was significant reduction in microorganisms in water, 10% glucose solution, and apple juice, and in yeast in beer. There was a slight decrease in microorganisms in tomato juice, pineapple juice, apple cider, and beer; and no effect in skim milk.

Influence of growth medium on thermal resistance of Pediococcus sp. NRRL B-2354 (formerly Micrococcus freudenreichii) in liquid foods.

Pediococcus sp. is a nonpathogenic heat-resistant spoilage organism that has been used as a test organism in milk pasteurization studies. These characteristics make this bacterium an attractive test organism to study the mode of bacterial thermal inactivation in a food pilot plant. We report here the effect of growth medium on the thermal D value of this organism in skim milk, whole liquid egg, 10% glucose solution, pineapple juice, apple juice, tomato juice, and water at 60 degrees C. Thermal inactivation was done in a submerged coil; D values were calculated from the linear portion of the survival curves by linear regression analysis. The range of D values of stationary-phase cells grown at 28 degrees C in tryptone glucose yeast extract (TGY) or tryptic soy broth (TSB) was 0.14 to 12.05 min in all heating menstrua tested. The TSB-grown cells exhibited the highest thermal resistance with skim milk and 10% glucose solution as the heating menstrua. Survival curves of the TGY-grown cells indicated the presence of a cell population heterogeneous in thermal resistance. The TSB-grown cells exhibited a cell population uniform in thermal resistance and with a lag time for thermal inactivation. When compared to TGY-grown cells, Pediococcus sp. grown in TSB showed a significant (P < 0.05) increase in D values by up to eightfold in all heating menstrua. Results from this study suggested that thermal inactivation of Pediococcus sp. was dependent on the growth medium and on the heating menstruum with respect to both pH and composition.

Critical role of anteiso-C15:0 fatty acid in the growth of Listeria monocytogenes at low temperatures.

Listeria monocytogenes is a food-borne pathogen capable of growth at refrigeration temperatures. Membrane lipid fatty acids are major determinants of a sufficiently fluid membrane state to allow growth at low temperatures. L. monocytogenes was characterized by a fatty acid profile dominated to an unusual extent (> 95%) by branched-chain fatty acids, with the major fatty acids being anteiso-C15:0, anteiso-C17:0, and iso-C15:0 in cultures grown in complex or defined media at 37 degrees C. Determination of the fatty acid composition of L. monocytogenes 10403S and SLCC 53 grown over the temperature range 45 to 5 degrees C revealed two modes of adaptation of fatty acid composition to lower growth temperatures: (i) shortening of fatty acid chain length and (ii) alteration of branching from iso to anteiso. Two transposon Tn917-induced cold-sensitive mutants incapable of growth at low temperatures had dramatically altered fatty acid compositions with low levels of i-C15:0, a-C15:0, and a-C17:0 and high levels of i-C14:0, C14:0, i-C16:0, and C16:0. The levels of a-C15:0 and a-C17:0 and the ability to grow at low temperatures were restored by supplementing media with 2-methylbutyric acid, presumably because it acted as a precursor of methylbutyryl coenzyme A, the primer for synthesis of anteiso odd-numbered fatty acids. When mid-exponential-phase 10403S cells grown at 37 degrees C were temperature down-shocked to 5 degrees C they were able, for the most part, to reinitiate growth before the membrane fatty acid composition had reset to a composition more typical for low-temperature growth. No obvious evidence was found for a role for fatty acid unsaturation in adaptation of L. monocytogenes to cold temperature. The switch to a fatty acid profile dominated by a-C15:0 at low temperatures and the association of cold sensitivity with deficiency of a-C15:0 focus attention on the critical role of this fatty acid in growth of L. monocytogenes in the cold, presumably through its physical properties and their effects, in maintaining a fluid, liquid-crystalline state of the membrane lipids.

Cold stress proteins induced in Listeria monocytogenes in response to temperature downshock and growth at low temperatures.

Listeria monocytogenes is a food-borne pathogen with the ability to grow at refrigerator temperatures. Twelve cold shock proteins (Csps) with apparent M(r)s of 48,600, 41,000, 21,800, 21,100, 19,700, 19,200, 18,800, 18,800, 17,200, 15,500, 14,500, and 14,400 were induced by cold shocking L. monocytogenes 10403S from 37 to 5 degrees C, as revealed by labeling with L-[35S]methionine followed by two-dimensional gel electrophoresis. Strain SLCC53 showed a similar response. Cold acclimation proteins were observed in cultures of strain 10403S growing at 5 degrees C, and four of these proteins, with apparent M(r)s 48,000, 21,100, 19,700, and 18,800, were also Csps. Two cold-sensitive transposon-induced mutants were labeled less efficiently than the parent strain, but the Csp response of the mutant examined was very similar to that of the parent strain.

Isolation and characterization of Clostridium acetobutylicum mutants with enhanced amylolytic activity.

Clostridium acetobutylicum mutants BA 101 (hyperamylolytic) and BA 105 (catabolite depressed) were isolated by using N-methyl-N'-nitro-N-nitrosoguanidine together with selective enrichment on the glucose analog 2-deoxyglucose. Amylolytic enzyme production by C. acetobutylicum BA 101 was 1.8- and 2.5-fold higher than that of the ATCC 824 strain grown in starch and glucose, respectively. C. acetobutylicum BA 105 produced 6.5-fold more amylolytic activity on glucose relative to that of the wild-type strain. The addition of glucose at time zero to starch-based P2 medium reduced the total amylolytic activities of C. acetobutylicum BA 101 and BA 105 by 82 and 25%, respectively, as compared with the activities of the same strains grown on starch alone. Localization studies demonstrated that the amylolytic activities of C. acetobutylicum BA 101 and BA 105 were primarily extracellular on all carbohydrates tested.

Regulation and localization of amylolytic enzymes in Clostridium acetobutylicum ATCC 824.

Amylolytic activity was primarily cell associated when Clostridium acetobutylicum was grown on glucose or maltose and primarily extracellular when grown on dextrin or starch. Total amylolytic activity decreased with increasing glucose concentration. When this microorganism was grown in P2 medium containing starch, the intracellular amylolytic activity was 90% membrane bound and 10% cytoplasmic in nature. The addition of 1% glucose to 2% starch-based P2 medium at different stages of growth indicated that this carbohydrate repressed the synthesis of C. acetobutylicum amylolytic enzymes at the level of transcription.