Kinetics and mechanism of bacterial inactivation by ultrasound waves and sonoprotective effect of milk components.

Inactivation of Escherichia coli and Listeria monocytogenes were investigated in buffer and milk upon treatment with ultrasound waves (USW). In addition, sonoprotective effect of milk components and ultrasound-induced changes in bacterial cells were investigated using scanning electron microscopy (SEM). Bacterial cells were added to phosphate buffer, whole milk, skim milk, or simulated milk ultrafiltrate (SMUF). To determine the sonoprotective effect of milk components, lactose (5%), casein (3%), or ß lactoglobulin (0.3%) was added to SMUF. Samples were sonicated with 24 kHz pulse USW while maintaining the system temperature between 30 to 35 °C. Aliquots were drawn at set times during sonication and bacteria were enumerated by surface plating appropriate dilutions on selective and nonselective media plates. Escherichia coli exhibited significantly higher D values in whole (2.43 min) and skim milk (2.41 min) than phosphate buffer (2.19 min). Listeria monocytogenes also showed higher D values in whole (9.31 min) and skim milk (8.61 min) compared to phosphate buffer (7.63 min). Data suggest that milk exerts a sonoprotective effect on these bacteria. Escherichia coli exhibited a log-linear inactivation kinetics followed by tailing whereas L. monocytogenes showed 1st-order kinetics throughout. Among the milk components tested, presence of lactose in SMUF resulted in significantly higher D values than SMUF for both organisms suggesting that lactose was exerting a protective effect on bacteria. SEM images showed that USW caused mechanical damage to the cell wall and cell membrane of bacteria leading to their inactivation.

Role of bacterial association and penetration on destruction of Escherichia coli O157:H7 in beef tissue by high pH.

This study was undertaken to determine if association with collagen enables Escherichia coli O157:H7 to resist high-pH treatments and to determine the effects of high pH on the survival of E. coli O157:H7 within different layers of beef tissue. E. coli O157:H7 was inoculated onto purified bovine type I collagen on 12-mm2 circular glass coverslips, plain 12-mm2 circular glass coverslips (control), and 12-mm2 irradiated (cobalt-60) lean beef tissue. The rates of destruction of E. coli O157:H7 inoculated on coverslips in pH 10.5 NaHCO3-NaOH buffer at 35 degrees C were determined at various sampling times. E. coli O157:H7 cells associated with collagen and treated in the same manner were also examined using scanning electron microscopy to determine if association with collagen enabled the organism to resist high-pH treatments. The inoculated tissue was treated in pH 13.0 NaHCO3-NaOH buffer at 25 degrees C, and penetrating cells of E. coli O157:H7 were recovered using a cryostat technique. There was no significant difference (P < 0.05) between the rates of destruction of collagen-associated E. coli O157:H7 and non-collagen-associated E. coli O157:H7 following exposure to high-pH treatments. Scanning electron micrographs showed that collagen-associated E. coli O157:H7 cells appeared physically damaged by exposure to high-pH treatments, and association of E. coli O157:H7 to collagen did not increase the resistance of the organism to destruction by high-pH rinses. No significant differences were seen between 20 ml of NaHCO3-NaOH buffer at pH 13.0 (treatment) and 20 ml of distilled water at pH 7.0 (control) when E. coli O157:H7 cells were recovered in beef tissue at depths of up to 2,000 microm (P < 0.05). The ability of E. coli O157:H7 to penetrate beef tissue may be an important factor in reducing the effectiveness of high-pH treatments in killing this organism on beef tissue. This finding should be considered in the future when designing treatments to decontaminate beef carcasses.

Destruction of Escherichia coli O157:H7 and Salmonella typhimurium in Lebanon bologna by interaction of fermentation pH, heating temperature, and time.

Fermented meats have caused food-borne illness due to enterohemorrhagic Escherichia coli. Consumption of Lebanon bologna was epidemiologically associated wit a recent outbreak of salmonellosis. The present study was conducted to determine the effect of pH (after the fermentation step), final heating temperature, and time on destruction of E. coli O157:H7 and Salmonella typhimurium in Lebanon bologna. Raw Lebanon bologna mix was inoculate with either of the pathogens (ca.10(8) CFR/g and fermented for 12 h at 80 degrees F (26.7 degrees C) and then at 100 degrees F (37.8 degrees C) unit the pH reached wither 5.2 or 4.7. The mix was then heated to 110, 115, or 120 degrees F (43.3, 46.1, or 48.9 degrees C). The bologna was sampled at various times, decimally diluted, and plated on either McConkey sorbitol agar or XLD agar to enumerate E. coli O157:H7 and S. typhimurium, respectively. Fermentation alone reduced populations of both pathogens by < 2 log units and heating alone reduced populations of E. coli O157:H7 by < 3 log units. A combination of fermenting to either pH 5.2 or 4.7, followed by heating at 110 degrees F (43.3 degrees C) for 20h, 115 degrees F (46.1 degrees C) for 10 h, or 120 degrees F (48.9 degrees C) for 3 h reduced populations of both pathogens by > 7 log units. Overall S. typhimurium cells were either equally or significantly less resistant (P < 0.01) than cells of E. coli O157:H7. Significantly interactions (P < 0.01) among the three factors for the destruction of E. coli O157:H7 were observed. A process-specific regression equation was developed to predict the destruction of E. coli O157:H7 in Lebanon bologna.

The role of defeathering in the contamination of turkey skin by Salmonella species and Listeria monocytogenes.

This study was undertaken to determine whether the incidence of either Salmonella spp. or Listeria monocytogenes on turkeys at three commercial processors could be related to the type of defeathering system: 1) conventional, 58 C common bath scald; 2) kosher, 7 C common bath scald; or 3) steam-spray, 62 C nonimmersion scald. Flocks were sampled before defeathering, after defeathering, and after chill at each facility. The incidence of Salmonella-positive turkeys significantly increased subsequent to conventional defeathering (10 positive out of 14) as compared with before defeathering (3/14). The number of Salmonella-positive carcasses following kosher (0/14) and steam-spray (2/14) defeathering were similar to the number of Salmonella-positive carcasses found prior to defeathering (1/14 and 3/14, respectively). The incidence of Salmonella-positive carcasses following chill was slightly lower, but not significantly different than the number of Salmonella-positive carcasses found immediately following defeathering at all processors (8/14, 0/14, 1/14 for conventional, kosher, and steam-spray processors, respectively). Although L. monocytogenes was detected on turkeys sampled before chilling (2/10, kosher) and after chilling (8/14, kosher; 1/14, conventional), no L. monocytogenes was detected on turkeys at any of the processors prior to the evisceration process. Flocks with high aerobic plate counts prior to processing were more likely to contain Salmonella-positive birds throughout processing. Aerobic plate counts of all flocks were similar after chill whether or not Salmonella spp. and L. monocytogenes were detected.

Effect of type of defeathering system on Salmonella cross-contamination during commercial processing.

The cross-contamination effects of three commercial defeathering systems were compared using turkeys from a single Salmonella-positive flock (< or = 15% cloacal-positive). Single or "common" flocks were used to control flock-to-flock variability. Thirty birds were mechanically defeathered in each system as the first flock of the day and compared with 30 hand-defeathered (control) birds. Three trials, each using a different common flock, were completed. In Trial 1, the incidence of Salmonella-positive birds decreased following mechanical defeathering at all three processors. The incidence of Salmonella-positive carcasses in test flocks increased following steam-spray (approximately 100%) and kosher (approximately 50%) defeathering in Trials 2 and 3, whereas no increase in Salmonella-positive carcasses resulted from conventional defeathering. The decrease in the number of Salmonella-positive birds as a result of defeathering observed in Trial 1, as compared to increases observed in Trials 2 and 3, may be related to the selection of feather-contaminated (Trial 1) vs intestinal-colonized (Trials 2 and 3) turkeys. Surface temperature of the carcasses and length of time required to defeather were monitored within each system. It is hypothesized that the increases in the number of Salmonella-positive birds following steam-spray and kosher defeathering in Trials 2 and 3 were a result of skin surface changes occurring during the defeathering process, which allowed increased adherence or entrapment of Salmonella spp. on or within remaining skin layers.

Enhanced thermal destruction of Listeria monocytogenes and Staphylococcus aureus by the lactoperoxidase system.

The lactoperoxidase system (LPS) enhanced thermal destruction of Listeria monocytogenes and Staphylococcus aureus. After LPS activation, biphasic survival curves were observed for L. monocytogenes at 57.8 degrees C and for S. aureus at 55.2 degrees C. The data were consistent with a model that assumed two bacterial populations differing in heat sensitivity. The more heat-sensitive fractions (93% of the L. monocytogenes, 92% of the S. aureus) were killed almost instantly. For these biphasic survival curves, D values were based on the much smaller, less-heat-sensitive fractions. For L. monocytogenes, the D52.2 degrees C values were 30.2 min (untreated milk) and 10.7 min (LPS activated); corresponding D55.2 degrees C values were 8.2 and 1.6 min; corresponding D57.8 degrees C values were 2.3 and 0.5 min. For S. aureus, the D52.2 degrees C values were 33.3 min (untreated milk) and 2.2 min (LPS activated), and the corresponding D55.2 degrees C values were 7.6 and 1.1 min, respectively. The most rapid killing of L. monocytogenes occurred when samples were heated soon after activation of the LPS. Activation of the LPS followed by heating can increase the margin of safety with respect to milkborne pathogens.

Nipple centralization for the correction of breast deformity from segmental mastectomy.

A breast deformed by lateral tissue deficiency and severe lateral displacement of the nipple, caused by the treatment of an early breast cancer with segmental mastectomy and radiotherapy, was corrected by a modification of techniques used commonly for mastopexy. The nipple and areola were moved medially on a central pedicle to create the illusion that the missing lateral tissues had been restored. Simultaneously, the opposite breast was reduced to improve symmetry. The result was a normalization of breast appearance without the need for distant tissue. It is possible that modifications of this approach could be used to treat similar deformities in other quadrants of the breast as well.

Acidification Process Technology to Control Thermophilic Spoilage in Canned Mushrooms.

Alternative processes involving the use of citric acid and/or EDTA in mushroom canning operations were evaluated and compared to a standard commercial process as means to control thermophilic spoilage. An average of 68% thermophilic spoilage was observed with the standard control process. Spoilage was reduced to an average of 23.9% by the addition of citric acid to the can brine, and to 16.8% when 500 ppm EDTA was also added to the can brine. However, the best results, 2.4% average spoilage, were observed when mushrooms were vacuum hydrated in a buffered citric acid solution (0.05M, pH 3.5) and EDTA was added to the can brine at 200 ppm equilibrium concentration. This treatment was as the Acid-Vacuum Hydration-Chelation (A-VH-C) Process. Bacteriological evaluation indicated that the A-VH-C Process caused no significant reduction in product spore load counts (after blanching) compared to the control, but did reduce spore load counts after thermal processing. However, cans from all treatments contained viable spores. Outgrowth studies conducted with spores that survived thermal processing and inoculated into Beef Extract Tryptone Iron (BETI) both indicated that spores from cans processed with the A-VH-C Process had the longest generation time. Similar experiments where the BETI broth was treated to simulate the conditions in the cans indicated that the addition of EDTA to the medium had the greatest effect on reducing outgrowth rate of surviving spores.

Prevalence and Distribution of Campylobacter jejuni and Campylobacter coli in Retail Meats.

Nine cooperating laboratories, distributed throughout the United States, determined the interlaboratory reproducibility of a sensitive, selective method for isolation of Campylobacter jejuni and Campylobacter coli from foods, and determined the prevalence and distribution of the organism in retail meats. A double-blind inoculated/recovery experiment demonstrated the ability to detect two cells of C. jejuni and C. coli per g of meat at a rate of 96% among the cooperating laboratories. However, a 7.5% false-positive rate for the presumptive detection of the organism was also reported. Samples of ground beef, beef flank steak, lamb stew meat, broiler chicken, pork sausage (without antimicrobials), and pork chops were selected to assess the presence of campylobacters. Each cooperator purchased five of each of the above samples from the refrigerated case of two retail outlets at quarterly intervals throughout the year. A total of 2,160 retail samples were analyzed for the presence of C. jejuni and C. coli . Results indicated that about 30% of the 360 chickens sampled yielded the organism. Analysis of 1,800 red meat products yielded campylobacters at a rate of about 5.1%. Pork samples yielded C. coli and other meats yielded C. jejuni . Higher numbers of isolations from the red meats were made during June and September (8.6%) as compared with December and March (4.2%). These results provide a baseline, for the prevalence of campylobacters in these selected foods, and also support epidemiologic data associating mishandled foods of animal origin as a potential vehicle in human gastroenteritis.

Selective method for the isolation of Sporolactobacillus from food and environment sources.

A detection procedure was developed in which a modified Lactobacilli MRS medium was used to enrich for Sporolactobacillus from a variety of foods, feed soil and environmental samples. Each sample was rinsed with 50 ml of a modified Lactobacilli MRS broth containing 1.0% (w/v) alpha-methyl glucoside 0.1% (w/v) potassium sorbate, 0.00224% (w/v) bromocresol green indicator, adjusted to pH 5.5 with acetic acid and incubated at 37 degrees C for 7 d under 5% CO2. Volumes of 2 ml from each sample were heat shocked at 80 degrees C for 5 min and 0.1 ml spread onto plates of Lactobacilli MRS agar (Difco), pH 5.5 and APT agar (BBL), pH 5.5. Plates were incubated for 5 d and suspect colonies were tested for catalase production, benzidine reaction, nitrate reduction, motility and Gram stain reaction. This method was demonstrated to be selective for Sporolactobacillus.

The microbiology of apples and apple products.

The apple industry has reached an annual production level of 8.5 billion pounds. CA storage of 25% of this crop has enabled a fresh market on a year-round basis. To achieve high quality in raw fruit and processed apple products, careful attention must be paid to maintaining a microbiologically stable environment. The ecology of the microflora associated with the apple is a reflection of the orchard, handling, harvesting, and storage practices. Yeasts predominate on orchard fruit, molds may become a storage problem, and bacteria cause spoilage, off flavors, and loss of quality in juice products. Despite the microbial problems inherent in producing of quality product, the apple industry is faced with the occurrence of patulin. Patulin, a mycotoxin produced by Penicillium and Aspergillus species, has been associated with damaged fruit. Decreased temperatures, coupled with CA storage; can deter mold growth and patulin production. Laboratory detection methods for derivations of patulin are able to detect microgram quantities. Means to eliminate patulin formed in apple products include addition of ascorbate and SO2, extending fermentation, or charcoal filtering. However, degradation products of patulin have not been evaluated toxicologically.

Occurrence ofVibrio and other bacteria on the sea nettle,Chrysaora quinquecirrha.

Sixty-two specimens of the sea nettle,Chrysaora quinquecirrha, were caught in the lower Chesapeake Bay, homogenized, and samples plated on a yeast extract-Bay water agar. Bacterial colonies were selected randomly, purified, and tested for 180 characteristics. Computer analysis permitted clustering of the 208 isolates into 15 groups (comprised of 133 strains) plus 75 nongrouped strains which failed to associate with any group at the 70% similarity level. The majority of the isolates (68.8%) wereVibrio species. These included 110 of the grouped strains (forming 12 of the 15 groups) and 33 of the nongrouped strains. The remainder of the isolates were distributed as follows:Pseudomonas (11.6%),Bacillus (8.2%),Flavobacterium (2.4%),Acinetobacter (2.4%),Moraxella (1.9%),Cytophaga (1.9%), Gram-positive cocci (1.4%), and miscellaneous (1.4%). All theBacillus were isolated from a group of moribund nettles and reflect an abnormal condition.Vibrio species predominated in the five "catches" of healthy nettles, but were distinctly different for each catch.