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Biomedical subjects

L R Beuchat

Publications and source records attributed to L R Beuchat.

At least 19 recordsLinked to original sources

Fermentation of low-salt miso as affected by supplementation with ethanol.

Steam-cooked soybeans and rice koji were combined (1:1, w/w), mixed with 5% (w/w) NaCl and ground into a fine paste. Samples (30 g) were deposited in nylon/polyethylene plastic bags and supplemented with 10 ml of aqueous ethanol solutions to give concentrations of 0, 2.5, 5, 7.5, 10, 15, 20, and 25% ethanol. Mixtures were homogenized, sealed, and incubated at 28 degrees C for eight weeks. Mold populations were less than 3 log10 CFU/g in all miso products after four weeks of fermentation. Yeast populations increased to 6.1 log10 CFU/g in the control (0% added ethanol) during the first week of fermentation and remained stable throughout the eight-week fermentation period. Yeasts were not detected in products containing 5-25% ethanol. Populations of lactic acid bacteria (LAB) increased to 6 log10 CFU/g after one week of fermentation in products containing 0 and 2.5% ethanol. However, after eight weeks of fermentation, LAB populations in all products were less than 4 log10 CFU/g. Rapid decreases in pH occurred only in products supplemented with 0 or 2.5% ethanol. Percentages of soluble protein in miso products containing various ethanol concentrations during the eight-week fermentation period revealed that protease activity was still active or not greatly inhibited in products supplemented with less than 10% ethanol. In comparison, koji enzymes were comparatively less affected by ethanol than were populations of molds, yeasts, and LAB. Total soluble carbohydrate and glucose contents were higher in products supplemented with 5, 7.5 and 10% ethanol than in other products. Discoloration (browning) during fermentation occurred most rapidly in products supplemented with 5 or 7.5% ethanol. Sensory evaluation of the low-salt (5%) product supplemented with 7.5% ethanol and fermented for eight weeks revealed normal or enhanced flavor ratings compared to ratings for a commercial product.

Aspergillus oryzae

Influence of temperature shifts on survival, growth, and toxin production by psychrotrophic and mesophilic strains of Bacillus cereus in potatoes and chicken gravy.

A study was done to determine the influence of temperature on growth and toxin production characteristics of psychrotrophic and mesophilic strains of Bacillus cereus when inoculated into mashed potatoes and chicken gravy containing various concentrations of sodium chloride and held at temperatures different from those at which cells had been cultured. Logarithmic growth phase cells (10 h, 30 degrees C) of psychrotrophic (F3802A/84) and mesophilic (B4ac-1) strains of Bacillus cereus were inoculated into rehydrated commercially processed instant mashed potatoes and chicken gravy supplemented with 0, 2, or 4% sodium chloride. Growth, survival, and diarrheal toxin production in potatoes and gravy held at 30, 37, and 10 degrees C (strain F3802A/84) or 30, 40, and 10 degrees C (strain B4ac-1) were monitored. Both strains grew in both foods containing no added sodium chloride or 2% sodium chloride when held at 30, 37, or 40 degrees C for 2 days. Strain B4ac-1 grew better than strain F3802A/84 in foods containing 4% sodium chloride. Maximum amounts of enterotoxin (1024 ng/g) were produced by strain B4ac-1 in chicken gravy held at 30 and 40 degrees C. Strain F3802A/84 grew to populations of 7 log10 CFU/g in foods containing no added sodium chloride or 2% sodium chloride at 10 degrees C. Strain F3802A/84 produced the highest amount of enterotoxin (1024 ng/g) at 30 degrees C in chicken gravy containing 0.7 or 2% sodium chloride; however, little or low amounts of toxin (4-16 ng/g) were produced in chicken gravy at 10 degrees C. Compared to strain B4ac-1, cells of strain F3802A/84 subjected to a downward shift in incubation temperature (10 degrees C) grew more rapidly in chicken gravy. Strain B4ac-1 produced the highest amount of toxin (1024 ng/g) at 30 degrees C in gravy containing 4% sodium chloride and at 40 degrees C in gravy containing 0.7% sodium chloride. Toxin was not detected in inoculated mashed potatoes. Results of this study indicate that shifts in incubation temperature influence growth and toxin production by psychrotrophic and mesophilic strains of B. cereux differently. It is important to store pasteurized, ready-to-eat foods at a temperature low enough to prevent the growth of B. cereus.

Animals

Tolerance of acid-adapted and non-adapted Escherichia coli O157:H7 cells to reduced pH as affected by type of acidulant.

A study was carried out to determine if three strains of Escherichia coli O157:H7 grown (18 h) in Tryptic Soy Broth (TSB) and TSB supplemented with 1.25% glucose (TSBG), i.e. unadapted and acid-adapted cells, respectively, exhibited changes in tolerance to reduced pH when plated on Tryptic Soy Agar (TSA) acidified (pH 3.9, 4.2, 4.5, 4.8, 5.1 and 5.4) with acetic, citric or malic acids. All test strains grew well on TSA acidified with acetic acid at pH > or = 5.4 or malic acid at pH > or = 4.5; two strains grew on TSA acidified with citric acid at pH > or = 4.5, while the third strain grew at pH > or = 4.8. Acid-adapted and control (unadapted) cells differed little in their ability to form visible colonies on TSA containing the same acid at the same pH. However, on plates not showing visible colonies, acid-adapted cells retained higher viability than unadapted cells when plated on acidified TSA. Growth of acid-adapted and control cells of E. coli O157:H7 inoculated into TSB containing acetic acid (pH 5.4 and 5.7) and citric or malic acids (pH 4.2 and 4.5) was also studied. There was essentially no difference in growth characteristics of the two types of cells in TSB acidified at the same pH with a given acid. Tolerance of acid-adapted and control cells on subsequent exposure to low pH is influenced by the type of acidulant. The order of sensitivity at a given pH is acetic > citric > malic acid. When performing acid challenge studies to determine survival and growth characteristics of E. coli O157:H7 in foods, consideration should be given to the type of acid to which cells have been exposed previously, the procedure used to achieve acidic environments and possible differences in response among strains. The use of strains less affected by pH than type of acidulant or vice versa could result in an underestimation of the potential for survival and growth of E. coli O157:H7 in acid foods.

Acids, Acyclic

Effect of shift in growth temperature on tolerance of psychrotrophic and mesophilic strains of Bacillus cereus to heat and sodium chloride.

A shift in growth temperature of a psychrotrophic (F3802A/84) strain and a mesophilic strain (B4ac-1) of Bacillus cereus grown at 30 degrees C for 10 h, then at 37 degrees C or 40 degrees C for 14 h, enhanced thermotolerance. Sodium chloride, at concentrations of 2.0 and 4.0% in brain heart infusion (BHI) broth, had no effect on thermotolerance of strain B4ac-1 heated at 50 degrees C, whereas the same concentrations of NaCl caused a decrease in thermotolerance of strain F3802A/84 heated at 48 degrees C. A downshift in growth temperature from 30 degrees C to 10 degrees C followed by incubation for 3 to 9 days increased thermotolerance of strain F3802A/84 but not strain B4ac-1 heated in BHI broth containing 2.0 or 4.0% NaCl compared to thermotolerance in BHI broth containing 0.5% NaCl. Protein analysis using one-dimensional gel electrophoresis revealed an increase in proteins with molecular weights of 54, 50, 44, and 42 kDa in cells of strain F3802A/84 and 83 and 69 kDa in cells of strain B4ac-1 subjected to an upshift in growth temperature from 30 degrees C to 37 degrees C or 40 degrees C, respectively. A downshift in growth temperature from 30 degrees C to 10 degrees C resulted in substantial amounts of proteins with molecular weights of 63, 40, and 29 kDa produced by strain F3802A/84 and 63 kDa to be produced by strain B4ac-1. Proteins produced in response to upshift or downshift in growth temperature are suspected to play an important role in heat resistance of the psychrotrophic and mesophilic strains of B. cereus examined in this study. Changes in resistance to heat or refrigeration temperatures, as well as tolerance to NaCl, as affected by previous exposure of cells to temperature shifts may influence the ability of B. cereus to grow in minimally processed foods during distribution and storage.

Animals

Comparison of chemical treatments to eliminate enterohemorrhagic Escherichia coli O157:H7 on alfalfa seeds.

The focus of this study was to determine the efficacy of various chemicals in eliminating 2.04 to 3.23 log10 CFU/g of Escherichia coli O157:H7 from alfalfa seeds and to determine the survivability of the pathogen on seeds stored for prolonged periods at three temperatures. Significant (P < or = 0.05) reductions in populations of E. coli O157:H7 on inoculated seeds were observed after treatments with 500 and 1,000 ppm of active chlorine (as Ca[OCl]2) for 3 but not 10 min and with > or =2,000 ppm of Ca(OCl)2 regardless of pretreatment with a surfactant. Treatment with 20,000 ppm of active chlorine failed to kill 2.68 log10 CFU/g of seeds. Acidified NaClO2 (500 ppm) was effective in reducing populations of the pathogen by >2 logs per g. Acidified ClO2 significantly reduced populations of E. coli O157:H7 on seeds at concentrations > or =100 ppm, and 500 ppm of ClO2 reduced the pathogen from 2.7 log10 CFU/g to <0.5 CFU/g. Chlorine (as NaOCl) was not effective at concentrations < or =1,000 ppm; significant reduction was achieved only after treatment with 2,000 ppm for 3 or 10 min. Notable reduction in populations was observed after treatment with 30 or 70% C2H3OH, but there was a dramatic decrease in germination percentage. Treatment with 0.2% H2O2 significantly reduced populations, and the organism was not detected by direct plating after treatment with > or =1% H2O2. Significant reduction in population of E. coli O157:H7 occurred after treatment with 1% trisodium phosphate, 40 ppm of Tsunami and Vortexx, and 1% Vegi-Clean. A significant decrease in the number of E. coli O157:H7 on dry seeds was observed within 1 week of storage at 25 and 37 degrees C, but not at 5 degrees C. Between 1 and 38 weeks, populations on seeds stored at 5 degrees C remained relatively constant. The pathogen was recovered from alfalfa seeds initially containing 3.04 log 10 CFU/g after storage at 25 or 37 degrees C for 38 weeks but not 54 weeks.

Calcium Compounds

Behavior of acid-adapted and unadapted Escherichia coli O157:H7 when exposed to reduced pH achieved with various organic acids.

A study was done to determine if various organic acids differ in their inhibitory or lethal activity against acid-adapted and unadapted Escherichia coli O157:H7 cells. E. coli O157:H7 strain EO139, isolated from venison jerky, was grown in tryptic soy broth (TSB) and in TSB supplemented with 1% glucose (TSBG) for 18 h at 37 degrees C, then plated on tryptic soy agar (TSA) acidified with malic, citric, lactic, or acetic acid at pH 5.4, 5.1, 4.8, 4.5, 4.2, and 3.9. Regardless of whether cells were grown in TSB or TSBG, visible colonies were not formed when plated on TSA acidified with acetic, lactic, malic, or citric acids at pH values of < or =5.4, < or =4.5, < or =4.2, or < or =4.2, respectively. Cells not adapted to reduced pH did not form colonies on TSA acidified with lactic acid (pH 3.9) or acetic acid (pH 3.9 and 4.2); however, a portion of acid-adapted cells remained viable on TSA containing lactic acid (pH 3.9) or acetic acid (pH 4.2) and could be recovered in TSB. Inactivation of acid-adapted cells was less than that of unadapted cells in TSB acidified at pH 3.9 with citric, lactic, or acetic acid and at pH 3.4 with malic acid. Significantly (P< or =0.05) higher numbers of acid-adapted cells, compared with unadapted cells, were detected 12 h after inoculation of TSB acidified with acetic acid at pH 3.9; in TSB containing lactic acid (pH 3.9), the number of acid-adapted cells was higher than the number of unadapted cells after 5 h. In TSB acidified at pH 3.9 with citric acid or pH 3.4 with malic acid, significantly higher numbers of acid-adapted cells survived. This study shows that organic acids differ in their inhibitory or lethal activity against acid-adapted and unadapted E. coli O157:H7 cells, and acid-adapted cells are more tolerant than unadapted cells when subsequently exposed to reduced pH caused by these acids.

Adaptation, Physiological

Microbiological quality and production of botulinal toxin in film-packaged broccoli, carrots, and green beans.

The production of toxin by a 10-strain mixture of proteolytic Clostridium botulinum in fresh produce packaged in polyethylene films with different oxygen permeability was determined. Broccoli florets, shredded carrots, and green beans inoculated with approximately 10(2) C. botulinum spores per g were placed in bags (1.4 kg per bag) composed of four films with different oxygen transmission rates (OTRs). Broccoli was packaged in bags with OTRs of 3 (7,000 cm3/m2/24 h) and 4 (16,000 cm3/m2/24 h), and green beans were packaged in bags with OTRs of 2 (6,000 cm3/m2/24 h) and 4. Broccoli and green beans in bags were compressed and heat-sealed. Shredded carrots were packaged in bags with OTRs of 1 (3,000 cm3/m2/24 h) and 3 and vacuum-sealed. Produce was stored at 4, 13, and 21 degrees C for up to 27 (broccoli) or 28 (carrots and green bean) days and analyzed periodically. At each sampling time, gas composition within the bags, pH of the produce microbial population (total aerobic and anaerobic microorganisms, lactic acid bacteria, psychrotrophic bacteria, yeasts, and molds), and the presence or absence of botulinal toxin were determined. Packaging material affected the quality of vegetables, especially broccoli stored at 4 and 13 degrees C. For example, broccoli was scored as "good" after 22 days at 4 degrees C when it was packaged in film with higher gas permeability (OTR of 4), whereas broccoli appeared to be in "poor" condition when packaged in film with lower gas permeability (OTR of 3). With the exception of lactic acid bacteria, packaging material did not noticeably influence the growth of microorganisms. Lactic acid bacteria grew better in broccoli packaged in bags with an OTR of 3 than in those with an OTR of 4 at all temperatures. Botulinal toxin was detected in broccoli packaged in bags with an OTR of 3 and stored at 13 degrees C for 21 days and in those with an OTR of 4 and 3 and stored at 21 degrees C for 10 days. All toxic samples were visibly spoiled. Toxin was not detected in produce packaged under any other test conditions.

Botulinum Toxins

Influence of temperature and pH on survival of Escherichia coli O157:H7 in dry foods and growth in reconstituted infant rice cereal.

Factors affecting the ability of Escherichia coli O157:H7 to survive in foods with a(w) less than required for growth have not been fully defined. This study was undertaken to determine the ability of E. coli O157:H7 to survive in a commercial dry infant rice cereal as affected by a(w) (0.35+/-0.04, 0.52+/-0.03 and 0.73+/-0.03), pH (4.0 and 6.8), and temperature (5, 25, 35 and 45 degrees C), and in nine other reduced-a(w) foods. Death of E. coli O157:H7 in cereal was enhanced with increased temperature and decreased pH during a 16- to 24-week storage period. Survival was enhanced at pH 6.8 compared to pH 4.0 in cereal at a(w) 0.34+/-0.04 during initial storage at 5 and 25 degrees C. The effect of temperature (8, 15, 21 and 30 degrees C) on survival and growth of acid-adapted cells of E. coli O157:H7 inoculated into cereal reconstituted with milk or apple juice at two inoculum levels (8.2-12.3 cfu/ml and 82-123 cfu/ml of slurry) was also studied. Growth occurred in cereal reconstituted with milk at all test temperatures and in cereal reconstituted with apple juice at 15, 21 and 30 degrees C. Populations increased by >1 log10 cfu/ml within 3-6 h at 21 and 30 degrees C. Acid-adapted and unadapted cells had similar growth patterns. The effects of temperature and acid adaptation on survival of E. coli O157:H7 in nine commercial foods and food ingredients with pH 4.07-6.49 and a(w) 0.17-0.82 were determined. The pathogen survived in these foods for various lengths of time, depending the storage temperature, with an order of survival of 5 degrees C >21 degrees C >37 degrees C. Survival appeared to be enhanced in foods with highest pH, and acid-adapted cells retained higher viability than unadapted cells in only two of the nine test foods. Of particular importance is the ability of E. coli O157:H7 to survive well in dry foods with a wide range in a(w) and pH, particularly at refrigeration temperature.

Animals

Influence of acid tolerance responses on survival, growth, and thermal cross-protection of Escherichia coli O157:H7 in acidified media and fruit juices.

A study was done to determine survival and growth characteristics of acid-adapted, acid-shocked, and control cells of Escherichia coli O157:H7 inoculated into tryptic soy broth (TSB) acidified with organic acids and three commercial brands of apple cider and orange juice. The three types of cells behaved similarly in TSB acidified with acetic acid; however, in TSB (pH 3.9) acidified with lactic acid, acid-adapted cells were more tolerant than acid-shocked cells which, in turn, were more tolerant than control cells. The ability of the three types of cells to grow after inoculation into acidified TSB, then plated on tryptic soy agar containing sodium chloride was determined. Tolerance of acid-adapted cells and, less markedly, acid-shocked cells to sodium chloride was diminished, compared to control cells. The pathogen showed extraordinary tolerance to the low pH of apple cider and orange juice held at 5 or 25 degrees C for up to 42 days. Growth occurred in one brand of apple cider (pH 3.98) incubated at 25 degrees C. Regardless of test parameters, there was no indication that cell types differed in tolerance to the acidic environment in apple cider or orange juice. Survival of control, acid-adapted, and acid-shocked cells heated in apple cider and orange juice was studied. Within each apple cider or orange juice, D(52 degrees C)-values of acid-adapted cells were considerably higher than those of acid-shocked or control cells, which indicates that heat tolerance can be substantially enhanced by acid adaptation compared to acid shock.

Beverages

Quality control of culture media--perspectives and problems.

The Working Party on Culture Media (WPCM) of the International Union of Microbiological Societies (IUMS) has promoted and facilitated the development of methods for quality control of culture media used for the detection and enumeration of food-borne microorganisms since the early 1980s. While progress has been made in establishing protocols to test for productivity and selectivity of media, problems associated with the influence of food constituents and background microflora, as well as the presence of sublethally injured cells in test foods, are yet to be fully addressed before optimum methods for assessing the quality of media can be defined. However, for various reasons, the development of standardised procedures which account for the influence of food constituents or state of debilitation of target microorganisms may not be practical or even desirable.

Culture Media

A collaborative study on media for the enumeration of yeasts in foods.

A collaborative study was made to evaluate the effectivity of a general purpose medium, tryptone glucose yeast extract (TGY) agar on the detection and enumeration of yeasts from food. Nine laboratories participated in the study and compared five media (four kinds of TGY with different concentrations of glucose, one of them without tryptone, and, for comparison, dichloran rose bengal chloramphenicol (DRBC) agar). Six food samples were investigated by each laboratory and 23 additional food samples were investigated individually by different laboratories. No difference was found in the performance of media with either the samples common to all laboratories or the various samples tested in different ones. A medium consisting of tryptone, glucose and yeast extract, at any concentration of glucose tested, appeared reliable for the detection and enumeration of yeasts from foods, and its performance did not differ from that of DRBC. Omission of tryptone as recommended by ISO provided an even simpler medium of equally good performance. TGY without chloramphenicol may result in higher total counts due to the development of bacteria. DRBC incubated in light results in lower counts compared to that incubated in the dark.

Aniline Compounds

Comparison of the SimPlate total plate count method with Petrifilm, Redigel, conventional pour-plate methods for enumerating aerobic microorganisms in foods.

The SimPlate Total Plate Count (TPC) method, developed by IDEXX Laboratories, Inc., is designed to determine the most probable number of aerobic microorganisms in foods. The 24-h test was compared to the conventional plate count agar (PCA) method, the Petrifilm Aerobic Count plates, and the Redigel Total Count procedure for enumerating microflora in 751 food samples. Results using the SimPlate TPC method were highly correlated (r > or = 0.96) with results from other test methods. Slopes (0.96-0.97) were not significantly different from 1, and y intercepts (-0.03-0.08) were not different from O. The SimPlate has a high counting range (> 1600 most probable number per single dilution), thus requiring fewer dilutions of samples compared to other methods evaluated. Some foods, e.g., raw liver, wheat flour, and nuts, contain enzymes that gave false-positive reactions on SimPlates. Overall, however, the SimPlate TPC method is a suitable alternative to conventional PCA, Petrifilm, and Redigel methods for estimating populations of mesophilic aerobic microorganisms in a wide range of foods.

Aerobiosis

Efficacy of spray application of chlorinated water in killing pathogenic bacteria on raw apples, tomatoes, and lettuce.

Washing whole and cut produce by dipping or submerging in chlorinated water has a sanitizing effect, although reduction in microbial populations is minimal and is usually less than 100-fold. A study was undertaken to evaluate the efficacy of a spray application of chlorine in killing Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, yeasts and molds, and total aerobic mesophilic microorganisms on whole apples, tomatoes, and lettuce leaves. Inoculated produce was treated (sprayed and then soaked) with water (control) or solutions containing 200 or 2,000 ppm of chlorine for 0, 1, 3, 5, or 10 min, rinsed with sterile water, and analyzed for populations (CFU/cm2) of target microorganisms. Compared to the control treatment, further reductions in numbers of pathogens of 0.35 to 2.30 log CFU/cm2 were achieved by treatment with chlorine. Chlorine was generally more effective at 2,000 ppm than at 200 ppm. Inactivation of microorganisms occurred essentially within 1 min after application of chlorine. These reductions are significant relative to populations of pathogenic microorganisms that may be present on produce. Spray application of chlorine to raw produce at food service or household levels may be a suitable, and more convenient, alternative to treatment by dipping or submersion.

Chlorine

Survival and growth of psychrotrophic Bacillus cereus in dry and reconstituted infant rice cereal.

The potential for growth of enterotoxigenic Bacillus cereus in reconstituted dry foods is a concern, especially when they are consumed by infants or the immunosuppressed. The ability of a four-strain mixture of spores or vegetative cells of psychrotrophic B. cereus to survive in a commercial, dry infant rice cereal as affected by water activity (a(w); 0.27 to 0.28, 0.52 to 0.55, and 0.75 to 0.78), pH (5.6 and 6.7), and temperature (5, 25, 35, and 45 degrees C) was investigated. The rate of death of vegetative cells in dry cereal stored for 36 weeks was not affected by a(w) or pH. Death of spores in cereal stored at 45 degrees C for up to 48 weeks was enhanced at a(w) 0.78 but was unaffected by pH; loss of viability at 5, 25, and 35 degrees C was largely unaffected by differences in a(w). The effect of temperature (8, 15, 21, and 30 degrees C) on outgrowth of spores of B. cereus inoculated at three levels (0.14, 14, and 133 CFU/g, dry weight basis) into cereal reconstituted with apple juice and commercial pasteurized milk (2% fat) was also studied. Outgrowth of spores did not occur in cereal reconstituted with apple juice. Cereal reconstituted with milk and inoculated with 0.14, 14, and 133 spores per g contained >3 log CFU/g within 24, 9, and 6 h, respectively, at 21 degrees C. Populations in cereal reconstituted with milk and inoculated with 133 CFU of B. cereus spores per g reached 7.11, 7.72, and 7.40 log CFU/g within 12, 48, and 72 h when stored at 30, 21, and 15 degrees C, respectively. The organism grew in cereal reconstituted with milk and held at 8 degrees C for 72 h; however, enterotoxin was not detected. In reconstituted cereal inoculated with 133 spores per g, enterotoxin was detected (detection limit 16 ng/g) after 24, 48, and 72 h at 30, 21, and 15 degrees C, respectively, when the population of B. cereus reached >7 log CFU/g. It is recommended that reconstituted infant foods be either consumed immediately or held at < or = 8 degrees C and consumed within 48 h after preparation.

Animals

Thermal inactivation of Escherichia coli O157:H7 isolated from ground beef and bovine feces, and suitability of media for enumeration.

Rates of thermal inactivation of five strains of Escherichia coli O157:H7 isolated from ground beef implicated in outbreaks of hemorrhagic colitis and five strains isolated from bovine feces were determined. Ground beef (22% fat, 10 g), inoculated with individual test strains at populations ranging from 6.85 to 7.40 log10 CFU g-1 of beef, was formed into patties (0.3 cm thick and 8.0 cm in diameter) and sealed in polyethylene bags. For each strain and treatment temperature (54.4, 58.9, 62.8, 65.6, or 68.3 degrees C), 6 bags were simultaneously immersed into a recirculating water bath. Viable cells in patties heated for various lengths of time were enumerated by plating diluted samples on sorbitol MacConkey agar supplemented with 4-methylumbelliferyl-beta-D-glucuronide (MSMA) and modified eosin methylene blue (MEMB) agar. Regardless of strain or treatment temperature, higher numbers of E. coli O157:H7 cells were generally recovered on MEMB agar than on MSMA, indicating the inferiority of MSMA as a recovery medium for quantitative determination of E. coli O157:H7 cells in heat-processed ground beef. Significantly (P < or = 0.05) higher D values when enumeration was done using MEMB agar compared with MSMA. Mean D values for combined strain data at 54.4, 58.9, 62.8, and 65.6 degrees C from cultures on MEMB agar were 123.90, 6.47, 0.62, and 0.20 min, respectively, whereas D values of 25.5, 5.21, and 0.18 min were obtained at the same temperatures from cultures on MSMA. Results suggest that cooking ground beef patties to an internal temperature of 68.3 degrees C for 40 s will inactivate at least 99.99% of E. coli O157:H7 cells; z values of 4.0 and 5.1 degrees C were calculated from mean D values obtained from MEMB agar and MSMA, respectively, as recovery media. Differences in D values existed among strains but rates of thermal inactivation do not appear to be correlated with the sources of the isolates.

Animals

Combined effects of pH, nisin, and temperature on growth and survival of psychrotrophic Bacillus cereus.

Growth of vegetative cells and outgrowth of spores of enterotoxigenic psychrotrophic Bacillus cereus in refrigerated minimally processed food products is a public health concern. A study was undertaken to determine the combined effects of pH, nisin, and temperature on growth and survival of 20 strains of B. cereus. The minimum growth temperatures in tryptic soy broth (pH 7.3) and brain heart infusion broth (BHI broth, pH 7.4) were 5 degrees C for two strains and 8 degrees C for five other strains. Vegetative cells of four of eight strains grew at 8 degrees C in BHI broth (pH 6.01 and 6.57) containing 10 micrograms of nisin per ml. At 15 degrees C, all strains grew at pH 5.53 to 6.57; three strains tolerated nisin at 50 micrograms/ml (pH 6.57), whereas two other strains had a maximum tolerance of 10 micrograms of nisin per ml. Tolerance of vegetative cells of B. cereus to nisin increased as the pH of the broth was increased from 5.53 to 6.01 and again to pH 6.57. Outgrowth of spores (six of six strains tested) was inhibited by 5 and 50 micrograms of nisin per ml at 8 and 15 degrees C, respectively. At 15 degrees C, outgrowth of spores of two strains occurred at pH 6.52 in BHI broth containing 10 micrograms of nisin per ml. The effectiveness of nisin in controlling the growth of psychrotrophic strains of B. cereus capable of causing human illness was more pronounced at 8 degrees C than at 15 degrees C and as the pH was decreased from 6.57 to 5.53. Studies to determine the effectiveness of nisin in controlling growth of psychrotrophic B. cereus in nonpasteurized foods held at refrigeration temperatures are warranted.

Anti-Bacterial Agents

Microbiological quality and the inability of proteolytic Clostridium botulinum to produce toxin in film-packaged fresh-cut cabbage and lettuce.

The production of toxin by a 10-strain mixture of proteolytic Clostridium botulinum in fresh produce packaged in polyethylene films having high (7,000 cc/m2/24 h; HOTR) and low (3,000 cc/m2/24 h; LOTR) relative oxygen permeability was determined. Shredded cabbage and lettuce inoculated with approximately 10(2) spores/g were placed in bags composed of the two films (1.4 kg/bag), and the bags were then vacuum sealed. Produce was stored at 4, 13, and 21 degrees C for up to 21 (cabbage) or 28 (lettuce) days and analyzed periodically. At each sampling time, the gas composition within the bags, pH of the produce, and microbial populations (total aerobic and anaerobic microorganisms, lactic acid bacteria, psychrotrophic bacteria, and yeasts and molds) were determined. In addition, the presence of botulinal toxin was determined using the standard U.S. Food and Drug Administration mouse bioassay protocol. Bags made of HOTR film prolonged sensory quality of cabbage and lettuce, especially at 13 and 4 degrees C. Packaging material had an effect on the growth of various groups of microorganisms; however, there was not a general trend. For example, lettuce packaged in HOTR bags had higher aerobic microbial populations than that packed in LOTR, but no significant difference (P < or = 0.05) was observed with cabbage. Growth of psychrotrophic bacteria was greater in vegetables packaged in HOTR film while growth of yeasts and molds was not affected by either packaging film. Most differences in microbial populations in produce packaged in LOTR and HOTR films were less than 1 log10 CFU/g. Botulinal toxin was not detected in cabbage or lettuce packaged in either film or stored under any test condition.

Animals