Recovery of heat-stressed yeasts in media containing plant oleoresins

C onner , D.E. & B euchat , L.R. 1985. Recovery of heat-stressed yeasts in media containing plant oleoresins. Journal of Applied Bacteriology 59 , 49–55.
Oleoresins from seven plants (allspice, cinnamon, clove, garlic, onion, oregano, and thyme) were tested for their effects on eight genera of sublethally heated food-spoilage and industrially important yeasts ( Candida lipolytica, Debaryomyces han-senii, Hansenula anomala, Kloeckera apiculata, Lodderomyces elongisporus, Rhodotorula rubra, Saccharomyces cerevisiae, and Torulopsis glabrata ). All heat-stressed yeasts had increased sensitivity to oleoresins in recovery media. Heated cells were most sensitive to cinnamon oleoresin, a lethal effect being observed to concentrations as low as 5 μg/ml. Results indicate that all eight yeasts underwent metabolic and/or structural injury as a result of exposure to elevated temperature. Whilst changes in sensitivity to oleoresins were not the same for all yeasts, it is concluded that the oleoresins may have a pronounced synergistic or additive effect on thermal inactivation of yeasts in foods. Furthermore, carryover of oleoresins in low dilutions of food to enumeration media could adversely influence the recovery of heat-stressed cells, thus resulting in apparent low populations.     

Efficacy of some methods and media for detecting and enumerating Campylobacter jejuni in frozen chicken meat

Four of five strains of Campylobacter jejuni survived in chicken meat stored at -18 degrees C for 12 months. Direct plating of samples was superior to the most probable number technique for enumerating C. jejuni. Enrichment culture using the Doyle & Roman enrichment method resulted in the highest rates of detection. Packaging under an atmosphere of CO2 did not substantially influence survival.     

Behavior of Listeria monocytogenes inoculated into raw tomatoes and processed tomato products

Rates of death and growth of Listeria monocytogenes inoculated onto raw whole and into chopped tomatoes stored at 10 and 21 degrees C were not influenced by prior treatment of tomatoes with chlorine or packaging under an atmosphere of 3% O2 and 97% N2. Growth of the pathogen occurred in whole tomatoes held at 21 degrees C but not at 10 degrees C, while death occurred in chopped tomatoes stored at these temperatures. Likewise, growth patterns of mesophilic aerobic microorganisms, psychrotrophic microorganisms, and yeasts and molds on whole and chopped tomatoes were essentially unaffected by chlorine and modified atmosphere packaging treatments. Populations of L. monocytogenes inoculated into commercially processed tomato juice and sauce and held at 5 degrees C remained constant for 14 days. A gradual decrease in the number of viable L. monocytogenes cells was observed in juice and sauce held at 21 degrees C. In contrast, the organism died rapidly when suspended in commercial tomato ketchup at 5 and 21 degrees C. Unlike low-acid raw salad vegetables such as lettuce, broccoli, asparagus, and cauliflower on which we have observed L. monocytogenes grow at refrigeration temperatures, tomatoes are not a good growth substrate for the organism. Nevertheless, L. monocytogens can remain viable on raw whole and chopped tomatoes and in commercial tomato juice and sauce for periods extending beyond their normal shelf-life expectancy.     

Combined effect of growth medium,age of cells and phase of sporulation on heat resistance and recovery of Hansenula anomala

Effects of two growth media, age of cells and phase of sporulation on heat resistance of Hansenula anomala were determined. Cells were grown on two solid media, McClary's acetate and V8 juice agars, at 21 ° C for 16 days. Heat resistance of cells was determined in 0.06 M potassium phosphate buffer at 48 ° C. Heat-stressed cells were plated on four recovery media: yeast extract-malt extract-peptone-glucose (YMPG), pH 7.0; YMPG, pH 3.5; YMPG containing 6% NaCl, pH 7.0; and YMPG containing 20% sucrose, pH 7.0. The composition of sporulation medium influenced the extent of sporulation and the relative heat resistance of sporulating cells. One-day-old cells were the most sensitive to heat. The heat resistance of cells was generally increased as the incubation time was extended to 16 days. Heat treatment caused a greater increase in sensitivity to NaCl than to sucrose or acid pH in recovery media. Young cells were more sensitive to NaCl than were older cells.     

Comparison of combinations of diluents and media for enumerating Zygosaccharomyces rouxii in intermediate water activity foods

Combinations of five diluents (0.1% peptone, 40 and 50% glucose, and 18 and 26% glycerol) and three enumeration media (tryptone glucose yeast extract, dichloran 18% glycerol and malt extract yeast extract 50% glucose (MY50G) agars) were evaluated for recovering a xerotolerant yeast, Zygosaccharomyces rouxii , from foods with intermediate water activity ( a _w). Combinations of 40% ( a _w, 0.936) or 50% ( a _w 0.898) glucose diluent and MY50G agar ( a _w 0.890) were superior in recovering highest populations. The type of solute in the diluent, as well as a reduced a _w, influences efficiency of recovering viable cells.     

Ingestion of Salmonella enterica Serotype Poona by a Free-Living Nematode, Caenorhabditis elegans, and Protection against Inactivation by Produce Sanitizers

Free-living nematodes are known to ingest food-borne pathogens and may serve as vectors to contaminate preharvest fruits and vegetables. Caenorhabditis elegans was selected as a model to study the effectiveness of sanitizers in killing Salmonella enterica serotype Poona ingested by free-living nematodes. Aqueous suspensions of adult worms that had fed on S. enterica serotype Poona were treated with produce sanitizers. Treatment with 20 μg of free chlorine/ml significantly (α = 0.05) reduced the population of S. enterica serotype Poona compared to results for treating worms with water (control). However, there was no significant difference in the number of S. enterica serotype Poona cells surviving treatments with 20 to 500 μg of chlorine/ml, suggesting that reductions caused by treatment with 20 μg of chlorine/ml resulted from inactivation of S. enterica serotype Poona on the surface of C. elegans but not cells protected by the worm cuticle after ingestion. Treatment with Sanova (850 or 1,200 μg/ml), an acidified sodium chlorite sanitizer, caused reductions of 5.74 and 6.34 log₁₀ CFU/worm, respectively, compared to reductions from treating worms with water. Treatment with 20 or 40 μg of Tsunami 200/ml, a peroxyacetic acid-based sanitizer, resulted in reductions of 4.83 and 5.34 log₁₀ CFU/worm, respectively, compared to numbers detected on or in worms treated with water. Among the organic acids evaluated at a concentration of 2%, acetic acid was the least effective in killing S. enterica serotype Poona and lactic acid was the most effective. Treatment with up to 500 μg of chlorine/ml, 1% hydrogen peroxide, 2,550 μg of Sanova/ml, 40 μg of Tsunami 200/ml, or 2% acetic, citric, or lactic acid had no effect on the viability or reproductive behavior of C. elegans. Treatments were also applied to cantaloupe rind and lettuce inoculated with S. enterica serotype Poona or C. elegans that had ingested S. enterica serotype Poona. Protection of ingested S. enterica serotype Poona against sanitizers applied to cantaloupe was not evident; however, ingestion afforded protection of the pathogen on lettuce. These results indicate that S. enterica serotype Poona ingested by C. elegans may be protected against treatment with chlorine and other sanitizers, although the basis for this protection remains unclear.     

Survival and growth of Escherichia coli O157:H7 in ground, roasted beef as affected by pH, acidulants, and temperature.

A study was undertaken to determine the fate of Escherichia coli O157:H7 in ground, roasted beef as influenced by the combined effects of pH, acidulants, temperature, and time. There was essentially no change in the viable population of E. coli O157:H7 when beef salads (pH 5.40 to 6.07) containing up to 40% mayonnaise were incubated at 5 degrees C for up to 72 h. At 21 and 30 degrees C, significant (P < or = 0.05) increases in populations of the organism occurred in salads containing 16 to 32% mayonnaise (pH 5.94 to 5.55) between 10 and 24 h of incubation. Death was more rapid as the pH of acidified beef slurries incubated at 5 degrees C was decreased from 5.98 to 4.70. E. coli O157:H7 grew in control slurries (pH 5.98) and in slurries containing citric and lactic acids (pHs 5.00 and 5.40) incubated at 21 degrees C for 24 h; decreases occurred in slurries acidified to pHs 4.70, 5.00, and 5.40 with acetic acid or pH 4.70 with citric or lactic acid. At 30 degrees C, populations decreased in slurries acidified to pHs 4.70 and 5.00 with acetic acid. Citric and lactic acids failed to prevent significant increases in populations in slurries at pH 4.70 to 5.40 between 10 and 24 h of incubation. The order of effectiveness of acidulants in inhibiting growth was acetic acid > lactic acid > or = citric acid. The same order was observed for inactivation of E. coli O157:H7 in acidified (pH 5.00) beef slurry heated at 54 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)     

Procedure for Evaluating the Effects of 2,450- Megahertz Microwaves upon Streptococcus faecalis and Saccharomyces cerevisiae

Modifications of a commercial 2,450-megahertz microwave oven were made so that 6 ml of microbial suspension could be exposed to the microwave field for various periods of time. The microorganisms were contained in the central tube of a modified Liebig condenser positioned in the approximate geometric center of the oven cavity. Kerosene at -25 C was circulated through the jacket of the condenser during microwave exposure permitting microwaves to reach the microbial suspension. Flow rates of the kerosene were varied to permit the temperature of the suspension to range from 25 to 55 C during microwave exposure. Conductive heating experiments using similar temperatures were also conducted. A thermocouple-relay system was employed to measure the suspension temperature immediately after the magnetron shutoff. Continuous application of microwaves to suspensions of 10(8) to 10(9)Streptococcus faecalis or Saccharomyces cerevisiae per ml appeared to produce no lethal effects other than those produced by heat. Respiration rates of microwave-exposed Scerevisiae were directly related to decreases in viable count produced by increased microwave exposure times.     

Efficacy of hyaluronic acid binding assay in selecting motile spermatozoa with normal morphology at high magnification

Background  

The present study aimed to evaluate the efficacy of the hyaluronic acid (HA) binding assay in the selection of motile spermatozoa with normal morphology at high magnification (8400x).     

Ethanol-Mediated Variations in Cellular Fatty Acid Composition and Protein Profiles of Two Genotypically Different Strains of Escherichia coli O157:H7

Two strains of Escherichia coli O157:H7 were grown in tryptic soy broth (TSB, pH 7.1) supplemented with 0, 2.5, 5.0, 7.5, and 10% ethanol at 30°C for up to 54 h. Growth rates in TSB supplemented with 0, 2.5, and 5.0% ethanol decreased with an increase in ethanol concentration. Growth was not observed in TSB supplemented with 7.5 or 10% ethanol. The pH of TSB containing 5.0% ethanol decreased to 5.8 within 12 h and then increased to 7.0 at 54 h. The ethanol content in TSB supplemented with 2.5 or 5.0% ethanol did not change substantially during the first 36 h of incubation but decreased slightly thereafter, indicating utilization or degradation of ethanol by both strains. Glucose was depleted in TSB supplemented with 0, 2.5, or 5.0% ethanol within 12 h. Cells grown under ethanol stress contained a higher amount of fatty acids. With the exceptions of cis-oleic acid and nonadecanoic acid, larger amounts of fatty acid were present in stationary-phase cells of the two strains grown in TSB supplemented with 5.0% ethanol for 30 h than in cells grown in TSB without ethanol for 22 h. The trans-oleic acid content was 10-fold higher in the cells grown in TSB with 5.0% ethanol than those grown in TSB without ethanol. In contrast, cis-oleic acid was not detected in ethanol-stressed cells but was present at concentrations of 0.32 and 0.36 mg/g of cells of the two strains grown in TSB without ethanol. Protein content was higher in ethanol-stressed cells than in nonstressed cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles varied qualitatively as affected by the strain and the presence of ethanol in TSB. An ethanol-mediated protein (28 kDa) was observed in the ethanol-stressed cells but not in control cells. It is concluded that the two test strains of E. coli O157:H7 underwent phenotypic modifications in cellular fatty acid composition and protein profiles in response to ethanol stress. The potential for cross protection against subsequent stresses applied in food preservation technologies as a result of these changes is under investigation.