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Biomedical subjects

R F McFeeters

Publications and source records attributed to R F McFeeters.

At least 19 recordsLinked to original sources

Protective effects of organic acids on survival of Escherichia coli O157:H7 in acidic environments.

Outbreaks of disease due to acid-tolerant bacterial pathogens in apple cider and orange juice have raised questions about the safety of acidified foods. Using gluconic acid as a noninhibitory low-pH buffer, we investigated the killing of Escherichia coli O157:H7 strains in the presence or absence of selected organic acids (pH of 3.2), with ionic strength adjusted to 0.60 to 0.68. During a 6-h exposure period in buffered solution (pH 3.2), we found that a population of acid-adapted E. coli O157:H7 strains was reduced by 4 log cycles in the absence of added organic acids. Surprisingly, reduced lethality for E. coli O157:H7 was observed when low concentrations (5 mM) of fully protonated acetic, malic, or l-lactic acid were added. Only a 2- to 3-log reduction in cell counts was observed, instead of the 4-log reduction attributed to pH effects in the buffered solution. Higher concentrations of these acids at the same pH aided in the killing of the E. coli cells, resulting in a 6-log or greater reduction in cell numbers. No protective effect was observed when citric acid was added to the E. coli cells. d-Lactic acid had a greater protective effect than other acids at concentrations of 1 to 20 mM. Less than a 1-log decrease in cell numbers occurred during the 6-h exposure to pH 3.2. To our knowledge, this is the first report of the protective effect of organic acids on the survival of E. coli O15:H7 under low-pH conditions.

Acetates↗

Determination of 5-log pathogen reduction times for heat-processed, acidified vegetable brines.

Recent outbreaks of acid-resistant food pathogens in acid foods, including apple cider and orange juice, have raised concerns about the safety of acidified vegetable products. We determined pasteurization times and temperatures needed to assure a 5-log reduction in the numbers of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella strains in acidified cucumber pickle brines. Cocktails of five strains of each pathogen were (separately) used for heat-inactivation studies between 50 and 60 degrees C in brines that had an equilibrated pH value of 4.1. Salmonella strains were found to be less heat resistant than E. coli O157:H7 or L. monocytogenes strains. The nonlinear killing curves generated during these studies were modeled using a Weibull function. We found no significant difference in the heat-killing data for E. coli O157:H7 and L. monocytogenes (P = 0.9709). The predicted 5-log reduction times for E. coli O157:H7 and L. monocytogenes were found to fit an exponential decay function. These data were used to estimate minimum pasteurization times and temperatures needed to ensure safe processing of acidified pickle products and show that current industry pasteurization practices offer a significant margin of safety.

Adaptation, Physiological↗

Independent effects of acetic acid and pH on survival of Escherichia coli in simulated acidified pickle products.

Our objective was to determine the effects of organic acids and pH on the rate at which selected strains of Escherichia coli O157:H7 die in acid solutions representative of acidified pickle products (pH < 4.6). We used gluconic acid/sodium gluconate (pKa = 3.7) as a noninhibitory buffer to maintain pH at selected values in the absence of other organic acids. This was possible because we found that the inhibitory effects of this acid on E. coli strains at pH 3.1 were independent of acid concentration over a range of 2 to 200 mM. By this method, the lethal effects of acetic acid solutions (100 to 400 mM) at selected pH values between 3.1 and 4.1 were compared with the effects of pH alone (as determined using gluconate buffer). We found D-values were two- to fourfold lower with acetic acid compared with the effect of pH alone for simulated pickle brines in this pH range. Glutamic acid, an amino acid that is known to enhance acid resistance in E. coli and is a component of pickle brines, protected the E. coli strains from the specific effects of acetic acid.

Acetic Acid↗

Effect of malic acid on the growth kinetics of Lactobacillus plantarum.

The fermentation kinetics of Lactobacillus plantarum were studied in a specially designed broth formulated from commercially available, dehydrated components (yeast extract, trypticase, ammonium sulfate) in batch and continuous culture. During batch growth in the absence of malic acid, the specific growth rate was 0.20 h(-1). Malic acid in the medium, at 2 mM or 10 mM, increased the specific growth rate of L. plantarum to 0.34 h(-1). An increase in the maximum cell yield due to malic acid also was observed. Malic acid in the medium (12 mM) reduced the non-growth-associated (maintenance energy) coefficient and increased the biomass yield in continuous culture, based on calculations from the Luedeking and Piret model. The biomass yield coefficient was estimated as 27.4 mg or 34.3 mg cells mmol(-1) hexose in the absence or presence of malic acid, respectively. The maintenance coefficient was estimated as 3.5 mmol or 1.5 mmol hexose mg(-1) cell h(-1) in the absence or presence of malic acid. These results clearly demonstrate the energy-sparing effect of malic acid on the growth- and non-growth-associated energy requirements for L. plantarum. The quantitative energy-sparing effect of malic acid on L. plantarum has heretofore not been reported, to our knowledge.

Cucumis sativus↗

Sulfite analysis of fruits and vegetables by high-performance liquid chromatography (HPLC) with ultraviolet spectrophotometric detection.

Free and total sulfite were analyzed in acidified vegetable products, instant mashed potatoes, and dried apples. Sulfite was separated by HPLC and quantified with a UV-vis detector. Resolution from components of food samples was achieved by varying the acid concentration of the eluant solution and by appropriate choice of the analytical wavelength. The minimum detectable levels for sulfite were 0.5 mg/L for a 10-cm analytical column and 1.5 mg/L for a 30-cm column. For analyses done with a 30-cm column, the coefficient of variation was <2% for analysis of free sulfite and total sulfite in acidified vegetables. For dried apples and instant potatoes, it ranged from 1 to 6.5%. The corresponding analytical errors were <4% and 1.2-5.6%, respectively, for the 10-cm column.

Chromatography, High Pressure Liquid↗

Solid-phase microextraction (SPME) technique for measurement of generation of fresh cucumber flavor compounds.

Investigations were carried out to determine whether flavor compounds characteristic for fresh cucumbers could be rapidly determined using a solid-phase microextraction (SPME) dynamic headspace sampling method combined with gas chromatography and flame ionization detection. Cucumbers were sampled, during blending, for fresh cucumber flavor compounds (E,Z)-2,6-nonadienal and (E)-2-nonenal. The GC was such that the two target compounds were separated and baseline-resolved. Relative standard deviations for analysis of both (E,Z)-2,6-nonadienal and (E)-2-nonenal using this SPME sampling method were +/-10%. Utility of the analytical method was demonstrated by determining the effect of heat treatments on the ability of cucumbers to produce these flavor impact compounds.

Chromatography, Gas↗

Novel quantitative assays for estimating the antimicrobial activity of fresh garlic juice.

Novel agar diffusion and broth dilution assays were developed for quantitatively estimating the antimicrobial activity of fresh garlic juice. Bacteria found to be inhibited by garlic juice in agar diffusion assay included two gram-positive and five gram-negative species. Leuconostoc mesenteroides was not inhibited. Escherichia coli B-103 (HB101, with pJH101, ampicillin resistant, 100 microg ml(-1)) was inhibited and chosen as the standard culture for quantitative assays. The agar diffusion assay was based on the slope ratio method, where the slope of dose response for garlic juice was divided by the slope of dose response for methylmethane thiosulfonate (MMTSO2). Juice from fresh garlic varied in activity between 1.76 and 2.31 microg of MMTSO2 per mg of garlic juice. The activity of juice decreased during 11 months of storage of garlic cloves at 5 degrees C from 2.31 to less than 0.1 microg of MMTSO2 per mg of juice. The broth dilution assay also used the E. coli B-103 culture, which permitted selective enumeration of this bacterium when 100 microg ml(-1) of ampicillin was incorporated into the enumerating agar. Selective enumeration was essential since the garlic juice was not sterile and, thus, contained natural flora. Growth of E. coli was unaffected by 0.1%, delayed by 0.25%, and completely inhibited at 0.5 and 2% garlic juice in broth during 24 h of incubation at 37 micro C. The minimum inhibition concentration of garlic juice by broth dilution assay was, thus, estimated to be 0.5%, which is equivalent to 3.46 microg of MMTSO2 per mg of garlic juice by the agar diffusion assay.

Bacteria↗

Development of oxidized odor and volatile aldehydes in fermented cucumber tissue exposed to oxygen.

Changes in volatile compounds in fermented cucumber tissue during exposure to oxygen were investigated by purge and trap sampling, followed by GC-MS. Hexanal and a series of trans unsaturated aldehydes, (E)-2-pentenal, (E)-2-hexenal, (E)-2-heptenal, and (E)-2-octenal, increased in fermented cucumber slurries exposed to oxygen. Sensory evaluation of oxidized odor was correlated with the increase in aldehyde concentrations. Other identified volatile components present after fermentation did not show major changes during exposure to oxygen. There was no decrease in the formation of aldehydes in fermented cucumber samples that were heated to inactivate enzymes before exposure to oxygen. These results indicated that the formation of aldehydes in oxygen was due to nonenzymatic reactions.

Aldehydes↗

Inhibition of formation of oxidative volatile components in fermented cucumbers by ascorbic acid and turmeric.

Two naturally occurring antioxidants, ascorbic acid and turmeric, were effective in inhibiting formation of hexanal, (E)-2-penenal, (E)-2-hexenal, (E)-2-heptenal, and (E)-2-octenal when slurries of fermented cucumber tissue were exposed to oxygen. Added ascorbic acid prevented formation of most of these oxidative aldehydes at 175 ppm or greater. Turmeric, which is used commercially as a yellow coloring in cucumber pickle products, was found to almost completely prevent aldehyde formation at 40 ppm.

Ascorbic Acid↗

Investigations into genotypic variations of peanut carbohydrates.

Carbohydrates are known to be important precursors in the development of roasted peanut quality. However, little is known about their genotypic variation. A better understanding of the role of carbohydrates in roasted peanut quality requires first an understanding of the genotypic variation in the soluble carbohydrate components. Ion exchange chromatography was used to isolate 20 different carbohydrate components in 52 genotypes grown in replicated trials at two locations. Inositol, glucose, fructose, sucrose, raffinose, and stachyose were quantitated, and 12 unknown peaks were evaluated on the basis of the peak height of the unknown relative to the cellobiose internal standard peak height. Peaks tentatively identified as verbascose and ajugose could not be properly integrated because of tailing. Of the 18 carbohydrates that were estimated, 9 exhibited significant variation between test environments, 5 among market types, 14 among genotypes within market types, and 11 exhibited some significant form of genotype x environment interaction. Genotypes accounted for 38-78% of the total variation for the known components, suggesting that broad-sense heritability for these components is high. The observed high genotypic variation in carbohydrate components is similar to the high genotypic variation observed for the sweetness attribute in roasted peanuts, which raises the question regarding possible interrelationships. The establishment of such interrelationships could be most beneficial to peanut breeding programs to ensure the maintenance of flavor quality in future peanut varieties.

Arachis↗

Relationships of sweet, bitter, and roasted peanut sensory attributes with carbohydrate components in peanuts.

Certain roasted peanut quality sensory attributes have been shown to be heritable. Currently the only means of measuring these traits is the use of a trained sensory panel. This is a costly and time-consuming process. It is desirable, from a cost, time, and sample size perspective, to find other methodologies for estimating these traits. Because sweetness is the most heritable trait and it has a significant positive relationship to the roasted peanut trait, the possible relationships between heritable sensory traits and 18 carbohydrate components (inositol, glucose, fructose, sucrose, raffinose, stachyose, and 12 unknown peaks) in raw peanuts from 52 genotypes have been investigated. Previously reported correlations among sweet, bitter, and roasted peanut attributes were evident in this study as well. Where there was positive correlation of total sugars with sweetness, there also was positive correlation of total sugars with roasted peanut attribute and negative correlation of total sugars with bitterness and astringency. The expected generalized relationship of total sugars or sucrose to sweetness could not be established because the relationship was not the same across all market-types. Further work is needed to determine the nature of the chemical components related to the bitter principle, which appear to modify the sweet response and interfere with the sensory perception of sweetness, particularly in the Virginia market-type. Also, certain carbohydrate components showed significant relationships with sensory attributes in one market-type and not another. These differential associations demonstrate the complexity of the interrelationships among sweet, bitter, and roasted peanut sensory attributes. Within two market-types it is possible to improve the efficiency of selection for sweetness and roasted peanut quality by assaying for total carbohydrates. On the basis of the regression values the greatest efficiency would occur in the fastigiate market-type and then the runner.

Arachis↗

Aerobic and anaerobic metabolism of Listeria monocytogenes in defined glucose medium.

A defined medium with glucose as the carbon source was used to quantitatively determine the metabolic end products produced by Listeria monocytogenes under aerobic and anaerobic conditions. Of 10 strains tested, all produced acetoin under aerobic conditions but not anaerobic conditions. Percent carbon recoveries of end products, typified by strain F5069, were as follows: lactate, 28%; acetate, 23%; and acetoin, 26% for aerobic growth and lactate, 79%; acetate, 2%; formate, 5.4%; ethanol, 7.8%; and carbon dioxide, 2.3% for anaerobic growth. No attempt to determine carbon dioxide under aerobic growth conditions was made. The possibility of using acetoin production to assay for growth of L. monocytogenes under defined conditions should be considered.

Acetoin↗

Kinetics and Modeling of Lactic Acid Production by Lactobacillus plantarum.

An unstructured model was developed to describe bacterial growth, substrate utilization, and lactic acid production by Lactobacillus plantarum in cucumber juice. Significant lactic acid production occurred during growth, as well as stationary phases. The percentage of acid produced after growth ceased was a function of the medium composition. Up to 51% of the lactic acid was produced after growth ceased when NaCl was not present in the medium, whereas not more than 18% of the total lactic acid was produced after the growth ceased in presence of NaCl, probably because of an increase in the cell death rate. An equation relating the specific death rate and NaCl concentration was developed. With the kinetic model proposed by R. Luedeking and E. L. Piret (J. Biochem. Microbiol. Technol. Eng. 1:393-412, 1958) for lactic acid production rate, the growth-associated and non-growth-associated coefficients were determined as 51.9 (+/-4.2) mmol/g of cells and 7.2 (+/-0.9) mmol/g of cells h respectively. The model was demonstrated for batch growth of L. plantarum in cucumber juice. Mathematical simulations were used to predict the influence of variations in death rate, proton concentration when growth ceased, and buffer capacity of the juice on the overall fermentation process.

Journal Article↗

A differential medium for the enumeration of homofermentative and heterofermentative lactic Acid bacteria.

A medium was developed for the differential enumeration of homofermentative and heterofermentative lactic acid bacteria. Essential components of the medium included fructose (14 mM), KH(2)PO(4) (18 mM), bromcresol green (as a pH indicator), and other nutrients to support growth. In agar medium, homofermentative colonies were blue to green, while heterofermentative colonies remained white. A total of 21 Lactobacillus, Pediococcus, Leuconostoc, and Streptococcus species were correctly classified with the medium.

Journal Article↗

Measurement of pectin methylation in plant cell walls.

A procedure was developed to measure the degree of pectin methylation in small samples of isolated cell walls from nonlignified plant tissues or pectin solutions. Galacturonic acid was determined colorimetrically with the 3,5-dimethylphenol reagent. Methylation was measured by base hydrolysis of galacturonic acid methyl esters, followed by gas chromatographic determination of released methanol. Estimates of the precision of analysis of pectin and cell wall samples were made. The coefficient of variation for estimates of the pectin esterification in cell walls isolated from 10-g samples of cucumber tissue ranged from 7.7 to 13.2%.

Cell Wall↗

Mutation and Selection of Lactobacillus plantarum Strains That Do Not Produce Carbon Dioxide from Malate.

A differential medium was developed to distinguish between malate-decarboxylating (MDC) and -non-decarboxylating (MDC) strains of Lactobacillus plantarum. MDC strains produced a visible acid reaction in the medium, whereas MDC strains did not. Use of the medium allowed for rapid screening and isolation of mutagenized cells that had lost the ability to produce CO(2) from malate.

Journal Article↗

Carbohydrate Changes during Maturation of Cucumber Fruit : Implications for Sugar Metabolism and Transport.

Changes in the carbohydrate profiles in the mesocarp, endocarp, and seeds of maturing cucumber (Cucumis sativus, L.) fruit were analyzed. Fruit maturity was measured by a decrease in endocarp pH, which was found to correlate with a loss in peel chlorophyll and an increase in citric acid content. Concentrations of glucose and fructose (8.6-10.3 milligrams per gram fresh weight, respectively) were found to be higher than the concentration of sucrose (0.3 milligrams per gram fresh weight) in both mesocarp and endocarp tissue. Neither raffinose nor stachyose were found in these tissues. The levels of glucose and fructose in seeds decreased during development, but sucrose, raffinose, and stachyose accumulated during the late stages of maturation. Both raffinose and stachyose were found in the seeds of six lines of Cucumis sativus L. This accumulation of raffinose saccharides coincided with an increase in galactinol synthase activity in the seeds. Funiculi from maturing fruit were found to be high in sucrose concentration (4.8 milligrams per gram fresh weight) but devoid of both raffinose and stachyose. The results indicated that sucrose is the transport sugar from the peduncle to seed, and that raffinose saccharide accumulation in the seed is the result of in situ biosynthesis and not from direct vascular transport of these oligosaccharides into the seeds.

Journal Article↗