Growth and aflatoxin production by Aspergillus parasiticus in a medium at different pH values and with or without pimaricin

A glucose-yeast extract-salt medium containing 0, 5, 7.5, 10, 15 or 20 micrograms pimaricin/ml with an initial pH of 3.5 or 5.5 was inoculated with Aspergillus parasiticus WB 108 and incubated at 15 degrees or 28 degrees C. The pH, weight of mycelium and amount of aflatoxin produced were determined after 3, 7, and 10 days and after 14, 21, and 30 days when incubation was at 28 degrees or 15 degrees C, respectively. Increasing the concentration of pimaricin in the medium with an initial pH of 5.5 decreased the amounts of aflatoxin B1 and G1 produced after 3 days of incubation. When the initial pH of the medium was 3.5, no growth or toxin production occurred after 3 days of incubation in the medium containing 7.5 micrograms or more of pimaricin/ml. The presence of 20 micrograms of pimaricin/ml inhibited growth and toxin production after 7 days of incubation. When cultures were incubated at 15 degrees C, there was a lag phase which extended from 9 to 16 days, and the amounts of aflatoxin produced decreased with an increasing concentration of pimaricin. Pimaricin did not completely inhibit the growth and aflatoxin production by A. parasiticus. Pimaricin, in combination with a low pH, low temperature or 4% or 6% NaCl, initially caused slow mycelial growth and low toxin production, but the mold overcame the inhibitory effects and produced substantial amounts of mycelium and toxin.     

Staphylococcus aureus growth and enterotoxin A production in an anaerobic environment.

The effects of strict anaerobic conditions on the growth of Staphylococcus aureus and the production of staphylococcal enterotoxin A (SEA) were studied. The growth of S. aureus, a facultative anaerobic bacterium, is slower anaerobically than aerobically. When grown on brain heart infusion broth at 37 degrees C, the anaerobic generation time at mid-log phase was 80 min, compared with 35 min for the aerobic control. In contrast to previous studies demonstrating that staphylococcal cell density was 9- to 17-fold greater in aerobic than in anaerobic cultures, data for a staphylococcal strain implicated in food poisoning showed that the cell density was only two to three times as great in aerobic cultures. Production of SEA was monitored by Western immunoblotting and shown to be growth dependent. With slower anaerobic growth, relatively less toxin was produced than under aerobic conditions. but in both cases SEA was detected after 120 min of incubation. The combined effects of temperature and aeration on S. aureus were also studied. Growth and toxin production of aerobic and anaerobic cultures at temperatures ranging from 14 to 37 degrees C were analyzed. Growth was still observed at low temperatures in both environments. A linear model for S. aureus aerobic or anaerobic growth as a function of incubation temperature was developed from these studies. The model was tested from 17 to 35.5 degrees C, and the results suggest that the model can accurately predict the S. aureus growth rate in this temperature range. The data suggest that anaerobic conditions are not an effective barrier against S. aureus growth.     

Growth and Toxin Production by Clostridium botulinum in Model Acldified Systems

TPGY medium was acidified to pH 4.20, 4.60, 5.00 and 5.40 with acetic or citric acid. The media were inoculated with spores from three strains of C. botulinum type A, B or E. Growth, pH, and toxin production under anaerobic conditions were monitored for 8 wk. Spores of C. botulinum types A and B were incapable of outgrowth and toxin production at pH 4.60 or below when incubated at 35°C. Spores of C. botulinum type E were capable of growth and toxin production at 26°C in citric acid acidified systems at pH 4.20. Growth and toxin production were not detected below pH 5.00 when acetic acid was used.     

Inhibitory effects of Enterococcus strains obtained from a probiotic product on in vitro growth of Salmonella enterica serovar enteritidis strain IFO3313

Enterococcus faecium and Enterococcus gallinarum strains were isolated from a commercial probiotic product and the effects of these strains on the growth of Salmonella enterica serovar Enteritidis strain IFO3313 were investigated. Viable cell counts of Salmonella Enteritidis in mixed cultures with the probiotic product isolate of E. faecium were significantly (P < 0.05) lower than those in pure cultures after 6, 8, and 24 h when the cultures were incubated in heart infusion broth at 37 and 41 degrees C. Significant differences in viable cell counts of Salmonella Enteritidis in mixed cultures with the probiotic product isolate of E. gallinarum and those in pure cultures were also observed after 8 and 24 h at 37 and 41degrees C. Similar observations were shown in mixed cultures of Salmonella Enteritidis with the reference strains of E. faecium GIFU8355 and E. gallinarum ATCC 49573. Significant differences in viable cell counts of these enterococcal strains were not shown among pure and mixed cultures with Salmonella Enteritidis. The pH values in pure and mixed cultures were 7.0 or 7.5 throughout the experiments. E. faecium strains were found to harbor the genes encoding enterocins A and B and showed inhibitory zones with a diameter of 4 to 6 mm against growth of Salmonella Enteritidis in the enterocin production assays. However, the E. gallinarum strains possessed neither of the enterocin genes tested and exhibited no inhibition zone in the enterocin production assays. These results indicated that enterococcal strains exhibit inhibitory effects on the growth of Salmonella Enteritidis and these effects were due to both enterocin and nonenterocin factors.     

THE INFLUENCE OF NUTRITIONAL FACTORS ON SYNTHESIS OF SALMONELLA TOXIN

The nutritional requirements for the synthesis of a cholera toxin-like toxin from Salmonella enteritidis serotype braenderup were examined. Toxin levels of both culture filtrates and sonicates were determined by the modified Chinese hamster ovary (CHO) floating cell assay. With few exceptions, most amino acids which contained charged polar R groups were stimulatory for Salmonella toxin synthesis. Glycerol enhanced toxin production to the largest extent; however, glucose was a poor carbon source suggesting that Salmonella toxin elaboration may be subject to catabolite repression. Biotin and Mn⁺⁺ were shown to increase the synthesis as well as release of this toxin. An ideal defined medium, supplemented with all the nutritional factors shown to stimulate toxin synthesis, was formulated. The amount of Salmonella toxin produced in this supplemented medium was about 82% of the total toxin elaborated when cultured in Casamino acids plus yeast extract.     

Influence of pH and temperature on the growth of and toxin production by neurotoxigenic strains of Clostridium butyricum type E

Strains of Clostridium butyricum that produce botulinal toxin type E have been implicated in outbreaks of foodborne botulism in China, India, and Italy, yet the conditions that are favorable for the growth and toxinogenesis of these strains remain to be established. We attempted to determine the temperatures and pH levels that are most conducive to the growth of and toxin production by the six strains of neurotoxigenic C. butyricum that have been implicated in outbreaks of infective and foodborne botulism in Italy. The strains were cultured for 180 days on Trypticase-peptone-glucose-yeast extract broth at various pHs (4.6, 4.8, 5.0, 5.2, 5.4, 5.6, and 5.8) at 30 degrees C and at various temperatures (10, 12, and 15 degrees C) at pH 7.0. Growth was determined by checking for turbidity; toxin production was determined by the mouse bioassay. We also inoculated two foods: mascarpone cheese incubated at 25 and 15 degrees C and pesto sauce incubated at 25 degrees C. The lowest pH at which growth and toxin production occurred was 4.8 at 43 and 44 days of incubation, respectively. The lowest temperature at which growth and toxin production occurred was 12 degrees C, with growth and toxin production first being observed after 15 days. For both foods, toxin production was observed after 5 days at 25 degrees C. Since the strains did not show particularly psychrotrophic behavior, 4 degrees C can be considered a sufficiently low temperature for the inhibition of growth. However, the observation of toxin production in foods at room temperature and at abused refrigeration temperatures demands that these strains be considered a new risk for the food industry.     

Effect of pH,aeration and feeding non‐sterilized agave juice in a continuous agave juice fermentation

BACKGROUND: Continuous cultures have been used since the 1950s in beer and wine fermentations due to their higher productivities compared to traditional batch systems; nevertheless, the tequila industry has not taken advantage of the possible improvements that continuous fermentations could offer. In this work, the effect of pH, aeration and feeding of non‐sterilized medium, on the fermentative capability of two Saccharomyces cerevisiae strains (S1 and S2) cultured in continuous fermentation, using agave juice as the fermentation medium, were studied. RESULTS: In continuous cultures, the control of the medium pH (set point at 4) did not have a significant effect on fermentation efficiency compared to fermentations in which the pH was not controlled (pH 2.5 ± 0.3). Conversely, aeration of the cultures of both strains improved biomass production and consumption of reducing sugars and ammonium. The aeration also significantly augmented ethanol production only for S1 cultures (P < 0.05). Furthermore, the feeding of medium, either sterilized or not, did not show significant differences on the production of ethanol for S1 cultures. Higher concentrations of acetoin, succinic acid and diacetyl were found in the cultures fed with non‐sterilized medium. CONCLUSIONS: Compared to S2, S1 has a better fermentative performance in continuous non‐sterilized medium fermentations. Not controlling the pH during the cultures could prevent the possibility of microbial contamination as a result of the extreme medium acidity (pH 2.5 ± 0.3). This work showed the possibility of scaling up agave juice continuous fermentation feeding non‐sterilized medium with no control of pH. Copyright © 2010 Society of Chemical Industry     

Acid tolerance in Salmonella typhimurium induced by culturing in the presence of organic acids at different growth temperatures

The influence of growth temperature and acidification of the culture medium up to pH 4.25 with acetic, citric, lactic and hydrochloric acids on the growth and subsequent acid resistance at pH 3.0 of Salmonella typhimurium CECT 443 was studied. The minimum pH value which allowed for S. typhimurium growth within the temperature range of 25–37 °C was 4.5 when the pH was reduced using citric and hydrochloric acids, and 5.4 and 6.4 when lactic acid and acetic acid were used, respectively. At high (45 °C) or low (10 °C) temperatures, the growth pH boundary was increased about 1 pH unit. The growth temperature markedly modified the acid resistance of the resulting cells. In all cases, D-values were lower for cells grown at 10 °C and significantly increased with increasing growth temperature up to 37 °C, at which D-values obtained were up to 10 times higher. Cells grown at 45 °C showed D-values similar to those found for cells grown at 25 °C. The growth of cells in acidified media, regardless of the pH value, caused an increase in their acid resistance at the four incubation temperatures, although the magnitude of the Acid Tolerance Response (ATR) observed depended on the growth temperature. Acid adapted cultures at 10 °C showed D-values ranging from 5.75 to 6.91 min, which turned out to be about 2 times higher than those corresponding to non-acid adapted cultures, while higher temperatures induced an increase in D-values of at least 3.5 times. Another finding was that, while at 10 and 45 °C no significant differences among the effect of the different acids tested in inducing an ATR were observed, when cells were grown at 25 and 37 °C citric acid generally turned out to be the acid which induced the strongest ATR. Results obtained in this study show that growth temperature is an important factor affecting S. typhimurium acid resistance and could contribute to find new strategies based on intelligent combinations of hurdles, which could prevent the development or survival of Salmonella spp. in foods. The fact that moderately low temperatures (10 °C) markedly decrease the acid resistance and increase the growth pH boundary of S. typhimurium suggests the convenience to control the temperature during food processing as a strategy to prevent the growth and survival of this pathogenic microorganism.     

Increased Aflatoxin Production by Aspergillus parasiticus Under Conditions of Cycling Temperatures

The effects of cycling temperatures (5°C for 12 hr and 25°C for 12 hr) on aflatoxin production by Aspergillus parasiticus NRRL 2999 in yeast extract sucrose (YES) medium were studied. Cycling temperatures, after preincubation at 25°C for various times, resulted in more aflatoxin B₁, G₁, and total aflatoxin production than did constant incubation at either 25°C, which is generally considered to be the optimum for aflatoxin production, or 15°C, which is the same total thermal input as the 5-25°C temperature cycling. With increased preincubation time at 25°C, toxin production increased and the lag phase of growth was shortened or not evident. Cultures that were preincubated at 25°C for 1, 2, and 3 days prior to onset of temperature cycling showed the greatest increase in maximum aflatoxin production over the 25°C and 15°C constant temperatures. Cultures that were not preincubated at 25°C but subjected to constantly fluctuating temperatures produced maximum amounts of aflatoxin equivalent to cultures incubated at a constant 25°C. The maximum aflatoxin production at all temperatures studied occurred during the late log phase of growth and at pH minimums. Aflatoxins were found in higher concentrations in the broth than the mycelia under temperature cycling conditions, at 15°C, and at 25°C during the first 21 days of incubation, whereas greater amounts of toxin were retained in mycelium at 25°C in the later incubation period (28-42 days).     

EFFECT OF GLYCERYL MONOLAURATE ON TOXIN PRODUCTION BY CLOSTRIDIUM BOTULINUM IN MEAT SLURRY

Glyceryl monolaurate when used in the proportion of 5 g per kg of meat slurry (pH 6.0–6.2) inhibited toxin production by Clostridium botulinum type A (strain 73A), type B (strain OKRA) and type E (strain RIV 2). The concentration of glycerol monolaurate required for inhibition was not reduced by addition of lactic acid to the meat slurry until the pH was lowered to pH 5.2 or below. In meat slurry (pH 6.0–6.2), potassium sorbate inhibited production of type B toxin at 32 g/kg. The addition of butylated hydroxy-anisole to glyceryl monolaurate had no effect upon the concentration needed for inhibition of botulinum toxin production.     

Effect of ethanol on the growth of Clostridium botulinum

Model broth studies were carried out to investigate the effect of ethanol on the growth of proteolytic (group I) strains of Clostridium botulinum. Ethanol extended the pathogen's lag phase, decreased its exponential growth rate, and decreased its final level of growth in the stationary phase. In all cases, botulinum neurotoxin production was associated with growth. Micrographs of C. botulinum cultures grown at 37 degrees C in trypticase peptone glucose yeast extract (TPGY) broths containing 2 and 4% ethanol showed elongation of vegetative cells and interference with cell division. The inhibition of growth and toxin production at the ethanol level predicted (5.5%, wt/wt) was confirmed by microscopy and by the mouse bioassay. A subsequent study was carried out to determine the combined effect of ethanol (0 to 8% [wt/wt]), water activity (aw; 0.953 to 0.997), and pH (6.2 to 8.2) on the probability of the growth of and neurotoxin production by proteolytic strains of C. botulinum (10(3) spores per ml). Growth and neurotoxin production occurred in 1 to 3 days in TPGY broths without ethanol (0%) and in 2 to 4 days in broths containing 2% ethanol regardless of the aw or pH levels (P < 0.005). Growth and neurotoxin production were delayed by an ethanol concentration of 4% ethanol and completely inhibited by a concentration of 6%. At an ethanol concentration of 4%, the probability of growth and toxin production over 365 days (Pt) was influenced by aw and pH. After 365 days, the maximum probability of growth and toxin production (Pmax) was 1 for all but one combination. However, tau, the time it took for 50% of all eventually positive replicates for any given combination of barriers to show growth and/or turbidity, ranged from <3 to 229 days. All tubes of TPGY broths that showed no growth after 365 days were subcultured in fresh TPGY broths. In all cases, growth and toxin production occurred within 24 h at 37 degrees C, indicating the reversible (sporostatic and/or bacteriostatic) effect of ethanol on C. botulinum.     

Response surface models for effects of temperature, pH, and previous growth pH on growth kinetics of Salmonella Typhimurium in brain heart infusion broth

Response surface models were developed for effects of temperature (15 to 40 degrees C), pH (5.2 to 7.4), and previous growth pH (5.7 to 8.6) on lag time (lambda) and specific growth rate (mu) of Salmonella Typhimurium in brain heart infusion broth (BHIB). Seventy-five growth curves for model development and 30 growth curves for model validation were fit to a two-phase linear growth model to obtain direct estimates of lambda and mu of Salmonella Typhimurium in BHIB. Response surface models for natural logarithm transformations of lambda and mu as a function of temperature, pH, and previous growth pH were obtained by regression analysis. Previous growth pH did not alter (P > 0.05) or interact with temperature or pH to alter subsequent growth kinetics of Salmonella Typhimurium. However, lambda and mu of Salmonella Typhimurium in BHIB were affected (P < 0.05) by linear and quadratic effects of temperature and pH. The models were validated against data not used in their development. Mean absolute relative error of predictions (model accuracy) was 7.8% for lambda and 6.6% for mu. Median relative error of predictions (model bias) was -1.8% for lambda and -2.8% for mu. Results of the current study indicated that the models developed accurately predicted growth kinetics of Salmonella Typhimurium in BHIB within the matrix of factors modeled and that the range of previous growth pH (5.7 to 8.6) investigated did not alter the subsequent growth kinetics of Salmonella Typhimurium in BHIB.     

Effect of the Parabens With and Without Nitrite on Clostridium botulinum and Toxin Production in Canned Pork Slurry

Antimiciobials were evaluated in thioglycollate broth at pH 6.5 for the ability to inhibit growth and toxin production by C. botulinum 12885A and ATCC 7949 (Type B). Methyl, ethyl, propyl, and butyl parabens (0.1%) and sorbic acid (0.2%) were effective in inhibiting growth of C. botulinum 12885A and ATCC 7949 in broth. Ethyl, propyl, and butyl parabens (0.1%) and sorbic acid (0.2%) inhibited toxin production by both strains in culture medium. Ethyl, propyl, butyl parabens (0.1%) and sorbic acid (0.2%) were individually added to a comminuted pork slurry having salt and sugar, with or without 40 ppm sodium nitrite. Cans were inoculated with a mixture of C. botulinum 12885A and ATCC 7949 spores. The canned product was abused by holding at 27°C and was observed over a 3-month period for swollen cans. Swollen cans were examined for botulinal toxin by the mouse bioassay. Propyl and butyl paraben did not inhibit or delay toxin production. Ethyl paraben with or without nitrite delayed toxin production for 4 wk. Sorbic acid inhibited toxin for 3 wk; when 40 ppm nitrite was added to the sorbic acid treatment, toxin production was delayed for 4 wk.     

Growth and survival of foodborne pathogens in beer

This work aimed to assess the growth and survival of four foodborne pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus) in beer. The effects of ethanol, pH, and storage temperature were investigated for the gram-negative pathogens (E. coli O157:H7 and Salmonella Typhimurium), whereas the presence of hops ensured that the gram-positive pathogens (L. monocytogenes and S. aureus) were rapidly inactivated in alcohol-free beer. The pathogens E. coli O157:H7 and Salmonella Typhimurium could not grow in the mid-strength or full-strength beers, although they could survive for more than 30 days in the mid-strength beer when held at 4°C. These pathogens grew rapidly in the alcohol-free beer; however, growth was prevented when the pH of the alcohol-free beer was lowered from the "as received" value of 4.3 to 4.0. Pathogen survival in all beers was prolonged at lowered storage temperatures.     

Effect of pH and CO2 on growth and toxin production by Clostridium botulinum in English-style crumpets packaged under modified atmospheres.

The effect of pH and CO2 on both growth of and toxin production by Clostridium botulinum in English-style crumpets, packaged under modified atmospheres was investigated using a 2 x 2 factorial experiment. English-style crumpets (water activity, 0.990; pH 6.5 and 8.3) were inoculated with C. botulinum spores types A and proteolytic B (500 spores/g), packaged in either 60% CO2 (balance N2) or 100% CO2, stored at ambient temperature (25 degrees C), and monitored daily for toxicity. Toxin was detected after 4 days in crumpets packaged in 60% CO2, irrespective of initial product pH. Toxin production was delayed 1.5 to 3 days in crumpets packaged under 100% CO2. Analysis of variance indicated a significant interaction effect of pH and %CO2 on time of earliest toxin detection. Delay of toxin production was greatest for high pH (8.3) crumpets. All products were organoleptically acceptable at the time of toxigenesis, and therefore, high moisture-high pH bakery products, if contaminated with spores of C. botulinum, could become hazardous if packaged in atmospheres containing CO2.     

Process optimization for enhanced production of diphtheria toxin by submerged cultivation

When stationary culture was replaced by submerged cultivation in a fermentor, a significant increase in the yield of diphtheria toxin in a short cultivation time (less than 48 h as against 7-8 d) was noted. It was found that under optimal conditions of temperature, vortex mixing and surface aeration, an alkaline pH favoured toxin release. Furthermore, to enhance the production volume, two-and three-step semicontinuous batch cultivations were carried out. The toxin produced was of good titre with an adequate antigenic purity. Under optimal conditions, the variation in the Limes of flocculation (Lf titre) was likely due to the quality of the production medium, which in turn depended on the quality of the raw materials used. The process was also optimized in a pilot-scale fermentor.     

Inactivation of ccpA and aeration affect growth, metabolite production and stress tolerance in Lactobacillus plantarum WCFS1

The growth of Lactobacillus plantarum WCFS1 and of its ΔccpA ery mutant, WCFS1-2, was compared in batch fermentations in a complex medium at controlled pH (6.5) and temperature (30°C) with or without aeration, in order to evaluate the effect of ccpA inactivation and aeration on growth, metabolism and stress resistance. Inactivation of ccpA and, to a lesser extent, aeration, significantly affected growth, expression of proteins related to pyruvate metabolism and stress, and tolerance to heat, oxidative and cold/starvation stresses. The specific growth rate of the mutant was ca. 60% of that of the wild type strain. Inactivation of ccpA and aerobic growth significantly affected yield and production of lactic and acetic acid. Stationary phase cells were more stress tolerant than exponential phase cells with little or no effect of inactivation of ccpA or aeration. On the other hand, for exponential phase cells inactivation of ccpA impaired both heat stress and cold/starvation stress, but increased oxidative stress tolerance. For both strains, aerobically grown cells were more tolerant of stresses. Evidence for entry in a viable but non-culturable status upon prolonged exposure to cold and starvation was found. Preliminary results of a differential proteomic study further confirmed the role of ccpA in the regulation of carbohydrate catabolism and class I stress response genes and allow to gain further insight on the role of this pleiotropic regulator in metabolism and stress. This is the first study in which the impact of aerobic growth on stress tolerance of L. plantarum is evaluated. Although aerobic cultivation in batch fermentations does not improve growth it does improve stress tolerance, and may have significant technological relevance for the preservation of starter and probiotic cultures.     

Effect of acid shock with hydrochloric, citric, and lactic acids on the survival and growth of Salmonella typhi and Salmonella typhimurium in acidified media

The effect of acid shock with hydrochloric, citric, or lactic acid on the survival and growth of Salmonella Typhi and Salmonella Typhimurium in acidified broth was evaluated. Salmonella serovars were acid shocked (1 h at 35 degrees C) in Trypticase soy broth acidified with hydrochloric, citric, or lactic acid at pH 5.5. Unshocked cells were exposed to the same media that had been neutralized before use to pH 7.0. Shocked and unshocked cells were inoculated into broth acidified with hydrochloric acid (pH 3.0), citric acid (pH 3.0), or lactic acid (pH 3.8), and growth and survival ability were evaluated. The acid shock conferred protection to Salmonella against the lethal effects of low pH and organic acids. The adaptive response was not specific to the anion used for adaptation. The biggest difference in reduction of survival between shocked and unshocked strains (approximately 2 log CFU/ml) was observed when the microorganisms were shocked with lactic acid and then challenged with citric acid. Salmonella Typhi was more tolerant of citric acid than was Salmonella Typhimurium, but Salmonella Typhimurium had higher acid tolerance in response to acid shock than did Salmonella Typhi. The acid shock decreased the extension of the lag phase and enhanced the physiological state values of Salmonella Typhi and Salmonella Typhimurium when the pH of growth was 4.5. This increased ability to tolerate acidity may have an important impact on food safety, especially in the case of Salmonella Typhi, given the very low infectious dose of this pathogen.     

EFFECT OF THE USE OF DIFFERENT LACTIC STARTERS ON THE MICROBIOLOGICAL AND PHYSICOCHEMICAL CHARACTERISTICS OF NATURALLY BLACK TABLE OLIVES OF GEMLIK CULTIVAR

The effect of different lactic acid bacteria cultures on the physicochemical and microbiological characteristics of brined black olives of Gemlik cultivar at low fermentation temperature was studied. Fermentation was carried out according to the traditional Gemlik method with modifications like low salt concentration and lactic starter addition. The brines with 7% salt concentration were inoculated with lactic acid bacteria ( Lactobacillus brevis, Leuconostoc cremoris and L. paramesenteroides ), which were previously isolated from olive fruits at low temperatures and a commercial strain of Lb. plantarum. Fermentation procedures were carried out at controlled temperatures (between 10–12C). Lactic acid bacteria survival was accompanied by yeast development, no Pseudomonas and Enterobacter species were detected in all treatments during fermentation. The highest total titratable acidity, lowest pH and least yeast growth were determined at the brines and fermentation products, which were inoculated with L. cremoris.

PRACTICAL APPLICATIONS

The use of suitable starter cultures is necessary to improve the microbiological control of the naturally black table olive process, help to standardize the fermentation, increase the lactic acid yield and accordingly provide the production of olives with high quality. The requirements mentioned for starter cultures include a rapid and predominant growth, homofermentative metabolism, tolerance to salt, acid and polyphenols, and few growth factor requirements. Especially at the regions where olives were picked later when environmental temperatures are lower, the use of a starter culture that has the ability to grow at low temperatures may be necessary. Use of such starter cultures may help to increase acidification, to control some types of spoilage and to shorten the fermentation process.     

Modeling Staphylococcus aureus growth and enterotoxin production in milk

Staphylococcus aureus growth and its enterotoxin production in sterilized milk were modeled with a modification of a new logistic model recently developed by us. The modified model and the Baranyi model described the early exponential phase of a growth curve more accurately than the previous model, at constant temperatures from 14 to 36.5 degrees C. The amount of toxin in milk increased linearly with time from the time the cell population reached about 10(6.5) cfu/ml. The rate of toxin production linearly increased at temperatures between 14 and 32 degrees C. From parameter values obtained at the constant temperatures, the model successfully predicted bacterial growth in the milk at a varying temperature. For toxin level estimation, we postulated that the rate of toxin production might be regulated with the temperature after the cell concentration reached 10(6.5) cfu/ml; the time point when the cell concentration reached that value was predicted with the modified growth model. Introduction of a correction factor in the toxin estimation successfully predicted the toxin level in milk at a varying temperature. These results showed that this prediction system consisting of the modified model and the toxin production algorithm might be a useful tool for modeling bacterial growth and its metabolite production in liquid foods.