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Effects of fermentation temperature on the strain population of Saccharomyces cerevisiae.

The influence of fermentation temperature (from 15 to 35 degrees C) on a mixed strain population was studied. Mitochondrial DNA analysis was used to differentiate Saccharomyces cerevisiae strains and the frequency of each strain during the alcoholic fermentation was determined. The chemical analyses of resulting wines were carried out. The temperature determined how Saccharomyces strains developed and how effectively they fermented. Some strains performed better at high temperatures and others at low temperatures. The maximal population size was similar at all temperatures. At low temperatures, however, it was reached later though it remained constant throughout the alcoholic fermentation. On the other hand, viable cells decreased at high temperatures, especially at 35 degrees C. Obviously, the composition of the wines changed as the temperature of fermentation changed. At low temperatures, alcohol yield was higher. Secondary metabolites to alcoholic fermentation increased as the temperature increased. Glycerol levels were directly affected by temperature.

Alcohols↗

Fermentation by Lactobacillus fermentum Ogi E1 of different combinations of carbohydrates occurring naturally in cereals: consequences on growth energetics and alpha-amylase production.

Glucose, fructose, sucrose and starch are naturally present in cereals. Fermentation of different combinations of these carbohydrates by Lactobacillus fermentum Ogi E1, a sourdough heterofermentative lactobacillus, was investigated to determine effects on fermentation kinetics, growth energetics and alpha-amylase production. Irrespective of the substrate combination, the strain was able to simultaneously produce alpha-amylase and consume starch, glucose, fructose and sucrose. In mixtures of starch with either sucrose or fructose or with both fructose and glucose, yields of alpha-amylase from biomass (Y(amy/x)) were similar to those observed for starch. However, for starch and glucose or starch, glucose, fructose and sucrose mixtures, both Y(amy/x) and the specific rate of alpha-amylase production decreased markedly. In fructose- or sucrose-containing mixtures, mannitol was formed stoichiometrically indicating that fructose served as electron acceptor, and acetate was produced at constant yield from biomass (Y(ac/x)) (1 g acetate g biomass(-1)). Acetate production was expected to confer to the strain a competitive advantage during natural fermentation by improving biomass formation and growth through an increase in the ATP gain. Y(ATP) varied depending on the carbohydrate mixture, indicating different effects of substrate mixtures on the efficiency in ATP coupling to biomass formation. Compared to starch fermentation, the highest value of Y(ATP) (29 g biomass mol ATP(-1)) was estimated for the starch/fructose mixture but no increase in mu(max) was observed. The lowest value (16 g biomass mol ATP(-1)) was obtained for the starch, glucose and fructose mixture, whereas for the mixture of all carbohydrates, Y(ATP) was similar to that obtained with starch alone (20 g biomass mol ATP(-1)) and it was intermediary for the starch and sucrose mixture (17 g biomass mol ATP(-1)). It is concluded that competitiveness of the strain cannot be based on expected energy gain in mixed substrate fermentation involving fructose and sucrose with glucose and starch, but rather on its ability to simultaneously use carbohydrates while producing alpha-amylase and to produce acetic acid. Acetic acid production could enhance the strain capacity to inhibit nonacid-tolerant, competitive microflora at the earlier stage of natural fermentation.

Acetic Acid↗

The effect of temperature on the growth and persistence of Salmonella in fermented liquid pig feed.

Two studies were conducted to investigate the effect of temperature on the fate of Salmonella enterica subsp. enterica serovar typhimurium DT104:30 in fermented liquid pig feed. These were (1) by co-inoculation of feed with S. typhimurium DT104:30 and Pediococcus pentosaceus, as the fermenting organism, and (2) by fermenting feed for 48, 72 or 96 h prior to inoculation with S. typhimurium DT104:30. In co-inoculated feed incubated at 20 degrees C, S. typhimurium DT104:30 persisted for at least 72 h. In contrast, in feed incubated at 30 degrees C, no S. typhimurium DT104:30 were detectable 48 h after inoculation. In prefermented feed, S. typhimurium DT104:30 died four to five times faster in feed maintained at 30 degrees C (D(value) 34-45 min) compared with feed maintained at 20 degrees C (D(value) 137-250 min). This was not entirely due to differences in lactic acid concentration as feed fermented for 72 or 96 h at 20 degrees C and feed fermented for 48 h at 30 degrees C contained similar concentrations of lactic acid (160-170 mM). Low numbers of S. typhimurium DT104:30 were still detectable in fermented feed 24 h after inoculation at 20 degrees C. In contrast, none were detectable 6-7 h after inoculation at 30 degrees C. The results of these studies indicate that it would be advisable for pig producers to control the temperature of liquid feed tanks to reduce the risk of Salmonella contamination.

Animal Feed↗

Proteolytic activities in togwa, a Tanzanian fermented food.

Proteolytic activities were investigated in sorghum-based togwa prepared by natural fermentation and using starter cultures previously isolated from the native product, i.e., Lactobacillus brevis, Lactobacillus cellobiosus, Lactobacillus fermentum, Lactobacillus plantarum, Pediococcus pentosaceus, and Issatchenkia orientalis in coculture with either L. brevis or L. plantarum. Both proteinase and aminopeptidase activities were substantially higher in naturally fermented togwa than in those with starters (14-30%, 12-70%, respectively). A variable but substantial part of the proteinase activity followed the particulate fraction of togwa; aminopeptidase activity was mainly in that fraction. The breakdown of relatively high molecular mass protein (64 kDa) in togwa was detected by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE); the products were mainly in the 14-30 kDa range. Reversed-phase fast-protein liquid chromatography (RP-FPLC)-protein/peptide patterns changed during fermentation with some variation between togwa of different cultures. Supplementation of gruel with malt increased the concentration of total protein [from 9.5% to 11.0% (w/w) on dry weight basis)] and of most of the free amino acids. Fermentation had no effect on total protein content; however, the concentration of most of the amino acids was reduced, except for the proline content that increased. Natural fermentation also increased the concentration of glutamic acid and ornithine. Fermentation by P. pentosaceus increased aspartic acid, while L. cellobiosus, L. fermentum, and L. brevis in coculture with I. orientalis increased the concentration of citrulline.

Aminopeptidases↗

Effects of fermentation temperature and Saccharomyces species on the cell fatty acid composition and presence of volatile compounds in wine.

Low temperature alcoholic fermentations are becoming more frequent due to the wish to produce wines with more pronounced aromatic profiles. However, their biggest drawback is the high risk of stuck and sluggish fermentations. Changes in the plasma membrane composition may be an adaptive response to low temperature fermentations. The production of volatile compounds and the changes in the membrane fatty acids were determined by GC to show the degree of cell adaptation and performance at low temperatures (13 degrees C) taking 25 degrees C as reference. The tests were done in two strains of Saccharomyces cerevisiae and one strain of Saccharomyces bayanus. Low temperatures restricted yeast growth and lengthened the fermentations. The analysis of plasma membrane fatty acids showed that dry yeasts had similar levels of unsaturation, between 70% and 80%, with no medium-chain fatty acids (MCFA). Long-chain saturated fatty acids (SFA) were the most frequent membrane fatty acids throughout the fermentations. Lipid composition changed with the growth temperature. The optimal membrane fluidity at low temperatures was modulated by changes in the unsaturation degree in S. cerevisiae strains. In S. bayanus, however, this change in the unsaturated fatty acid (UFA) percentage was not observed at different growth temperatures but the concentration of MCFA at low fermentation temperatures was higher. Concentrations of volatile compounds were higher in wines produced at lower temperatures and depended on the strain.

Chromatography, Gas↗

Changes in wine yeast storage carbohydrate levels during preadaptation, rehydration and low temperature fermentations.

The metabolism of glycogen and trehalose was analysed in a wine yeast strain fermenting at 25 and 13 degrees C. Trehalose and glycogen degradation were completed during the lag phase of fermentation. Ammonia was taken up rapidly and once it had been reduced to negligible amounts, the synthesis of trehalose started. Glycogen followed a similar pattern. If trehalose synthesis was taken as a stress indicator, the fermentation at 13 degrees C could not be considered stressful because the maximum concentrations are similar at both temperatures. In industrial fermentations, and after a preadaptation in grape must for several hours at 18 degrees C, the lag phase was reduced significantly, and this may be why trehalose and glycogen were completely depleted at the beginning of the low temperature fermentation. Various preadaptation conditions were tested so that their influence on trehalose and glycogen degradation could be determined. The presence of fermentable carbon sources, such as glucose or fructose, triggered the mobilisation and use of trehalose. However, just increasing the osmotic pressure did not reduce the trehalose content. No such differences were observed in glycogen metabolism.

Fermentation↗

Survival and growth of acid-adapted and unadapted Shigella flexneri in a traditional fermented Ghanaian weaning food as affected by fortification with cowpea.

Shigellae are among the major causes of diarrheal diseases in infants and young children in developing countries. We conducted a study to determine the effect of fermentation of corn and corn/cowpea doughs used to prepare a traditional weaning porridge on the survival and growth characteristics of acid-adapted and unadapted Shigella flexneri. Porridges were prepared from doughs fermented for 0, 24, and 48 h at 30 degrees C. Four-strain mixtures of acid-adapted and unadapted S. flexneri cells were separately inoculated (10(4)-10(5) and 10(6)-10(7) cfu/ml) into porridges made from unfermented (pH 5.74-6.05) and fermented (pH 4.07-4.38) doughs. Viability of acid-adapted cells was retained at higher levels in porridge made from fermented dough, compared to unfermented dough, regardless of composition of the porridge or incubation temperature. Cells inoculated into the porridges containing fermented dough were not detected (<1 cfu/ml) within 4 h at 48 degrees C. Results indicate that prior exposure of cells to acid stress renders them more resistant to subsequent acidic conditions. The addition of cowpea flour to corn dough followed by fermentation had little effect on the survival of S. flexneri in porridges made from the dough.

Adaptation, Physiological↗

Processing and storage of lye-treated carrots fermented by a mixed starter culture.

A mixed culture of Lactobacillus plantarum and Saccharomyces cerevisiae was compared with a single culture of L. plantarum as starter for the fermentation of lye-treated carrots. Using the mixed culture, more than 95% of glucose, fructose and malic acid was consumed after 7 days of fermentation in a brine containing 2.5% w/v NaCl and 0.7% acetic acid, but only 54% of sucrose was degraded. The fermentation products quantified were lactic acid, ethanol and acetic acid and carbon recovery was 88%. Using the single culture of L. plantarum, substrate consumption was lower, and carbon recovery almost 100%. In uninoculated lye-treated carrots, heterofermentative bacteria grew, with the production of considerable amounts of mannitol. In all cases, the stability, sensorial characteristics and carotenoid content of the packed product were studied during 9 months of storage at 30 degrees C, and two different preservation systems were compared: addition of preservatives, approximately 500 and 1000 mg/kg of sorbic and benzoic acid, respectively, and pasteurization at 80 degrees C for 10 min. The pasteurized samples were microbiologically stable during the storage period, while lactic acid bacteria grew in the samples with preservatives. Storage time significantly (P < 0.05) affected the texture and colour parameters (L*, a*, b*) of the carrots, but not the amounts of alpha- and beta-carotene. The type of fermentation had no significant effect on texture, colour parameters or carotenoid content. The preservation method had no significant effect on texture or carotenoid content, but did affect colour (parameter a*). The flavour of carrots fermented by the mixed culture was significantly (P < 0.05) preferred to that of those fermented with a single culture of L. plantarum.

Daucus carota↗

Influence of sodium chloride concentration on the controlled lactic acid fermentation of "Almagro" eggplants.

The effect of a commercial Lactobacillus starter and sodium chloride concentration on the fermentation of "Almagro" eggplants (Solanum melongena L. var. esculentum depressum) was studied. The results of fermentation using added starter and varying salt concentrations (4, 6, and 10% w/v) in brine were compared with the results of spontaneous fermentation taking place in brine with a salt concentration of 4%. Fresh fruits, medium in size (34-44 g), were used in all cases; all fruits were blanched under identical conditions. Temperature in the fermenters was 32+/-2 degrees C. The results obtained indicate that addition of a suitable starter shortened the fermentation process, provided the salt concentration in the brine did not exceed 6%. In the conditions tested, the eggplants obtained after fermentation were found to be of good quality though somewhat bitter which may explained by the starter employed.

Dose-Response Relationship, Drug↗

A review of traditional fermented foods and beverages of Zimbabwe.

Several traditional fermented foods and beverages are produced at household level in Zimbabwe. These include fermented maize porridges (mutwiwa and ilambazi lokubilisa) fermented milk products (mukaka wakakoralamasi and hodzeko) non-alcoholic cereal-based beverages (mahewu, tobwa and mangisi) alcoholic beverages from sorghum or millet malt (doroluthwala and chikokivana) distilled spirits (kachasu) and fermented fruit mashes (makumbi). There are many regional variations to the preparation of each fermented product. Research into the processing technologies of these foods is still in its infancy. It is, therefore, important that the microbiology and biochemistry of these products, as well as their technologies be studied and documented in order to preserve them for future generations. This article reviews the available information regarding traditional fermented foods in Zimbabwe and makes recommendations for potential research areas.

Animals↗

Growth of bifidobacteria in soymilk and their survival in the fermented soymilk drink during storage.

Growth of Bifidobacterium infantis CCRC 14633 and B. longum B6, in soymilk was investigated in the present study. It was found that soymilk could support the growth of both organisms tested. B. infantis grew better than B. longum in soymilk. Supplementation of bifitose (isomaltooligosccharie), glucose, lactose or galactose to soymilk increased the growth of B. infantis and B. longum as determined after 48 h of fermentation. On the other hand, addition of yeast extract, peptone, tryptone, casitone or N-Z-Case plus to soymilk enabled B. infantis to reach its maximum population in a shorter cultivation time of 24 h. Acid production by B. longum and B. infantis in soymilk was mainly non-growth associated, while in the yeast extract-supplemented soymilk, acid produced by B. infantis was found to be growth-associated. Populations of B. longum reduced more than did B. infantis in the prepared fermented soymilk drink during storage period. Viable population of both test organisms reduced less in the fermented drink held at 5 degrees C than at 25 degrees C. After a 10-day storage at 5 degrees C, viable B. infantis and B. longum reduced by 0.44 and 3.18 log CFU/ml, respectively, in the fermented drink. Addition of sucrose to the fermented drink resulted in an increased reduction of viable bifidobacteria during the storage period. This phenomenon was most prominent with B. infantis in the fermented drink held at 25 degrees C.

Animals↗

Role of fermentable carbohydrate supplements with a low-protein diet in the course of chronic renal failure: experimental bases.

During the past few years, considerable attention has been given to the impact of nutrition on kidney disease. The question arises of whether the effect of a moderate dietary protein restriction could be reinforced by enrichment of the diet with fermentable carbohydrates. Feeding fermentable carbohydrates may stimulate the extrarenal route of nitrogen (N) excretion through the fecal route. Such an effect has been reported in several species, including healthy humans and patients with chronic renal failure (CRF). Furthermore, studies of these subjects show that the greater fecal N excretion during the fermentable carbohydrate supplementation period was accompanied by a significant decrease in plasma urea concentration. In animal models of experimental renal failure, the consumption of diets containing fermentable carbohydrates results in a greater rate of urea N transfer from blood to the cecal lumen, where it is hydrolyzed by bacterial urease before subsequent microflora metabolism and proliferation. Therefore, this results in a greater fecal N excretion, coupled with a reduction in urinary N excretion and plasma urea concentration. Because elevated concentrations of serum urea N have been associated with adverse clinical symptoms of CRF, these results suggest a possible usefulness of combining fermentable carbohydrates with a low-protein diet to increase N excretion through the fecal route. Further investigations in this population of patients of whether fermentable carbohydrates in the diet may be beneficial in delaying or treating the symptoms and chronic complications of CRF will certainly emerge in the future. This should be realized without adversely affecting nutritional status and, as far as possible, by optimizing protein intake for the patients without being detrimental to renal function.

Ammonia↗

Polysaccharide production by kefir grains during whey fermentation.

Fermentation of deproteinised whey with kefir grains CIDCA AGK1 was studied focusing on polysaccharide production from lactose. Kefir grains were able to acidify whey at different rates depending on the grain/whey ratio. During fermentation, kefir grains increased their weight and a water-soluble polysaccharide was released to the media. Exopolysaccharide concentration increased with fermentation time, reaching values of 57.2 and 103.4 mg/l after 5 days of fermentation in cultures with 10 and 100 g kefir grains/l, respectively. The polysaccharide fraction quantified after fermentation corresponded to the soluble fraction, because part of the polysaccharide became a component of the grain. Weight of kefir grains varied depending on the time of fermentation. Polysaccharide production was affected by temperature. Although the highest concentration of polysaccharide in the media was observed at 43 degrees C at both grain/whey ratios, the weight of the grains decreased in these conditions. In conclusion, kefir grains were able to acidify deproteinised whey, reducing lactose concentration, increasing their weight and producing a soluble polysaccharide.

Animals↗

Immunomodulating effects of milks fermented by Lactobacillus helveticus and its non-proteolytic variant.

The effect of milks fermented by Lactobacillus helveticus and its non-proteolytic variant on mucosal and tumoral immunity was studied. Milks fermented by Lb. helveticus wild type or its non-proteolytic variant were administered orally to mice for different periods (3, 5 and 7 d). The immune response was assessed by analysing the activity of the peritoneal macrophages, the number of cells secreting IgA associated with the gut-associated lymphoid tissue and with the bronchial-associated lymphoid tissue. The number of cells was determined by direct immunofluorescence. The antitumour activity was monitored by studying the regression of the subcutaneously implanted fibrosarcomas. After 3 d feeding of milk fermented by Lb. helveticus wild type, the number of sIgA increased significantly at both the intestinal and bronchial levels, indicating that a cellular migration had occurred. This effect was not noticeable when milk fermented by Lb. helveticus Protease (-) was orally administered. Both fermented milks (wild type or its variant) exhibited an effect on the activity of the peritoneal macrophages, which might be indirectly correlated to the regression of the fibrosarcoma. Although the mechanism by which the lactic acid bacteria enhance the immune system is not clear, this study clearly suggests that the bioactive compounds released during milk fermentation might contribute to the immunoenhancing properties of these products. By releasing biopeptide, lactic acid bacteria have important implications in modulation of the host's immune response, more specifically its cellular immune response.

Adjuvants, Immunologic↗

Field-flow fractionation as analytical technique for the characterization of dry yeast: correlation with wine fermentation activity.

Important oenological properties of wine depend on the winemaking yeast used in the fermentation process. There is considerable controversy about the quality of yeast, and a simple and cheap analytical methodology for quality control of yeast is needed. Gravitational field flow fractionation (GFFF) was used to characterize several commercial active dry wine yeasts from Saccharomyces cerevisiae and Saccharomyces bayanus and to assess the quality of the raw material before use. Laboratory-scale fermentations were performed using two different S. cerevisiae strains as inocula, and GFFF was used to follow the behavior of yeast cells during alcoholic fermentation. The viable/nonviable cell ratio was obtained by flow cytometry (FC) using propidium iodide as fluorescent dye. In each experiment, the amount of dry wine yeast to be used was calculated in order to provide the same quantity of viable cells. Kinetic studies of the fermentation process were performed controlling the density of the must, from 1.071 to 0.989 (20/20 density), and the total residual sugars, from 170 to 3 g/L. During the wine fermentation process, differences in the peak profiles obtained by GFFF between the two types of commercial yeasts that can be related with the unlike cell growth were observed. Moreover, the strains showed different fermentation kinetic profiles that could be correlated with the corresponding fractograms monitored by GFFF. These results allow optimism that sedimentation FFF techniques could be successfully used for quality assessment of the raw material and to predict yeast behavior during yeast-based bioprocesses such as wine production.

Cell Culture Techniques↗

Dietary modulation of bacterial fermentative capacity by edible seaweeds in rats.

The effect of edible seaweeds [nori (Porphyra tenera) and wakame (Undaria pinnatifida)] on the modulation of colonic microbiota was studied in adult male Wistar rats. Each alga was fed to rats as the only source of dietary fiber and compared with cellulose. After 12 days, animals were sacrificed and cecal contents used as inoculum to ferment lactulose, citrus pectin, cellulose, nori, and wakame in vitro. Dietary treatment did not affect food intake or food efficiency, yet alga caused a significant increase in cecal weight. Nori and wakame were poorly fermented by the cellulose inoculum, with intermediate substrate degradation (76 and 57% for nori and wakame, respectively) and low metabolism to short-chain fatty acids (SCFA) (30% fermentability compared with lactulose). Cecal contents from rats fed nori and wakame showed a reduced ability to ferment all of the studied substrates compared with the cellulose inoculum, causing a reduction in SCFA production and dry matter disappearance. Only nori induced a bacterial adaptation that brought about a higher fermentation of this substrate. The different behaviors of the two algae could be due to their distinct chemical compositions. In conclusion, nondigestible components of edible seaweeds modified the metabolic activity of intestinal microflora, leading to a reduction of its fermentative capacity.

Animals↗

Isoflavone transformation during soybean koji preparation and subsequent miso fermentation supplemented with ethanol and NaCl.

Soybeans were soaked with water for 4 h, steam-cooked, inoculated with the conidia of Aspergillus oryzae, and incubated for 3 days for koji preparation. The koji was then mixed with water-soaked and steam-cooked soybeans (1:2, w/w), ground into paste, and supplemented with 15% ethanol and 12.5% NaCl or 3% ethanol and 6% NaCl for miso fermentation at 30 degrees C. Daidzin, genistin, daidzein, and genistein contents were extracted from the lyophilized and pulverized soybean powder or from the miso homogenate by a developed one-tube procedure and analyzed with an HPLC. After water soaking, daidzein and genistein contents increased markedly, whereas daidzin and genistin contents decreased. Further increases of daidzein and genistein contents and decreases of daidzin and genistin contents were observed after koji mold growth. During fermentation, fungal and lactic acid bacterial (LAB) growth in the miso products was inhibited, whereas soluble protein contents increased much more rapidly in the low-salt miso products supplemented with 3% ethanol and 6% NaCl than the other products. When the 4- and 8-week-fermented miso products were cooked with tofu for sensory evaluation, flavor ratings of the low-salt products were higher than that of a popular commercial product. In both products, the most daidzins and genistins were hydrolyzed after 4 weeks of fermentation. The hydrolytic enzymes contributing to isoflavone transformation originated from soybeans after water soaking and from koji with mold growth. It was of merit that the low-salt fermented products were fairly acceptable in flavor rating and rich in daidzein and genistein contents after 4 weeks of fermentation.

Aspergillus oryzae↗

Carotenoid content, physicochemical, and sensory qualities of deep-fried carrot chips as affected by dehydration/rehydration, antioxidant, and fermentation.

Carrot slices were subjected to one of the following experiments prior to deep-frying: (A) dehydration/rehydration, (B) soaking in different antioxidants, and (C) fermentation with/without blanching. There were no significant differences (P > or = 0.05) in carotenoid contents among carrot chips treated with/without dehydration. Soaking in sodium metabisulfite resulted in the highest carotenoid content and lightness (L), redness (a), and yellowness (b) values among the antioxidant treatments. Fermentation without blanching significantly decreased (P < 0.05) carotenoid content, vitamin A activity, and fat content. Dehydration and fermentation with blanching significantly increased (P < 0.05) the lightness (L), redness (a), and yellowness (b) values of the chips. Dehydration/rehydration, but not antioxidant and fermentation, significantly decreased (P < 0.05) the water activity of the chips. The textural values of carrot chips prepared using sodium metabisulfite, without dehydration and without fermentation, were the lowest among other treatments which suggests the crispiest. Carrot chips prepared using sodium metabisulfite, without dehydration and without fermentation, had the highest carotenoid content and retention, and the highest overall acceptability score.

Antioxidants↗