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Isolation and characterization of microorganisms associated with the traditional sorghum fermentation for production of sudanese kisra.

Sorghum flour obtained from Sudan was mixed with water in a 1:2 (wt/vol) ratio and fermented at 30 degrees C for 24 h. The bacterial populations increased with fermentation time and reached a plateau at approximately 18 h. At the end of 24 h, sorghum batter pH had dropped from 5.95 to 3.95 and the batter had a lactic acid content of 0.80%. The microbial population during the 24 h of fermentation consisted of bacteria (Pediococcus pentosaceus, Lactobacillus confusus, Lactobacillus brevis, Lactobacillus sp., Erwinia ananas, Klebsiella pneumoniae, and Enterobacter cloacae), yeasts (Candida intermedia and Debaryomyces hansenii), and molds (Aspergillus sp., Penicillium sp., Fusarium sp., and Rhizopus sp.). P. pentosaceus was the dominant microorganism at the end of the 24-h fermentation. When three consecutive fermentations using an inoculum from the previous fermentation were carried out, the bacterial population increase plateaued at 9 h. The microbial populations in these fermentations were dominated by P. pentosaceus.

Journal Article↗

In Vitro Stimulation of Forage Fiber Degradation by Ruminal Microorganisms with Aspergillus oryzae Fermentation Extract.

Aspergillus oryzae fermentation extract (Amaferm) was evaluated for its ability to influence degradation of brome grass and switchgrass fiber fractions by mixed ruminal microorganisms in vitro. Addition of Amaferm at a concentration of 0.067 mg/ml, which is approximately the concentration found in the rumen ecosystem (0.06 mg/ml), increased the degradation of brome grass neutral detergent fiber (NDF) by 28% after fermentation for 12 h (P < 0.01), but had no effect after fermentation for 24 or 48 h. The levels of degradation of both the cellulose and hemicellulose fractions were increased after fermentation for 12 h (P < 0.01). Additions of 0.08 and 8% (vol/vol) Amaferm filtrate (12.5 g/100 ml) stimulated degradation of switchgrass NDF by 12 and 24% (P < 0.01), respectively, after fermentation for 12 h; when 80% filtrate was added, degradation was decreased by 38%. The concentrations of total anaerobes in culture tubes containing 80% filtrate were 5 times greater than the concentrations in the controls; however, the concentrations of cellulolytic organisms were 3.5 times lower than the concentrations in the controls (P < 0.05). These results suggested that the filtrate contained high concentrations of soluble substrate which did not allow the cellulolytic organisms to compete well with other populations. The remaining concentrations of esterified p-coumaric and ferulic acids were lower at 12 h in NDF residues obtained from fermentation mixtures supplemented with Amaferm. Because the total anaerobes were not inhibited in fermentation mixtures containing Amaferm, antibiotics are unlikely to be involved as a mode of action for increasing NDF degradation. The possibility that Amaferm contains enzymes (possibly esterases) that may play a role in stimulating the rate of fiber degradation by mixed ruminal microorganisms by removal of plant cell wall phenolic acid esters is discussed.

Journal Article↗

Fermentation and Sulfur Reduction in the Mat-Building Cyanobacterium Microcoleus chthonoplastes.

The mat-building cyanobacterium Microcoleus chthonoplastes carried out a mixed-acid fermentation when incubated under anoxic conditions in the dark. Endogenous storage carbohydrate was fermented to acetate, ethanol, formate, lactate, H(inf2), and CO(inf2). Cells with a low glycogen content (about 0.3 (mu)mol of glucose per mg of protein) produced acetate and ethanol in equimolar amounts. In addition to glycogen, part of the osmoprotectant, glucosyl-glycerol, was degraded. The glucose component of glucosyl-glycerol was fermented, whereas glycerol was released into the medium. Cells with a high content of glycogen (about 2 (mu)mol of glucose per mg of protein) did not utilize glucosyl-glycerol. These cells produced more acetate than ethanol. M. chthonoplastes was also capable of using elemental sulfur as the electron acceptor during fermentation, resulting in the production of sulfide. With sulfur present, acetate production increased whereas ethanol production decreased. Also, less formate was produced and the evolution of hydrogen ceased completely. In general, the carbon recoveries were satisfactory but the oxidation-reduction balances were too high. The latter could be explained by assuming the reduction of ferric iron, which is associated with the cells, mediated by the oxidation of formate. The switch from photoautotrophic to fermentative metabolism did not require de novo protein synthesis, and fermentation started immediately upon transfer to dark anoxic conditions. From the molar ratios of the fermentation products and from measurement of enzyme activities in cell extracts, we concluded that glucose derived from glycogen and glucosyl-glycerol is degraded via the Embden-Meyerhof-Parnas pathway.

Journal Article↗

Pure culture fermentation of green olives.

The method previously developed by us for the pure-culture fermentation of brined cucumbers and other vegetables has been applied successfully to Manzanillo variety olives. Field-run grade fruit was processed first by conventional procedures to remove most of the bitterness. Then the relative abilities of Lactobacillus plantarum, L. brevis, Pediococcus cerevisiae, and Leuconostoc mesenteroides to become established and produce acid in both heat-shocked (74 C for 3 min) and unheated olives, brined at 4.7 to 5.9% NaCl (w/v basis), were evaluated. The heat-shock treatment not only proved effective in ridding the fruit of naturally occurring, interfering, and competitive microbial groups prior to brining and inoculation, but also made the olives highly fermentable with respect to growth and acid production by the introduced culture, particularly L. plantarum. Of the four species used as inocula, L. plantarum was by far the most vigorous in fermentation ability. It consistently produced the highest levels of brine acidity (1.0 to 1.2% calculated as lactic acid) and the lowest pH values (3.8 to 3.9) during the fermentation of heat-shocked olives. Also, L. plantarum completely dominated fermentations when used in two-species (with P. cerevisiae) and three-species (with P. cerevisiae and L. brevis) combinations as inocula. In contrast, when L. plantarum was inoculated into the brines of unheated olives it failed to become properly established; the same was true for the other species tested, but even to a more pronounced degree. L. brevis was the only species used that failed to develop in brines of both heat-shocked and unheated olives. Modification of the curing brine by the addition of lactic acid at the outset, either with or without dextrose, led to a much earlier onset of fermentation with accompanying acid development, as compared to treatments with dextrose alone or nonadditive controls. Reasons for the marked improvement of the fermentability of Manzanillo olives receiving the prebrining heat-shock treatment are discussed.

Journal Article↗

Dietary fermented ammoniated condensed whey and postprandial effects on blood and urine acid-base metabolites in lactating cows.

Effects of a fermented whey product containing 38% lactic acid, time post-feeding, and temperature on acid-base metabolism were studied with four lactating Jersey cows in a 4 X 4 Latin square. Treatments included 0, 33, 66, and 100% replacement in soybean meal with fermented whey in a 60% corn silage, 40% concentrate diet (dry basis). Increasing fermented whey to 100% reduced intake and mild production, 27 and 32% compared to animals fed 0 and 33% fermented whey diets. Fermented whey had no effect on blood pH, carbon dioxide tension, bicarbonate, and potassium whereas increasing fermented whey about 33% replacement reduced glucose and sodium in plasma 15 and 40 mg/100 ml from controls. Increasing consumption of fermented whey to 100% replacement increased urinary ammonium excretion 1365/meq per day and slightly decreased urine pH, which suggested changes in acid excretion. Effects of time postfeeding on blood traits were measured from samples taken at 0, 1, 2, 3, 4, 6, and 8 h. Blood pH was higher at 1 to 3 h as compared with 4 to 8 h postfeeding, whereas carbon dioxide tension and bicarbonate increased 10.6 mm mercury and 6.3 meq/liter at 1 h and declined afterward. Period effects for blood acid-based characteristics showed decreasing blood carbon dioxide tension with increasing ambient temperature.

Acid-Base Equilibrium↗

[Suppression of allergy development by habitual intake of fermented milk foods, evidence from an epidemiological study].

BACKGROUND: The interest in anti-allergic immunoregulation by lactic acid bacteria (LAB) has been growing over the last few decades. However, little is known about the effect of habitually intake of fermented milk on allergy development. METHODS: An epidemiological study was carried out on the first-year junior high school students in Wakayama Prefecture. Analyses were performed to investigate the relationships among eating habits of fermented milk or fermented soybean foods and the presence of atopic diseases. Serum levels of total IgE values, specific IgE to house dust mite and Japanese cedar pollen in these subjects were evaluated to clarify atopic status. Analyses were performed on the data of 134 subjects. RESULTS: Serum total IgE levels were found to be significantly lower in those subjects habitually eating yogurt and/or fermented milk drinking, in comparison with those who do not habitually eat such fermented milk foods. Subjects with habitual intake of these fermented milk foods were significantly lower in having various allergy diseases compared with those without such an eating habit. However, no difference was found on the total IgE titers and having allergy diseases between subjects with or without habitual intake of Natto, a fermented soybean food. CONCLUSION: These findings supported the idea that intestinal bacteria such as lactic acid bacteria and bifidobacteria involve in regulation of allergy development. Our results indicated the need of further scale-up epidemiological study to verify the present finding.

Child↗

Aerobic fermentation and the depletion of the amino acid pool in yeast cells.

The amino acid pool of yeast cells, Saccharomyces cerevisiae, incubated with galactose remains at a constant level for 100 minutes. This is 30 minutes beyond the time at which the oxidative phase of the induced-enzyme formation begins. Washed yeast cells, the pools of which have been depleted 60 per cent by incubation with glucose, do not replenish their pools as do washed cells incubated without a substrate. These facts indicate that the induced enzymes are formed at least partially from pool-replenishing amino acids. The time of onset of pool depletion is the time at which the aerobic fermentation phase of induced-enzyme formation begins for cells incubated with galactose. With 0.1 per cent galactose the respiratory phase begins at 100 minutes but no aerobic fermentation nor pool depletion occurs. The rates of respiration and aerobic fermentation are constant for four glucose concentrations from 0.1 to 1.0 per cent. The amount of aerobicfermentation is proportional to the initial concentration of glucose. Amino acid pool depletion occurs for all concentrations but depletion ceases and is followed by pool replenishment after aerobic fermentation is complete. Ultraviolet radiations, which delay the appearance of the respiratory phase of induced-enzyme formation, completely eliminate both the appearance of aerobic fermentation and pool depletion. The results indicate an intimate association between aerobic fermentation and amino acid pool depletion.

Amino Acids↗

Yeast Strain Development and Process Intensification in High-Gravity Fermentation.

High- and very-high-gravity (HG/VHG) fermentation increases substrate loading and product titers, thereby improving fermenter utilisation and potentially reducing water use and downstream processing requirements. Initially developed for brewing and fuel ethanol production, these approaches are now applied more broadly in food, beverage, and bioproduct manufacturing. This MiniReview summarises operational definitions and industrial drivers of HG/VHG fermentation and examines the associated constraints in rheology, mass and heat transfer, osmotic and ethanol stress, nutrient availability, and oxidative damage. Yeast improvement strategies are reviewed, including adaptive laboratory evolution, mutagenesis, genome shuffling, multiplex genome editing, non-conventional yeasts, and multi-omics-guided selection. Process developments such as no-cook simultaneous liquefaction, saccharification and fermentation (SLSF), enzyme formulation, nutrient management, and in situ product recovery are considered together with applications in alcoholic beverages, organic acids, microbial lipids, and other value-added products. The review also discusses coproduct valorisation and the need to integrate strain development with process design. Current evidence supports HG/VHG fermentation as a useful process-intensification platform, although performance and sustainability depend strongly on feedstock, operating conditions, product requirements, and the basis used to report fermentation outcomes.

circular bioeconomy↗

Observations on a laboratory method for submerged acetic fermentation.

Submerged acetic fermentation experiments were performed for the purpose of determining the conditions under which this type of fermentation should be conducted under laboratory conditions. The apparatus used consisted of a set of glass tubes provided with air spargers. Acetobacter acetigenum was found to be the most suitable bacterium among six Acetobacter compared under submerged acetic fermentation conditions in a synthetic medium. Statistically significant different rates of fermentation were observed in acetators that were identical in construction, fermentation medium, and aeration characteristics. Extremely long growth lag periods and complete absence of growth were often observed when starting fermentations. The causes of this behavior were investigated. It was found that it was not produced by lack of nutrients or by presence of a bacteriophage. Different kinds of bacterial starters were studied and compared. Cultures maintained in a liquid medium were reliable starters with a short growth lag period. Liquid medium cultures maintained their good starter characteristics after periods of storage of up to 11 weeks at 40 F (4 C).

Acetobacter↗

Regulation of alcoholic fermentation in batch and chemostat cultures of Kluyveromyces lactis CBS 2359.

Kluyveromyces lactis is an important industrial yeast, as well as a popular laboratory model. There is currently no consensus in the literature on the physiology of this yeast, in particular with respect to aerobic alcoholic fermentation ('Crabtree effect'). This study deals with regulation of alcoholic fermentation in K. lactis CBS 2359, a proposed reference strain for molecular studies. In aerobic, glucose-limited chemostate cultures (D = 0.05-0.40 h-1) growth was entirely respiratory, without significant accumulation of ethanol or other metabolities. Alcoholic fermentation occurred in glucose-grown shake-flask cultures, but was absent during batch cultivation on glucose in fermenters under strictly aerobic conditions. This indicated that ethanol formation in the shake-flask cultures resulted from oxygen limitation. Indeed, when the oxygen feed to steady-state chemostat cultures (D = 0.10 h-1) was lowered, a mixed respirofermentative metabolism only occurred at very low dissolved oxygen concentrations (less than 1% of air saturation). The onset of respirofermentative metabolism as a result of oxygen limitation was accompanied by an increase of the levels of pyruvate decarboxylase and alcohol dehydrogenase. When aerobic, glucose-limited chemostat cultures (D = 0.10 h-1) were pulsed with excess glucose, ethanol production did not occur during the first 40 min after the pulse. However, a slow aerobic ethanol formation was invariably observed after this period. Since alcoholic fermentation did not occur in aerobic batch cultures this is probably a transient response, caused by an imbalanced adjustment of enzyme levels during the transition from steady-state growth at mu = 0.10 h to growth at mu max. It is concluded that in K. lactis, as in other Crabtree-negative yeasts, the primary environmental trigger for occurrence of alcoholic fermentation is oxygen limitation.

Aerobiosis↗

Design of metabolic feed controllers: application to high-density fermentations of Pichia pastoris.

High-density cultures of the methylotrophic yeast Pichia pastoris were found to exhibit oscillatory metabolic behavior when fed methanol under closed-loop operations using a dissolved oxygen-based bioreactor feed controller (DOstat). This behavior, if left unattended, led to the irreversible loss of culture productivity, 1 to 2 days after growth on methanol commenced, presumably through the accumulation of incompletely oxidized intermediates. To provide insights into how fermentation operation conditions and strain variations might contribute to this phenomenon a theoretical study was initiated. In this article, a simple mathematical model of the closed-loop DOstat is developed and analyzed with the goal of deriving theoretical stability criteria applicable to the design of metabolic feed controllers during high-cell-density fermentations. The model consists of a system of differential, integral, and algebraic equations describing the biological process and the components of the standard proportional-integral (PI) feedback controller. Inputs into the process model include metabolic pathway information, oxidative metabolism stoichiometry, and substrate uptake kinetics. Frequency-response analysis and the Bode stability criterion are applied to derive controller stability criteria with particular emphasis on elucidating the role(s) that model parameters, both biological and operational, have on system stability. The results of this analysis are used to construct a closed-loop fermentation-operating diagram relating fermentation operating parameters to the oxidative capacity of the culture. The utility of this analysis is demonstrated through the application of the results to the design and stabilization of the DOstat during high-density fermentations of P. pastoris growing on methanol or glycerol. From this analysis, it is possible to conclude that, when the rate of oxygen transfer approaches in magnitude the rate of oxygen utilization, the potential for controller destabilization is greatest. Under these conditions, the region of parameter space associated with stable controller operations is further constrained. Because the only system-specific experimental inputs into the model are the routinely measured residual substrate and dissolved oxygen concentrations, the framework presented should provide simple and practical theoretical guidelines relevant to the design of similar industrial fermentation feed controllers independent of the specifics of the biological system at hand.

Biotechnology↗

Utilization of Lactobacillus sp. for steroid glycoalkaloids degradation by lactic acid fermentation.

The degradation fo steroid glycoalkaloids (SGAs) has been studied in model solutions. The number of colony forming units (CFU) was determined using a nondirect (cultivation) method during all stages of fermentation. The changes in SGAs content were observed by HPLC on the Supelcosil LC-NH2 column. The changes in alpha-tomatine concentration added to fermented Lactobacillus MRS broth have been studied. A mathematical model of steroid glycoalkaloids degradation during lactic fermentation was proposed. The mathematical model was based on the experimental data of SGA and glucose concentration and should be used for study and prediction of SGA concentration changes of fermented samples. The ratio of SGA degradation rate by fermentation and by lactic acid hydrolysis was calculated. The experimental data evaluated by proposed mathematical model for the selected strain of Lactobacillus plantarum 976H show real feasibility for SGA degradation by lactic acid fermentation.

Alkaloids↗

Kinetic model for nitrogen-limited wine fermentations.

A physical and mathematical model for wine fermentation kinetics has been developed to predict sugar utilization curves based on experimental data from wine fermentations with various initial nitrogen and sugar concentrations in the juice. The model is based on: (1) yeast cell growth limited by nitrogen; (2) sugar utilization rates and ethanol production rates proportional solely to the number of viable cells; and (3) a death rate for cells proportional to alcohol content. All but one parameter in the model can be estimated from existing data. However, experiments to find this final parameter, a constant describing cell death, indicate that cell death may not be the critical factor in determining fermentation kinetics as cell viability remains significant until sugar utilization has ceased. The model, nevertheless, predicts a transition from normal to sluggish to stuck fermentations as initial nitrogen levels decrease. It also predicts that fermentations with high initial Brix levels may go to completion when supplemented with nitrogen in the form of ammonia. Therefore, we hypothesize that the model is valid but that ethanol causes the yeast cells to become inactive while remaining viable. Experimental verification of the model has been performed using flask-scale experiments. The model has also been used to evaluate the possibility of using nitrogen or viable cell additions to avoid or correct problem (i.e., sluggish or stuck) fermentations.

Carbohydrate Metabolism↗

Effect of wood ash treatment on improving the fermentability of wood hydrolysate.

Softwood hydrolysates were overlimed with wood ash to improve the fermentability of hydrolysates. It could be demonstrated in fermentation tests that wood ash treatment increases fermentability compared to the hydrolysates untreated and treated with alkaline compounds such as Ca(OH)(2), NaOH, and KOH, which are commonly used for overliming. The enhanced fermentability of the hydrolysate treated with wood ash is due to the reduction of the inhibitors of the fermentation such as furan and phenolic compounds and to nutrient effects of some inorganic components from the wood ash on the fermentation.

Bioreactors↗

Application of dynamic calorimetry for monitoring fermentation processes.

The rate of heat evolution (kcal/liter-hr) in mycelial fermentations for novobiocin and cellulase production with media containing noncellular solids was measured by an in situ dynamic calorimetric procedure. Thermal data so obtained have proved significant both in monitoring cell concentration during the trophophase (growth phase) and in serving as a physiological variable in the fermentation process. The validity of this technique has been demonstrated by closing the overall material and energy balances. The maintenance energy in a batch fermentation can be calculated by integrating heat evolution data. This integration method is applicable to a fermentation lacking a precise cell growth curve. The maintenance coefficient, obtained for the novobiocin fermentation by Streptomyces niveus, is equal to 0.028 g glucose equivalent/g cell-hr. The production of novobiocin in the idiophase (production phase) also correlates well with the amount of energy catabolized for maintenance and this results in an observed conversion yield of glucose to novobiocin of 11.8 mg of novobiocin produced per gram of glucose catabolized. A new physiological variable, kilocalories of heat evolved per millimole of oxygen consumed, has been proposed to monitor the state of cells during the fermentation. This method may provide a simple way to monitor on-line shifts in the efficiency of cell respiration and changes in growth yields during a microbial process.

Calorimetry↗

Proteolytic degradation of ewe milk proteins during fermentation of yoghurts and storage.

Yoghurts are mostly produced from cow milk and to a very limited extent from ewe milk. The evolution of caseins and whey proteins in ovine milk submitted to different thermal treatments (63 degrees C/30 min; 73 degrees C/15 min; 85 degrees C/10 min or 96 degrees C/5 min) was followed during fermentation of yoghurts and during their storage up to 14 days, using two different sets of starters. One set of starter LAB was a "ropy" culture (YC-191), which is a well-defined mixed strain culture containing Streptococcus thermophilus ST-143 and Lactobacillus delbrueckii subsp. bulgaricus (LB-18 and LB-CH2). The other set of starter bacteria (YC-460) was a standard yoghurt culture("non-ropy") containing mixed strain culture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. Contents of free amino groups in produced yoghurts increased gradually during the fermentation, up to a maximal value obtained after 4 h fermentation, then they did not change significantly during storage of yoghurt produced with YC-191 starter. In contrary, a large drop in the amount of free amino groups was observed in the first 24 h of storage in the case of yoghurt made with YC-460 indicating that microorganisms continue still to grow in low temperatures. During fermentation and storage of both yoghurt types, alpha-lactalbumin was hydrolyzed to a slightly bigger extent than beta-lactoglobulin. During fermentation, beta-casein was slightly more degraded than alpha(s)-caseins; however, the opposite was observed during storage up to 14 days. Generally, a more intense heat pretreatment led to a higher degradation of whey proteins and caseins during fermentation and storage. Differences in proteolytic activity between the two starters used (whey proteins more degraded by YC-191; caseins more degraded by YC-460) may lead to improvement in production and formulation of yoghurts differing in their physicochemical and rheological properties.

Amino Acids↗

Fermented dairy products, dietary calcium and colon cancer: a case-control study in The Netherlands.

To examine whether the consumption of fermented dairy products or the dietary intake of calcium decreases colon cancer risk, a case-control study was conducted in the The Netherlands. Dietary patterns were assessed in detail (for cases before diagnosis or symptoms occurred) using a structured dietary history questionnaire. After adjustment for potential confounding variables, consumption of fermented dairy products, hard cheese and unfermented dairy products was not significantly associated with risk of colon cancer: an odds ratio (OR) of 1.1 was found for individuals consuming more than one serving of fermented dairy products per day as compared to those consuming less than 10% of one serving a day. Adjustment for dietary calcium attenuated the associations. Total dietary calcium was positively but non-significantly associated with colon cancer risk after adjustment for age, gender, urbanization level and total energy intake. Additional adjustment for a positive family history of colorectal cancer, cholecystectomy and energy-adjusted intake of total fat, dietary fibre, vitamin C and alcohol increased the association. No differences were observed between calcium from fermented and from unfermented dairy sources. The observed associations for fermented dairy products and dietary calcium differed between men and women: positive significant associations were observed in men, while in women non-significant inverse associations were found. Our results do not support the hypothesis that an increased intake of commercially available, commonly used fermented dairy products or dietary calcium decreases the risk of colon cancer.

Aged↗

Effects of the nematode Gyrinicola batrachiensis on development, gut morphology, and fermentation in bullfrog tadpoles (Rana catesbeiana): a novel mutualism.

We describe a novel mutualism between bullfrog tadpoles (Rana catesbeiana) and a tadpole-specific gastrointestinal nematode (Gyrinicola batrachiensis). Groups of tadpoles were inoculated with viable or nonviable nematode eggs, and development, morphology, and gut fermentation activity were compared between nematode-infected and uninfected tadpoles. Nematode infection accelerated tadpole development; the mean time to metamorphosis was 16 d shorter and the range of times to metamorphosis was narrower in nematode-infected tadpoles than in uninfected tadpoles. At metamorphosis, infected and uninfected bullfrogs did not differ in body size or condition. Colon width, wet mass of colon contents, and concentrations of most fermentation byproducts (short-chain fatty acids: SCFAs) in the hindgut were greater in infected tadpoles. Furthermore, in vitro fermentation yields for all SCFAs combined were over twice as high in infected tadpoles than in uninfected tadpoles. One explanation for accelerated development in infected tadpoles is the altered hindgut fermentation associated with the nematodes. Energetic contributions of fermentation were estimated to be 20% and 9% of the total daily energy requirement for infected and uninfected tadpoles, respectively. Infection by G. batrachiensis nematodes potentially confers major ecological and evolutionary advantages to R. catesbeiana tadpoles. The mutualism between these species broadens our understanding of the taxonomic diversity and physiological contributions of fermentative gut symbionts and suggests that nematodes inhabiting the gut regions of other ectothermic herbivores might have beneficial effects in those hosts.

Animals↗