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Results for “In vitro fermentation”

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PubMed1984

Screening of sugars inhibitory against sucrose-dependent synthesis and adherence of insoluble glucan and acid production by Streptococcus mutans.

Various sucrose derivatives and a sucrose analogue were enzymatically synthesized on the assumption that structural similarities of the sugars to sucrose would endow them with an ability to inhibit the cariogenicity of sucrose. The effect of these sugars on sucrose-dependent synthesis and adherence of insoluble glucan and on acid fermentation of sucrose by Streptococcus mutans was examined in vitro. Although none of them inhibited the acid fermentation, several sugars not only inhibited the synthesis and adherence of insoluble glucan, but were themselves only slightly fermented as well.

Acids↗

PubMed1987

Ethanol production during batch fermentation with Saccharomyces cerevisiae: changes in glycolytic enzymes and internal pH.

During batch fermentation, the rate of ethanol production per milligram of cell protein is maximal for a brief period early in this process and declines progressively as ethanol accumulates in the surrounding broth. Our studies demonstrate that the removal of this accumulated ethanol does not immediately restore fermentative activity, and they provide evidence that the decline in metabolic rate is due to physiological changes (including possible ethanol damage) rather than to the presence of ethanol. Several potential causes for the decline in fermentative activity have been investigated. Viability remained at or above 90%, internal pH remained near neutrality, and the specific activities of the glycolytic and alcohologenic enzymes (measured in vitro) remained high throughout batch fermentation. None of these factors appears to be causally related to the fall in fermentative activity during batch fermentation.

Ethanol↗

PubMed1996

Phytic acid, in vitro protein digestibility, dietary fiber, and minerals of pulses as influenced by processing methods.

The objective of this project was to determine the effect of various types of processing on selected nutrition related parameters of commonly consumed Indian pulses and soybean. Germination reduced the phytic acid content of chickpea and pigeonpea seeds by over 60%, and that of mung bean, urd bean, and soybean by about 40%. Fermentation reduced phytic acid contents by 26-39% in all these legumes with the exception of pigeonpea in which it was reduced by more than 50%. Autoclaving and roasting were more effective in reducing phytic acid in chickpea and pigeonpea than in urd bean, mung bean, and soybean. Germination and fermentation greatly increased the in vitro protein digestibility (IVPD). IVPD was only slightly increased by roasting and autoclaving of all legumes. Germination and fermentation also remarkably decreased the total dietary fiber (TDF) in all legumes. Autoclaving and roasting resulted in slight increases in TDF values. All the processing treatments had little effect on calcium, magnesium and iron contents.

Dietary Fiber↗

PubMed2001

Influence of supplementary fibrolytic enzymes on the fermentation of corn and grass silages by mixed ruminal microorganisms in vitro.

This study was done to determine the effectiveness of supplementary enzymes at increasing the fiber digestion by ruminal microorganisms and to assess whether enzyme activity limits the rate of fiber digestion in ruminal digesta. In vitro comparisons of enzyme activities in two feed enzyme preparations (A and B) with enzyme activities extracted from ruminal fluid indicated that the addition of fibrolytic enzymes at the application rates recommended by the manufacturers would not be expected to increase significantly glycanase and polysaccharidase activities in ruminal fluid. Preparations A and B both increased (P < 0.001) the rate of gas production from freeze-dried corn and grass silages in in vitro incubations with ruminal fluid, but only at concentrations much higher than recommended application rates. Autoclaved controls had little or no effect. Ultrafiltration of enzyme B indicated that most stimulation was due to components >100 kDa, which is consistent with the cause of the stimulation being enzyme activity. Fibrolytic enzymes from other sources were also able to stimulate gas production: increased rates of gas production were observed in seven out of eight combinations of "cellulase" and corn or grass silage (P < 0.05). The comparison of glycanase and polysaccharidase activities with gas-stimulatory activity in the different enzyme preparations indicated that the highest correlation was between increased gas production and enzyme activity against microgranular cellulose (P < 0.05). In a wider range of fibrolytic enzyme preparations, those with endo-(beta-1,4)- or exo-(beta-1,4)-xylanase activity equal to that of preparation A did not produce similar increased rates of fermentation of corn silage when glucanase activity was low (P > 0.05). In contrast, preparations with glucanase activity similar to enzyme A gave at least as great (P < 0.05) an improvement in gas production than enzyme A, irrespective of xylanase activity. It was concluded that enzyme activity, probably a type of endo-(beta-1,4)-glucanase activity, limits the rate of fermentation of corn and grass silage in the rumen. Enzyme supplements of the type used in these experiments are unlikely to possess sufficient activity to overcome this limitation by direct application to ruminal digesta, implying that treatment of the ration prefeeding will be key to harnessing the potential of exogenous fibrolytic enzymes in ruminant nutrition.

Animal Feed↗

PubMed1998

Small-intestinal digestion of partially resistant cornstarch in healthy subjects.

The aims of this study were to measure the amount of starch from partially resistant starches (retrograded and complexed high-amylose cornstarches) escaping small-intestinal digestion in healthy humans by use of an intubation method and to compare these data with data obtained by indirect in vitro methods. Experiments were carried out in vivo in 6 healthy humans by using ileal intubation and stool analysis and in vitro by using 3 different methods for analyzing resistant starch. In intubated subjects, 51 +/- 2% of the retrograded and 21 +/- 2% of the complexed starch was delivered to the ileum and was fermented almost completely in the colon. In vitro estimates of the absorption of resistant starch were much lower. We conclude that technologically modified starches may substantially increase the amount of carbohydrate available for colonic fermentation in humans, but that in vitro measurements of resistant starch are inaccurate for predicting malabsorption in healthy humans.

Adult↗

PubMed1988

Short-chain fatty acid production from mono- and disaccharides in a fecal incubation system: implications for colonic fermentation of dietary fiber in humans.

An in vitro fecal incubation system was used to demonstrate how lactose, lactulose and monosaccharides (mainly constituents of dietary fiber) influence short-chain fatty acid production in colon. Short-chain fatty acids were formed from all mono- and disaccharides tested (except L-glucose): D-glucose, D-galactose, D-fructose, D-mannose, L-rhamnose, D-sorbitol, D-arabinose, D-xylose, D-ribose, D-galacturonate, D-glucuronate, lactose and lactulose. All saccharides increased acetate formation; propionate production was increased from rhamnose, arabinose, xylose, ribose, galacturonic and glucuronic acid, whereas the synthesis of butyrate was elevated in assays incubated with sorbitol, galacturonic and glucuronic acid, and to a lesser degree ribose. Isobutyrate, valerate, isovalerate and hexanoate were produced in increased amounts in assays incubated with albumin, but in fact decreased in many incubations with saccharides. It is speculated that saccharide fermentation always results in formation of acetate, and that the relative production of acetate, propionate and butyrate is related to the monosaccharide composition of dietary fiber available for colonic bacteria. However, the production of isobutyrate, valerate, isovalerate and hexanoate is probably not due to saccharide fermentation, but is rather of polypeptide origin.

Acetates↗

PubMed1963

Maintenance of the rumen microbial population in continuous culture.

The design and use of an apparatus for continuous in vitro cultivation of the mixed bovine rumen microbial population are described. Data relative to the concentrations and proportions of volatile fatty acids produced, methanogenesis, carbon dioxide production, and survival of bacteria and protozoa indicated that an essentially normal in vivo fermentation pattern was maintained in vitro for experimental periods of from 3 to 10 days. The continuous cultures were responsive to major changes in type of feed intake. A change from grain to hay resulted in increased acetate and decreased propionate production, whereas decreased acetate and increased propionate resulted when feed intake was changed from hay to grain. Methanogenesis, volatile fatty acid production, and the numbers of microorganisms in the cultures were proportional to the amount of feed materials added up to levels calculated to approximate the in vivo maintenance requirement of the adult bovine.

Acetates↗

PubMed1989

In vitro degradation of choline from selected feedstuffs and choline supplements.

The objective of this experiment was to determine the extent of in vitro degradation of choline from barley, corn, corn gluten meal, cottonseed meal, fish meal, soybean meal, alfalfa hay, timothy hay, choline chloride, and choline stearate. During four individual fermentation runs, samples were incubated in vitro for .25, 1, 3, 6, 12, 18, 24, 36, and 48 h with an inoculum mixture containing rumen fluid obtained from a rumen-fistulated dairy cow fed 17.5% corn silage and 28.7% grass silage and 53.8% concentrate diet. Because of their low choline content (less than .68 mg/g) corn, corn gluten meal, alfalfa hay, and timothy hay gave erratic values for choline disappearance for different fermentation runs and times of incubation although disappearance tended to increase with time. Data for the rest of the feeds and choline supplements were analyzed using nonlinear regression procedure to obtain estimates of potentially degradable choline, rapidly degradable choline, and the rate of choline degradation in vitro. The mean estimates of rumen degradable choline (%) were 79.4, 84.7, 82.9, 83.8, 98.0, and 98.6 for barley, cottonseed meal, fish meal, soybean meal, choline stearate, and choline chloride, respectively. The results suggest that incorporating choline-rich feedstuffs in diets can only marginally increase the postruminal flow of choline in dairy cows.

Animal Feed↗

PubMedDate not supplied

Correlation of in vitro cytotoxicity with preclinical in vivo antitumor activity.

Several human and murine tumor cell lines were evaluated in an in vitro cytotoxicity assay as prescreens for fermentation extracts and pure materials subsequently tested in vivo against P388 leukemia or B16 melanoma. Each material, regardless of its in vitro cytotoxicity, was evaluated in vivo. At the criteria levels of in vitro positivity and in vivo activity invoked, a highly significant relationship between these two endpoints was demonstrated for each cell line. When cell lines were compared, most of them performed in a similar manner, with HCT-116 human colon carcinoma cells providing a modest advantage predicting for P388 activity in some comparisons. Using the data from any two cell lines in concert did not improve the acuity of the prescreen beyond that associated with the better cell lines used singularly and only a minority of active materials was predicted for uniquely. Overall, the in vitro cytotoxicity assay provided a useful prescreen for selecting P388 and B16 in vivo active materials.

Animals↗

PubMed2004

Antioxidant activity of fermented soybean extract.

Free radicals are considered to be important causative factors in the development of cancer and cardiovascular diseases. This relationship has led to interest in evaluating the antioxidant capacities of many dietary supplements. Fermented soybean extract is produced by symbiotic fermentation of organic soybean with 20 types of Lactobacillus and yeast. In vitro and in vivo models are used in this study to evaluate the antioxidant effect of fermented soybean extract. Several in vitro models are used to detect the antioxidant capacity of the fermented soybean extract, which is compared to vitamin C and Trolox. The results demonstrate that the fermented soybean extract has strong antioxidant activity against unsaturated fatty acid peroxidation compared to vitamin C and Trolox. By the means of the test system developed by Y. Toshiki et al., it is shown that the fermented soybean extract can function both as an antioxidant and as a free radical acceptor that can convert free radicals into harmless substances through an energy-decreasing procedure. An in vivo study examines the effects of fermented soybean extract on the activity of antioxidant enzymes. The activities of the antioxidant enzymes (AOE) including total superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) are measured in liver, kidney, and brain from male Sprague-Dawley rats. The activities of CAT, SOD, and GPX are increased in the liver. However, the SOD activity is decreased in the kidney. SOD and GPX activities are decreased in the brain. These results lead to the conclusion that fermented soybean extract not only has antioxidant activity but also has an effect on the activity of antioxidant enzymes in liver.

Animals↗

PubMed1996

The lactulose breath hydrogen test and small intestinal bacterial overgrowth.

OBJECTIVES: To i) document the sensitivity and specificity of a combined scintigraphic/lactulose breath hydrogen test for small intestinal bacterial overgrowth and ii) investigate the validity of currently accepted definitions of an abnormal lactulose breath hydrogen test based on "double peaks" in breath hydrogen concentrations. METHODS: Twenty-eight subjects were investigated with culture of proximal small intestinal aspirate and a 10-g lactulose breath hydrogen test combined with scintigraphy. Gastroduodenal pH, the presence or absence of gastric bacterial overgrowth, and the in vitro capability of overgrowth flora to ferment lactulose were determined. RESULTS: Sensitivity (16.7%) and specificity (70.0%) of the lactulose breath hydrogen test alone for small intestinal bacterial overgrowth were poor. Combination with scintigraphy resulted in 100% specificity, because double peaks in serial breath hydrogen concentrations may occur as a result of lactulose fermentation by cecal bacteria. Sensitivity increased to 38.9% with scintigraphy, because a single rise in breath hydrogen concentrations, commencing before the test meal reaches the cecum, may occur in this disorder. Sensitivity remained suboptimal irrespective of the definition of small intestinal bacterial overgrowth used, the nature of the overgrowth flora, favorable luminal pH, the presence of concurrent gastric bacterial overgrowth, or the in vitro ability of the overgrowth flora to ferment lactulose. CONCLUSIONS: Definitions of an abnormal lactulose breath hydrogen test based on the occurrence of double peaks in breath hydrogen concentrations are inappropriate. Not even the addition of scintigraphy renders this test a clinically useful alternative to culture of aspirate for diagnosing small intestinal bacterial overgrowth.

Bacterial Infections↗

PubMed1997

Mistletoe extract-induced effects on immunocompetent cells: in vitro studies.

Cytotoxic as well as immunomodulatory effects of mistletoe extracts and their components have been described and seem to depend upon the host tree, the manufacturing process and the composition of the different components present in the extracts. In vitro studies showed that a fermented mistletoe extract derived from Viscum album L. grown on pine trees was less cytotoxic to peripheral blood mononuclear cells (PBMC) than other preparations. This finding could be related to its very low content of mistletoe lectins. Furthermore, this extract stimulated PBMC from healthy and especially allergic donors who had never received any mistletoe treatment. By analysing these in vitro reactions, an involvement of CD4+ T helper cells and CD14+ monocytes/macrophages was observed, suggesting an interaction of the specific and nonspecific immune system. In the supernatants of stimulated PBMC from healthy individuals, type-1 (interferon-gamma and interleukin-2) and type-2 (interleukin-4 and interleukin-5) associated cytokines were detected in about 20%. In patients with colorectal tumours, however, reduced frequency, suggesting a functional impairment of certain immunocompetent cells in these patients. These studies may help to evaluate properties of the natural and the specific immune system.

Adjuvants, Immunologic↗

PubMed1990

Sorbitol adaptation of dental plaque in people with low and normal salivary-secretion rates.

Caries-related bacteriological and biochemical factors were studied in 12 persons with low and 11 persons with normal salivary-secretion rates before and after a four-week period of frequent mouthrinses with 10% sorbitol solution (adaptation period). After the adaptation period, an increase in total numbers of sorbitol-fermenting bacteria and mutans streptococci was found in plaque samples from both groups. The pH drop in dental plaque in situ after a mouthrinse with sorbitol was more pronounced after the adaptation period, especially in the group with the low salivary-secretion rate. Increased acid production in plaque from sorbitol was also observed in the two groups in vitro. The composition of the fermentation end-products was affected by the sorbitol exposure. It is suggested that in subjects with diminished natural protection against caries development, such as patients with low salivary-secretion rate, a cariogenic potential of sorbitol should be considered when this polyol is included in products that might be used frequently.

Adaptation, Physiological↗

PubMed1989

[Rusitec--fermentation equipment for the study of digestive processes in ruminants].

The fermentation equipment RUSITEC (Rumen Simulation Technique) is a simplified model of the rumen which stimulates the conditions inside the rumen, enables to keep the normal microbial population of the rumen for a long time under strictly controllable conditions, and enables a permanent monitoring of fermentation processes. Trials were conducted to see whether this model of "artificial rumen" can be used to determine the nutritive value of untreated beech sawdust and beech sawdust treated hydrobarothermically at the pressure of 1.4 MPa and impregnated with 1% H2SO4. The digestibility of the dry matter and fibre components was higher in the treated beech sawdust than in the untreated sawdust. In both reaction vessels, the amounts of acetic, propionic and butyric acids as well as the total VFA, were higher during the fermentation of treated beech sawdust, as compared with the diet with raw beech sawdust. The increased output of VFA, especially the acetic and butyric acids, resulted in a proportional increase in the output of gases, especially carbon dioxide and methane, as compared with the production of gases in rumens digesting untreated beech sawdust. In comparison with other in-vitro methods, this model of fermentation equipment characterizes the nutritive values of various kinds of feed in a more thorough way: using this equipment, the simulation technique has proceeded much closer to reality.

Animals↗

PubMed2006

Effects of nonstructural carbohydrates and protein sources on intake, apparent total tract digestibility, and ruminal metabolism in vivo and in vitro with high-concentrate beef cattle diets.

To investigate the effects of synchronizing nonstructural carbohydrate (NSC) and protein degradation on intake and rumen microbial fermentation, four ruminally fistulated Holstein heifers (BW = 132.3 +/- 1.61 kg) fed high-concentrate diets were assigned to a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments studied in vivo and in vitro with a dual-flow continuous culture system. Two NSC sources (barley and corn) and 2 protein sources [soybean meal (SBM) and sunflower meal (SFM)] differing in their rate and extent of ruminal degradation were combined resulting in a synchronized rapid fermentation diet (barley-SFM), a synchronized slow fermentation diet (corn-SBM), and 2 unsynchronized diets with a rapidly and a slowly fermenting component (barley-SBM, and corn-SFM). In vitro, the fermentation profile was studied at a constant pH of 6.2, and at a variable pH with 12 h at pH 6.4 and 12 h at pH 5.8. Synchronization tended to result in greater true OM digestion (P = 0.072), VFA concentration (P = 0.067), and microbial N flow (P = 0.092) in vitro, but had no effects on in vivo fermentation pattern or on apparent total tract digestibility. The NSC source affected the efficiency of microbial protein synthesis in vitro, tending to be greater (P = 0.07) for barley-based diets, and in vivo, the NSC source tended to affect intake. Dry matter and OM intake tended to be greater (P > or = 0.06) for corn- than barley-based diets. Ammonia N concentration was lower in vitro (P = 0.006) and tended to be lower in vivo (P = 0.07) for corn- than barley-based diets. In vitro, pH could be reduced from 6.4 to 5.8 for 12 h/d without any effect on ruminal fermentation or microbial protein synthesis. In summary, ruminal synchronization seemed to have positive effects on in vitro fermentation, but in vivo recycling of endogenous N or intake differences could compensate for these effects.

Animal Feed↗

PubMed1999

Ileal effluent as a fermentation substrate: implications for butyrate production in the colon.

Fermentation of fiber can lead to an enhanced production of short chain fatty acids (SCFA) and, hence, contribute to the proposed anticarcinogenic properties of butyrate in the colon. The fermentation of fiber isolates and the corresponding ileal effluents has been compared under in vitro conditions. Yield of SCFA per gram of substrate fermented was similar for isolates and fiber-enriched effluents (approximately 4.9 mmol/g) and it could be inferred that nonfiber components of effluent also generated SCFA. Butyrate production was highest for glucan-based polymers (approximately 30% total SCFA) and, from the measured acidogenic profile, production of SCFA will occur mainly in the proximal colon. The buffering capacity of ileal effluents during fermentation restrict the potential for a reduction in pH during acidogenesis compared to fiber isolates. This buffering capacity could limit the bioavailability of butyrate in the colon and, hence, the ability to satisfy the proposed antineoplastic properties of butyrate in the colon.

Anticarcinogenic Agents↗

PubMed1988

Effects of alkaline hydrogen peroxide treatment on in vitro degradation of cellulosic substrates by mixed ruminal microorganisms and Bacteroides succinogenes S85.

The effects of sodium hydroxide (NaOH) and alkaline hydrogen peroxide (AHP) treatments on wheat straw (WS) and various cellulosic substrates were determined by measuring susceptibility to degradation by mixed ruminal organisms or Bacteroides succinogenes S85. In vitro incubations were used to measure differences in fermentation resulting from each successive step in the AHP treatment process. In vitro incubations through 48 or 108 h were conducted to measure these differences. The AHP treatment of WS increased (P less than 0.05) dry matter, neutral detergent fiber, and acid detergent fiber degradation over control WS when these substrates were incubated with mixed ruminal microorganisms or B. succinogenes S85. Fermentations containing AHP-treated WS had greater (P less than 0.05) microbial purine (RNA) and volatile fatty acid concentrations by 12 h compared with those containing untreated or NaOH-treated WS. Xylose in AHP-treated WS was utilized more extensively (P less than 0.05) by 12 h compared with the xylose of untreated or NaOH-treated WS. Treatment with AHP removed 23% of the alkali-labile phenolic compounds from WS. When substrates with high levels of crystalline cellulose (raw cotton fiber, Solka floc, and Sigmacell-50) were treated with NaOH or AHP and incubated for 108 h with B. succinogenes S85, extent of acid detergent fiber degradation of cotton fiber and Sigmacell-50 was similar to that of their respective controls. Sodium hydroxide and AHP treatments were effective in increasing acid detergent fiber degradation of the Solka floc which contained, on average, 3.3 and 4.8 percentage units more acid detergent lignin and hemicellulose, respectively, than cotton fiber and Sigmacell-50.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2002

Structure, stereochemistry, and biological activity of integramycin, a novel hexacyclic natural product produced by Actinoplanes sp. that inhibits HIV-1 integrase.

[structure: see text] HIV-1 integrase is a critical enzyme for viral replication, and its inhibition is an emerging target for potential antiviral chemotherapy. We have discovered a novel inhibitor, integramycin, from screening of fermentation extracts using an in vitro assay. Integramycin possesses a hexacyclic ring system and exhibited an IC50 value of 4 microM against HIV-1 integrase (strand transfer). The isolation, structure elucidation, stereochemistry, conformation, and biological activity has been described.

Fermentation↗