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Anaerobic fermentation of woody biomass pretreated with supercritical ammonia.

The degradability of ground hardwood by thermophilic anaerobic bacteria (Clostridium thermocellum with or without Thermoanaerobacter strain B6A) was greatly enhanced by pretreatment of the substrate with supercritical ammonia. Relative to C. thermocellum monocultures, cocultures of C. thermocellum and Thermoanaerobacter strain B6A degraded 1.5-fold more pretreated soft maple but produced 2- to 5-fold more fermentation endproducts because Thermoanaerobacter sp. removed reducing sugars produced by C. thermocellum during the fermentation. Dry weight losses were not totally accounted for in end products, due to formation of partially degraded material (<0.4 mum diameter wood particles) during the fermentation. One pretreated hardwood, Southern red oak, was fermented poorly because it released soluble inhibitors at the 60 degrees C incubation temperature. Considerable (6- to 11-fold) increases in substrate degradability were also noted for supercritical ammonia-pretreated wood materials fermented in an in vitro rumen digestibility assay. Degradation of pretreated softwoods by either thermophilic or mesophilic fermentation was not measurable under the conditions tested.

Journal Article↗

Effect of ratio of roughage concentrate on glucose-induced heat production in sheep rumen fluid in vitro.

The effect of ration on heat of glucose fermentation in sheep rumen fluid was investigated. Heat production was measured in a semiadiabatic calorimeter. In trial 1, the effect of glucose (.4 to 6.4 mg) on fermentative heat production was determined in rumen fluid from sheep fed 25 or 100% roughage diet. Heat of glucose fermentation decreased with increase in glucose dose in both diets. Maximal heat of glucose fermentation in both diets agreed with stoichiometric calculations. However, at 6.4 mg glucose, maximal heat was 18 kcal/mol in the 25% roughage diet and 14 kcal/mol in the 100% roughage diet. Purine N and maximal rate of heat production were not affected by diet type. In trial 2, the effect of glucose (1.6 and 6.4 mg) on fermentative heat production was determined in rumen fluid from sheep fed 25, 50, 75, and 100% roughage. In addition, fermentation pattern was measured in donors of the rumen fluid. Heat of glucose fermentation was positively correlated with organic matter digestibility and negatively correlated with rumen pH and acetate concentration. These observations indicate that in addition to the effect of roughage on the fermentation pattern, supplemental adaptation may occur, as indicated by the reduction in the heat of glucose fermentation.

Animals↗

In situ study of the glycolytic pathway in Saccharomyces cerevisiae.

1. The problem of the influence of protein concentration on the kinetic parameters of enzymes has been approached studying the glycolytic enzymes from Saccharomyces cerevisiae in permeabilized cells (in situ). 2. The values of Km and Vmax for the different enzymes were essentially the same in dilute solutions of protein and in concentrated ones (in situ) except in the case of enolase where some differences were observed. 3. Functioning of the whole glycolytic pathway was compared in situ and in vitro measuring the rate of the fermentation of glucose. The rate of fermentation in situ was two fold higher than in vitro and the lag before active fermentation was also much shorter. 4. An unidentified phosphorylated compound, possibly polyphosphate, accumulates during the fermentation of glucose under in situ conditions.

Carbohydrate Epimerases↗

Short-chain fatty acid production and fiber degradation by human colonic bacteria: effects of substrate and cell wall fractionation procedures.

Three dietary fiber sources (corn fiber, oat bran, wheat bran) were analyzed for chemical composition and potential fermentation by human colonic bacteria in vitro. Total dietary fiber (TDF) concentration of substrates was 64.3, 11.1 and 50.4 g/100 g dry matter for corn fiber, oat bran and wheat bran, respectively. Original material (ORIG), TDF fractions and simulated (SIM) cell wall fractions (produced by combining cellulose, hemicelluloses and pectic substances in proportions they represented in the cell wall) from each substrate were fermented in vitro for 6, 12, 18, 24 or 48 h using inoculum prepared from freshly voided feces from each of three human volunteers. Substrate dry matter remaining after 48 h of fermentation was 87.8, 39.8 and 73.5% for TDF fractions of corn fiber, oat bran and wheat bran, respectively. Disappearance of ORIG fractions was considerably greater than that of TDF due to fermentation of nonfibrous material. Disruption of cell wall structure during isolation of polysaccharide fractions allowed for dramatically increased fermentability of SIM relative to TDF. Averaged across all treatments, production of the short-chain fatty acids, acetate, propionate and butyrate, occurred in the molar ratio 63:21:16; however, profiles of short-chain fatty acids produced were influenced by both treatment and inoculum source. Extent of substrate fermentation varied among inoculum donors, implying that colonic microbial activities differ among individuals. Potential colonic fermentability of fiber sources was influenced by substrate, method of fiber preparation and inoculum source.

Adult↗

The effect of milk fermentation by Lactobacillus helveticus on the release of peptides during in vitro digestion.

This study evaluated the effect of protein hydrolysis by lactic acid bacteria during milk fermentation on the release of amino acids and peptides duing subsequently simulated peptic and pancreatic digestion. After digestion with trypsin, we compared the elution patterns of proteins and peptides obtained from unfermented milk and from milk fermented by Lactobacillus helveticus under pH control, using HPLC gel filtration and reverse-phase HPLC. The results indicate that milk fermentation affects the release of some amino acids during simulated gastrointestinal digestion and has a major impact on the modification of protein elution profiles obtained after digestion with trypsin. We conclude that proteolysis during fermentation may lead to the formation of novel peptides during gastrointestinal digestion.

Amino Acids↗

In vitro effects of the thiopeptide A10255 on ruminal fermentation and microbial populations.

Experiments used unadapted mixed cultures of ruminal microorganisms in batch or continuous culture fermentation to investigate the effect of a thiopeptide, A10255, on ruminal fermentation and microbial populations. After 24 h of fermentation in batch culture, addition of A10255 (.5 to 20 ppm of the culture fluid) to 0, 45, 60, and 75% concentrate diets had no effect on total VFA but increased molar proportion of propionate and decreased butyrate. The molar proportion of acetate was decreased by treatment only in the 0 and 75% concentrate diets. The increase in molar proportion of propionate by 20 ppm of A10255 was less than the increase caused by a similar concentration of monensin. The same concentration of A10255 (20 ppm) decreased ADF digestion less than 20 ppm of monensin. In continuous culture, A10255 (33 mg/kg of dietary DM) did not affect total VFA concentration, culture pH, OM digestion, or ADF digestion. Ruminal bacterial populations of total anaerobes and lactate-producing, lactate-utilizing, cellulolytic and amylolytic bacteria were unaffected by treatment. However, molar proportions of acetate, butyrate, and isovalerate were decreased, and propionate was increased, by addition of A10255.

Acetates↗

Influence of different concentrations of disodium fumarate on methane production and fermentation of concentrate feeds by rumen micro-organisms in vitro.

Batch cultures of mixed rumen micro-organisms were used to study the effects of different concentrations of disodium fumarate on the fermentation of five concentrate feeds (maize, barley, wheat, sorghum and cassava meal). Rumen contents were collected from four Merino sheep fed lucerne hay ad libitum and supplemented with 300 g concentrate/d. Disodium fumarate was added to the incubation bottles to achieve final concentrations of 0, 4, 7 and 10 mM-fumarate. In 17 h incubations, the final pH and total volatile fatty acid production increased (P<0.001) linearly for all substrates as fumarate concentration increased from 0 to 10 mm. Propionate and acetate production increased (P<0.05), while the value of the acetate:propionate ratio decreased (P<0.05) linearly with increasing doses of fumarate. In contrast, l-lactate and NH3-N concentrations in the cultures were not affected (P>0.05) by the addition of fumarate. For all substrates, fumarate treatment decreased (P<0.05) CH4 production, the mean values of the decrease being 2.3, 3.8 and 4.8 % for concentrations of 4, 7 and 10 mM-fumarate respectively. Addition of fumarate did not affect (P>0.05) the total gas production. If the results of the present experiment are confirmed in vivo, fumarate could be used as a feed additive for ruminant animals fed high proportions of cereal grains, because it increased pH, acetate and propionate production and it decreased CH4 production.

Acetates↗

H2 and acetate transfers during xylan fermentation between a butyrate-producing xylanolytic species and hydrogenotrophic microorganisms from the human gut.

The aim of this work was to investigate in vitro interrelationships during xylan fermentation between an H2 and butyrate-producing xylanolytic species recently isolated in our laboratory from human faeces and identified as Roseburia intestinalis and the H2-utilizing acetogen Ruminococcus hydrogenotrophicus or the methanogen Methanobrevibacter smithii. H2 transfer between M. smithii or Ru. hydrogenotrophicus and the xylanolytic species was evidenced, confirming the great potential of these H2-consuming microorganisms to reutilize fermentative H2 during fibre fermentation in the gut. In addition, acetate transfer was demonstrated between the xylanolytic Roseburia sp. and the acetogenic species, both metabolites transfers leading to butyric fermentation of oat xylan without production of H2.

Acetates↗

Effect of manganese on preferential degradation of lignin by Pleurotus ostreatus during solid-state fermentation.

Practical utilization of the polysaccharides in the lignocellulosic complex is limited because of the high lignin content of the complex. In this study we focused on the effect of Mn on lignin and cellulose biodegradation during solid-state fermentation by the edible mushroom Pleurotus ostreatus. Preferential degradation of lignin was enhanced by the addition of Mn(II) to cotton stalks at concentrations ranging from 30 to 620 micrograms of Mn per g. This effect was most apparent when we compared mineralization rates of [14C] lignin with mineralization rates of [14C] cellulose. Enhanced selectivity was also observed when we analyzed residual organic matter at the end of the fermentation period by using crude fiber analysis. The cellulose fraction in the original material was 1.8 times larger than the cellulose fraction of lignin. The cellulose/lignin ratio increased during 32 days of solid-state fermentation from 2.5 in the control to 3.3 following the addition of Mn to the medium. The in vitro digestibility value for fermented cotton stalks was 53% of the dry matter. Addition of 600 micrograms of Mn per g to the cotton stalks resulted in a digestibility value of 65.4%. Enhancement of preferential lignin degradation could be result of either increased activity of the ligninolytic enzymes or production of Mn (III), which might preferentially degrade aromatic structures in the lignocellulosic complex.

Biodegradation, Environmental↗

Effect of a simethicone-containing tablet on colonic gas elimination in breath.

The effect of a tablet containing the antiflatulent, simethicone, on intestinal hydrogen (H2) elimination in breath was studied. In three trials, normal subjects (age 12-52 years) received, on subsequent days, lactulose or lactulose with two tablets of either simethicone or placebo in randomized order. Breath samples were collected over 210 min and analyzed by gas chromatography for H2. The time course of H2 expiration above baseline levels was calculated and compared for the three tests. No significant differences in transit time were found. Cumulative H2 expiration was significantly lower after simethicone compared to placebo. H2 production from stool incubated with simethicone or placebo indicated that the drug had no effect in reducing the fermentative production of H2 in vitro. Interestingly, the vehicle present in the tablets could be fermented by intestinal bacteria. Simethicone reduced the amount of H2 eliminated in breath, but this effect was offset partially by H2 production from the fermentation of unabsorbable substances used in the formulation of the tablets.

Adolescent↗

Fermentation by the human large intestine microbial community in an in vitro semicontinuous culture system.

A semicontinuous culture of the microbial community of the human large intestine that was maintained over 81 days is described. The initial inoculum was feces, and about 200 ml of nutrient suspension was fed to 500 ml of fermentor contents once or twice daily. The nutrient suspension contained comminuted fibrous food, sodium deoxycholate, urea, acid-hydrolyzed casein, vitamins, and salts. The fermentation was monitored, and the major products were acetate, propionate, butyrate, methane, hydrogen, and carbon dioxide. The concentration of anaerobic bacteria was 2 X 10(9) per ml of culture contents and was 100 times that of fecal coliforms. When the nutrient suspension contained lettuce, celery, carrots, and unsweetened applesauce, the predominant nonsporeforming anaerobes isolated were Bacteroides species. When carrots and applesauce were omitted, the predominant nonsporeforming isolates were Fusobacterium species. On both diets, clostridia were isolated that resembled Clostridium clostridiiforme. The fermentation and bacteriological analyses indicated that the in vitro ecosystem appears to be a reasonable facsimile of the large intestine ecosystem.

Bacteria↗

Degradation of quillaja saponins by mixed culture of rumen microbes.

Quillaja saponin (QS) was incubated at 39 degrees C in an in vitro medium containing rumen liquor from a cow fed a roughage diet. No degradation of QS was observed up to 6 h of fermentation. Incubation for 9, 12 and 24 h decreased the content of QS by 16%, 45% and 100%. The content of QS did not decrease when incubated for 24 h in the medium containing autoclaved rumen liquor, suggesting that rumen microbes have enzyme(s) capable fo degrading QS. The fate of QS will help gain a better understanding of mechanisms of action of QS on rumen fermentation, and its beneficial effects mediated by binding to ammonia.

Animals↗

Short chain fatty acids in inflammatory bowel disease. The effect of bacterial fermentation of blood.

An in vitro faecal incubation system was used to investigate how blood added to faeces influences short chain fatty acid (SCFA) production. The result was a change in SCFA pattern from one largely dominated by acetate and propionate to a pattern less dominated by these two acids but with greater amounts of longer and branched SCFA (butyrate, isobutyrate, valerate and isovalerate). Patients with active ulcerative colitis revealed variable concentrations of SCFA in their individual stool specimens, 66% of the samples being outside the 95% confidence interval set by a control group and without any specific trend. The SCFA concentrations were normal in patients with Crohn's disease of the colon. The study concludes that the changes in SCFA pattern seen elsewhere in studies on ulcerative colitis could be due to bacterial fermentation of blood either in the colon or in the stools after passing. It cautions against using faecal concentrations in this disease without due regard to the phenomenon of dilution or pollution of the colonic chymus by colonic effusion of blood.

Blood↗

Study of changes of properties in thermically and hydrothermically treated feeds. 2. N fractions in fermentation medium at the incubation of treated feeds under in vitro conditions.

In the extensive experiments in vitro degradation ability of hydrothermically and thermically treated feeds have been studied. The feeds used were soybean meal, horse bean, alfalfa meal, field pea. The feeds have been treated at 90, 110 and 130 degrees C for 30, 60 and 90 min. The treated feeds have been tested both for microbial degradation ability due to rumen microflora and enzymatic degradation ability via pepsin and trypsin. The fact has been regarded as an important finding that the degradation ability of nitrogenous compounds had been restricted effectively by hydrothermic treatment that was manifested by low ammonia-N levels in fermentation medium (11.3; 8.8; 15.9 and 1.1% out of nitrogen contents in the different feeds). On the other hand improved protein enzymatic digestibility have been recorded in treated feeds as compared with native ones.

Animal Feed↗

In vitro and in vivo lactose and lactulose effects on colonic fermentation and portal-systemic encephalopathy parameters.

Lactose intolerance occurs in the majority of human groups, excluding people from Northern Europe. Because its effect is similar to that of lactulose, lactose seems to be an alternative treatment for patients with portal-systemic encephalopathy (PSE) and lactase deficiency. The mechanism of action of lactose is similar to that of lactulose. In vivo, lactose improves PSE parameters and causes acidic diarrhea. We performed in vitro studies in a fecal incubation system to investigate the biochemical and bacteriological effects induced by different substances customarily used for the treatment of patients with PSE (lactose, lactulose and Neomycin). In vitro experiments showed that lactose and lactulose decreased aerobic flora counts and reduced the pH of fecal incubation. Both disaccharides reduced the ammonia concentration in the incubation system.

Anti-Bacterial Agents↗