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PubMed1989

Effect of human faecal donor on in vitro fermentation variables.

Homogenized and diluted human faeces (66.6 g/l) collected from six individuals were incubated with four standard substrates (oat bran, wheat bran, kidney beans (Phaseolus vulgaris), and guar gum) for 4, 8, 12, and 24 h. Neutral detergent fibre (NDF) digestibility coefficients, short-chain fatty acid (SCFA) production (mmol/g organic matter (OM], and gas production (ml/g OM) were significantly affected by donor, substrate, and donor X substrate interactions, within an incubation time, but substrate fermentability rankings were similar at 24 h for all six donors. Substrate differed in NDF digestibilities at all incubation times, whereas SCFA and gas productions differed only at 8, 12, and 24 h. The results indicate that inoculum source (donor) can influence in vitro fermentation variables but not the substrate fermentability ranking at 24 h. It is recommended that several donors, at least three, be used to improve the accuracy of in vitro estimates of colonic fermentation.

Colon↗
PubMedDate not supplied

In vitro fermentation of high-amylose cornstarch by a mixed population of colonic bacteria.

BACKGROUND: Malabsorbed starch is probably the most important substrate for bacterial fermentation in the human large intestine. Fermentability of starch may depend on the composition of the colonic flora and its adaptation to the substrate supply. METHODS: Ten healthy volunteers were fed a controlled diet containing either 7.0 to 8.3 or 50.7 to 59.7 g/d of resistant starch (Hylon VII) for 4 weeks. At the end of each diet period, fecal starch concentrations were measured. Fecal samples were incubated in 48-hour batch cultures containing 10 g/L Hylon VII or digestible Lintner's starch. Bacterial breakdown of starch and short-chain fatty acid concentrations were measured at 0, 3, 6, 12, 24, and 48 hours. RESULTS: Fecal starch concentrations were higher during the Hylon VII period (35.7 +/- 16 vs 8.9 +/- 3.3 mg/g). Starch was fermented rapidly and completely in vitro in all but two subjects. Fermentability of resistant starch was comparable to that of digestible starch. No differences were found between the dietary periods. Fermentation of resistant starch produced higher rates of n-butyrate. Two subjects had substantially higher fecal starch concentrations. In vitro starch breakdown in these subjects was slow and incomplete. CONCLUSIONS: Fermentation of resistant starch by the colonic microflora was rapid and complete in 8 of 10 subjects. No adaptation of the fermentation capacity was observed after 4 weeks of dietary resistant starch supplementation. Fermentation of resistant starch increased the proportion of n-butyrate in vitro. In two subjects, fecal starch concentrations were substantially higher than in the other subjects and in vitro starch fermentation was slow and incomplete.

Adult↗
PubMed2000

Glucose-based oligosaccharides exhibit different in vitro fermentation patterns and affect in vivo apparent nutrient digestibility and microbial populations in dogs.

To evaluate the potential of indigestible oligosaccharides (OS) to serve as "dietary fiber-like" ingredients, it is necessary to determine their extent of indigestibility. In vitro fermentation characteristics of two novel OS, alpha-glucooligosaccharides (GOS) and a maltodextrin-like OS (MD), were compared to those of fructooligosaccharides (FOS), gum arabic (GA), guar gum (GG) and guar hydrolysate (GH). Total short-chain fatty acid (SCFA) production (micromol/g dry matter) as a result of MD fermentation was higher initially compared with GA (P<0.01), but GA was more extensively fermented at 24 h (P<0.01). Total SCFA production for GOS was similar to that for FOS, GG, GH and GA. In the second experiment, GOS and MD were added at 6% to an enteral formula control diet (Control) and fed to ileal-cannulated dogs in a 3x3 replicated Latin-square design. Ileal digestibility of glucose was lower (P<0.05) and carbohydrate (CHO) numerically lower (P = 0.08) for both GOS and MD compared with the Control. Total tract digestibility of CHO and glucose was lower only for MD (P<0.01) compared with the Control. Total fecal weights were higher (P<0.01) for both GOS and MD treatments. Fecal concentration of bifidobacteria was numerically increased by GOS and MD supplementation (P = 0.13 and 0.23, respectively). Thus, GOS and MD are indigestible yet fermentable OS, and may act as "dietary fiber-like" ingredients.

Animals↗
PubMed1987

Effect of human faecal inoculum on in vitro fermentation variables.

1. Homogenized and diluted faeces (66.6 g/l) collected from one human source on three different months was incubated with four standard substrates (oat bran, wheat bran, red kidney beans (Phaseolus vulgaris) and guar gum) for 4, 8, 12 and 24 h. 2. Neutral-detergent fibre and organic matter (OM) digestibility measurements and gas production (ml gas/g OM) were influenced by substrate and incubation period but not by day of collection. 3. Production of short-chain fatty acids (SCFA) (mmol/g OM) was a function of substrate and incubation period but not day of collection at 4, 8 and 12 h. Rapidly fermentable substrates such as red kidney beans and guar gum did not ferment beyond 12 h and SCFA values were not different at 24 h. 4. Substrates differed in amount, rate and type of SCFA produced. 5. The results indicate that human faeces collected on different occasions were sufficiently uniform to yield similar in vitro fermentation findings among collections. Therefore, it is concluded that human faecal material is a practical source of micro-organisms to evaluate the fermentation properties of substrates.

Dietary Fiber↗
PubMed1995

Dietary fiber for cats: in vitro fermentation of selected fiber sources by cat fecal inoculum and in vivo utilization of diets containing selected fiber sources and their blends.

Two experiments were conducted to evaluate the addition of single sources and blends of dietary fibers to cat diets. In Exp. 1, fermentability of selected fibrous substrates by cat fecal microflora was evaluated. After 24 h of fermentation, OM disappearance (OMD) and total short-chain fatty acid (SCFA) production were greatest (P < .05) for citrus pectin, guar gum, and locust bean gum, whereas Solka Floc resulted in the least (P < .05) OMD and total SCFA production. In Exp. 2, six diets were formulated based on results of Exp. 1. The highest (P < .05) digestibilities of DM and OM occurred when cats consumed the diet with no supplemental fiber, and the lowest (P < .05) digestibilities occurred when cats consumed the SCFA blend (SC) diet. Nitrogen and lipid digestibilities also were lowest (P < .05) for cats consuming the SC diet, whereas total dietary fiber (TDF) digestibility (P < .05) was greatest for cats consuming the beet pulp, SC, and combination blend diets. Fecal consistency scores were highest (P < .05) for cats consuming the SC diet, indicating liquid, unformed stools. In conclusion, the in vitro fermentation technique was reasonably accurate in predicting in vivo digestion of fiber. The SC diet, which contained the most fermentable fibers, severely decreased nutrient digestibility and resulted in poor stool characteristics. Diets that contain moderately fermentable fiber provide fermentation end products that may be important in maintaining the health of the gastrointestinal tract of the cat.

Animal Feed↗
PubMed2002

Fecal inoculum can be used to determine the rate and extent of in vitro fermentation of dietary fiber sources across three lemur species that differ in dietary profile: Varecia variegata, Eulemur fulvus and Hapalemur griseus.

To estimate fermentative capacity among lemur species, four fiber substrates were tested across three species, Eulemur fulvus, Hapalemur griseus and Varecia variegata. The substrates, cellulose, beet pulp, citrus pulp and citrus pectin, ranged in composition from completely insoluble fiber (IF) to completely soluble fiber (SF), respectively. The lemurs consumed a nutritionally complete biscuit formulated for primates [85 g/100 g diet dry matter (DM)] and locally available produce (15 g/100 g diet DM). Feces were then collected and used to inoculate fermentation tubes prefilled with fiber substrates and an anaerobic growth medium. Dry matter disappearance (DMD), and acetate, propionate, butyrate, and total short-chain fatty acid (SCFA) production were measured in tubes subjected to 6, 12, 24 or 48 h of fermentation. Results were fitted to a logistic growth model. The maximal production (MP) time at which production or disappearance is at one-half maximum (t(50)) and the fermentation rate at 3 h were calculated. The maximal disappearance of DM differed among substrates (citrus pectin > citrus pulp > beet pulp; P < 0.0001) and species (E. fulvus > H. griseus > V. variegata; P < 0.001). V. variegata reached t(50) for acetate and total SCFA production faster than H. griseus or E. fulvus (P < 0.02). Three-hour production rates of acetate and total SCFA were also greater for V. variegata for citrus pulp and citrus pectin (P < 0.01). Few species differences were observed for beet pulp. Results provide evidence for differences in fermentative capacity and suggest that fiber solubility and fermentability should be considered when assessing the nutritional management of lemurs.

Animal Feed↗
PubMed2001

Effects of rumen fluid collection site on microbial population structure during in vitro fermentation of the different substrates quantified by 16S rRNA hybridisation.

Rumen fluid samples from a cow were withdrawn manually from the feed mat (solid phase) or the liquid phase below this mat and incubated in vitro with wheat straw, sorghum hay and a concentrate mixture. From the inoculum and several samples collected during in vitro incubation RNA was extracted to assess microbial population size and structure. RNA content recovered from the solid phase rumen fluid was significantly higher than from the liquid phase. The composition of the microbial population in the solid phase material was characterised by a high proportion of Ruminococci. Neither the proportion of other cell wall degrading organisms (Fibrobacter and Chytridiomycetes) nor the Eukarya and Archaea populations differed between the two sampling sites. Gas production was higher when substrates were incubated with solid phase than with liquid phase rumen fluid regardless of sampling time. However, the higher level of gas production was not accompanied by a corresponding increase in true digestibility. The RNA probes showed that during in vitro incubation with liquid phase rumen fluid, the eukaryotic population was inactive no matter which substrate was used and the activity of methanogens (Archaea) was lower than with solid phase rumen fluid. The population pattern of the cell wall degrading organisms was influenced mainly by the substrate fermented, and to a smaller extent by the inoculum used for in vitro fermentation.

Animals↗
PubMed2000

Changes to the quantity and processing of starchy foods in a western diet can increase polysaccharides escaping digestion and improve in vitro fermentation variables.

This study investigated how readily achievable changes to the quantity and processing of starchy foods in a typical Western diet: (1) were reflected in levels of resistant starch (RS) and NSP excreted from the small intestine; and (2) more favourable profiles of butyrate, NH3 and phenol production. Two diets, a low-starch diet (LSD) and a high-starch, low-fat diet (HSLFD) were compared. The LSD with 20% total energy (%E) from starch was based on a 'typical' Australian diet, while the HSLFD (40%E as starch) was the same Australian diet modified by an increased content of legumes, starchy foods and coarsely-ground cereals and by a reduced fat content. Four subjects with iliostomies consumed each diet for 2 d, with ileal effluent collection on the second day. On the HSLFD compared with the LSD, RS in ileal effluent increased from from 0.49 to 1.7 g/MJ per d (P < 0.005) while ileal NSP excretion increased from 2.0 to 3.3 g/MJ per d (P < 0.05). Ileal effluents obtained after each diet were incubated for 24 h in vitro with a human faecal innoculum. After fermentation, ileal effluent from the HSLFD produced more butyrate relative to other short-chain fatty acids (17.5 v. 15.8 molar %, P < 0.005) and less phenol (2.3 v. 5.7 mg/l, P < 0.05) and NH3 (20.3 v. 23.1 mmol/l, P < 0.005) than the LSD diet. The HSLFD also generated a lower pH (6.15 v. 6.27, P < 0.05). On a wt/wt basis, RS was 2.3-fold higher in the HSLFD effluent while NSP did not increase, suggesting that the change in RS largely contributed to the fermentation effects. Changes in in vitro variables when the HSLFD ileal effluent was ground before fermentation indicated the importance of physical structure in determining ileal excretion of RS. We conclude that: (1) readily achievable modifications to the amount and processing of starchy foods in an Australian diet would produce potential benefits for in vitro fermentation variables; and (2) the physical structure of grains and cereals is important in determining access by colonic bacteria to a carbohydrate substrate.

Adolescent↗
PubMed1997

In vitro fermentation of carbohydrate by breast fed and formula fed infants.

Unabsorbed carbohydrates are fermented by colonic bacteria to short chain fatty acids (SCFA) which are rapidly absorbed, salvaging energy and reducing stool output. There are marked differences between the faecal flora and SCFA of breast fed (BF) and formula fed (FF) infants which may be related to the higher incidence of diarrhoea in FF infants. Part of this effect may be caused by a difference in the ability of the microflora to ferment carbohydrate. To test the hypothesis that BF and FF have different fermentation capacities for simple and complex carbohydrates, in vitro cultures of faeces from healthy infants (2-10 weeks; 11 BF, 11 FF) containing glucose, lactose, raftilose (a fructo-oligosaccharide), or soybean polysaccharide were incubated anaerobically. Results were compared with those of adult faecal cultures using the same carbohydrates. Cultures of faeces from BF and FF infants produced comparable amounts of total SCFA in all cultures. These cultures produced less SCFA than those from adult faeces and produced very little SCFA from complex carbohydrate. BF cultures produced more acetic acid than FF in all cultures, whereas FF cultures produced more propionate with sugars and more butyrate with raftilose. Both groups of infants produced less butyrate than adults in all cultures. Thus it is unlikely that a lower ability to ferment carbohydrate is a major cause of increased risk of diarrhoea in FF fed infants but individual SCFA production may be important.

Bottle Feeding↗
PubMed1993

Sugar composition of dietary fibre and short-chain fatty acid production during in vitro fermentation by human bacteria.

The aim of the present study was to assess the relationship between the disappearance of dietary fibre sugars and the production of individual short-chain fatty acids (SCFA). The bacterial degradation of five dietary fibres whose sugars were quantified was investigated in vitro using a human faecal inoculum. Involvement of the main fibre sugars in SCFA production was evaluated by a stepwise multiple linear regression. The results show first that the nature and chiefly the associations between the fibre sugars were key variables in the fermentability. Second, the nature and the amounts of SCFA produced were closely related to the in vitro fermentation of the main sugars available: uronic acids seemed to be principally involved in the production of acetic acid whereas the production of propionic acid could be promoted by the fermentation of glucose and, to a lesser extent, by that of xylose and arabinose. Xylose tended to have a greater impact than uronic acids and glucose on the production of butyric acid. Thus, it would be possible to predict which SCFA could be specifically produced during the fermentation of a fibre, as far as the chemical composition and structure of this fibre are known.

Acetates↗
PubMed2001

In vitro fermentation characteristics of native and processed cereal grains and potato starch using ileal chyme from dogs.

Two in vitro experiments were conducted to evaluate the ability of small intestinal bacteria of dogs to ferment native and extruded cereal grains and potato starch and cereal grain and potato flours. Substrates included barley, corn, potato, rice, sorghum, and wheat. In addition to testing native grains and flours, extruded substrates also were tested. Substrates were extruded at low temperatures (LT; 79 to 93 degrees C) and high temperatures (HT; 124 to 140 degrees C) using a Wenger extruder (model TX-52). Substrates varied widely in concentrations of rapidly digestible starch (RDS), slowly digestible starch (SDS), resistant starch (RS), and total starch (TS). Extrusion of most substrates at HT vs LT resulted in increased RDS and decreased RS concentrations. Organic matter disappearance (OMD) values attributed to microbial fermentation for a 5-h period were as high as 27% for native extruded substrates (LT potato starch) and 39% for potato flour. Average OMD was higher for cereal and potato flours than for native extruded substrates (29.9 vs 25.4%). Average molar percentages of short-chain fatty acids produced from all substrates fermented for 5 h were 73, 14, and 13% (acetate, propionate, and butyrate, respectively). Average lactate production for substrates ranked as follows: flours > native and extruded cereal grains and potato starch (0.33 and 0.18 mmol/g OM, respectively). In vitro microbial fermentation of starches by ileal bacteria can be substantial and is affected by differences in starch source, fraction, and processed form.

Animals↗
PubMedDate not supplied

Effect of silicate minerals (zeolite, bentonite, kaolin, granite) on in vitro fermentation of amorphous cellulose, meadow hay, wheat straw and barley.

The objective of the present experiment was to determine the effects of addition of silicate minerals, zeolite (Z), bentonite (B), kaolin (K), granite (G) on the rumen fermentation parameters, total gas, methane, total and individual volatile fatty acids (VFA) and hydrogen recovery in rumen fluid inoculum from sheep. Different materials (0.25 g) meadow hay (MH), wheat straw (WS), barley (BA) and amorphous cellulose (AC) were used as substrates. Silicate minerals (0.1 g) were added to the fermentation bottles containing substrates and rumen fluid inoculum and incubated for 72 h in vitro. The gas production technique simulates fermentation in the rumen was used to determine fermentation parameters. The total gas production was significantly higher compared to control for MH plus B (MHB), MH plus G (MHG), WS plus Z (WSZ), WS plus B (WSB), WS plus K (WSK), WS plus G (WSG), AC plus B (ACB), AC plus G (ACG), BA plus Z (BAZ), BA plus B (BAB), BA plus K (BAK), BA plus granite (BAG). Significant differences of the methane production were found between the controls, WSG, BAB and BAK. The total VFA concentration was increased in ACG (83.1 mM). The acetate: propionate (A:P) ratio of the control and additives ranged between 3.1 and 3.6 for MH, 2.7 and 3.5 for WS, 1.6 and 1.8 for AC and 2.3 and 2.9 for BA. It was concluded that the silicate minerals had no appreciable effect on the methane production, however, they support the microbial metabolism by influencing (bentonite, granite) and slightly influencing (zeolite, kaolin) the rumen fermentation.

Animal Feed↗
PubMed2005

Tea saponins affect in vitro fermentation and methanogenesis in faunated and defaunated rumen fluid.

The effect of tea saponins (TS) on rumen fermentation and methane emission was examined using an in vitro gas production technique named Reading Pressure Technique. Three levels of TS addition (0, 0.2, 0.4 mg/ml) were evaluated in the faunated and defaunated rumen fluid. Compared to the control, TS addition decreased the 24 h gas production in the faunated rumen fluid, but had a minor effect on gas yield in the defaunated rumen fluid. The TS significantly reduced methane production in vitro. In the faunated rumen fluid, 0.2 or 0.4 mg/ml TS decreased the 24 h methane emission by 12.7% or 14.0%, respectively. Rumen fluid pH value was affected neither by TS addition nor by defaunation. The TS addition had only minor effects on volatile fatty acids, but the yield and pattern of volatile fatty acids were greatly affected by defaunation. While the molar proportion of acetate was not affected by defaunation, the propionate was significantly increased and the butyrate significantly decreased. Ammonia-N concentration and microbial protein yield were influenced by TS inclusion and defaunation. Inclusion of 0.4 mg/ml TS increased the microbial protein mass by 18.4% and 13.8% and decreased the ammonia-N concentration by 8.3% and 19.6% in the faunated and defaunated rumen fluid, respectively. Protozoa counts were significantly reduced by TS inclusion. The current study demonstrated the beneficial effect of TS on methane production and rumen fermentation, and indicated that this may be due to the effect of the associated depression on protozoa counts.

Animals↗
PubMed1996

In vitro fermentation by human faecal bacteria of total and purified dietary fibres from brown seaweeds.

The in vitro degradation of dietary fibre from three brown seaweeds (Himanthalia elongata, Laminaria digitata and Undaria pinnatifida) was studied, using human faecal flora. Two sets of fibre were tested: (1) total algal fibres extracted from the whole algae, mainly composed of alginates, and (2) purified fibres (sulphated fucans, Na-alginates and laminarans) representative of those contained in the whole brown algae. Mannuronate, one algal component, was also investigated. Substrate disappearance and short-chain fatty acid (SCFA) production were monitored after 6, 12 and 24 h fermentation. Gas production was followed hourly during the first 9 h and then at 12 and 24 h. Sugarbeet fibre was used as a fermentation reference substrate. According to the fermentative indices used, most of each of the total algal fibres disappeared after 24 h (range 60-76%) but, unlike the reference substrate, they were not completely metabolized to SCFA (range 47-62%). Among the purified algal fibres, disappearance of laminarans was approximately 90% and metabolism to SCFA was approximately 85% in close agreement with the fermentation pattern of reference fibres. Sulphated fucans were not degraded. Na-alginates exhibited a fermentation pattern quite similar to those of the whole algal fibres with a more pronounced discrepancy between disappearance and production of SCFA: disappearance was approximately 83% but metabolism was only approximately 57%. Mannuronate was slowly fermented but its metabolism corresponded to its disappearance from the fermentative medium. Thus, the characteristic fermentation pattern of the total fibres from the three brown algae investigated was attributed to the peculiar fermentation of alginates, and mannuronate was shown not to be directly involved.

Alginates↗
PubMed2001

In vitro fermentation of cellulosis amorphous and meadow hay in experimentally Ascaris suum-infected lambs.

The objective of this experiment was to determine the effect of rumen inocula from experimentally Ascaris suum (AS)-infected lambs on cellulose amorphous (CA) and meadow hay (MH) used as substrates during 72h incubation in vitro. The rumen inocula were obtained post-mortem from eight lambs that had been experimentally infected with 1000 AS eggs per lamb daily for 3 weeks. Samples of rumen inocula were obtained from the lambs weekly throughout the experiment for 8 weeks. Two lambs were kept as uninfected control animals. The substrates were incubated together with buffered rumen fluid in sealed fermentation bottles. In vitro dry matter degradability (IVDMD), total gas, methane and total and individual fatty acid (VFA) for both incubated substrates were measured and compared by the pressure transducer technique. Comparison of the values for the controls, total gas, methane and VFA revealed significant differences (P<0.05, P<0.01, P<0.001, respectively) for both substrates. Pronounced differences (P<0.001) were also found between CA and MH especially for IVDMD, total gas and total VFA production. A decrease (P<0.001) of IVDMD, total gas, methane and total VFA was observed from week 6 to week 8. Restriction of fermentation was evident from week 7 to week 8. The results suggest that the functional damage arising from pathological lesions within the parasite-infected organs considerably affected the fermentation parameters of the incubated substrates.

Journal Article↗