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PubMed2006

In vitro fermentation of mixed linkage glucooligosaccharides produced by Gluconobacter oxydans NCIMB 4943 by the human colonic microflora.

The aim of this study was to develop selectively fermented (prebiotic) carbohydrate molecules which would also result in the generation of butyric acid. Gluco-oligosaccharides produced by Gluconobacter oxydans NCIMB 4943 from various types of maltodextrins were evaluated for their fermentation by mixed cultures of human colonic microflora. The selectivity of growth of desirable bacteria (bifidobacteria, lactobacilli) was studied in stirred pH-controlled (6.8) batch cultures. Bacterial populations were enumerated using fluorescent in situ hybridization (FISH). Gluco-oligosaccharides resulted in significantly (P<0.05) increased numbers of bifidobacteria and lactobacilli within 24 hours. Bacteroides, clostridial and eubacterial populations were slightly decreased at 48 h. There was very little difference in selectivity between the maltodextrin substrates and the products, although maltodextrin displayed a slightly less selective fermentation than the gluco-oligosaccharide products, also stimulating the growth of bacteroides, clostridia and eubacteria. Gluco-oligosaccharides, produced from G19 maltodextrin, resulted in the best prebiotic effect with the highest prebiotic index (PI) of 5.90 at 48 hours. Acetate, propionate and butyrate were all produced from gluco-oligosaccharides, derived from G19 maltodextrin, at 48 hours but no lactate or formate were detected.

Acetic Acid↗
PubMed1983

Effects of diaryliodonium chemicals on rumen fermentation in vitro and in vivo.

In vitro fermentation studies demonstrated that diaryliodonium chemicals were inhibitors of amino acid utilization. Only small differences were found in comparative effectiveness of different analogs and salt forms of chemicals tested. Protection against degradation and(or) uptake was greatest for valine, methionine, isoleucine, leucine and phenylalanine. Although changes were not as large, animals fed 4,4'-dimethyldiphenyliodonium chloride for 112 d had higher ruminal concentrations of amino acids and lower concentrations of ammonia. Decreased production of acetate in conjunction with increased production of propionate and butyrate resulted in improved fermentation efficiencies. Fermentative shifts in vivo were not as large as those observed in vitro and additional investigations are needed to determine whether feeding diaryliodonium chemicals increases the transfer of energy in digested carbohydrates to volatile fatty acids.

Amino Acids↗
PubMed2002

Effect of vitamin E on ruminal fermentation in vitro.

The effects of vitamin E on pH value, total protozoa counts, volatile fatty acid (VFA), ammonia nitrogen and lactate levels were examined using an in vitro ruminal incubation system. The ruminal fluid (100 ml) of the first and second group was supplemented with 0.4 mg or 0.8 mg of vitamin E, respectively. Samples were taken immediately before and following 3, 6, 12 and 24 h of incubation at 39 degrees C and analysed for the total protozoa counts, the pH and the levels of ammonia nitrogen, lactate and VFA. Levels of propionate at 24 h and ammonia nitrogen at 12 and 24 h were significantly higher in the second group than in the control. In contrast, the levels of butyrate at 6, 12 and 24 h and lactate at 6, 12 and 24 h were lower in the second group than in the control. Propionate at 24 h, acetate levels at 6, 12 and 24 hand ammonia nitrogen levels at 6, 12 and 24 h and total rumen protozoa counts at 6, 12 and 24 h were significantly higher in the second group as compared with control. In contrary, butyrate levels at 6, 12 and 24 h, lactate levels at 6, 12 and 24 h were lower in second group than in control. There was no statistically significant difference among the groups in the pH values. In conclusion, the addition of vitamin E to in vitro ruminal fluid was found to increase the concentrations of acetate and propionate, total counts of protozoa, levels of ammonia nitrogen, but to decrease the butyrate and lactate levels of the ruminal aliquots in in vitro ruminal fermentation.

Ammonia↗
PubMed1987

Effect of sugar fatty acid esters on rumen fermentation in vitro.

1. The effect of sugar fatty acid esters (SFEs; currently used as food additives for human consumption) on rumen volatile fatty acids (VFA) and gas production was studied with sheep rumen contents in vitro. 2. Some SFEs having monoester contents of more than 70% increased the molar proportion of propionate in conjunction with reduction in the acetate:propionate ratio when the individual SFE was added to rumen contents in a final concentration of 4 g/l. Laurate sugar ester was the most potent propionate enhancer and rumen gas depressor, the effective dose being as low as 1 g/l in a final concentration. Fatty acid esters other than SFEs had little, if any, effect on rumen VFA production and their molar proportions. 3. Approximately 50% of laurate sugar ester was hydrolysed by in vitro incubation with rumen fluid for 2 h. The addition of fatty acids and sucrose was also effective in the alterations of rumen VFA and gas production. However, the effect of SFEs on in vitro rumen fermentation was significantly greater than that of their constituent fatty acids or sucrose, or both. Accordingly, the effect appeared to be ascribed to the complex action of SFE itself and to its constituents, free fatty acids and sucrose. 4. SFEs, at the level of 4 g/l, reduced substantially the froth formation (ingesta volume increase) and seemed to be effective for the prevention of bloat.

Animals↗
PubMed2003

Dietary galactooligosaccharides affect ileal and total-tract nutrient digestibility, ileal and fecal bacterial concentrations, and ileal fermentative characteristics of growing pigs.

The objective of this study was to evaluate dietary galactooligosaccharide (Gal OS) addition on swine nutrient digestibility, ileal and fecal bacterial populations, and ileal short-chain fatty acid (SCFA) production, and to determine their impact on ileal fermentative characteristics in vitro. Twelve T-cannulated pigs (BW = 25 kg) were fed a diet free of Gal OS for 21 d. On d 22, ileal digesta samples were collected for an in vitro fermentation experiment (Exp. 1). Substrates included: raffinose/stachyose combination (R + S), soy solubles (SS), and transgalactooligosaccharides (TOS). Also included were the non-OS components of SS and TOS. Nine pigs (three donors per treatment) served as ileal effluent donors. Each substrate was fermented in vitro for 6 h, and pH and SCFA and gas production were determined. Pigs then were allotted to three treatments: a Gal OS-free control diet and the control diet with either 3.5% added Gal OS from SS or TOS. Diets, feces, and digesta samples collected weekly for 6 wk on d 6 (feces) and 7 (digesta) were analyzed for DM, OM, CP, and chromic oxide concentrations. Feces and ileal digesta were analyzed for bifidobacteria and lactobacilli populations. Ileal digesta samples were analyzed for SCFA. On d 64, a second in vitro fermentation experiment (Exp. 2) was conducted using ileal effluent from three pigs per treatment and the same substrates used in Exp. 1. In vivo results showed that ileal and total tract DM and OM digestion were decreased (P < 0.05) by addition of both SS and TOS to the diet. Ileal and total-tract N digestibilities were decreased (P < 0.05) by dietary addition of SS. Fecal bifidobacteria and lactobacilli were increased (P < 0.05) by addition of SS and TOS to the diet. Ileal propionate and butyrate concentrations were greater (P < 0.05) for pigs fed diets containing both sources of Gal OS. In vitro results showed that fermentation data were not affected by donor animal adaptation to treatment. For both in vitro experiments, gas and SCFA production were higher (P < 0.05) for R + S than for SS or TOS. Fermentation of R + S resulted in a higher pH (P < 0.05) than did SS or TOS. Fermentation of non-OS components of SS and TOS resulted in more (P < 0.05) gas and SCFA production, and pH values that did not differ (P > 0.05) compared to SS and TOS. The Gal OS used in this study were prebiotics, increasing beneficial bacteria in vivo and SCFA concentrations both in vivo and in vitro.

Animal Feed↗
PubMed1994

Evaluation of fermentability of acid-treated maize husk by rat caecal bacteria in vivo and in vitro.

Fermentable energy in insoluble dietary fibre (DF) sources was evaluated by in vivo and in vitro methods using rats. Test diets contained 50 and 100 g maize husk or organic-acid-treated maize husk/kg diet. Soluble fractions were removed from both the DF sources by washing. The acid treatment increased digestibility by a microbial hemicellulase from 12.7% to 32.6%. The fermentability of DF was evaluated by measurement of the production rate of short-chain fatty acids (SCFA) in a short-term in vitro incubation of the caecal contents of rats fed on test diets for 22 d. The production rates of the major SCFA, acetic, propionic and butyric acids, were increased by feeding both DF sources, and these production rates in the acid-treated DF group were significantly higher than those in the untreated DF group. The production rate of a minor SCFA, isovaleric acid, was decreased by feeding both diets. The production rate of total SCFA in rats given the acid-treated maize husk was 32.6% higher than that in rats given the untreated maize husk. The fermentable energy in DF was estimated in vivo by subtracting the faecal excretion of DF energy from ingested DF energy. The fermentable energy in DF was increased by the acid treatment (32.5% in maize husk and 63.4% in acid-treated maize husk), which agreed with the SCFA production rate predicted in the caecum. These results indicate that a short-term incubation of caecal contents is a useful method for evaluation of the fermentability of DF sources, and that acid treatment can increase the fermentability of an insoluble DF source.

Acids↗
PubMed1981

Effect of lasalocid or monensin on lactate production from in vitro rumen fermentation of various carbohydrates.

Lasalocid and monensin effectively reduced the lactate produced during in vitro fermentation of various sugars and ground grains with rumen fluid from either hay- or grain-fed cattle. The minimum effective dose was 6 micrograms/ml. Both lasalocid and monensin at 6 micrograms/ml reduced the fermentation rate of glucose, fructose, galactose, sucrose, lactate, mannose, ground corn, ground sorghum, and ground wheat, and raised the pH and substantially lowered lactate concentration compared with controls. Although both antibiotics decreased total lactate production, the proportion of D(-) lactate to L(+) lactate increased. Apparently both antibiotics were less inhibitory to the formation of D(-) than to L(+) lactate isomer. Lasalocid was more effective than monensin in inhibiting lactate production except when rumen fluid from grain-fed cattle was used in fermentation.

Animal Feed↗
PubMed1988

Comparison of ileal effluents, dietary fibers, and whole foods in predicting the physiological importance of colonic fermentation.

An in vitro fermentation system that simulates the human colon was used in conjunction with the human ileostomy model to determine whether whole foods or dietary fiber isolates from the same foods could be used in lieu of ileal effluent to estimate the daily colonic production of short chain fatty acids (SCFA). A basal diet and a test food were fed for 3 days to a healthy ileostomate, and the ileal effluent was collected. Dietary fiber intake significantly increased ileal dry matter, ash, protein, and available carbohydrate (total carbohydrate minus dietary fiber) (p less than 0.05). Basal diet, test foods, ileal effluents, and dietary fiber isolates then were fermented in vitro with mixed human fecal microflora for 24 h, and SCFA production (mmol SCFA/g organic matter) was measured. SCFA production from ileal effluent significantly correlated with that from dietary fiber isolates (r = 0.86, p less than 0.05), but not with that from whole foods. Therefore, dietary fiber isolates, rather than whole foods, can provide the closest estimates of colonic SCFA production when ileostomates are not available. Test foods significantly changed molar ratios of individual SCFA within the ileal effluent and dietary fiber isolate groupings (p less than 0.05). In view of the different ileal loads arriving in the colon, we conclude that large differences in SCFA production probably depend on the food consumed.

Colon↗
PubMed1993

Effects of Aspergillus oryzae fermentation extract on in vitro equine cecal fermentation.

The objective of this study was to examine the effects of Aspergillus oryzae fermentation extract on the in vitro equine cecal fermentation of soluble starch, amino acids/peptides, coastal bermudagrass hay, and alfalfa hay. Cecal contents were obtained from a cecally fistulated Quarter Horse gelding fed coastal bermudagrass and grain (70:30) either unadapted or adapted to dietary A. oryzae supplementation (2 g/d). Mixed cecal microorganisms were incubated in anaerobic media for either 24 h (soluble starch, amino acids) or 48 h (bermudagrass hay, alfalfa hay). A. oryzae was added to the incubation bottles (n = 4) at concentrations of 0, .07, or .7 g/L. Fermentation of soluble starch in the presence of .7 g/L of A. oryzae resulted in increased concentrations of acetate, propionate, NH3, and L-lactate and decreased final pH. Addition of .7 g/L of A. oryzae to amino acid/peptide fermentations decreased final pH and increased concentrations of H2, acetate, propionate, butyrate, and total VFA. When alfalfa hay or bermudagrass hay was fermented with .7 g/L of A. oryzae, CH4, IVDMD, and digestion of NDF and ADF decreased. When adapted mixed cecal microorganisms were used, .7 g/L of A. oryzae did not inhibit methanogenesis or fiber digestion and L-lactate concentrations were not increased with soluble starch as the substrate. These results suggest that cecal microorganisms exposed to A. oryzae via the diet may adapt to the product. Incorporation of A. oryzae into in vitro incubations at concentrations similar to current recommended usage levels resulted in little change in final pH and fermentation products.

Animal Feed↗
PubMed2026

Insights into specific and nonspecific butyrate-producing pathways during the in vitro fecal fermentation of butyrylated starch.

Butyrylated starch is a special type-4 resistant starch with butyrate-carrying attribute. In this study, the unique butyrate-producing capability of butyrylated starch was deeply investigated by focusing on its specific and nonspecific butyrate-producing pathways, respectively, using specially designed substrates as controls. In vitro fermentation studies revealed that butyrylated and isobutyrylated starches generated high levels of butyrate and isobutyrate, respectively, highlighting the role of butyryl group metabolism in the specificity of butyrate production. Carboxylesterase assays have demonstrated that butyryl group metabolism is primarily facilitated by carbohydrate esterases expressed in the gut microbiota. Combined with 16S rRNA sequencing and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, it was found that butyrylated starch fermentation did not significantly enhance traditional butyrate synthesis pathways but modified the balance between the butyryl-CoA:acetyl-CoA transferase and butyrate kinase pathways by altering the gut microbiota composition, specifically by upregulating the relative abundance of indicator species such as Bacteroides, the Lachnospiraceae_NK4A136_group, and Parabacteroides. These insights offer theoretical guidance for designing butyrylated starch structures and regulating intestinal health.

Starch↗
PubMed2006

Effects of tea saponins on in vitro ruminal fermentation and growth performance in growing Boer goat.

Two experiments were conducted to investigate the effects of tea saponins (TS) on in vitro ruminal fermentation and growth performance in growing Boer goats. In Experiment 1, the Reading Pressure Technique (RPT) system was used to investigate the effect of addition of TS (0, 0.2, 0.4 and 0.8 mg/ml) on the ruminal fermentation in vitro. The 24h gas production and methane emission were significantly decreased when 0.4 or 0.8mg TS was included, suggesting that the TS could inhibit the release of methane. Compared to the control, the TS had little effect on pH values and the concentration of total volatile fatty acids in the ruminal fluids. However, the fermentation patterns were changed, with lower acetate and higher proportions of propionate when TS was added. Ammonia-N concentration and protozoal counts were significantly reduced, while microbial protein yield was increased by the TS addition, suggesting that the TS could modify the ruminal fermentation. In Experiment 2, 27 growing Boer goats were used to evaluate the effects of the TS addition on growth performance. The animals received the same basal diets, and added TS at levels of 0 (C), 3 g (T1) and 6 g (T2) per day. The experiment lasted for 60 days with the first 15 days for adaptation. Blood samples were obtained by jugular venipuncture before the morning feeding on the final day of the experiment. During the whole periods, dry matter intake, average daily gain and feed efficiency in T1 were higher than in the other two. Serum total protein, albumin, high density lipoprotein cholesterol, Ca and P and alkaline phosphatase levels were higher in group T1 than in C and T2, whereas the blood urea nitrogen, creatinine and total cholesterol were lower in the TS-added groups. The concentrations of glucose, glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase were not affected by the TS. From the results obtained in this study, it is inferred that the TS could modify the ruminal fermentation and that proper doses of TS may have potential in improving the animal growth performance, whereas at high doses, it may have adverse effects on animal production.

Animals↗
PubMed2004

Effects of a twin strain of saccharomyces cerevisiae live cells on mixed ruminal microorganism fermentation in vitro.

This experiment was designed to investigate the effects of different concentrations (0, 0.33, 0.66, 0.99, and 1.32 g/L) of a twin-strain of Saccharomyces cerevisiae live cells on in vitro mixed ruminal microorganism fermentation of corn starch, soluble potato starch, and sudangrass hay (60.5%, DM basis) plus concentrate mixture (39.5%, DM basis). Ruminal fluid was collected from two dairy cows, mixed with phosphate buffer (1:2), and incubated (30 mL) anaerobically at 38 degrees C for 6 and 24 h with or without yeast supplement, using 200 mg (DM basis) of each substrate. Medium pH, ammonia-N, and numbers of protozoa were unaffected (P = 0.38) by yeast cells in all substrates. Molar proportion of acetate was unchanged (P = 0.56) with cornstarch and soluble potato starch, but increased quadratically (P = 0.02) with hay plus concentrate by treatment. Addition of yeast cells caused a linear increase of total VFA (P = 0.008) in all substrates. Excluding the soluble potato starch, supplementation of S. cerevisiae resulted in a quadratic increase of propionate (P = 0.01), with a quadratic decrease (P = 0.04) of acetate:propionate. When soluble potato starch was used as a substrate, a linear increase (P = 0.006) of the molar proportion of propionate and a quadratic decrease (P = 0.007) in acetate:propionate was observed by treatment. Molar proportion of butyrate was unchanged (P = 0.35) with cornstarch and soluble potato starch, whereas it decreased linearly (P = 0.007) with hay plus concentrate by yeast cell supplementation. When cornstarch and soluble potato starch were used as a substrate, minor VFA were decreased (P = 0.05) by treatment. Accumulation of lactate was linearly decreased by treatment (P = 0.007) in all substrates. During incubation with hay plus concentrate, IVDMD was linearly increased (P = 0.006), whereas production of methane (linear; P = 0.02) and accumulation of hydrogen was decreased (quadratic; P = 0.005) by treatment after 24 h. These results showed that a twin strain of S. cerevisiae live cells stimulated in vitro mixed ruminal microorganism fermentation with decreased lactate, and a small decrease of methane and hydrogen with hay plus concentrate.

Ammonia↗
PubMed1993

Effects of aibellin, a novel peptide antibiotic, on rumen fermentation in vitro.

A new icosapeptide, aibellin, markedly modified rumen fermentation in vitro. Batch culture experiments with mixed rumen microorganisms showed that 12.5 to 25 mg/L of aibellin enhanced propionate production and reduced methanogenesis without significantly affecting production of total VFA, protozoal survival, or cellulose digestion. Aibellin had essentially the same effects in continuous culture with hay powder and concentrate. Monensin (5 mg/L) had similar effects on propionate production and methanogenesis, but total VFA, protozoa, and cellulolysis were decreased even by this low concentration of monensin. Commercially available peptide antibiotics also were compared with aibellin. Of the antibiotics examined, only graminicidin D (7.5 to 15 mg/L) enhanced propionate production and reduced methanogenesis. However, gramicidin D decreased total VFA, protozoa, and cellulolysis even at 7.5 mg/L. Alamethicin (7.5 to 15 mg/L), which resembles aibellin in its structure, did not increase propionate production but raised the percentage of propionate because of reduced production of total VFA. Alamethicin depressed methanogenesis but also decreased protozoal survival and cellulose digestion. These in vitro experiments indicate that aibellin could be a useful and potent modifier of rumen fermentation.

Alamethicin↗
PubMed1998

Inhibition of sulfate reduction to sulfide by 9,10-anthraquinone in in vitro ruminal fermentations.

We studied the effects of sulfur and 9,10-anthraquinone on in vitro ruminal fermentation and production of hydrogen sulfide. A complete, pelleted diet containing 26.8% acid detergent fiber, 15.9% crude protein, and 0.25 to 0.29% sulfur was used as the basal substrate. Fermentations were conducted using the basal substrate and various sulfur additions (elemental sulfur, thiosulfate, calcium sulfate, and sodium sulfate) with or without varying amounts of 9,10-anthraquinone. An increase in the sulfur content of the substrate to > 1.0% with various sources of sulfur had minimal effects on concentrations of volatile fatty acids, but the production of hydrogen sulfide increased. High amounts of 9,10-anthraquinone (10 and 25 ppm of fluid) decreased the molar proportion of acetate and decreased the production of methane and hydrogen sulfide. However, 9,10-anthraquinone increased the molar proportions of propionate and butyrate. Approximately 70% of 9,10-anthraquinone was recovered after 24 h of in vitro ruminal fermentation. These findings suggest that 9,10-anthraquinone has the potential to reduce the production of methane and hydrogen sulfide in ruminal fermentations.

Ammonia↗
PubMed1995

Ileal recovery of starch from whole diets containing resistant starch measured in vitro and fermentation of ileal effluent.

Six subjects with ileostomies consumed five diets containing 61-164 g starch/d of which 0.4-34.8 g was resistant starch (RS). Ileal excretion of starch was 97% of that measured as dietary RS in vitro with no significant difference between RS fed and starch recovered on any of the test diets. Variation in starch excretion between subjects was partly due to differences in mouth-to-stoma transit time. In vitro fermentation of ileal effluent from RS-supplemented diets produced significantly more short-chain fatty acids, a higher molar proportion of butyrate (17% compared with 12%), and a lower concentration of ammonia compared with control subjects. These results indicate that the amount of starch that reaches the large intestine can be predicted from measurements in vitro for a wide range of RS intakes under normal eating conditions. They also support the hypothesis that RS, through fermentation, has distinctive influences on the colonic environment.

Adult↗
PubMed1989

Influence of phospholipids on ruminal fermentation in vitro and on nutrient digestion and serum lipids in sheep.

Phospholipid supplements were incubated in vitro or fed to sheep to determine how they affected ruminal fermentation, nutrient digestion and serum lipid concentration. In vitro, deoiled soybean lecithin added to hay increased (P less than .05) total VFA concentration but had no effect on fiber digestion in four of five trials. Purified phospholipid (synthetic phosphatidylethanolamine), however, had little effect on in vitro hay fermentation when tested at four levels (0, 10, 20 or 30 mg) in three separate trials. Deoiled soybean lecithin disappeared in vitro at two rates, 12.9 and .66%/h for the fast- and slow-degrading fractions, respectively. Compared with 2.4% corn oil supplements, 5.2% soybean lecithin added to sheep diets (56% concentrate) reduced (P less than .05) energy, fiber and nitrogen digestibilities. Crude lecithin reduced (P less than .05) energy digestibility more than deoiled lecithin. Both types of lecithin increased (P less than .05) serum nonesterified fatty acid concentrations but had no effect on serum glucose, cholesterol or triglyceride concentrations compared with the corn oil diet. In summary, phospholipids had different effects on ruminal fermentation, depending on their source. None had a beneficial effect on fiber digestion. Phospholipids are degraded in the rumen and inhibit digestion in a manner similar to that of the commercial fats and oils.

Animals↗