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PubMed2002

Large bowel fermentation of maize or sorghum-acorn diets fed as a different source of carbohydrates to Landrace and Iberian pigs.

Twenty-four finishing pigs (twelve Iberian and twelve Landrace) were used in a growing and slaughtering experiment. Animals were fed two diets differing in their ingredients, maize (diet C) or sorghum-acorn (diet A). At an average weight of 107.0 kg pigs were slaughtered and hindgut digesta sampled to study the effect of breed and diet on large bowel fermentation. Flows of digesta to the hindgut compartment were estimated based on an indigestible flow marker (Cr2O3) and were higher in Iberian than in Landrace pigs (P<0.001), and higher in animals fed diet A than diet C (P=0.07). The higher flows in Iberian pigs were mainly associated with a higher voluntary feed intake (3.50 v. 2.70 kg/d, P<0.01) and lower ileal digestibility of NSP (-12.8 v. 47.8, P<0.01). Differences between diets were mainly associated with a lower ileal digestibility of starch from diet A (89.2 v. 96.9 %, P=0.06), although no differences in the resistant starch content were observed in vitro. Fermentation of different carbohydrates through the large bowel showed that NSP-glucose had lower digestibility in Iberian than in Landrace pigs (62.5 v. 94.2 %, P<0.001), but no differences were observed in starch, or other NSP-fibre fractions (arabinose, xylose and galactose). The type and amount of carbohydrates reaching the large bowel were related to the diet but also to breed, and promoted differences in the fermentative activity associated with different volatile fatty acid patterns and changes in microbial enzymic activity.

Animal Feed↗

PubMed1977

[Estimation of the net energy using volatile acids produced in the process of in-vitro digestion with ruminal fluid].

47 types of green feeds and roughages were subjected to an in-vitro fermentation with dilute ruminal fluid. The volatile fatty acids produced in this process were determined quantitatively in accordance with the method of digestibility estimation proposed by Tilley and Terry (1963). An average of 5 Mol FFS (FFS=volatile fatty acids) was found per kg of dried feed, a value is also reported in the literature. The ratios of acetate to propionate to n-butyrate to iso-butyrate to n-valerianate were 64.2 : 25.4 : 6.6 : 0.8 : 1.5 : 1.5. In this ratio, propionic acid predominated so that acetic acid and n-butyric acid were misrepresented compared with the data of in-vivo measurements made for the corresponding foodstuffs. Consequently, it is only within certain limits that values of FFS concentrations obtained in vitro may be used for estimating net energy data, disregardful of the fact that the FFS are the main source of metabolizable energy in ruminants. The reliability index for an estimation of Starch Equivalents and NEFr based on the above-mentioned method was found to be considerable lower (0.71 and 0.80) than that based on in-vitro digestibility measurements (0.86).

Animal Feed↗

PubMed1975

In vitro study of the influence of virginiamycin and spiramycin on the composition and biochemical activities of the gastrointestinal flora of piglets. II. Influence on the biochemical activities of the microflora.

Chemical analysis was carried out on 54 in vitro fermentations to study the influence of nutritional doses of Virginiamycin and Spiramycin on the metabolic activity of the gastro-intestinal flora of 18 in vivo treated piglets. Changes in the decomposition of glucose and the production of NH3, volatile acids and lactic acid are recorded in function of time. These results are in good accordance with changes noted in the composition of the bacterial flora, such as a marked decrease in the production of lactic acid, caused by Virginiamycin, combined with a decrease in the number of Lactobacilli and Enterococci. Virginiamycin caused marked changes in the metabolic activity of certain groups of bacteria, while there was only a slight effect noted for Spiramycin, although in vivo treatments with this antibiotic still show up in the results of the in vitro incubations. The overall results indicate that both antibiotics have marked influences on the intestinal flora of the pig, either by selective inhibiting of the flora resulting in a change of the metabolities formed, either by a change in the metabolic activity of some bacterial groups.

Acetates↗

PubMed1994

Degradation of polyphenols (catechin and tannic acid) in the rat intestinal tract. Effect on colonic fermentation and faecal output.

Low- and intermediate-molecular-weight polyphenols are usually extracted by using different solvents (e.g. water, methanol, aqueous acetone). The aim of the present work was to study the possible effects of some extractable polyphenols (EPP) on fat and protein digestibilities and on the colonic microflora. Degradability of these compounds through the intestinal tract was also studied. Catechin and tannic acid (TA) were chosen as representatives of the most common basic structures of EPP (flavonoids and gallic acid respectively). Three groups of eight male Wistar rats were given either a control diet free of EPP, or diets containing 20 g/kg dry matter of catechin and TA. Body-weight and food intake were monitored during a 3-week experimental period. Faeces and urine were collected daily during the third experimental week. EPP and fat were determined in faeces, and N in both urine and faeces. Only 3.1 and 4.6% of the ingested catechin and TA respectively were excreted in faeces, indicating that absorption and/or degradation of these EPP had occurred. HPLC analysis of the polyphenolic content of faeces showed qualitative differences between groups. A significant increase of total faecal weight as well as water, fat and N excretion was produced by TA. Catechin only caused an increase in fat excretion. In vitro fermentation assays were also performed to study the effect of EPP on the colonic microflora. Both catechin and TA affected the yield of end-products of fermentation, and were also degraded during the fermentation process.

Animals↗

PubMed2005

Influence of disodium malate on microbial growth and fermentation in rumen-simulation technique fermenters receiving medium- and high-concentrate diets.

Two incubation trials were carried out with the rumen-simulation technique (RUSITEC). In each trial, four vessels received a diet of grass hay and concentrate (600 and 400 g/kg DM, respectively; diet F), and the other four were fed a diet composed of concentrate and barley straw (900 and 100 g/kg DM, respectively; Diet C). Vessels were given 20 g of the corresponding diet daily, and half of them were supplemented with disodium malate to achieve a final concentration of 6.55 mM. There were no effects (P>0.05) of malate either on pH or on the daily production of NH3-N, but malate treatment increased (P<0.05) DM, neutral detergent and acid detergent fibre disappearance after 48 h incubation. The daily production of propionate and butyrate increased (P<0.001), and the ratio CH4:volatile fatty acids decreased (P<0.001) by supplementing both diets with malate. Whereas adding malate to the F diet produced an increase in acetate production (P=0.011) and the growth of solid-associated micro-organisms (P=0.037), no effects (P>0.05) were observed for diet C. For both diets, there were no differences (P>0.05) between treatments in the daily flow of liquid-associated micro-organisms measured using (15)N as a microbial marker. These results indicate that malate stimulated the in vitro fermentation of both diets by increasing the apparent disappearance of the diet and decreasing the ratio of CH4:volatile fatty acids, but a greater response was observed with diet F. If these results are confirmed in vivo, malate could be used as a feed additive for ruminants fed diets containing medium proportions of forage (i.e. dairy animals) and not only in animals fed high-concentrate diets, as has so far been proposed.

Acetates↗

PubMed1995

Comparison of lactulose and inulin as reference standard for the study of resistant starch fermentation using hydrogen breath test.

Disaccharide lactulose is commonly used as a standard to quantitate the colonic fermentation of undigested sugars by means of H2 breath measurements. However, its high hydrogen production rate during fermentation may make it inappropriate for mimicking the fermentation of more complex carbohydrates, such as starch. Indigestible carbohydrates with a higher molecular weight might be more suitable than lactulose as a standard in H2 breath studies of starch digestibility. To test this hypothesis, we measured H2 breath in 8 healthy volunteers after a standard meal supplemented with 5 g or 10 g of lactulose or inulin, an indigestible oligosaccharide with an average degree of polymerization 4.5 times higher than that of lactulose. The results were then compared with those obtained after a standard meal containing a known amount (6.1 g) of resistant starch from high-amylose corn starch. Median H2 breath excretion per gram of reference carbohydrate was lower after the 5 g dose of inulin than after the 5 g dose of lactulose (19.1 vs 26.6 ppm x h x g-1; Wilcoxon's rank test p = 0.021) but similar after the two 10 g doses (inulin 22.4; lactulose 23.6; p = 0.234). Median H2 breath excretion per gram of resistant starch was significantly lower than that for both lactulose and inulin (p < 0.02), being 4.7 ppm x h x g-1. In vitro fermentation for 8 hrs with fecal homogenate showed similar mean hydrogen production rates for inulin and lactulose (30.5 vs 27.7 mL/mg fermented carbohydrate), and a significantly lower rate for starch (9.1 mL/mg) (n = 7; ANOVA p = 0.0007).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1996

Effects of organic acid and monensin treatment on in vitro mixed ruminal microorganism fermentation of cracked corn.

The objective of this study was to determine the effects of organic acids and monensin on the in vitro fermentation of cracked corn by mixed ruminal microorganisms. Ruminal fluid was collected from a steer fed 36.3 kg of wheat silage and 4.5 kg of concentrate supplement once daily. Mixed ruminal microorganisms were incubated in anaerobic media that contained 20% (vol/vol) ruminal fluid and .4 g of cracked corn. Incubations were carried out in batch culture for 24 h at 39 degrees C. Organic acids (L-aspartate, fumarate, and DL-malate) were added to serum bottles (n = 4) to achieve final concentrations of 0, 4, 8, or 12 mM. Monensin, dissolved in ethanol, was included in serum bottles at a final concentration of 0 or 5 ppm of culture fluid. The addition of 8 and 12 mM organic acids to cracked corn fermentations increased final pH (P < .05), tended to increase total gas production and CO2 concentration, and decreased the acetate:propionate ratio (P < .05). Organic acids tended to decrease methane concentrations and hydrogen concentration was not altered. DL-Malate addition at all levels reduced (P < .05) lactate accumulation. Additive effects of monensin and organic acids were observed in some fermentations. In conclusion, organic acid addition to in vitro mixed ruminal microorganism fermentations yielded beneficial results independent of monensin treatment by decreasing the acetate: propionate ratio and increasing final pH.

Acetates↗

PubMed2006

Nutritional significance of the selective ingestion of Albizia zygia gum exudate by wild chimpanzees in Bossou, Guinea.

The selective ingestion of plant gum exudates by chimpanzees has been frequently observed at various study sites. At Bossou, Guinea, chimpanzees also frequently ingest Albizia zygia gum exudate. A functional explanation for this behavior is lacking, so we evaluated its possible contribution of energy in the form of short-chain fatty acids (SCFA) as well as minerals. An in vitro fermentation study of A. zygia gum using the fecal bacteria of a Bossou chimpanzee showed that carboxylic acids were produced with a 6-hr lag phase up to 44 mmol/l by 18 hr of incubation. Acetate was the most abundant acid produced, followed by lactate and propionate. The energy supplied from the fermentation of a piece of gum exudate (20-30 g) was negligible in comparison with the estimated daily energy requirements of chimpanzees in the wild. However, A. zygia gum exudate (20-30 g) can supply sufficient amounts of calcium, manganese, magnesium, and potassium to fulfill the daily requirements for these minerals in chimpanzees.

Albizzia↗

PubMed1995

Colonic lactate metabolism and D-lactic acidosis.

D-Lactic acidosis is seen in patients with intestinal bypass or short bowels in whom colonic produced D-lactate accumulates. An intestinal bypassed patient with D-lactic acidosis had higher fecal D-lactate (122.4 mmol/liter) and L-lactate (90.1 mmol/liter) than described before in humans. D-Lactate fluctuated between 0.5 and 3.1 mmol/liter in plasma (normal < 0.1 mmol/liter) and between 1.1 and 52.8 mmol/liter in urine (normal < 0.7 mmol/liter) within a few hours, indicating that the human organism do metabolize and excrete D-lactate. The patient with D-lactic acidosis had a 10-fold increased DL-lactate production from glucose in fecal homogenates compared to 14 healthy controls and a patient with intestinal bypass, who did not have D-lactic acidosis. A 67% carbohydrate (starch)-enriched diet resulted in a minor elevation of fecal and plasma lactate, whereas 50 + 100 + 150 g of ingested lactose increased D-lactate in feces (84.0 mmol/liter) and plasma (2.3 mmol/liter) considerably in the patient with D-lactic acidosis. Intestinal prolongation (22 cm ileum) had a temporary effect on fecal and plasma D-lactate, but intestinal continuity was reestablished 26 months later because D-lactic acidosis recurred (plasma 8.6 mmol/liter, urine 101.3 mmol/liter). Large amounts of lactulose (160 g/day) to 12 normal individuals increased D-lactate to 13.6 +/- 3.5 mmol/liter in feces, but never increased D-lactate in plasma or urine. The in vitro fermentation of glucose in fecal homogenates increased DL-lactate, which disappeared after complete metabolization of the glucose. L-Lactate was converted to D-lactate and vice versa, and both were degraded to the short-chain fatty acids acetate, propionate, and butyrate. An infrequent, but elevated ability of the colonic flora to produce lactate may be a prerequisite for D-lactic acidosis to occur and may explain why the syndrome is so seldom seen even in patients with intestinal bypass or short bowels. The suggestion that D-lactate is not metabolized and hence accumulates is probably not valid.

Acidosis, Lactic↗

PubMed2000

Effects of thymol on ruminal microorganisms.

Thymol (5-methyl-2-isopropylphenol) is a phenolic compound that is used to inhibit oral bacteria. Because little is known regarding the effects of this compound on ruminal microorganisms, the objective of this study was to determine the effects of thymol on growth and lactate production by the ruminal bacteria Streptococcus bovis JB1 and Selenomonas ruminantium HD4. In addition, the effect of thymol on the in vitro fermentation of glucose by mixed ruminal microorganisms was investigated. Neither 45 nor 90 microg/ml of thymol had any significant effect on growth or lactate production by S. bovis JB1, but 180 microg/ml of thymol completely inhibited growth and lactate production. In the case of S. ruminantium HD4, 45 microg/ml of thymol had little effect on growth and lactate production; however, 90 microg/ml of thymol completely inhibited growth of S. ruminantium HD4. Thymol also decreased glucose uptake by whole cells of both bacteria. When mixed ruminal microorganisms were incubated in medium that contained glucose, 400 microg/ml of thymol increased final pH and the acetate to propionate ratio and decreased concentrations of methane, acetate, propionate, and lactate. In conclusion, thymol was a potent inhibitor of glucose fermentation by S. bovis JB1 and S. ruminantium HD4. Even though thymol treatment decreased methane and lactate concentrations and increased final pH in mixed ruminal microorganism fermentations of glucose, concentrations of acetate and propionate were also reduced.

Animals↗

PubMed2006

Interactions between Euphorbia esula toxins and bovine ruminal microbes.

Cattle generally avoid grazing leafy spurge (LS; Euphorbia esula), whereas sheep and goats will often eat it. Understanding metabolism of toxic phytochemicals in LS by bovine rumen microflora may help explain why cattle often develop aversions to LS after initially eating it. Toxicity of LS compounds after in vitro fermentation with normal vs. antibiotic-modified bovine rumen digesta was evaluated at different lengths of fermentation. Levels of toxic and aversion-inducing ingenols were determined for fermented and nonfermented mixtures of LS and bovine rumen digesta, and the toxicity of an aversion-inducing extract of LS to rumen microbial species that are common in cattle, sheep, and goats was evaluated. Fermentation of LS with bovine digesta increased the toxicity of extracted compounds. Introduction of neomycin (an antibiotic that preferentially inhibits gram-negative bacteria) into the LS and bovine rumen digesta mixtures did not appear to affect toxicities regardless of fermentation length. Levels of ingenol were observed in LS and bovine digesta mixtures (both fermented and nonfermented) that were consistent with levels of ingenols reported for LS. Finally, a toxic extract of LS had little or no negative effect on the growth of several common species of rumen bacteria. The results indicate that LS is not generally toxic to the ruminal bacteria, but that microbial activity in the rumen may be responsible for enhancing LS toxicity to cattle.

Animals↗

PubMed2006

Microbial metabolism of caffeic acid and its esters chlorogenic and caftaric acids by human faecal microbiota in vitro.

Caffeic acid and its esters, chlorogenic and caftaric acids, are major dietary polyphenols present in various foods and beverages. Although caffeic acid is easily absorbed in the small intestine, its esterification with quinic acid, as in chlorogenic acid, decreases its gut absorption and increases the quantities reaching the colon and its microbiota. The microbial conversion of caftaric acid, the tartaric acid ester of caffeic acid, has not been studied earlier. In this work we compared the direct action of a human faecal microbiota on the metabolism of caffeic, chlorogenic and caftaric acids in an in vitro fermentation model. All substrates disappeared quickly and none of the free acids (caffeic, quinic or tartaric acids) were detected after 2 hours of incubation. Two major microbial metabolites were identified by HPLC-ESI-MS-MS as 3-hydroxyphenylpropionic (3-HPP) and benzoic acids (BA). Maximal levels of 3-HPP were reached after 2 h of fermentation and accounted for 9-24% of the dose of caffeic acid and its esters. BA was formed steadily throughout the incubation, accounting for 4-5% of the initial dose of the substrates after 24 h of incubation. The similarities in the metabolic patterns observed for caffeic, chlorogenic and caftaric acids suggest that esterification does not influence the metabolism of caffeic acid by the gut microbiota.

Bacteria↗

PubMed2005

Carbon dioxide stimulates the production of amylovorin L by Lactobacillus amylovorus DCE 471, while enhanced aeration causes biphasic kinetics of growth and bacteriocin production.

The effects of both oxygen and carbon dioxide on growth of and product formation by Lactobacillus amylovorus DCE 471, a promising new sourdough starter culture, were assessed through controlled, in vitro fermentation experiments, using a temperature of 37 degrees C and a constant pH of 5.4. It was seen that aeration affected both cell growth and amylovorin L production. At aeration rates of 1 l min(-1) and more, the bacterial population was subjected to oxidative stress as reflected by biphasic growth patterns. During the first growth phase, the maximum specific growth rate increased with increasing aeration rates stabilizing at the highest oxygen concentrations. The maximum obtainable cell yields decreased. During the second growth phase, the amylovorin L production was stimulated at the highest aeration. However, amylovorin titers were never higher in the presence of oxygen compared with the anaerobic fermentations. Carbon dioxide did not influence cell growth of L. amylovorus DCE 471. The maximum specific growth rate and the biomass concentrations were merely affected. On the other hand, the maximum soluble bacteriocin titers coincided with the highest carbon dioxide flow rates. These results indicate that mild aeration of type II sourdoughs might enhance both cell yield and amylovorin L production by L. amylovorus DCE 471, thereby contributing to the competitiveness of the strain. Growth in an ecosystem together with yeasts producing carbon dioxide might exert a positive effect on the production of amylovorin L as well.

Bacteriocins↗

PubMed2026

A streamlined workflow for high throughput metaproteomic analysis of the rumen microbiome.

Metaproteomics can provide direct functional insights into complex microbial communities, yet its application in rumen research remains limited due to labor-intensive and low-throughput sample preparation workflows before the MS analysis. This work aimed to develop and characterize a streamlined, high throughput metaproteomic workflow optimized for rumen samples. Key steps, including microbial cell extraction, cell lysis, protein digestion, and LC-MS/MS acquisition, were systematically assessed and optimized to reduce hands-on time while maintaining deep proteome coverage. The optimized workflow integrates a minimized cell extraction protocol using 0.5 g starting material and in-solution tryptic digestion. Application of the final workflow to 72 samples from in vitro fermentation revealed that biological variability between inocula dominated technical variability, which remained moderate (median CV of 21-24% across batches). Overall, the optimized workflow supports robust taxonomic and functional characterization of the rumen microbiome with improved scalability. These advances provide a foundation for applying metaproteomics to larger experimental designs, including nutritional trials and cohort studies, thereby enabling broader functional interrogation of rumen microbial ecosystems. SIGNIFICANCE: This study addresses current limitations in the application of metaproteomics to rumen microbiome research by developing a streamlined and scalable sample preparation workflow. By optimizing key steps and reducing sample input while maintaining reproducibility and proteome coverage, this work enables more efficient processing of larger sample sets. These advances support the broader use of metaproteomics in rumen studies and facilitate functional investigations relevant to animal nutrition and sustainable livestock production.

Animals↗

PubMed2000

Estimating the extent of degradation of ruminant feeds from a description of their gas production profiles observed in vitro: derivation of models and other mathematical considerations.

Equations to describe gas production profiles, obtained using manual or automated systems for in vitro fermentation of ruminant feeds, were derived from first principles by considering a simple three-pool scheme. The pools represented were the potentially degradable and undegradable feed fractions, and accumulated gases. The equations derived and investigated mathematically were the generalized Mitscherlich, generalized Michaelis-Menten, Gompertz, and logistic. They were obtained by allowing the fractional rate of degradation to vary with time. The equations permit the extent of ruminal degradation (hence the supply of microbial protein to the duodenum) to be evaluated, thus linking the gas production technique to animal production.

Animal Feed↗

PubMed2002

Characterization of resistant starch type III from banana (Musa acuminata).

Banana starch (Musa acuminata var. Nandigobe) was evaluated for its use in generating resistant starch (RS) type III. Structural, physicochemical, and biological properties of these products were analyzed. The investigated process includes debranching of the native starch and retrogradation under different storage temperatures and starch concentrations. After enzymatic debranching, a high amount of low-molecular-weight polymers with a degree of polymerization between 10 and 35 glucose units beside a higher molecular weight fraction were found. The resulting products comprised RS contents of about 50%. After heat-moisture treatment, the RS yield increased up to 84%. Peak temperatures of about 145 degrees C found in DSC measurements pointed to a high thermal stability of the RS products. In vitro fermentations of the RS products, carried out with intestinal microflora of healthy humans, resulted in a molar ratio of acetate:propionate:butyrate of about 49:17:34. The established method allowed the production of a high-quality RS with prebiotic properties for health preventing applications.

Amylose↗

PubMed2004

Human fecal metabolism of soyasaponin I.

The metabolism of soyasaponin I (3-O-[alpha-L-rhamnopyranosyl-beta-D-galactopyranosyl-beta-D-glucuronopyranosyl]olean-12-ene-3beta,22beta,24-triol) by human fecal microorganisms was investigated. Fresh feces were collected from 15 healthy women and incubated anaerobically with 10 mmol soyasaponin I/g feces at 37 degrees C for 48 h. The disappearance of soyasaponin I in this in vitro fermentation system displayed apparent first-order rate loss kinetics. Two distinct soyasaponin I degradation phenotypes were observed among the subjects: rapid soyasaponin degraders with a rate constant k = 0.24 +/- 0.04 h(-)(1) and slow degraders with a k = 0.07 +/- 0.02 h(-)(1). There were no significant differences in the body mass index, fecal moisture, gut transit time, and soy consumption frequency between the two soyasaponin degradation phenotypes. Two primary gut microbial metabolites of soyasaponin I were identified as soyasaponin III (3-O-[beta-D-galactopyranosyl-beta-D-glucuronopyranosyl]olean-12-ene-3beta,22beta,24-triol) and soyasapogenol B (olean-12-ene-3beta,22beta,24-triol) by NMR and electrospray ionized mass spectroscopy. Soyasaponin III appeared within the first 24 h and disappeared by 48 h. Soyasapogenol B seemed to be the final metabolic product during the 48 h anaerobic incubation. These results indicate that dietary soyasaponins can be metabolized by human gut microorganisms. The sugar moieties of soyasaponins seem to be hydrolyzed sequentially to yield smaller and more hydrophobic metabolites.

Adolescent↗

PubMed2005

Physicochemical characterization and biological effects of inulin enzymatically synthesized from sucrose.

We first developed the method to produce inulin from sucrose using an enzyme from Bacillus sp. 217C-1. Synthesized inulin consists of a linear polymer having beta(2-1) linkages of d-fructose with one terminal glucose. The synthesized inulin has similar properties (pH and thermal stability, Maillard reaction, and in vitro fermentation) to plant-derived inulin. The marked difference is the polydispersity of the inulin chain length. Synthesized inulin with a narrow degree range of fructose polymerization shows better solubility in water than plant-derived inulin. Synthesized inulin (5%, w/w) treatment for 12 weeks reduced the elevation in body weight and serum and liver lipids in rats fed high fat- and high sucrose-supplemented diets, and blood glucose in rats fed a standard diet. Synthesized inulin (15%, w/w) significantly suppressed the elevation in blood glucose of human healthy subjects after dextrin loading. These results suggest that daily intake of synthesized inulin modulates carbohydrate and lipid metabolism.

Animals↗