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PubMed1988

The relation between bacterial degradation of viscous polysaccharides and stool output in human beings.

1. The relation between bacterial degradation of three viscous polysaccharides (guar gum, ispaghula and xanthan gum) by colonic bacteria in vitro and their effects on colonic function were investigated by comparing the results of anaerobic in vitro incubations with fresh faeces from seven healthy volunteers (measuring viscosity, pH and gas production) with the effects of feeding all three polysaccharides to the same volunteers for 1 week each (14-15 g/d) on faecal mass and whole-gut transit time. 2. Guar gum was rapidly fermented in vitro by faecal bacteria from all volunteers with concomitant loss of viscosity, reduction in pH and generation of gases. Ispaghula maintained its viscosity during incubation, but the pH fell significantly. The results of xanthan gum incubations showed considerable individual variation. 3. Only ispaghula significantly increased faecal mass, whilst none of the gums significantly affected stool frequency or transit time. Statistical analysis of the pooled results showed that although transit time and faecal output were inversely related, feeding viscous polysaccharides could influence these indices independently. Stool frequency was significantly correlated with the transit time, but not the faecal output. 4. Transit time was reduced by gum feeding to a significantly greater extent in those subjects whose faecal bacteria reduced or removed the viscosity of that gum, than in those subjects where the viscosity was maintained. In contrast, there was a smaller increase in faecal mass when the viscosity of the appropriate cultures was removed than when it was maintained or reduced. Increases in stool frequency were significantly associated with hydrogen production from in vitro cultures.

Adult↗

PubMed1996

In vitro reassembly of the malolactic fermentation pathway of Leuconostoc oenos (Oenococcus oeni).

The mechanism of metabolic energy generation by malolactic fermentation was studied with artificial membrane vesicles of Leuconostoc oenos (Oenococcus oeni). (Note that although L. oenos was recently reclassified as O. oeni [L. M. T. Dicks, F. Dellaglio, and M. D. Collins, Int. J. Syst. Bacteriol. 45:395-397, 1995], the old designation was kept in the present work.) Purified malolactic enzyme was entrapped in artificial membrane vesicles prepared from L. oenos cells able to transport L-malate. We show that the in vitro reconstituted system, including an electrogenic L-malate carrier and the decarboxylating malolactic enzyme, generated a proton motive force that was able to drive intravesicular accumulation of leucine.

Biological Transport↗

PubMed2004

Reduction of alpha-galactooligosaccharides in soyamilk by Lactobacillus fermentum CRL 722: in vitro and in vivo evaluation of fermented soyamilk.

AIMS: Consumption of soya-derived products has been hampered by the presence of alpha-galactooligosaccharides (alpha-GOS) because mammals lack pancreatic alpha-galactosidase (alpha-Gal) which is necessary for their hydrolysis. These sugars thus reach the large intestine causing gastrointestinal disorders in sensitive individuals. The use of lactic acid bacteria (LAB) expressing alpha-Gal is a promising solution for the degradation of alpha-GOS in soyamilk. METHODS AND RESULTS: The capacity of the LAB Lactobacillus fermentum CRL 722 to properly degrade alpha-GOS was studied in vitro using controlled fermentation conditions and in vivo using a rat model. Lactobacillus fermentum CRL 722 was able to grow on commercial soyamilk and completely eliminated stachyose and raffinose during fermentation because of its high alpha-Gal activity. Rats fed soyamilk fermented by this LAB had smaller caecums compared with rats fed unfermented soyamilk. CONCLUSIONS: Soyamilk fermentation by Lact. fermentum CRL 722 results in the reduction of alpha-GOS concentrations in soyamilk, thus eliminating possible undesirable physiological effects normally associated with its consumption. SIGNIFICANCE AND IMPACT OF THE STUDY: Fermentation with Lact. fermentum CRL 722 could prevent gastrointestinal disorders in sensitive individuals normally associated with the consumption of soya-based products. This LAB could thus be used in the elaboration of novel fermented vegetable products which better suit the digestive capacities of consumers.

Animals↗

PubMed1978

Effects of heavy metals and other trace elements on the fermentative activity of the rumen microflora and growth of functionally important rumen bacteria.

The inhibitory effects of high concentrations of essential and non-essential trace elements were tested on the rumen microflora using the rate of fermentation in vitro as the assay. The elements (and the concentration causing 50% inhibition) in decreasing order of toxicity were Hg2+ (20 microgram/ml), Cu2+ (21 microgram/ml), Cr6+ (70 microgram/ml), Se4+ (73 microgram/ml), Ni2+ (160 microgram/ml), Cd2+ (175 microgram/ml), As3+ (304 microgram/ml) and As5+ (1610 microgram/ml). The elements tested that were either weak or noninhibitory at concentrations greater than 400 microgram/ml included Zn2+, Cr2+, Fe2+, Mn2+, Pb2+, and Co2+. Methylmercury was as inhibitory as mercuric chloride to the fermentation. When the inhibitory effect of Cd2+ was tested on separated bacterial and protozoal fractions, it was more inhibitory to the bacteria. The inhibitory effects of trace elements were also determined for a number of axenic cultures of rumen bacteria. The bacteria which most frequently exhibited the greatest sensitivity were Bacteroides succinogenses, Ruminococcus albus, Bacteroides amylophilus, and Eubacterium ruminantium. Those often exhibiting intermediate sensitivities included Butyrivibrio fibrisolvens, Selenomonas ruminantium, and Megasphera elsdenii, while Streptococcus bovis was very refractory to all elements tested. Rumen fluid provided a modest protective effect for the bacteria.

Animals↗

PubMed2006

Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems.

Two experiments were conducted to determine the effects of several essential oil active compounds on rumen microbial fermentation. In the first experiment, 4 doses (5, 50, 500, and 5,000 mg/L) of 5 essential oil compounds were evaluated using in vitro 24-h batch culture of rumen fluid with a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were control (CON), eugenol (EUG), guaiacol, limonene, thymol (THY), and vanillin. After 24 h, the pH was determined, and samples were collected to analyze ammonia N and volatile fatty acids (VFA). The highest dose of all compounds decreased total VFA concentration and increased the final pH. Eugenol at 5 mg/L tended to reduce the proportion of acetate and the acetate to propionate ratio, at 50 and 500 mg/L tended to reduce ammonia N concentration, and at 500 mg/L reduced the proportion of propionate and branched-chain VFA concentration, without affecting total VFA concentration. All other treatments had minor effects or changes occurred only after total VFA concentration decreased. In the second experiment, 8 dual-flow continuous culture fermenters (1,320 mL) were used in 3 replicated periods (6 d of adaptation and 3 d of sampling) to study the effects of THY and EUG on rumen microbial fermentation. Fermenters were fed 95 g/d of DM of a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were CON, 10 mg/L of monensin (positive control), and 5, 50, or 500 mg/L of THY and EUG, and were randomly assigned to fermenters within periods. During the last 3 d of each period, samples were taken at 0, 2, 4, and 6 h after the morning feeding and analyzed for peptides, amino acids, and ammonia N concentrations, and total and individual VFA concentrations. Monensin changed the VFA profile as expected, but inhibited nutrient digestion. Eugenol and THY decreased total VFA concentration and changed the VFA profile, and only 5 mg/L of THY tended to reduce the proportion of acetate, increased the proportion of butyrate, and increased the large peptides N concentration without decreasing total VFA concentration. Most of these essential oil compounds demonstrated their antimicrobial activity by decreasing total VFA concentration at high doses. However, EUG in batch fermentation and 5 mg/L of THY in continuous culture modified the VFA profile without decreasing total VFA concentration, and EUG in batch fermentation decreased ammonia N concentration.

Acetates↗

PubMed2001

Fermentation properties of gentio-oligosaccharides.

AIMS: To investigate the fermentation properties of gentio-oligosaccharides (GOS), as compared to fructo-oligosaccharides (FOS) and maltodextrin in mixed faecal culture. METHODS AND RESULTS: The substrates were incubated in 24 h batch culture fermentations of human faecal bacteria. Fluorescent in situ hybridization was used to determine changes in populations of bifidobacteria, lactobacilli, clostridia, bacteroides, streptococci and Escherichia coli. Gas and short-chain fatty acid (SCFA) production was also measured. GOS gave the largest significant increases in bifidobacteria, lactobacilli and total bacterial numbers during the incubations. However, FOS appeared to be a more selective prebiotic as it did not significantly stimulate growth of bacterial groups which were not probiotic in nature. GOS and maltodextrin produced the highest levels of SCFA. Lowest gas production was seen with GOS and highest with FOS. CONCLUSIONS: GOS possessed bifidogenic activity in vitro. Although fermentation of GOS was not as selective as FOS, gas production was lower. Gas production is often seen as an undesirable side effect of prebiotic consumption. SIGNIFICANCE AND IMPACT OF THE STUDY: The study has provided the first data on fermentation of GOS in mixed faecal culture. The study has also used molecular microbiology methods (FISH) to quantify bacterial groups. The data extend our knowledge of the selectivity of fermentation of oligosaccharides by the gut microflora.

Bacteria↗

PubMed2002

A poorly fermented gel from psyllium seed husk increases excreta moisture and bile acid excretion in rats.

Psyllium seed husk (PSH) increases stool output and lowers blood cholesterol levels in humans. PSH and three fractions isolated from it were meal-fed to colectomized rats and fermented in vitro to test the hypothesis that viscous, gel-forming fraction B was responsible for these physiological actions. Control rats were fed 50 g/kg cellulose. The concentration of each PSH fraction in the test meals was equivalent to its concentration in PSH. Yields of the fractions were: A, 171; B, 575; and C, 129 g/kg of PSH. The wet weight and moisture content of ileal excreta (IE) from rats fed test meals containing PSH or fraction B were greater than those measured in excreta from rats fed meals containing cellulose or the other two PSH fractions. Total bile acids in IE did not differ between rats fed PSH or fraction B and were greater in these groups than in the other groups. Fraction A was not fermented during 3 d of incubation; fraction B was poorly fermented, with approximately 30% of the constituent sugars disappearing; and fraction C was rapidly and nearly completely fermented. These results indicate that the gel-forming fraction we isolated from PSH is the physiologically active component of the husks.

Animals↗

PubMedDate not supplied

Potential water-holding capacity and short-chain fatty acid production from purified fiber sources in a fecal incubation system.

Liquid enteral diets supplemented with dietary fiber, typically soy fiber, are in widespread use to increase stool bulking and maintain gastrointestinal function. Yeast cells, containing beta-glucans, represent another commercially available fiber source that could be added to enteral diets. The objective of this study was to compare commercially available purified fiber sources, i.e., mixed fiber (Fibrad), psyllium hydrocolloid (Metamucil), and yeast cell walls (Fibercel), for potential use in liquid diets in terms of potential water-holding capacity (PWHC) and SCFA production (mmol SCFA/g organic matter [OM]). Fiber samples were fermented in vitro with human fecal microbiota from three donors for 24 h, and SCFA production was greatest from beta-glucan (8.24 mmol/g OM), followed by psyllium (5.57 mmol/g OM) and mixed fiber (4.50 mmol/g OM) (p less than 0.0001). WHCs of the original materials (g H2O/g OM) differed (p less than 0.02) at 4.59, 5.79, and 8.63 for beta-glucan, mixed fiber, and psyllium, respectively. PWHC (g H2O/g OM), an in vitro measure of stool bulking, differed (p less than 0.0001) with psyllium (4.48) greater than mixed fiber (3.52) greater than beta-glucan (2.22). These three fiber systems appear to have distinctly different physiological effects in vivo. Of the three products, mixed fiber was the most inert. Psyllium seems to enhance both SCFA production and stool bulk relative to mixed fiber, whereas beta-glucan most significantly increases SCFA production with a minimal effect on stool bulk.

Body Water↗

PubMed1980

Inhibition of ruminal degradation of L-tryptophan to 3-methylindole, in vitro.

Available evidence supports the view that acute bovine pulmonary edema and emphysema (ABPE) is related to ruminal production of 3-methylindole (3MI) from L-tryptophan (TRP). Ruminal production of 3MI is a two-step process involving the conversion of TRP to indoleacetic acid (IAA) followed by decarboxylation of IAA and 3MI. Reduction in ruminal 2MI production by the inhibition of either of these processes may prevent the onset of ABPE. A closed-system, in vitro ruminal fermentation technique was used to screen 27 compounds for their ability to reduce the conversion of TRP to 3MI. Several compounds tended to reduce 3MI production at both 25 and 5 micrograms/ml. Desoxysalinomycin, X-206, chloral hydrate, nigericin, lasalocid, monensin, narasin and salinomycin all reduced 3MI production by more than 80% at 5 micrograms/ml without reducing total VFA production. All of these compounds, except chloral hydrate, are polyether antibiotics. At least part of the inhibition due to monensin and narasin occurs at the level of TRP conversion to IAA.

Animals↗

PubMed1981

Antibacterial activity of moxalactam (LY-127935) compared with cefotaxime and other beta-lactam antibiotics against clinical isolates of enterobacteriacea and non-fermenters.

The in vitro activity of moxalactam, a new semisynthetic 1-oxa-beta-lactam, was compared to those cefotaxime, cefuroxime, cephalothin, piperacillin and tobramycin against more than 500 clinical isolates of Enterobacteriaceae and non-fermenters. The geometric mean MIC against 450 Enterobacteriaceae in microgram/ml was 0.09 for moxalactam, 0.08 for cefotaxime, 5.3 for cefuroxime, 22.9 for cephalothin, 3.5 for piperacillin and 0.72 for tobramycin. The geometric mean MIC in microgram/ml against 60 P. aeruginosa strains was 12.7 for moxalactam, 22.9 for cefotaxime, 6.8 for piperacillin, 1.5 for tobramycin and 2.9 for cefsoludin. The minimum inhibitory and the bactericidal concentrations of moxalactam were almost the same in most species. The effect of the inoculum on the bactericidal concentration was slight, between 10(3) and 10(7) CFU/ml for the E. coli and the Klebsiella strains. In isolates of S. marcescens and P. aeruginosa the bactericidal concentrations increased by 4 to 5 log2 and in isolates of P. mirabilis the increased by 9 log2 with the largest inoculum.

Anti-Bacterial Agents↗

PubMed2005

Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture.

Eight continuous culture fermentors inoculated with ruminal liquor from heifers fed a 50:50 alfalfa hay:concentrate diet (17.6% crude protein, 28.0% neutral detergent fiber) were used in 3 replicated periods to study the effects of cinnamaldehyde (CIN) and garlic oil (GAR) on rumen microbial fermentation. Treatments were no additive (negative control); 1.25 mg/L (MON) and 12.5 mg/L (MON10) of the ionophore antibiotic monensin (positive control); 31.2 mg/L CIN (CIN) and 312 mg/L (CIN10) of CIN; and 31.2 mg/L GAR (GAR) and 312 mg/L (GAR10) of GAR (Allium sativa). The MON10 caused expected changes in microbial fermentation patterns (a decrease in fiber digestion, ammonia N concentration, and proportions of acetate and butyrate; an increase in the proportion of propionate; and a trend to increase small peptide plus AA N concentration). The CIN decreased the proportion of acetate and branch-chained volatile fatty acids (VFA) and increased the proportion of propionate; CIN10 decreased the proportion of acetate and increased the proportion of butyrate compared with the control. The GAR10 increased the proportion of propionate and butyrate and decreased the proportion of acetate and branch-chained VFA compared with the control. The GAR10 also increased the small peptide plus amino acid N concentration, although no effects were observed on large peptides or ammonia N concentrations. The CIN and GAR10 resulted in similar effects as monensin, with the exception of the effects on the molar proportion of butyrate, which suggests that they might have a different mode of action in affecting in vitro microbial fermentation.

Acetates↗

PubMed2004

L-Rhamnose increases serum propionate in humans.

BACKGROUND: Acetic and propionic acids are produced by colonic bacterial fermentation of unabsorbed carbohydrates and are absorbed into the portal circulation. From there, they travel to the liver, where acetate is a lipogenic substrate and propionate can inhibit lipogenesis. The extent to which peripheral blood short-chain fatty acid concentrations reflect differences in colonic fermentation is uncertain. The unabsorbed sugar lactulose produces mainly acetate when fermented in vitro, whereas L-rhamnose yields propionate. OBJECTIVE: The objective of the study was to ascertain whether ingestion of L-rhamnose and lactulose would have different acute effects on peripheral acetate and propionate concentrations and on breath hydrogen and methane concentrations. DESIGN: Twenty-two subjects were fed 25 g L-rhamnose, lactulose, or glucose on 3 separate occasions in a randomized crossover design. Blood and breath samples were collected hourly for 12 h. RESULTS: Serum propionate was significantly higher with ingestion of L-rhamnose than with that of lactulose or glucose (P < 0.001). The area under the curve for serum acetate was significantly higher with ingestion of lactulose than with that of glucose (P < 0.03). The ratio of serum acetate to propionate was significantly higher with ingestion of lactulose than with that of glucose or L-rhamnose (P < 0.01). Breath hydrogen was significantly higher with ingestion of lactulose than with that of L-rhamnose or glucose (P < 0.0001). CONCLUSIONS: The selective increases in serum acetate and propionate concentrations in humans were obtained by feeding specific fermentable substrates. Presumably, these changes in serum concentrations reflect changes in colonic production. Selective alteration of colonic fermentation products could yield a new mechanism for modifying blood lipids.

Acetates↗

PubMed1986

[In vitro study of ionophore antibiotics and various derivatives. I. Action on fermentation products in the rumen].

An in vitro study was conducted to test the action of different ionophore antibiotics and some of their derivatives on the end-products of rumen fermentation. Nigericin and narasin, like monensin and lasalocid, increased the molar proportion of propionate in the V.F.A. mixture and decreased the proportion of acetate and especially that of butyrate. They had no action on total V.F.A. production. The effect observed with derivatives (lasalocid O-acetyl or nigericin-O-acetyl) was generally less. Gas production, chiefly methane, decreased with the addition of antibiotics. This result agrees with the stoichiometric reactions of carbohydrate fermentation in the rumen. The amount of ammonia nitrogen fixed by bacteria was generally lowered by the addition of five antibiotics (nigericin, narasin, monensin, grisorixin and lasalocid) or two derivatives (lasalocid O-acetyl and nigericin O-acetyl), indicating a decrease in bacterial synthesis. In contrast, calcimycin improved the butyrate production at the expense of propionate, and had no effect on gas composition or bacterial synthesis; X 14547 A and alborixin had only a small effect on rumen fermentations. These results, however, must be interpreted with care since the inocula used came from non-adapted sheep.

Animals↗

PubMed1980

In vitro activity of netilmicin, gentamicin, tobramycin and amikacin against glucose fermenting and nonfermenting bacteria.

The in vitro activity of netilmicin was compared to gentamicin, tobramycin and amikacin against 461 strains of glucose fermenting and nonfermenting bacteria. The minimum inhibitory concentrations of netilmicin, gentamicin and tobramycin against the majority of Enterobacteriaceae, Staphylococcus and Streptococcus were quite similar. Gentamicin, however, was approximately fourfold more active against strains of S. marcescens. Amikacin was the most effect antibiotic against strains of fermenting and nonfermenting bacilli resistant to at least one aminoglycoside. Many gentamicin-resistant species of nonfermenting bacilli, however, remain highly resistant to all four aminoglycosides tested.

Amikacin↗

PubMed1998

Ensiling effects of the ethanol fractionation of forages using gas production.

We studied the use of gas curve subtraction to distinguish between two fractions soluble in neutral detergent solution. Samples of unfractionated (whole) forage, residue insoluble in 90% ethanol, and isolated NDF were fermented in vitro, and gas production was monitored. The gas volume associated with the ethanol solubles (A fraction) was determined as the difference between the gas from the whole forage and from the ethanol residue. The gas yield associated with the fraction insoluble in 90% ethanol but soluble in neutral detergent solution (B fraction) was determined by subtracting the isolated NDF gas curve from the corresponding ethanol residue curve. This experiment included three forages (alfalfa, bromegrass, and orchardgrass) harvested at two maturities and preserved by freeze-drying or ensiling to form a 3 x 2 x 2 factorial arrangement. Ensiling reduced the rate of gas formation from the A fraction by approximately 30% (P < .01). Ensiling increased (P < .05) the size of the A fraction (2 to 10% of DM) but did not change the volume of gas produced (P > .05). The gas yield from the B1 fraction was reduced 40% (P < .05) by ensiling with no significant change in rate. Curve subtraction of gas production profiles may be used to obtain rate estimates for multiple neutral detergent soluble pools. The separation of the neutral detergent solubles into two pools clarified the effects caused by ensiling. Changes due to ensiling on the rate of gas produced were associated with the A fraction, and the effects on final gas volume were associated with the B1 fraction.

Animals↗

PubMed1995

Effect of oxygen tension on killing of Escherichia coli by bovine polymorphonuclear neutrophil leucocytes in vitro.

A batch fermenter modified to simulate the physical conditions of an inflamed v. uninflamed mammary gland was utilized to evaluate the effect of oxygen tension on killing of Escherichia coli P4 by bovine polymorphonuclear neutrophil leucocytes. Leucocytosis was simulated in vitro 4 h after inoculation with Escherichia coli by adding bovine neutrophils isolated from peripheral blood to the culture medium. Experiments were conducted at 39 degrees C in UHT treated milk. At micro-aerophilic oxygen tension (oxygen partial pressure, 3.11 kPa), bacterial numbers declined during the first hour following addition of the neutrophils. Oxygen tension declined rapidly following PMN addition. Once oxygen was depleted, neutrophil activity was presumably diminished and Esch. coli numbers began to increase. Under anaerobic conditions (oxygen partial pressure, 0.17 kPa), no reduction in population was observed. Photomicrographs taken at the time of neutrophil addition and at subsequent time intervals demonstrated a specific association between neutrophils and the pathogen. Subsequent lysis of neutrophils associated with Esch. coli growth was seen coincident with oxygen depletion.

Animals↗

PubMed2005

Streptococcus macedonicus ACA-DC 198 produces the lantibiotic, macedocin, at temperature and pH conditions that prevail during cheese manufacture.

Streptococcus macedonicus ACA-DC 198, a natural cheese isolate, produces the anticlostridial bacteriocin, macedocin. Bacteriocin activity was detected from the mid-exponential growth phase and remained constant during the stationary phase. A secondary model was setup to describe the influence of temperature (20-45 degrees C) and pH (5.1-6.9) on cell growth of and bacteriocin production by S. macedonicus ACA-DC 198 during in vitro laboratory fermentations. The optimum temperature for bacteriocin production (20-25 degrees C) was markedly lower than the optimum growth temperature (42.3 degrees C). In contrast, the specific macedocin production was maximal around pH 6.0, whereas growth was optimal at pH 6.4. Consequently, the maximum bacteriocin activity was reached between pH 6.0 and 6.5.

Anti-Bacterial Agents↗

PubMed1997

In vitro 14C-amino acid fermentation by CF3, a characterized continuous-flow competitive exclusion culture of caecal bacteria.

Competition for growth-limiting nutrients by broiler caecal bacteria and Salmonella has been suggested as one factor associated with decreased Salmonella caecal colonization. The amino acids arginine, aspartic acid, serine and threonine have been indicated as Salmonella growth-limiting nutrients. Broiler caecal bacteria maintained in a continuous-flow culture (CF3) were used as inoculum for media containing 14C-arginine, 14C-aspartic acid, 14C-serine or 14C-threonine. The 14C-labelled amino acids, except 14C-arginine, were metabolized to lactic, formic, acetic, propionic and butyric acids. In glucose-based media, 72%, 72% and 33% of the radiolabel from aspartic acid, serine and threonine, respectively, were detected in organic acids. Fermentation without glucose resulted in 48%, 50% and 71% of the radiolabel from aspartic acid, serine and threonine, respectively, being detected in organic acids. Results indicated that the early establishment of CF3 in young chicks may result in the depletion of growth-limiting amino acids and, therefore, reduction of Salmonella colonization.

Animals↗