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Effect of plant oils and aspartate on rumen fermentation in vitro.

The effect of plant oils and aspartate (ASP) on rumen fermentation in vitro. The objective of this study was to determine the effect of plant oils (rapeseed - RO, sunflower - SO; linseed - LO; 10% wt/wt) and 8 mmol sodium aspartate on rumen fermentation of a diet (250 mg) consisting of hay, barley and sugar beet molasses (60 : 30 : 10). Rumen fluid was collected from two Slovak Merino sheep fed the same diet twice daily. Mixed ruminal micro-organisms were incubated in fermentation fluid (40 ml) containing rumen fluid and McDougall's buffer (1 : 4). Incubations were carried out in batch cultures for 72 h at 39 degrees C two times in a 3-week intervals. When compared to the control, all supplemented diets (RO, SO, LO, ASP) significantly increased the pH, the mol% of propionate (LO + 8.7%; SO + 10.12%; RO + 8.65%; ASP + 5.86%) and the acetate : propionate ratio and numerically decreased methane production (SO -32.8%; LO, RO -30.08%; ASP -21.56%). Lactate production was also significantly decreased. Addition of plant oils to aspartate-treated incubations partly inhibited the decrease of n-butyrate, lactate and the increase of pH and in vitro dry matter digestibility (IVDMD) caused by ASP treatment. The effect of combined additives (RO + ASP, SO + ASP, LO + ASP) on methane production SO + ASP (-19.23%) and mol% propionate SO + ASP (+2.66%), LO + ASP (+4.28%) was less effective. All combined additives caused a significant decrease in digestibility of the given feeds. No effect of plant oils and ASP could be observed on the parameters of rumen fermentation (mainly methane and propionate).

Animal Feed↗

Evaluation of rare-earth markers using an in vitro ruminal fermentation system and effect of processing method on ruminal turnover of sized corn particles.

A two-stage in vitro ruminal fermentation experiment was conducted to determine the extent of marker migration and the effects of marker addition on digestibility. Fermentation studies were terminated either at the end of stage I, the fermentation phase, or stage II, the pepsin digestion phase. Treatment diets contained ytterbium-marked corn plus chopped Coastal bermudagrass hay and nonmarked corn plus chopped hay. After stage I of the in vitro ruminal fermentation, 90% of ytterbium was recovered. Of this, 8.8% of the ytterbium had migrated to the hay, 2% was recovered in the supernatant and 89.2% remained bound to the corn. After completion of both stages, 72% was recovered, of which 7.9% was bound to corn, 33.6% was bound to the hay, 1.1% was recovered in the stage I supernatant and 57.4% was associated with the acid pepsin supernatant. Dry matter digestion was not affected after stage I but after stage II, digestibility was decreased 7% due to marker addition. A ruminal-cannulated Holstein steer weighing approximately 680 kg was used to determine turnover rates from the rumen of liquid and five different particle sizes of corn. Corn from each of two processing methods was fractionated into five particle sizes by dry-sieving. The corn particles included steam-rolled corn (SRC) retained on a 4-mm sieve and marked with dysprosium, SRC retained on a 2-mm sieve and marked with erbium, cracked corn (CC) retained on a 4-mm sieve and marked with yttrium, CC retained on a 2-mm sieve and marked with ytterbium, and CC retained on a 1-mm sieve and marked with samarium.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

In vitro ruminal fermentation of organic acids common in forage.

Mixed rumen bacteria from cows fed either timothy hay or a 60% concentrate were incubated with 7.5 mM citrate, trans-aconitate, malate, malonate, quinate, and shikimate. Citrate, trans-aconitate, and malate were fermented at faster rates than malonate, quinate, and shikimate. Acetate was the primary fermentation product for all six acids. Quinate and shikimate fermentations gave rist to butyrate, whereas malate and malonate produced significant amounts of propionic acid. High-pressure liquid chromatography of fermentation products from trans-aconitate incubations revealed a compound that was subsequently identified as tricarballylate. As much as 40% of the trans-aconitate acid was converted to tricarballylate, and tricarballylate was fermented slowly. The slow rate of tricarballylate metabolism by mixed rumen bacteria and its potential as a magnesium chelator suggest that tricarballylate formation could be an important factor in the hypomagnesemia that leads to grass tetany.

Animals↗

[Effects of orally administered substances on rumen fermentation patterns in cattle (in vitro). 7. Effect of dried yeast in chronic ruminal acidosis].

The influence of dried yeast on the in-vitro-fermentation of chronic acidotic bovine rumen fluid was investigated using the longterm rumen simulation technique (RUSITEC). Five investigation periods keeping 21 days each were carried out. After a control period of eight days a five days lasting test phase followed. During this time 2 g of dried yeast were added to two reaction vessels and two reaction vessels remained as controls. The following effects of dried yeast during the investigation period on rumen fluid and rumen gas, respectively, could be noted: negligible increase of pH for 0.1 units (ns) increase of short volatile fatty acids concentrations up to 10% (p < 0.01), (acetate +1.5% [ns], propionate +6.0% [ns], i-butyrate up to +180% [p < 0.001], n-butyrate 0%, i-valerinate +39%, [p < 0.05], n-valerinate +17.6% [p < 0.001], hexanat +33% [p < 0.001]) increase of gas production up to 29% (p < 0.001) decrease of methane production up to 30% (ns) increase of hydrogen production up to 10% (ns) increase of protein concentrations up to 23% (p < 0.05) increase of ammonia concentrations up to 385% (p < 0.001). The fermentation patterns became normal during 6 days after withdrawal of the yeast.

Acidosis↗

Effects of whole cottonseed, niacin, and niacinamide on in vitro rumen fermentation and on lactating Holstein cows.

In Experiment 1, effects of whole cottonseed (0, 5, 15, or 30% of the total ration DM) on in vitro ruminal fermentation showed increased ruminal pH and ammonia concentration but lowered microbial protein. Acetic acid concentration was greatest with diets of 15 and 30% whole cottonseed, but propionate and total VFA concentrations were reduced by increasing whole cottonseed from 0 to 30%. In Experiment 2, neither niacin nor niacinamide (0, 100, 200, or 400 ppm) altered substantially fermenter pH or ammonia concentration. Both niacin and niacinamide increased synthesis of microbial protein. Acetate and propionate concentrations were not altered by treatment. Total VFA concentration tended to be lower as concentration of niacin and niacinamide increased. In Experiment 3, 28 Holstein cows were used to determine the effects of supplemental niacin on feed intake, milk yield, and composition. Cows were fed individually complete mixed diets ad libitum containing either: 1) 0; 2) .015; 3) .03; or 4) .06% niacin. There was a trend for lower milk fat test with niacin supplementation. Milk protein percentage was higher without niacin than with niacin at .015 or .03% in the diet, but daily milk and protein yields were higher with .06% versus .015% of niacin. Supplemental niacin did not affect casein nitrogen, lactose or minerals percentage, or concentrations of plasma glucose and insulin.

Animals↗

Influence of sodium fumarate addition on rumen fermentation in vitro.

The influence of sodium fumarate on rumen fermentation was investigated in vitro using batch and semi-continuous cultures of mixed rumen micro-organisms taken from three sheep receiving a basal diet of hay, barley, molasses, fish meal and a mineral-vitamin supplement (500, 299.5, 100, 91 and 9.5 g/kg DM respectively). Batch cultures consisted of 10 ml strained rumen fluid in 40 ml anaerobic buffer containing 200 mg of the same feed given to the sheep. Sodium fumarate was added to achieve a final concentration of 0, 5 or 10 mmol/l, as a result of the addition of 0, 250 or 500 mumol, equivalent to 0, 200 and 400 g/kg feed. CH4 production at 24 h (360 mumol in the control cultures) fell (P < 0.05) by 18 and 22 mumol respectively (SED 7.5). Total gas production was increased by the addition of fumarate without significant accumulation of H2. Substantial increases in acetate production (92 and 194 mumol; SED 26.7, P < 0.01) were accompanied by increases in propionate formation (212 and 396 mumol; SED 13.0, P < 0.001). Longer-term effects of fumarate supplementation on ruminal fermentation and CH4 production were investigated using the rumen simulation technique (Rusitec). Eight vessels were given 20 g basal diet/d, and half of them received a supplement of fumarate (disodium salt) over a period of 19 d. The response to the daily addition of 6.25 mmol sodium fumarate was a decrease in CH4 production of 1.2 mmol (SED 0.39, P < 0.05), equivalent to the consumption of 4.8 mmol H2, and an increase in propionate production of 4.9 mmol (from 10.4 to 15.3 (SED 1.05) mmol/d, P < 0.01). The inhibition of CH4 production did not decline during the period of time that fumarate was added to the vessels. Thus, the decrease in CH4 corresponded well to the fraction of the fumarate that was converted to propionate. Fumarate had no significant (P > 0.05) effect on total bacterial numbers or on the number of methanogenic archaea, but numbers of cellulolytic bacteria were increased (8.8 v. 23.9 (SED 2.49) x 10(5) per ml, P < 0.01). Fumarate also increased DM digestibility of the basal diet after 48 h incubation (0.476 v. 0.508 (SED 0.0123), P < 0.05). Thus, it was concluded that sodium fumarate may be a useful dietary additive for ruminants, because it diverts some H2 from CH4 production and because it is able to stimulate proliferation of cellulolytic bacteria and digestion of fibre.

Acetates↗

Studies on the effects of selenium on rumen microbial fermentation in vitro.

The effects of selenium (Se) on ruminant microbial fermentation were investigated in vitro using rumen microflora collected from a rumen-fistulated dairy cow. First, the effects of L-selenomethionine (SeMet; at 0.2 or 2 ppm Se) in the presence or absence of wheat bran (WB, 500 mg per incubation flask) were evaluated. Second, the effects of several forms of Se (elemental Se: 50 ppm Se; sodium selenite: 2 ppm Se; SeMet: 2 ppm Se) were compared. Results showed that the amounts of short-chain fatty acids (SCFAs) tended to be increased by SeMet treatment, whereas SeMet in the presence of WB transiently suppressed fermentation. The addition of SeMet tended to increase the production of acetate while reducing the production of butyrate with the without WB supplementation. Among the different Se compounds tested, the amounts of SCFAs were greater with SeMet treatment, which yielded a higher proportion of acetate compared to other treatments. Selenite did not influence the total SCFAs concentrations; however, it increased the relative proportion of butyrate at the expense of acetate. Elemental Se did not significantly affect fermentation. Higher bacterial Se concentrations were observed for selenite than for SeMet. It was concluded that Se supplementation can influence rumen microbial fermentation and that Se compounds differ in this regard.

Acetic Acid↗

Molecular weight of guar gum affects short-chain fatty acid profile in model intestinal fermentation.

Dietary fiber exerts many beneficial physiological effects; however, not all types of dietary fiber display the same effects. Partially hydrolyzed guar gum (PHGG), a lower molecular weight form of guar gum, is more easily incorporated into food, but may have less pronounced physiological effects than the native form. The aim of this study was to identify differences in intestinal fermentability based on the molecular weight of guar gum. Guar gum of four molecular masses (15, 20, 400, and 1,100 kDa) was fermented using a batch in vitro fermentation system. Human fecal inoculum was the source of microbes. The 400-kDa fraction produced the greatest concentrations of total short-chain fatty acid (SCFA) at 8 h and the highest amounts of butyrate at 24 h. At 24 h, the 400-kDa fraction produced more total SCFA and propionate than the 15 kDa, but was not different than 20 kDa or 1,100 kDa fractions. The molecular weight of guar gum was positively correlated with acetate production and negatively correlated with propionate production. This study concludes that 400-kDa guar gum may be optimal for intestinal fermentability. In conclusion, the molecular weight of guar gum affects in vitro fermentability and should be considered when adding to a food or beverage.

Acetic Acid↗

Coffee grounds. I. Effects of coffee grounds on ration digestibility and diuresis in cattle, on in vitro rumen fermentation, and on rat growth.

Holstein steers were used to compare rations in which coffee grounds replaced grain at rates of 0, 5, 10, or 20%. The coffee grounds analyzed (%) dry matter 91.1, protein 11.8, fat 23.1, fiber 42.5, ash .7, and caffeine .13. Digestibility coefficients for dry matter, crude protein, and energy decreased, and ether extract increased progressively as concentrations of coffee grounds in the ration were increased. There was a significant decrease in nitrogen retention and utilization when the rations contained 10 or 20% coffee grounds. Coffee grounds depressed feed intake, increased urinary output, and increased water intake (based on dry matter intake). Coffee grounds caused diuresis and renal, urethral, and bladder irritation. Adding coffee grounds up to 35% of the substrate did not depress rumen microbial fermentation, as measured by in vitro gas production. Weight gain of rats fed rations containing 22.5% untreated coffee grounds was reduced significantly. Water extraction removed some undersirable factors in coffee grounds for rats, and ether extraction removed most of them. Incremental additions of coffee oil (1, 3, or 5%) to the basal diet resulted in incremental decreases in rat weight gain.

Animal Feed↗

[The effect of virginiamycin on rumen fermentation in vitro after adaptation of donors to the inoculum].

Virginiamycin is an antibiotic active against grampositive bacteria in the alimentary tract, which is also suitable for supplementation of diets of growing and finishing ruminants. The aim of this work was to specify the effect of virginiamycin on some parameters of rumen fermentation in vitro with inoculi taken from wethers adapted or non-adapted to the virginiamycin intake. Incubations were performed anaerobically at 39 degrees C in serum bottles closed with Bunsen valves. Virginiamycin was added at 0 or 10 mg/l to the rumen fluid diluted with McDougall buffer. Virginiamycin significantly decreased production and utilization of lactic acid, production of methane and decomposition of casein when rumen fluid was taken from non-adapted wethers. Most of its effects disappeared when rumen fluid was sampled from wethers adapted to the virginiamycin intake (100 mg per head daily for 2 months). Adaptation of wethers to virginiamycin was further confirmed by analyses of the rumen fluid which was used for inoculation of in vitro cultures. Molar percentages of acetate, propionate, butyrate and valerate were the same before and after the adaptation. Therefore it can be concluded that the effects of virginiamycin on rumen parameters are not stable and its addition to ruminant diets cannot be recommended, with exception of the milk nutrition period. In the last experiment the stability of virginiamycin in the rumen fluid of adapted wethers was investigated.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗

Effects of methionine and lysine on fermentation in vitro and in vivo, nutrient flow to the intestine, and milk production.

Three experiments were conducted to investigate the feasibility of using crystalline methionine and lysine as protein supplements for lactating Holstein cows. In the first experiment, Met (dl-methionine) and Lys (l-lysine-HCl) were added to diets used in continuous culture bioreactors to estimate optimal concentrations for use in subsequent in vivo experiments. The second experiment measured ruminal fermentation and nutrient flow to the small intestine when Met and Lys were top-dressed on diets fed to nonlactating cows. The third experiment measured lactation performance when Met and Lys were added to diets fed to late-lactation cows. Providing 0.29 and 2.27% of dry matter as Met and Lys, respectively, provided the largest improvement in fermentation in vitro and these concentrations were used in subsequent experiments. When Met and Lys were top-dressed on diets fed to nonlactating cows, no changes in total tract N digestion were observed. No changes in microbial protein production or ruminal fermentation were observed. Adding Met and Lys did not change production or efficiency of production of milk or milk components by late lactation cows. These data indicate that providing supplemental Met and Lys during late lactation does not significantly improve the protein status of the cow and therefore may not improve milk production.

Ammonia↗

Effect of short-chain fatty acids on the human colonic mucosa in vitro.

Fermentable dietary fiber components are known to stimulate colonic crypt proliferation. As these compounds are rapidly degraded to short-chain fatty acids (SCFAs) by the anaerobic microflora, the hypothesis was tested that this trophic effect of fiber may be mediated by SCFAs. Biopsies were taken from normal cecal mucosa of 45 individuals during routine colonoscopy. They were incubated for 3 hours with sodium salts of SCFAs at physiological concentrations (three SCFAs = acetate 60 mmol/L + propionate 25 mmol/L + butyrate 10 mmol/L; acetate 60 mmol/L; propionate 25 mmol/L; butyrate 10 mmol/L) or equimolar NaCl (control). Cell proliferation was measured autoradiographically by subsequent pulse labeling with [3H]thymidine (1 hour). The labeling index (number of labeled cells divided by the total number of cells) was computed for the crypt as a whole and for five equal crypt compartments (compartment 1 = crypt base, compartment 5 = crypt surface). Cecal crypt proliferation was raised significantly in all incubation experiments with SCFAs. Butyrate (10 mmol/L, increase + 89%) and propionate (25 mmol/L, + 70%) were as effective in stimulating proliferation as the combination of three SCFAs (+103%), although the effect of acetate (+31%) was minor. Increasing the butyrate concentration to 25 mmol/L or 60 mmol/L did not result in a further increase of cell labeling. SCFAs stimulated proliferation in the basal three crypt compartments only. An expansion of the proliferative zone to compartments 4 and 5 was not observed. SCFAs, especially butyrate and propionate, are luminal trophic factors for the cecal epithelium.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗

In vitro lactose fermentation by human colonic bacteria is modified by Lactobacillus acidophilus supplementation.

Adaptation of the colonic flora to lactose may contribute to lactose digestion in lactose maldigesters, and supplementation with Lactobacillus acidophilus may modify colonic fermentation of lactose and short-chain fatty acid production. We evaluated the capability of colonic bacteria to ferment lactose and the ability of L. acidophilus to modify lactose fermentation by the colonic microflora in vitro. An anaerobic continuous culture was established and inoculated with fresh samples of human feces. Lactose infusion was maintained at 25 g/d and pH at 6.7. L. acidophilus strain LA-1 (1.5 x 10(10) cells) was introduced into the fermenter on d 0 or added daily on d 0 through 4. The control was the continuous culture without the addition of lactobacilli. Rapid adaptation of colonic bacteria to lactose occurred within 1-2 d, with a significant decrease in lactose concentration and increase in beta-galactosidase activity, and lactose concentrations fell below 3 mmol/L by d 7. Supplementation with strain LA-1 resulted in a significantly greater decrease in lactose concentration and greater increase in acetate and propionate production within the first day compared with the control group. However, there was no significant difference between the fermentation treated with L. acidophilus daily and the control after the first day. These data suggest that the colonic bacteria adapt quickly to lactose, causing efficient utilization of lactose. L. acidophilus supplementation may enhance lactose fermentation during early periods when the adaptation is not established in this model.

Adult↗