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Response of forage fiber degradation by ruminal microorganisms to branched-chain volatile fatty acids, amino acids, and dipeptides.

This study evaluated the effect of branched-chain volatile fatty acids (VFA; isobutyric acid, isovaleric acid), amino acids (valine, leucine), and dipeptides (valine-valine, leucine-leucine) on neutral detergent fiber (NDF) degradation by rumen microorganisms in vitro. The CP (%) and in situ NDF degradation rate (%/h) for alfalfa, bermudagrass, and pangolagrass hays, and napiergrass silage were 17.2 and 7.5, 4.7 and 3.1, 8.3 and 5.3, and 9.6 and 3.4, respectively. In vitro NDF digestibility was the lowest for bermudagrass; alfalfa and napiergrass were the highest. When the incubation contained more ammonia initially, digestibilities increased, but relative differences among forages were unchanged. Adding branched-chain VFA (2 mM) to incubations increased digestibilities more than controls on 15 out of 16 occasions. The effectiveness varied with isoacids and forages used. Amino acid (2 mM) or dipeptide (1 mM) addition consistently increased digestibility over controls. Amino acids further increased digestibility over corresponding isoacids on 14 occasions. Improvement in digestibility over control by leucine appeared to be greater than that by valine. Digestibilities with dipeptides were always greater than those with isoacids, except for one case. Dipeptide addition further increased digestibility significantly over corresponding amino acids on only six occasions, while percent improvement in digestibility numerically by dipeptides occurred in 10 cases. Valine-valine seemed to exert different effect than leucine-leucine, depending on initial ammonia availability. The results indicate that dipeptides could be more effective than isoacids and amino acids in improving NDF digestion. Forages with high CP content or rapid NDF degradation rate appeared to respond to additives to smaller degrees.

Amino Acids↗

Branched chain amino acids as source of specific branched chain volatile fatty acids during the fermentation process of fish sauce.

The source of the formation of branched chain volatile fatty acids (VFA) in fish sauce was investigated. Certain branched VFA were derived from the degradation of specific amino acids as iso-butyric acid from valine and iso-valeric acid from leucine. Short and long straight chain VFA were significantly higher in the linoleic acid added sample than in the control but did not significantly bring changes to the branched chain VFA. It is suggested that straight chain VFA developed from fish fats. Alanine and isoleucine did not have a clear influence on the production of volatile fatty acids.

Amino Acids, Branched-Chain↗

Ruminal lactic acidosis: forestomach epithelial receptor activation by undissociated volatile fatty acids and rumen fluids collected during loss of reticuloruminal motility.

Forty-three reticuloruminal epithelial receptors, with excitatory receptive fields within the reticulum, were isolated in 19 anaesthetised sheep. The responsiveness of these receptors to acetic, propionic, butyric and DL-lactic acids were assessed as well as their ability to be activated by rumen fluids obtained from sheep with induced ruminal lactic acidosis. Eighteen (41.9 per cent) receptors were excited by all three volatile fatty acids (VFAs) and 17 (39.5 per cent) responded to two VFAs. Butyric acid was the most potent volatile fatty acid eliciting responses in 41 (95.3 per cent) receptors. Acetic acid activated 33 (76.7 per cent) receptors and propionic acid excited 22 (53.5 per cent) receptors. DL-lactic acid only activated 13 (30.2 per cent) receptors, six of which required concentrations of 200 mM to be activated. Rumen fluids obtained when forestomach motility was impaired, activated 37 (81.1 per cent) of the 43 receptors, whereas rumen fluids obtained when forestomach motility was normal did not activate any receptors. Despite the high levels of lactic acid in rumen fluids which activated epithelial receptors, this acid was not responsible for receptor excitation.

Acetates↗

[Intravenous infusion of volatile fatty acids as a metabolic loading test for the evaluation of the possible effects of butafosfane on the energy metabolism of cattle].

In 10 German Black and White heifers metabolic stress reactions were induced by intravenous infusion of propionate and butyrate to demonstrate possible effects of prior medication with butafosfane. This investigation included a preliminary feeding period, followed by 18 hours of feed withdrawal, premedication with butafosfane or placebo and infusion of volatile fatty acids. Reactions were assessed by monitoring biochemical and clinical parameters. A total of 19 blood samples per series were taken in which the concentrations of glucose, free fatty acids, volatile fatty acids, beta-hydroxybutyrate, cortisol and insulin were measured. Clinical parameters recorded included heart rate and respiratory rate. During loading with propionate effects of butafosfane on plasma levels of cortisol and insulin could be demonstrated. The differences noted in the other parameters were only insignificant. The loading test chosen is suitable for the demonstration of effects of butafosfane on metabolic parameters which could hardly be influenced by this substance in unstressed animals.

Animals↗

[Direct gas chromatography analysis of volatile fatty acids and lactic acids in microbiological diagnosis].

The gas chromatographic analysis of short-chain fatty acids is of great significance in the rapid diagnosis of anaerobic infections and bacterial meningitis. A new analytical method is presented detecting lactic acid without prior derivatisation in addition to volatile fatty acids in the aqueous phase directly. Lactic acid levels greater than 30 mg/dl are measured in cerebrospinal fluids of patients with acute bacterial meningitis. The sole detection of acetic acid in concentrations higher than 20 mg/dl indicates aerobes in clinical materials. A high correlation exists between the determination of n-butyric acid as well as iso-valeric acid and the occurrence of anaerobic bacteria.

Bacteria, Anaerobic↗

Analysis by gas liquid chromatography of production of volatile fatty acids by anaerobic bacteria grown on solid medium.

Volatile fatty acids produced in Robertson's cooked meat medium by a range of clinically relevant anaerobes were compared by gas liquid chromatography with those produced in blood agar. The same volatile fatty acid profiles were obtained in both media, although the concentration of acids was lower in blood agar. We conclude that detection of volatile fatty acids from a pure culture of an organism on solid medium is practicable and offers advantages over the conventional technique.

Acetates↗

Impact of concentration, temperature, and pH on inactivation of Salmonella spp. by volatile fatty acids in anaerobic digestion.

It is known that the presence of volatile fatty acids may play a role in the inactivation of pathogens for systems that employ an acid phase reactor. This study was conducted to investigate the influence of volatile fatty acids on the inactivation of Salmonella spp. over a range of digestion temperatures. In this study, digesters that were treating municipal wastewater treatment plant sludges were operated at temperatures that ranged from 35 to 49 degrees C and had a solids residence time of 15 days. Samples collected from the effluent of the digesters were dosed with solutions containing acetic, propionic, and butyric acids alone and in mixtures, and the dosed effluents were analyzed for Salmonella spp. over time. In the first round of testing, the digester effluents were dosed with individual organic acids and also a mixture containing all three volatile fatty acids over a range of concentrations from 750 to 6000 mg/L, and the pH of the samples was fixed at a value of 5.5. In the second round of testing, the sample sludges were spiked with a fixed amount of organic acid mixture, and the pH was varied from 4.5 to 7.5. The reduction of Salmonella spp. in digester effluents, when dosed with volatile organic acids, was found to depend on pH, temperature, the chain length of the acids, and the concentration and composition of the acids present. Increases in temperature appeared to increase the inhibitory effects of the volatile organic acids. At mesophilic temperatures, acidic pHs resulted in a greater inhibition of Salmonella spp.; whereas at higher temperatures neutral pHs were found to be more inhibitory. The results suggest that acid phase digesters that operate at elevated temperatures and low pH can achieve substantial reduction of Salmonella spp.

Anaerobiosis↗

Effect of volatile fatty acid infusion on blood plasma free amino acid profiles in growing lambs.

Five ram lambs (average body mass: 25 kg) were given, through a catheter inserted into the left ruminal vein, a total of 28.8 mM sodium acetate, 14.4 mM sodium propionate and 4.8 mM sodium butyrate per kg body mass as a 2-hour infusion. During and at 0, 1, 2, 4, 6, 10 and 24 h after the infusion blood samples were taken from the jugular vein and the blood plasma was assayed for free amino acid (FAA) and immunoreactive insulin (IRI) concentrations. Volatile fatty acid (VFA) infusion significantly decreased the blood plasma concentrations of all FAA but cystine. The lowest FAA concentrations were measured in plasma samples taken at the end of the 2-h infusion. Subsequently the level of all amino acids rose and by 24 h after the infusion the blood plasma concentration of all FAA came close to the preinfusion value. The largest differences were observed in the concentration of glutamate, glycine, leucine and isoleucine. In contrast to FAA, IRI concentration was increased significantly (almost fivefold) by VFA infusion. By 10 h after the infusion IRI concentration returned to the initial level. The results reported here indicate that energy supply given in the form of VFA infusion significantly affects blood plasma FAA profiles, supposedly as a result of changes induced in protein synthesis in tissues. Insulin presumably plays a role in the regulation of these changes.

Amino Acids↗

Volatile fatty acids, lactic acid, and pH in the stools of breast-fed and bottle-fed infants.

We measured fecal pH, volatile fatty acids (VFA), and lactic acid concentrations in 67 healthy infants ranging in age from 1 to 5 months. The babies were divided into five groups according to their diet: group 1: human milk, with subgroups of 1 month of age (n = 8), and 2-5 months (n = 12); group 2: formula milk, with subgroups of 1 month of age (n = 9) and 2-5 months (n = 10); group 3: cow's milk, one group of 2-5 months of age (n = 12); group 4: human milk supplemented with formula milk, 2-5 months of age (n = 8); and group 5: human milk supplemented with cow's milk, 2-5 months of age (n = 8). Fecal VFA concentration and pH of bottle-fed infants (groups 2 and 3) were significantly higher and lactic acid concentrations significantly lower than those of the breast-fed group. Groups 4 and 5 showed significantly higher pH and propionic acid concentration, and lower lactic acid concentration than Group 1 did. There were no significant differences in the data on pH, VFA, and lactic acid concentrations between the feces of formula and cow's milk groups. On the other hand, the addition of formula or cow's milk to the human milk was sufficient to alter the profile of VFA, the pH, and the lactic acid concentration of breast-fed infants stools. Low pH, low VFA, and high lactic acid concentrations make the colonic content of breast-fed babies an environment favorable for Bifidobacteria growth, and this is probably the main protective factor against gastrointestinal infections.

Acetates↗

[Gas chromatographic determination of the volatile fatty acids (C1-C5) including lactic acid after conversion into their benzyl esters by phenyldiazomethane. Determination of the acids in silages (author's transl)].

A gas chromatographic method for the determination of the volatile fatty acids (C1-C5) including lactic acid is described. The acids are converted into their benzyl esters by means of phenyldiazomethane without previous purification of the reagent by distillation. The benzyl esters are well separated from the solvent as well as from one another. Silicone SE-30 is used as the stationary phase. An application of the method on silage acids is demonstrated.

Animal Feed↗

Effect of ruminal infusion of glucose, volatile fatty acids and hydrochloric acid on mineral metabolism in sheep.

Two experiments were conducted to study the effects of alterations in ruminal pH and volatile fatty acid (VFA) concentrations on utilization of Mg and other minerals. In Exp. 1, two metabolism trials were conducted with 12 ruminally cannulated crossbred wethers fed 800 g/d of orchard-grass (Dactylis glomerata, L.) hay. After each feeding, wethers were ruminally infused with 500 ml (4.2 ml/min) or either 1) deionized water, 2) 40% (w/v) glucose solution, 3) .26 M propionic and .17 M butyric acid solution or 4) .35 M HCl. The pH of the VFA solution was adjusted to 6.8 with 10N NaOH. In Exp. 2, a metabolism trial was conducted with 12 ruminally cannulated crossbred wethers fed 600 g of orchard-grass hay and infused with a buffered VFA solution prepared as in Exp. 1 or with an unbuffered solution. In both experiments each trial consisted of a 5-d adaption period followed by four 5-d collections of feed, feces and urine. Compared with the glucose treatment, infusion of the buffered VFA solution produced similar acetic and propionic and higher (P less than .05) butyric acid concentrations (Exp. 1). The HCl solution produced changes in ruminal and pH values similar to those of the glucose infusion. In Exp. 1, apparent absorption of Mg was increased over twofold by the glucose infusion (P less than .05), but the other infusions had no effect. Apparent absorption of P was decreased (P less than .05) by HCl infusion, and K absorption was decreased by HCl and glucose infusions. In Exp. 2, infusion of the unbuffered VFA solution decreased apparent Mg absorption by 15.7%, compared with infusion of the buffered solution. These experiments suggest that the increased Mg absorption observed with carbohydrate supplementation is not due to alterations in ruminal pH or VFA levels.

Animals↗

Effect of diet and infusion of volatile fatty acids into the rumen on the concentration of plasma free amino acids in sheep.

The effect of supplementing barley diets with urea (U), extracted decorticated groundnut meal (GNM) or Peruvian fish meal (PFM) on plasma free amino acid concentrations in sheep have been examined and the first limiting amino acid has been indicated by measuring the changes in the concentration of the plasma essential amino acids (PEAA) during a rumen infusions of a volatile fatty acid (VFA) mixture. 2. Three wethers fitted rumen and re-entrant duodenal cannulas were given isonitrogenous, isoenergetic diets containing (g/kg dry matter (DM) U 20, GNM 106 and PFM 78, the crude protein (nitrogen x 6.25) contents being 139, 145 and 148 respectively. The sheep were fed hourly, the mean daily DM intake being 0.634 kg. 3. Plasma concentrations of valine, threonine, lysine, isoleucine and leucine were linearly related to their concentrations in duodenal digesta. 4. A VFA mixture was infused into the rumen for 6 h to supply (mmol/min) acetate 1.47, propionate 0.22 and n-butyrate 0.27. Blood samples were taken 6 h before, during and 12 h after the end of the infusion. 5. The concentration of all PEAA decreased relative to the pre-infusion and post-infusion controls but there were no significant differences between diets. 6. The mean decrease in concentration averaged over all three diets showed that the decrease in concentration of methionine (41.5%) was far greater than for any other essential amino acid suggesting that under these conditions methionine was the first limiting amino acid.

Amino Acids, Essential↗

Net volatile fatty acid absorption in nonlactating holstein cows.

Net absorption of volatile fatty acids was measured in four nonlactating Holstein cows fed orchardgrass-clover silage ad libitum and 50 g trace mineralized salt daily. Cows ranged in age from 2 to 4 yr and in body weight from 326 to 525 kg. Portal blood flow and volatile fatty acid concentrations of portal and arterial plasma were determined at 30-min (three cows) or 90-min (one cow) intervals for about 12 h. Mean portal blood flow was 836 liters/h. Mean volatile fatty acid concentrations of portal plasma and differences of portal-arterial concentrations were (mM): acetate 2.00, .67; propionate .264, .228; isobutyrate .022, .017; n-butyrate .057, .038; 2-methylbutyrate .017, .014; 3-methylbutyrate .007, .005; and n-valerate .012, .008. Mean net volatile fatty acid absorption was 682 mmol/h. Acetate and propionate accounted for 91% of net volatile fatty acid absorption on a molar basis and 85% on an energy basis. Net energy absorbed as volatile fatty acid was 5.43 Mcal/cow per day, which was about 35% of calculated metabolizable energy intake. Portal blood flow was maximal 1.5 h postfeeding, and net volatile fatty acid absorption was maximal 2.5 h postfeeding.

Animals↗

Effect of volatile fatty acids on blood flow and oxygen uptake by the dog colon.

The effects of volatile fatty acids on colonic hemodynamics and oxygen uptake were examined in a denervated autoperfused dog colon preparation. While measuring arterial pressure, colonic blood flow, venous pressure, and arteriovenous O2 difference, a Tyrode's solution containing acetic (75 mM), propionic (30 mM), and butyric acids (30 mM) was placed into the colon lumen. The volatile fatty acid mixture increased colonic blood flow (23.5%) and oxygen uptake (18.4%), yet did not affect oxygen extraction. Of the three fatty acids, only acetic acid (75 mM) could mimic the effects of the entire mixture on colonic hemodynamics and oxygen uptake. Butyric (30 mM) and propionic (30 mM) acids were without appreciable effects on colonic blood flow and oxygen uptake. The vehicle for the volatile fatty acids, Tyrode's did not alter blood flow, yet increased colonic oxygen uptake (18.7%) through an increase in oxygen extraction (18.3%). When compared with Tyrode's, acetic acid increased blood flow, yet did not alter oxygen uptake, while butyric and propionic acids did not alter colonic blood flow, yet inhibited colonic oxygen uptake. Close intraarterial infusion of acetic acid increased colonic blood flow and oxygen uptake. These data suggest that physiologic concentrations of volatile fatty acids (particularly acetic acid) significantly alter colonic flow.

Acetates↗

Determination of volatile fatty acids in plasma after ethanolic extraction.

1. A new rapid micro-method for measuring plasma volatile fatty acids is described. The volatile fatty acids are extracted from plasma with ethanol in the presence of a known quantity of internal standard (sodium isobutyrate). After evaporation of the ethanolic solution of the sodium salts, the residue is dissolved in a dilute solution of orthophosphoric acid to permit analysis by g.l.c. 2. A technique of g.l.c. analysis is described which permits the separation of all the volatile fatty acids from the other plasma constituents at temperatures below 100 degrees C in 5 min. 3. Steam-distillation techniques are unsatisfactory when the acetic acid concentrations in the plasma are below 0.2mm. Heating of a number of plasma constituents in acid conditions gives rise to acetic acid. 4. The binding of volatile fatty acids to plasma proteins was studied; this binding is negligible for acetic acid, but increases with the length of the fatty acid carbon chain. 5. The limits of use of the method and the physiological implications are discussed.

Animals↗

Inhibition of reticulo-ruminal motility by volatile fatty acids and lactic acid in sheep.

1. A study was made of the influence on reticulo-ruminal motility, recorded by electromyography, of ruminal infusions of volatile fatty acids (VFAs) and lactic acid in twenty-four sheep maintained by intragastric infusion of a complete liquid diet, in three sheep fed grass pellets, and in nine chronically vagotomized sheep; abomasal and duodenal infusions of VFA and lactic acid were tested in five sheep fed grass pellets. 2. Ruminal infusions of VFAs and lactic acid progressively inhibited the amplitude of the reticulo-ruminal contractions. In many experiments there was no effect on contraction frequency until the cessation of all reticulo-ruminal contractions at which point the maximal concentration of VFA recorded in the abomasum was 28 mM, and that of lactic acid was 20 mM. 3. The concentrations of undissociated VFAs causing cessation of reticulo-ruminal contractions in the vagus-intact sheep were very similar to the concentrations causing abolition of the organized intrinsic motility of the chronically vagotomized sheep. 4. The inhibition of reticulo-ruminal motility with ruminal infusions of mixtures of VFAs and of lactic acid together with VFAs could largely be explained by the sum of the effects of the individual acids present. 5. Abomasal infusion of VFA or lactic acid inhibited the amplitude of ruminal, especially primary ruminal, contractions at concentrations of undissociated acid of 60 mM and above and increased the frequency of reticulum and primary ruminal contractions at about 80 mM. 6. Duodenal infusion of VFAs and lactic acid (100 mM, 5 ml/min) strongly inhibited abomasal motility without affecting reticulo-ruminal motility, and at a higher rate (100 mM, 10 ml/min) abolished motility and inhibited both the amplitude and frequency of reticulo-ruminal contractions. 7. It is concluded that the initial inhibition of reticulo-ruminal motility in ruminal acidosis is unlikely to involve any significant influence from duodenal, or abomasal receptors. The final cessation of reticulo-ruminal motility with ruminal acidosis could involve local effects of VFAs in the reticulo-rumen as well as through excitation of acid-sensitive reticulo-ruminal receptors.

Action Potentials↗

EFFECT IN THE COW OF INTRARUMINAL INFUSIONS OF VOLATILE FATTY ACIDS AND OF LACTIC ACID ON THE SECRETION OF THE COMPONENT FATTY ACIDS OF THE MILK FAT AND ON THE COMPOSITION OF BLOOD.

1. The effects in the cow of intraruminal infusions of acetic acid, propionic acid or butyric acid on the secretion of the component fatty acids of the milk fat, and of these acids and of lactic acid on the composition of the blood plasma of the jugular vein, have been studied. 2. The infusion of acetic acid or butyric acid increased the yield of the C(4)-C(16) acids of milk fat but decreased the yield of C(18) acids. The infusion of propionic acid decreased the yields of all major component acids except palmitic acid and possibly lauric acid. 3. The changes in the concentrations in blood plasma of glucose and of ketone bodies were consistent with the glucogenic effect of propionic acid and the ketogenic effects of butyric acid and acetic acid. The effects of lactic acid were not consistent from cow to cow. Only with the infusion of acetic acid was a significant increase in the concentration of total volatile fatty acids in blood plasma found. Infusions of butyric acid and of propionic acid tended to depress the concentration of citric acid in the blood plasma and infusion of acetic acid increased it. No consistent effects of the infused acids on the concentration in blood plasma of esterified cholesterol, free cholesterol, triglyceride or phospholipid were observed. 4. The possibility is discussed that the effects of the infused acids on milk-fat secretion are caused through an alteration of the concentrations of precursors of milk fat in mammary arterial blood.

Acetates↗