Presence Of Propionate Research Articles

Effect of propionate toxicity on methanogen-enriched sludge, Methanobrevibacter smithii, and Methanospirillum hungatii at different pH values

The effect of propionate toxicity at different pH values (6.5, 7.0, and 8.0) on methanogen-enriched sludge. Methanobrevibacter smithii, and Methanospirillum hungatii was studied. Organisms were grown in Balch medium 3 in Hungate tubes, and toxicity was characterized by a decrease in production of methane and in bacterial numbers. Propionate inhibited bacterial growth and cumulative methane production at concentrations as low as 20 mM. In the absence of propionate, the methanogen-enriched sludge and M. smithii showed better cumulative methane production at pH 6.5 and 7.0 than at pH 8.0. However, in the presence of propionate, these organisms showed better cumulative methane production at pH 8.0. M. hungatii differed in its behavior; the best values of cumulative methane production for this organism occurred at pH 7.0. Bacterial numbers reflected the microbial response to the presence of propionate. The highest counts of methanogenic bacteria were observed at pH 6.5 and 8.0. The numbers of methanogens were affected by the presence of propionate even at concentrations as low as 20 or 30 mM; at propionate concentrations above 80 mM, the methanogen count was affected by at least 2 orders of magnitude. Upon comparison of the responses of the pure cultures and the methanogen-enriched sludge to increasing propionate concentrations, it was found that the sensitivity of the pure cultures was similar to that of the methanogens in the sludge.

Formation of poly(3-hydroxyalkanoates) by phototrophic and chemolithotrophic bacteria

The formation of poly(3-hydroxyalkanoic acid), PHA, by various strains of chemolithotrophic and phototrophic bacteria has been examined. Chemolithotrophic bacteria were grown aerobically under nitrogen-limiting conditions on various aliphatic organic acids. Phototrophic bacteria were grown anaerobically in the light on a nitrogen-rich medium and were subsequently transferred to a nitrogen-free medium containing acetate, propionate, valerate, heptanoate or octanoate as carbon source. All 41 strains investigated in this study were able to synthesize and accumulate PHA. All 11 strains of chemolithotrophic bacteria and all 15 strains belonging to the non-sulfur purple bacteria synthesized a polymer, which contained 3-hydroxy-valerate (3HV) beside 3-hydroxybutyrate (3HB), if the cells were cultivated in the presence of propionate, valerate or heptanoate. Many non-sulfur purple bacteria synthesized copolyesters of 3HB and 3HV even with acetate as carbon source. In contrast, most sulfur purple bacteria did not incorporate 3HV at all. Among 15 strains tested, only Chromatium vinosum strain 1611, C. purpuratum strain BN5500 and Lamprocystis roseopersicina strain 3112 were able to synthesize polyesters containing 3HV with propionate, valerate or heptanoate as carbon source.

Effects of Propionate on Lipid Biosynthesis in Isolated Rat Hepatocytes

The effects of propionate, a product of intestinal fiber fermentation, on fatty acid and sterol synthesis were studied in isolated rat hepatocytes. Fatty acid synthesis, as measured by tritium incorporation from 3H2O, was inhibited in the presence of 1 mmol/L propionate with no substrate additions or additions of acetate, butyrate, lactate or oleate. Incorporation of [1-14C]acetate into fatty acids was also inhibited in the presence of propionate. Although propionate markedly depressed [1-14C]acetate incorporation into sterols in hepatocyte preparations, tritium incorporation from 3H2O into sterols was not inhibited, indicating that overall sterol synthesis was not affected. Thus, in vitro, the effect of propionate on lipid metabolism is apparently limited to inhibition of de novo fatty acid synthesis.

Effect of propionate on the utilization of nitrogen from 15NH 4Cl for urea synthesis in hepatocytes isolated from sheep liver

1. 1. The effect of omithine (2.0 mM) and propionate (5.0 mM) on the utilization of N from 15NH 4Cl (5.0 mM) for urea synthesis in hepatocytes isolated from sheep liver was investigated. 2. 2. The capacity of sheep hepatocytes to utilize [ 15N]ammonia in the absence of the other exogenous substrates was very low and amounted 132 ± 37.3 μmol/hr per 1 g dry wt. 3. 3. Omithine failed to affect the total [ 15N]ammonia uptake and total urea synthesis, but at the same time it markedly increased the utilization of [ 15N]ammonia for ureagenesis and diminished the rate of urea synthesis from endogenous sources. 4. 4. Propionate markedly increased total [ 15N]ammonia utilization and total urea formation; this increase resulted from the rise of ammonia utilization for urea synthesis and it was similar in the presence or absence of omithine. 5. 5. The capacity of sheep liver cells to utilize ammonia in the presence of propionate (in the presence or absence of omithine) amounted to 256 μmol/hr per 1 g dry wt, thus being similar to the values in vivo. 6. 6. It is concluded that in sheep hepatocytes both omithine and propionate stimulate the utilization of ammonia for urea synthesis and these effects take place independently and occur by different mechanisms.

Non-hormonal and hormonal control of glycogen metabolism in isolated sheep liver cells

1. 1. Control of glycogen metabolism by various substrates and hormones was studied in ruminant liver using isolated hepatocytes from fed sheep. 2. 2. In these cells glucose appeared uneffective to stimulate glycogen synthesis whereas fructose and propionate activated glycogen synthase owing to (i) a decrease in phosphorylase a activity and (ii) changes in the intracellular concentrations of glucose 6-phosphate and adenine nucleotides. 3. 3. The activation of hepatic glycogenolysis by glucagon and α 1-adrenergic agents was associated with increased phosphorylase a and decreased glycogen synthase activities. 4. 4. The simultaneous changes in these two enzyme activities suggest that in sheep liver, activation of phosphorylase a is not a prerequisite step for synthase inactivation. 5. 5. In sheep hepatocytes, in the presence of propionate and after a lag period, insulin activated glycogen synthase without affecting phosphorylase a. 6. 6. This latter result suggests that the direct activation of glycogen synthase by insulin is mediated by a glycogen synthase-specific kinase or phosphatase. Insulin also antagonized glucagon effect on glycogen synthesis by counteracting the rise of cAMP.

Poly(β-hydroxyalkanoate) biosynthesis and accumulation by differentRhizobiumspecies

Abstract Synthesis and accumulation of poly(β-hydroxy-alkanoate) in Rhizobium leguminosarum , R. leguminosarum biovar. trifolii, Rhizobium ‘galega’, Rhizobium ‘hedysarum’ and R. meliloti were studied. Poly(β-hydroxybutyrate) is accumulated up to 55% of the cell dry weight. At 30–50% air saturation R. meliloti accumulates 50% of its biomass (5.5 g·1 −1 dry weight) as PHB after 90 h of batch fermentation. At 90% air saturation maximum accumulation (37.5%) of PHB occurs after 70 h cultivation. R. meliloti strain 41 is able to synthesize the copolymer poly(β-hydroxybutyrate-co-β-hydroxyvalerate) at concentrations up to 58% of the biomass dry weight, containing up to 22 mol% β-hydroxyvalerate. Different concentrations of both copolymer and hydroxyvalerate were obtained when the microorganism was grown, in batch culture, in the presence of propionate or valerate and with glucose, sucrose or succinate as main carbon source.

The influence of pH on membrane conductance and intercellular resistance in the rat lens.

1. The conductance of the rat lens was measured using a two-internal-microelectrode technique. The voltage response to a step of current consisted of two components arising from bulk and membrane resistance respectively. 2. The potassium permeability was calculated by applying Goldman theory to 86Rb+ efflux data. 3. The internal pH (pHi) and internal free calcium (pCai) were measured directly using single- and double-barrelled ion-sensitive microelectrodes. 4. Lens pHi was 6.9 in control solution (external pH, pHo = 7.3) and was reduced on lowering pHo. The presence of propionate or 100% CO2 in the external solution accentuated this effect. 5. Internal acidification was accompanied by a depolarization of membrane potential, an increase in membrane and cell-to-cell resistance and a decrease in potassium permeability. The acidification had no effect on pCai. 6. The intracellular pH was increased by perifusing with trimethylamine or NH4Cl. Both treatments induced a membrane depolarization with little change in potassium permeability. Subsequent removal of NH4Cl led to a sustained decrease in pHi. 7. In every case where pHi decreased, the changes in membrane potential and conductance could be explained largely on the basis of a decrease in potassium permeability. The concomitant increase in cell-to-cell resistance was less pronounced and probably insufficient to uncouple the lens system.

Effects of propionate and carnitine on the hepatic oxidation of short- and medium-chain-length fatty acids.

Accumulation of propionate, or its metabolic product propionyl-CoA, can disrupt normal cellular metabolism. The present study examined the effects of propionate, or propionyl-CoA generated during the oxidation of odd-chain-length fatty acids, on hepatic oxidation of short- and medium-chain-length fatty acids. In isolated hepatocytes, ketone-body formation from odd-chain-length fatty acids was slow as compared with even-chain-length fatty acid substrates, and increased as the carbon chain length was increased from five to seven to nine. In contrast, rates of ketogenesis from butyrate, hexonoate and octanoate were all approximately equal. Propionate (10 mM) inhibited ketogenesis from butyrate, hexanoate and octanoate by 81%, 53% and 18% respectively. Addition of carnitine had no effect on ketogenesis from the even-chain-length fatty acids, but increased the rate of ketone-body formation from pentanoate (by 53%), heptanoate (by 28%) and from butyrate or hexanoate in the presence of propionate. The inhibitory effect of propionate could not be explained by shunting acetyl-CoA into the tricarboxylic acid cycle, as CO2 formation from butyrate was also decreased by propionate. Examination of the hepatocyte CoA pool during oxidation of butyrate demonstrated that addition of propionate decreased acetyl-CoA and CoA as propionyl-CoA accumulated. Addition of carnitine decreased propionyl-CoA by 50% (associated with production of propionylcarnitine) and increased acetyl-CoA and CoA. Similar changes in the CoA pool were seen during the oxidation of pentanoate. These results demonstrate that accumulation of propionyl-CoA results in inhibition of short-chain fatty acid oxidation. Carnitine can partially reverse this inhibition. Changes in the hepatocyte CoA pool are consistent with carnitine acting by generating propionylcarnitine, thereby decreasing propionyl-CoA and increasing availability of free CoA. The data provide further evidence of the potential cellular toxicity from organic acid accretion, and supports the concept that carnitine's interaction with the cellular CoA pool can have a beneficial effect on cellular metabolism and function under conditions of unusual organic acid accumulation.

Interactions of propionate and carnitine metabolism in isolated rat hepatocytes

Accumulation of propionate and its metabolic products propionyl-CoA and methylmalonyl CoA results in disruption of normal hepatic metabolism. Carnitine can partially restore normal cellular function in the presence of propionate. This beneficial effect of carnitine has been hypothesized to result from removal of propionyl groups in the form of propionylcarnitine. The present study was designed to elucidate the kinetics of propionylcarnitine formation in isolated rat hepatocytes, and the consequences of propionylcarnitine formation on propionate and carnitine metabolism. 14C-Propionate was converted to CO 2, glucose, and propionylcarnitine in the hepatocyte system. Rates of CO 2 production plateaued at propionate concentrations above 0.5 to 1.0 mmol/L, while in contrast, rates of glucose production declined as the propionate concentration was increased from 1.0 to 10.0 mmol/L. Increasing concentrations of carnitine up to 10.0 mmol/L resulted in increased production of propionylcarnitine. Despite formation of propionylcarnitine, propionate conversion to CO 2 and glucose was unaffected by addition of carnitine. Thus, 10 mmol/L carnitine was able to increase total propionate metabolism (conversion to CO 2, glucose and propionylcarnitine) by 40%. Hepatocyte metabolism of propionate was associated with a decrease in carnitine concentration and an increase in short chain acylcarnitines. This decrease in carnitine concentration was also seen in the presence of 150 μmol/L added carnitine, and was greater with propionate as a substrate as compared to butyrate. High performance liquid chromatography was used to permit specific quantification of propionylcarnitine. This technique confirmed that in the presence of propionate, propionylcarnitine was the major acylcarnitine generated and was responsible for the depletion of free carnitine from the system. The present study demonstrates that carnitine can significantly increase total propionate metabolism by hepatocytes. This effect requires large concentrations of carnitine and results from propionylcarnitine generation. Propionylcarnitine production may also deplete carnitine availability for other cellular processes.

Mechanisms of Growth Inhibition by Propionate and Restoration of the Growth by Sodium Bicarbonate or Acetate in Rhodopseudomonas sphaeroides S

Mechanisms of growth inhibition by propionate on the growth of Rhodopseudomonas sphaeroides were studied. Partially purified pyruvate dehydrogenase complex (PDC) from R. sphaeroides was inhibited by propionyl-CoA, one of the metabolic intermediates of propionate, while propionate itself did not inhibit the enzyme. This suggests that the inhibitor of the growth in vivo is not propionate but propionyl-CoA. The inhibition by propionyl-CoA was competitive with respect to coenzyme A concentration. The K1 value for propionyl-CoA was 0.84 mM. Addition of NaHCO3, which restored the growth of this bacterium in the presence of propionate, increased the rate of propionate incorporation by 1.7-fold and decreased the intracellular level of propionyl-CoA by half. These findings suggest that HCO3-ion lowers the level of propionyl-CoA by accelerating its carboxylation reaction, which is catalyzed by propionyl-CoA carboxylase. Effects of NaHCO3 and acetate on the growth restoration were also studied by the use of propionyl-CoA carboxylase-deficient mutants. NaHCO3 did not restore the growth of the mutants, indicating an essential role of propionyl-CoA carboxylase on the restoration of growth by NaHCO3 as suggested above. Addition of acetate restores the growth of the mutants in the presence of propionate. Acetate probably restores the growth by supplying acetyl-CoA.

Effect of acetate and octanoate on tricarboxylic acid cycle metabolite disposal during propionate oxidation in the perfused rat heart.

Tricarboxylic acid cycle pool size is determined by anaplerosis and metabolite disposal. The regulation of the latter during propionate metabolism was studied in isolated perfused rat hearts in the light of the characteristics of NADP-linked malic enzyme, which is inhibited by acetyl-CoA. The acetyl-CoA concentration was varied by infusions of acetate and octanoate, and the rate of metabolite disposal was calculated from a metabolic balance sheet compiled from the relevant metabolic fluxes. Propionate addition increased the tricarboxylic acid cycle pool size 4-fold and co-infusion of acetate or octanoate did not change it further. Propionate caused a decrease in the CoA-SH concentration and a 10-fold increase in the propionyl-CoA concentration. A paradoxical increase in the CoA-SH concentration was observed upon co-infusion of acetate in the presence of propionate, an effect probably caused by competitive inhibition of propionate activation. A more pronounced decline in the propionyl-CoA concentration was observed upon the co-infusion of octanoate. In a metabolic steady state, acetate and octanoate reduced propionate disposal only slightly, but did not increase the tricarboxylic acid cycle pool size. The results are in accord with the notion that the tricarboxylic acid pool size is mainly regulated by the anaplerotic mechanisms.

Effects of volatile fatty acids and ketone bodies on lipolysis in isolated adipocytes from Norwegian reindeer (Rangifer tarandus).

The effect of volatile fatty acids (acetate and propionate) and ketone bodies (acetoacetate and DL-3-hydroxybutyrate) on adrenaline stimulated lipolysis have been studied in isolated adipocytes from Norwegian reindeer. All four substances caused a strong (63-90%) inhibition of lipolysis, measured as glycerol release, at low concentrations of adrenaline (5 nM). At higher adrenaline concentrations (50 nM) inhibition only took place in the presence of propionate (30% inhibition) or DL-3-hydroxybutyrate (43% inhibition). DL-3-hydroxybutyrate caused a marked reduction in adrenaline stimulated glycerol release, even at very low concentrations. The effective dose causing 50% inhibition (ED50) was 1.5 mM. It is suggested that both volatile fatty acids and ketone bodies can act as important volatile regulators of lipopolysis in this high arctic ruminant undergoing great seasonal changes in volatile fatty acid and supposedly ketone body production.

The effect of implantation of anabolic steroids into suckling and ruminating lambs on the metabolism of alanine in livers perfused in the presence or absence of volatile fatty acids

SUMMARYThe effect of implantation of trenbolone acetate and oestradiol into lambs soon after birth on the metabolism of alanine in the presence and absence of propionate has been studied in isolated perfused livers obtained from suckling and from ruminating lambs. The effect of butyrate on the metabolism of alanine in the presence and absence of propionate has been studied in livers obtained from the steroid implanted lambs.Glucose production in the presence of alanine by livers from non-implanted suckling lambs was more than 3 times greater than that by livers from non-implanted ruminating lambs. Steroid implantation did not affect alanine uptake by the livers but metabolism within the livers was altered. Significant interactions were observed between the age of the lambs and whether or not they had been implanted with steroids for the rates of net glucose production in the absence of propionate (P< 0·01) and for the rates of release of urea by the livers in the presence of (P< 0·05) and absence of propionate (P< 0–01).Propionate did not affect alanine uptake by livers obtained from either suckling or ruminating lambs, but there was a significant interaction (P< 0·05) between the age of the lambs and whether or not butyrate was infused, indicating that butyrate depressed alanine uptake by livers obtained from implanted ruminating lambs perfused in the absence of propionate. Despite this depression of alanine uptake the rates of net glucose production and urea production were unchanged, indicating that butyrate modified the metabolism of alanine in the livers of implanted ruminating lambs in that a greater proportion of the alanine carbon taken up must have been converted to glucose. Butyrate stimulated net glucose production in the presence of propionate and alanine in livers from suckling (P< 0·05) but not ruminating lambs. Butyrate was ketogenic and the release of 3-hydroxybutyrate accounted for 45 and 30% of the carbon taken up as butyrate by livers from suckling and ruminating lambs respectively perfused in the absence of propionate. Propionate was antiketogenic. Propionate inhibited urea formation from alanine by livers from implanted suckling (P< 0·05) and implanted ruminating (P< 0·001) lambs. This effect was diminished by the presence of butyrate. Lactate was released in the presence of propionate and accounted for approximately 12–5% of the carbon taken up as propionate.It may be concluded that implantation of lambs with anabolic steroids maintains hepatic gluconeogenesis from alanine at rates more closely resembling those found in younger suckling lambs. The effects of the volatile fatty acids on alanine metabolism are likely to be of physiological significance when sheep are fed diets which result in a markedly altered pattern of rumen fermentation.

The role of chloride in acetylcholine metabolism.

The chloride dependence of acetylcholine (ACh) synthesis and release and of choline uptake was studied in synaptosomal preparations from rat brain. The substitution of propionate for chloride, in the presence of 35 mM-potassium, lowered the ACh content of the synaptosomes. However, in the presence of 5 mM-potassium, the ACh level in synaptosomes was reduced, but significantly less so. Propionate had no effect on choline acetyltransferase (EC 2.3.1.6) activity when measured in a standard chloride-containing medium. In the presence of propionate, the spontaneous release of ACh was unchanged, but potassium-stimulated release of ACh was markedly reduced as compared with a chloride-containing medium. The synthesis of ACh, as measured by the net increase in the amount of ACh in the synaptosomes and that released to the medium, was reduced with propionate at 5 mM-potassium and was totally inhibited when the potassium concentration was increased to 35 mM. Choline uptake studies revealed that with propionate only a low-affinity component of the choline transport system existed. Further, the Vmax was markedly reduced when the potassium concentration was increased to 35 mM. The results suggest that under certain conditions choline transported by a low-affinity system might provide a substantial source of choline for ACh synthesis.

Nicotinic acid metabolism: stereochemical course of the (2R,3S)-2,3-dimethylmalate lyase reaction.

1. The enantiomers of chiral propionate were prepared by cleavage of (2R,3S)-2,3-dimethylmalate and of (2R,3S)-[3-3H1]-2,3-dimethylmalate with the corresponding lyase in tritiated and in ordinary water, respectively. 2. Using commercially available citrate lyase, an enzymic method was elaborated for the formation of (3S)-citryl-CoA from acetyl-CoA and citrate in a preparative scale (mumol). The product was used in the presence of propionate (or acetate) and partially inactivated citrate lyase to prepare propionyl-CoA (or acetyl-CoA). The method was applied to the chiral propionates as well as symmetrically labelled [2-3H2]propionate and is suggested to be superior to others for the preparation of propionyl-CoA (or acetyl-CoA) from mumol of radioactively labelled acid. 3. On configurational determination with propionyl-CoA carboxylase the [2-3H12]propionyl-CoA derived from unlabelled dimethylmalate retained nearly all of the tritium label, whereas that derived from the labelled substrate lost the tritium label completely; [2-3H2]propionyl-CoA lost 50% of its label. 4. It was concluded from these results that the cleavage reaction on (2R,3S)-2,3-dimethylmalate lyase proceeds with inversion of configuration at C-3 of the substrate which becomes C-2 of propionate.

The stimulation of pyruvate appearance in the rabbit oviduct lumen by sodium propionate.

Sodium propionate stimulated the appearance of pyruvte in saline medium recirculated through the lumen of the rabbit oviduct for 4 h in vivo. The propionate was equally effective whether added to the lumen or infused into the bloodstream. The final pyruvate concentration in the lumen in the presence of propionate was 1.3 times that in the plasma. Propionate increased the pyruvate content of oviduct tissue slices incubated in vitro, indicating that a proportion of the propionate-stimulated pyruvate appearance could be due to the metabolic conversion of propionate into pyruvate.

Nonequilibration of Acetyl Units from Fatty Acids and Glucose in Mammary Glands of Lactating Mice

Lipid synthesis by lactating mouse mammary gland slices from [1-carbon-14] propionate, [1-carbon-14] valerate, and [1-carbon-14] heptanoate, in the presence of glucose and from [uniformly labeled carbon-14] glucose in the presence of propionate, valerate, and heptanoate was studied. Regardless of the labeled substrate, almost all of the radioactivity incorporated in the total lipid fraction was in triglycerides. In experiments with labeled glucose and either the propionic. the valeric, or the heptylic acid, the carbon-14 fatty acids in triglycerides were almost exclusively composed of even-chain fatty acids. When the substrate was either [1-carbon-14] propionate, [1-carbon-14] valerate, or [1-carbon-14] heptanoate, in the presence of glucose, the pattern changed to one in which a larger proportion of the radioactivity was in the odd-chain fatty acids. These findings suggest that in mouse mammary gland, acetyl units produced via β-oxidation of fatty acids may not equilibrate with those formed from oxidation of pyruvate which is derived from glucose.

Activation of volatile fatty acids in bovine liver and rumen epithelium. Evidence for control by autoregulation.

1. The total capacities of homogenates of bovine liver and rumen epithelium to activate acetate, propionate and butyrate were determined. 2. Activating capacities were assayed by measuring the rate of formation of the corresponding CoA esters. The methods used for determining the concentrations of the CoA esters allowed the CoA esters of acetate, propionate and butyrate to be distinguished. It was thus possible to investigate the effect of the presence of a second volatile fatty acid on the rate at which a given volatile fatty acid was activated. 3. The propionate-activating capacity in rumen epithelium was decreased by about 87% in the presence of butyrate, the acetate-activating capacity in liver was decreased by about 55% in the presence of either propionate or butyrate, and the butyrate-activating capacity in liver was decreased by about 40-50% in the presence of propionate. 4. All three activating capacities in liver appeared to be located in the mitochondrial matrix and membrane. The three activating capacities had similar locations to each other in rumen epithelium as well, although in this case activity was more evenly divided between the mitochondria and the cytoplasm. 5. The relative activating capacities towards the volatile fatty acids in the two tissues, together with the ability of one volatile fatty acid to inhibit the activation of another volatile fatty acid, appear to ensure that butyrate is mainly metabolized in the rumen epithelium and that propionate is metabolized in the liver.

A sensitive assay for the determination of propionyl-CoA in biological material

A method has been developed for the determination of low amounts of propionyl-CoA in biological material. The method involves 14CO 2 fixation by propionyl-CoA in the presence of purified propionyl-CoA carboxylase. Values for propionyl-CoA in rat liver in vivo and in isolated rat livers perfused in the presence of propionate are reported.

Production within the rumen and removal from the blood-stream of volatile fatty acids in sheep given a diet deficient in cobalt

1. The production and metabolism of volatile fatty acids were studied in sheep offered a cobalt-deficient diet.2. The molar proportions of acetic (60%), propionic (26%) and butyric (14%) acids in the rumen fluids of sheep given the Co-deficient diet, but whose stores of vitamin B12were adequate, were similar before and after administration of an oral supplement of Co.3. In pair-fed sheep, one member of which was vitamin B12-deficient and the other (control) treated with vitamin B12parenterally, the concentrations after feeding of both total and individual volatile fatty acids in the blood tended to be higher in deficient than in control sheep.4. Following injection of the respective salts of individual volatile fatty acids into the blood-stream, formate clearance was apparently not affected, whereas that of acetate was slightly, and that of propionate very significantly, delayed in vitamin B12-deficient sheep compared with pair-fed control animals.5. Acetate metabolism was retarded in the presence of propionate; the effect was greater in deficient than in pair-fed control sheep.6. The hypothesis is advanced that it is the failure to metabolize propionate at the normal rate that leads to the progressive loss of appetite in vitamin B12-deficient sheep.