The iron-regulatory hormone hepcidin is transcriptionally up-regulated by gluconeogenic signals. Recent evidence suggeststhat increases in circulating hepcidin may decrease dietary iron absorption following prolonged exercise, however evidence is limited on whether gluconeogenic signals contribute to post-exercise increases in hepcidin. Mice with genetic knockout of regulated in development and DNA response-1 (REDD1) display greater glycogen depletion following exercise, possibly indicating greater gluconeogenesis. The objective of the present study was to determine liver hepcidin, markers of gluconeogenesis and iron metabolism in REDD1 knockout and wild-type mice following prolonged exercise. Twelve-week-old male REDD1 knockout and wild-type mice were randomised to rest or 60 min treadmill running with 1, 3 or 6 h recovery (n = 5–8/genotype/group). Liver gene expression of hepcidin (Hamp) and gluconeogenic enzymes (Ppargc1a, Creb3l3, Pck1, Pygl) were determined by qRT-PCR. Effects of genotype, exercise and their interaction were assessed by two-way ANOVAs with Tukey's post-hoc tests, and Pearson correlations were used to assess the relationships between Hamp and study outcomes. Liver Hamp increased 1- and 4-fold at 3 and 6 h post-exercise, compared to rest (P-adjusted < 0⋅009 for all), and was 50% greater in REDD1 knockout compared to wild-type mice (P = 0⋅0015). Liver Ppargc1a, Creb3l3 and Pck1 increased with treadmill running (P < 0⋅0001 for all), and liver Ppargc1a, Pck1 and Pygl were greater with REDD1 deletion (P < 0⋅02 for all). Liver Hamp was positively correlated with liver Creb3l3 (R = 0⋅62, P < 0⋅0001) and Pck1 (R = 0⋅44, P = 0⋅0014). In conclusion, REDD1 deletion and prolonged treadmill running increased liver Hamp and gluconeogenic regulators of Hamp, suggesting gluconeogenic signalling of hepcidin with prolonged exercise.

The liver is a key metabolic organ that undertakes a multitude of physiological processes over the course of a day, including intrahepatic lipid and glucose metabolism which plays a key role in the regulation of systemic lipid and glucose concentrations. It serves as an intermediary organ between exogenous (dietary) and endogenous energy supply to extrahepatic organs. Thus, perturbations in hepatic metabolism can impact widely on metabolic disease risk. For example, the accumulation of intra-hepatocellular TAG (IHTG), for which adiposity is almost invariably a causative factor may result in dysregulation of metabolic pathways. Accumulation of IHTG is likely due to an imbalance between fatty acid delivery, synthesis and removal (via oxidation or export as TAG) from the liver; insulin plays a key role in all of these processes.

To unveil the adaptation of Litopenaeus vannamei to elevated ambient ammonia-N, crustacean hyperglycaemic hormone (CHH) was knocked down to investigate its function in glucose metabolism pathway under ammonia-N exposure. When CHH was silenced, haemolymph glucose increased significantly during 3–6 h, decreased significantly during 12–48 h and recovered to the control groups’ level at 72 h. After CHH knock-down, dopamine (DA) contents reduced significantly during 3–24 h, which recovered after 48 h. Besides, the expressions of guanylyl cyclase (GC) and DA1R in the hepatopancreas decreased significantly, while DA4R increased significantly. Correspondingly, the contents of cyclic AMP (cAMP), cyclic GMP (cGMP) and diacylglycerol (DAG) and the expressions of protein kinase A (PKA), protein kinase G (PKG), AMP active protein kinase α (AMPKα) and AMPKγ were significantly down-regulated, while the levels of protein kinase C (PKC) and AMPKβ were significantly up-regulated. The expressions of cyclic AMP response element-binding protein (CREB) and GLUT2 decreased significantly, while GLUT1 increased significantly. Moreover, glycogen content, glycogen synthase and glycogen phosphorylase activities in hepatopancreas and muscle were significantly increased. Furthermore, the levels of key enzymes hexokinase, pyruvate kinase and phosphofructokinase in glycolysis (GLY), rate-limiting enzymes citrate synthase in tricarboxylic acid and critical enzymes phosphoenolpyruvate carboxykinase, fructose diphosphate and glucose-6-phosphatase in gluconeogenesis (GNG) were significantly decreased in hepatopancreas. These results suggest that CHH affects DA and then they affect their receptors to transmit glucose metabolism signals into the hepatopancreas of L. vannamei under ammonia-N stress. CHH acts on the cGMP-PKG-AMPKα-CREB pathway through GC, and CHH affects DA to influence cAMP-PKA-AMPKγ-CREB and DAG-PKC-AMPKβ-CREB pathways, thereby regulating GLUT, inhibiting glycogen metabolism and promoting GLY and GNG. This study contributes to further understand glucose metabolism mechanism of crustacean in response to environmental stress.

Since nutritional requirements are increased at the end of gestation to meet the demands of the pregnant uterus, pregnant beef cows are susceptible to mobilization of body reserves (mainly fat and amino acids (AAs)) and to alter the metabolism of nutrients in the liver and muscle to support such demands. The objective of this study was to evaluate the effect of CP supplementation on maternal nutrient metabolism in the late gestation of beef cows grazing a low-quality pasture. Forty-three pregnant Nellore cows gestating male fetuses (average age = 6 years; average weight = 544 kg) at 193 ± 30 (mean ± SD) days (d) of gestation were divided into eight groups (experimental units, with four to five cows each). Treatments were (1) control (CON, n = 4): pasture-based (PB) diet without CP supplementation and (2) supplemented (SUP, n = 4): PB diet daily supplemented with 2 g/kg of BW of a 43.5% CP supplement. Liver and skeletal muscle biopsies were performed at 265 days of gestation and samples were collected for mRNA expression. On day 280 of gestation, blood samples were collected to assess plasma levels of AA. The CON-fed cows tended to have greater (P = 0.057) total circulating AA than SUP-fed cows. The circulating glycogenic AA was greater (P = 0.035) in CON than in SUP cows. CON cows was greater for histidine (P = 0.015), methionine (P = 0.007) and alanine (P = 0.036) than SUP cows. The CON- and SUP-fed showed no differences for gluconeogenesis, fatty acid transport and signaling axis markers in the liver. The mRNA expression of markers for skeletal muscle synthesis, p7056k (P = 0.060) and GSK3B (P = 0.096), tended to be greater in cows from CON than SUP group. No differences were found for mRNA expression of markers for skeletal muscle degradation. We conclude that CP supplementation to CP-restricted late-pregnant beef cows reduces the maternal tissue mobilization and changes the profile of plasma circulating AA and the mRNA expression of markers for the synthesis of skeletal muscle tissue.

For their glucose supply, ruminants are highly dependent on the endogenous synthesis in the liver, but despite the numerous studies that evaluated hepatic glucose production, very few simultaneously measured hepatic glucose production and uptake of all precursors. As a result, the variability of precursor conversion into glucose in the liver is not known. The present study aimed at investigating by meta-analysis the relationships between hepatic glucose net release and uptake of precursors. We used the FLuxes of nutrients across Organs and tissues in Ruminant Animals database, which gathers international results on net nutrient fluxes at splanchnic level measured in catheterized animals. Response equations were developed for intakes up to 41 g DM intake/kg BW per day of diets varying from 0 to 100 g of concentrate/100 g DM in the absence of additives. The net hepatic uptake of propionate, α-amino-N and l-lactate was linearly and better related to their net portal appearance (NPA) than to their afferent hepatic flux. Blood flow data were corrected for lack of deacetylation of the para-aminohippuric acid, and this correction was shown to impact the response equations. To develop response equations between the availability of precursors (portal appearance and hepatic uptake) and net glucose hepatic release, missing data on precursor fluxes were predicted from dietary characteristics using previously developed response equations. Net hepatic release of glucose was curvilinearly related to hepatic supply and uptake of the sum of precursors, suggesting a lower conversion rate of precursors at high precursor supply. Factors of variation were explored for the linear portion of this relationship, which applied to NPA of precursors ranging from 0.99 to 9.60 mmol C/kg BW per h. Hepatic release of glucose was shown to be reduced by the portal absorption of glucose from diets containing bypass starch and to be increased by an increased uptake of β-hydroxybutyrate indicative of higher body tissue mobilization. These relationships were affected by the physiological status of the animals. In conclusion, we established equations that quantify the net release of glucose by the liver from the net availability of precursors. They provide a quantitative overview of factors regulating hepatic glucose synthesis in ruminants. These equations can be linked with the predictions of portal absorption of nutrients from intake and dietary characteristics, and provide indications of glucose synthesis from dietary characteristics.

An oral starch administration trial was used to evaluate glucose homoeostasis in grass carp (Ctenopharyngodon idella) and Chinese longsnout catfish (Leiocassis longirostris Günther). Fish were administered with 3 g of a water and starch mixture (with 3:2 ratio) per 100 g body weight after fasting for 48 h. Fish were sampled at 0, 1, 3, 6, 12, 24 and 48 h after oral starch administration. In grass carp, plasma levels of glucose peaked at 3 h but returned to baseline at 6 h. However, in Chinese longsnout catfish, plasma glucose levels peaked at 6 h and returned to baseline at 48 h. The activity of intestinal amylase was increased in grass carp at 1 and 3 h, but no significant change in Chinese longsnout catfish was observed. The activity of hepatic glucose-6-phosphatase fell significantly in grass carp but change was not evident in Chinese longsnout catfish. The expression levels and enzymic activity of hepatic pyruvate kinase increased in grass carp, but no significant changes were observed in the Chinese longsnout catfish. Glycogen synthase (gys) and glycogen phosphorylase (gp) were induced in grass carp. However, there was no significant change in gys and a clear down-regulation of gp in Chinese longsnout catfish. In brief, compared with Chinese longsnout catfish, grass carp exhibited a rapid increase and faster clearance rate of plasma glucose. This effect was closely related to significantly enhanced levels of digestion, glycolysis, glycogen metabolism and glucose-induced lipogenesis in grass carp, as well as the inhibition of gluconeogenesis.

Feeding a high concentrate (HC) diet is a widely used strategy for supporting high milk yields, yet it may cause certain metabolic disorders. This study aimed to investigate the changes in milk production and hepatic metabolism in goats fed different proportions of concentrate in the diet for 10 weeks. In total, 12 mid-lactating goats were randomly assigned to an HC diet (65% concentrate of dry matter, n=6) or a low concentrate (LC) diet (35% concentrate of dry matter, n=6). Compared with LC, HC goats produced greater amounts of volatile fatty acids and produced more milk and milk lactose, fat and protein (P<0.01). HC goats showed a greater concentration of ATP, NAD, plasma non-esterified fatty acids and hepatic triglycerides than LC goats (P<0.05). Real-time PCR results showed that messenger RNA (mRNA) expression of gluconeogenic genes, namely, glucose-6-phosphatase, pyruvate carboxylase and phosphoenolpyruvate carboxykinase were significantly up-regulated and accompanied greater gluconeogenic enzyme activities in the liver of HC goats. Moreover, the expression of hepatic lipogenic genes including sterol regulatory element-binding protein 1c, fatty acid synthase and diacylglycerol acyltransferase mRNA was also up-regulated by the HC diet (P<0.05). HC goats had greater hepatic phosphorylation of AMP-activated protein kinase than LC (P<0.05). Furthermore, histone-3-lysine-27-acetylation contributed to this elevation of gluconeogenic gene expression. These results indicate that lactating goats fed an HC diet for 10 weeks produced more milk, which was associated with up-regulated gene expression and enzyme activities involved in hepatic gluconeogenesis and lipogenesis.

At the crossroad between nutrient supply and requirements, the liver plays a central role in partitioning nitrogenous nutrients among tissues. The present review examines the utilisation of amino acids (AA) within the liver in various physiopathological states in mammals and how the fates of AA are regulated. AA uptake by the liver is generally driven by the net portal appearance of AA. This coordination is lost when demands by peripheral tissues is important (rapid growth or lactation), or when certain metabolic pathways within the liver become a priority (synthesis of acute-phase proteins). Data obtained in various species have shown that oxidation of AA and export protein synthesis usually responds to nutrient supply. Gluconeogenesis from AA is less dependent on hepatic delivery and the nature of nutrients supplied, and hormones like insulin are involved in the regulatory processes. Gluconeogenesis is regulated by nutritional factors very differently between mammals (glucose absorbed from the diet is important in single-stomached animals, while in carnivores, glucose from endogenous origin is key). The underlying mechanisms explaining how the liver adapts its AA utilisation to the body requirements are complex. The highly adaptable hepatic metabolism must be capable to deal with the various nutritional/physiological challenges that mammals have to face to maintain homeostasis. Whereas the liver responds generally to nutritional parameters in various physiological states occurring throughout life, other complex signalling pathways at systemic and tissue level (hormones, cytokines, nutrients, etc.) are involved additionally in specific physiological/nutritional states to prioritise certain metabolic pathways (pathological states or when nutritional requirements are uncovered).

In the present study, the effects of partial substitution of dietary protein by digestible starch on endogenous glucose production were evaluated in European seabass (Dicentrarchus labrax). The fractional contribution of dietary carbohydrates v. gluconeogenesis to blood glucose appearance and hepatic glycogen synthesis was quantified in two groups of seabass fed with a diet containing 30 % digestible starch (DS) or without a carbohydrate supplement as the control (CTRL). Measurements were performed by transferring the fish to a tank containing water enriched with 5 % 2H2O over the last six feeding days, and quantifying the incorporation of 2H into blood glucose and hepatic glycogen by 2H NMR. For CTRL fish, gluconeogenesis accounted for the majority of circulating glucose while for the DS fish, this contribution was significantly lower (CTRL 85 (sem 4) % v. DS 54 (sem 2) %; P< 0·001). Hepatic glycogen synthesis via gluconeogenesis (indirect pathway) was also significantly reduced in the DS fish, in both relative (CTRL 100 (sem 1) % v. DS 72 (sem 1) %; P< 0·001) and absolute terms (CTRL 28 (sem 1) v. DS 17 (sem 1) μmol/kg per h; P< 0·001). A major fraction of the dietary carbohydrates that contributed to blood glucose appearance (33 (sem 1) % of the total 47 (sem 2) %) had undergone exchange with hepatic glucose 6-phosphate. This indicated the simultaneous activity of hepatic glucokinase and glucose 6-phosphatase. In conclusion, supplementation of digestible starch resulted in a significant reduction of gluconeogenic contributions to systemic glucose appearance and hepatic glycogen synthesis.

In strict carnivorous domestic cats, a metabolic competition arises between the need to use amino acids for gluconeogenesis and for protein synthesis both in health and disease. The present study investigated the amino acid-sparing potential of propionic acid in cats using dietary propionylated starch (HAMSP) supplementation. A total of thirty cats were fed a homemade diet, supplemented with either HAMSP, acetylated starch (HAMSA) or celite (Control) for three adaptation weeks. Propionylated starch was hypothesised to provide propionic acid as an alternative gluconeogenic substrate to amino acids, whereas acetic acid from HAMSA would not provide any gluconeogenic benefit. Post-adaptation, a 5-d total faecal collection was carried out to calculate apparent protein digestibility coefficients. Fresh faecal and blood samples were collected to analyse fermentation endproducts and metabolites. The apparent protein digestibility coefficients did not differ between supplements (P = 0·372) and were not affected by the protein intake level (P = 0·808). Faecal propionic acid concentrations were higher in HAMSP than in HAMSA (P = 0·018) and Control (P = 0·003) groups, whereas concentrations of ammonia (P = 0·007) were higher in HAMSA than in HAMSP cats. Tendencies for or higher propionylcarnitine concentrations were observed in HAMSP compared with HAMSA (P = 0·090) and Control (P = 0·037) groups, and for tiglyl- + 3-methylcrotonylcarnitine concentrations in HAMSP as compared with Control (P = 0·028) cats. Methylmalonylcarnitine concentrations did not differ between groups (P = 0·740), but were negatively correlated with the protein intake level (r –0·459, P = 0·016). These results suggest that HAMSP cats showed more saccharolytic fermentation patterns than those supplemented with HAMSA, as well as signs of sparing of valine in cats with a sufficient protein intake.

The review is based on a compiled data set from studies quantifying liver release of glucose concomitant with uptake of amino acids (AA) and other glucogenic precursors in periparturient dairy cows. It has become dogma that AAs are significant contributors to liver gluconeogenesis in early lactation, presumably accounting for the observed lack of glucogenic precursors to balance estimated glucose need. Until recently, there has been paucity in quantitative data on liver nutrient metabolism in the periparturient period. Propionate is the quantitatively most important glucogenic precursor throughout the periparturient period. However, the immediate post partum increment in liver release of glucose is not followed by an equivalent increment in propionate uptake, because of the lower rate of increment in feed intake compared with the rate of increment in requirements for milk synthesis. The quantitative data on liver metabolism of AA do not support the hypothesis that the rapid post partum increase in net liver release of glucose is supported by increased utilisation of AA for gluconeogenesis. Only alanine is likely to contribute to liver release of glucose through its role in the inter-organ transfer of nitrogen from catabolised AA. AAs seem to be prioritised for anabolic purposes, indicating the relevance of investigating effects of supplying additional protein to post partum dairy cows. Combining data from quantitative and qualitative experimental techniques on L-lactate metabolism point to the conclusion that the quantitatively most important adaptation of metabolism to support the increased glucose demand in the immediate post partum period is endogenous recycling of glucogenic carbon through lactate. This is mediated by a dual site of adaptation of metabolism in the liver and in the peripheral tissues, where the liver affinity for L-lactate is increased and glucose metabolism in peripheral tissues is shifted towards L-lactate formation over complete oxidation.

Increased gluconeogenesis (GNG) has been suggested to contribute to protein-induced satiety via modulation of glucose homoeostasis. The objective was to determine GNG and appetite in healthy human subjects after a high-protein v. a normal-protein diet and to assess whether GNG contributes to protein-induced satiety. A total of twenty-two healthy subjects (ten men and twelve women: age 23 (sem 1) years, BMI 22·1 (sem 0·5) kg/m2) received an isoenergetic high-protein (30/0/70 % of energy from protein/carbohydrate/fat) or normal-protein diet (12/55/33 % of energy from protein/carbohydrate/fat) for 1·5 d in a randomised cross-over design. Appetite ratings were measured using visual analogue scales (VAS); endogenous glucose production and GNG were measured via infusion of [6,6-2H2]glucose and ingestion of 2H2O. Moreover, fasting glucose and β-hydroxybutyrate concentrations were measured. Glycogen stores were lowered at the start with a glycogen-lowering exercise test. During the high-protein compared with the normal-protein diet, GNG was increased and appetite was suppressed (GNG: 148 (sem 7) v. 133 (sem 6) g/24 h, P < 0·05; and 24 h area under the curve for hunger: 694 (sem 46) v. 1055 (sem 52) mm VAS × 24 h, P < 0·001; fullness: 806 (sem 59) v. 668 (sem 64) mm VAS × 24 h, P < 0·05; desire to eat: 762 (sem 48) v. 1004 (sem 66) mm VAS × 24 h, P < 0·001). There was no correlation between appetite ratings and GNG. Glucose concentration was lower (4·09 (sem 0·10) v. 4·89 (sem 0·06) mmol/l, P < 0·001) and β-hydroxybutyrate concentration was higher (1349 (sem 139) v. 234 (sem 25) μmol/l, P < 0·001) after the high-protein compared with the normal-protein diet. In conclusion, after a high-protein diet, GNG was increased and appetite was lower compared with a normal-protein diet; however, these were unrelated to each other. An increased concentration of β-hydroxybutyrate may have contributed to appetite suppression on the high-protein diet.

N balance and postprandial acylcarnitine profile following intestinal fermentation of oligofructose and inulin were investigated in healthy cats. Two diets were tested in a crossover design: a commercial high-protein cat food supplemented with 4 % DM oligofructose and inulin (spectrum: degree of polymerisation (DP) 2–10: 60 (se 5) % DM; DP>10: 28 (se 5) % DM) as high-fermentable fibre (HFF) diet, and the same commercial diet supplemented with 4 % DM cellulose as low-fermentable fibre diet. Eight adult cats were randomly allotted to each of the two diets at intervals of 4 weeks. At the end of each testing period, faeces and urine were collected over a 5-d period, and blood samples were obtained before and at the selected time points postprandially. No differences were found for N intake, N digestibility and faecal N excretion, whereas urinary N excretion was lower when the HFF diet was fed (P = 0·044). N balance was positive in all the cats, and tended to be increased when the HFF diet was fed (P = 0·079). Propionylcarnitine concentrations (P = 0·015) and their area under the curve (AUC) (P = 0·013) were increased when the HFF diet was fed, revealing a more pronounced production and absorption of propionate. Yet, methylmalonylcarnitine concentrations and concurrent AUC were not elevated when the HFF diet was fed, indicating reduced amino acid catabolism. 3-Hydroxy-3-methylglutarylcarnitine concentrations (P = 0·026) and their AUC (P = 0·028) were also reduced when the HFF diet was fed, implying diminished use of branched-chain amino acids as well. In healthy cats, oligofructose and inulin added to a high-protein diet were suggested to reduce postprandial amino acid-induced gluconeogenesis by substitution with propionate.

The activity of the gluconeogenic enzyme, alanine-amino-transferase (ALT), in a preparation from the liver of rats was studied by means of an in vitro assay throughout the course of a primary infection of Nippostrongylus brasiliensis, established by a subcutaneous injection of approximately 4000 3rd-stage larvae. The activity was measured on days 1–14 p.i. in both uninfected and infected rats and a marked pattern in the enzyme's activity was observed. In infected rats, the activity increased from 1·46±0·19 U/g liver on day 1 p.i. to a peak on day 4 p.i. of 10·75±1·62 U/g liver, then decreased to a trough of 0·44±0·18 U/g liver on day 10 p.i. before returning to original levels by day 14 p.i., by which time the infection had been largely eliminated. In uninfected rats the activity of the liver enzyme remained constant throughout this period with a value of 2·54±0·12 U/g liver. The activity of the enzyme in vitro was found to be related to the size of the inoculum on days 4 and 10 p.i. It was proposed that these observations could be due to either (1) a direct effect of the parasite, or (2) a consequence of the host immune response to the infection. In order to investigate the second proposition more fully, liver ALT activity was investigated by in vitro assay on selected days p.i. in rats experiencing a secondary N. brasiliensis infection. In these rats the liver ALT activity was observed to reach a peak on day 2 p.i., with an activity of 3·87 ± 0-28 U/g liver, and a trough on day 4 p.i. with an activity of 0·11 ±0·03 U/g liver, returning to similar levels to those measured in uninfected rats by day 7 p.i. When serum prepared from rats having secondary N. brasiliensis infections collected on day 4 p.i. was added to the assay, a reduction in the activity of liver ALT activity from both the infected and uninfected rats was measured by in vitro assay. The results are discussed in relation to protein metabolism and gluconeogenesis in rats infected with N. brasiliensis, and also in relation to the host’s immune response to the parasitic infection.

The activities of glucose-6-phosphatase (G6Pase), frucrose-1,6-bisphosphatase (FBPase), phosphoenolpyruvate carboxykinase (PEPCK) and pyruvate carboxylase (PC) were determined in homogenates of adult Schistosoma mansoni worms and compared with the activities in homogenates of rat liver and rat skeletal muscle, tissues with a high and a low gluconeogenic capacity, respectively. All four gluconeogenic enzymes were present in S. mansoni. The enzymes were less active than in rat liver, but the activities of G6Pase, PEPCK and PC were at least an order of magnitude higher than in rat skeletal muscle whereas FBPase was approximately equally active in S. mansoni and in rat muscle. Experiments with 14C-labelled substrates or [14C]NaHCO3 failed to demonstrate the actual occurrence of gluconeogenesis in S. mansoni. Some possible other functions of the gluconeogenic enzymes were investigated. Experiments with inhibitors of PEPCK gave no indications that this enzyme was involved in the degradation of glucose. This was confirmed by 13C-NMR experiments which indicated that lactate was formed from phosphoenolpyruvate via the actions of pyruvate kinase and lactate dehydrogenase, and that PEPCK did not participate in the formation of lactate. Substrate cycling between fructose-6-phosphate and fructose-1,6-bisphosphate was demonstrated to occur in adult S. mansoni. This shows that FBPase participates in the glucose metabolism of this parasite.

The contribution of gluconeogenesis to glucose production is estimated from the enrichment of the H bound to C-5 of glucose relative to either that bound to C-2 of glucose or the enrichment in body water on ingesting 2H2O in the fasted state. Contributions of all gluconeogenic substrates are included in the estimate and the limitation of an uncertain precursor enrichment removed. The half-life of 2H2O in body water precludes a repeat study for many weeks. Glycogen cycling could result in underestimation, but there is evidence that glycogen cycling does not occur in liver in the fasted state. Gluconeogenesis has been estimated by mass-isotopomer-distribution analyses, usually by administering 13C-labelled glycerol. Underestimates emphasize the major limitation of the method, i.e. the need to assume a single enrichment of the precursor pool. Estimates of gluconeogenesis from isotopomer distribution in arterial-blood glucose and lactate on infusing [U-13C6] glucose are unreliable, as a proportion of the glucose is formed from glycerol and from amino acids not converted to glucose via pyruvate. Loss of label in the Krebs cycle and relying on enrichment of arterial-blood lactate as a measure of hepatic pyruvate further add to the uncertainty. Estimates of the rate of gluconeogenesis by NMR are obtained by subtraction of the rate of glycogenolysis determined by NMR from the rate of glucose production. Estimates are then the mean rate for the period over which glycogen contents are measured. Technical considerations can limit the accuracy of analyses and result in overestimates.

The objective was to measure the activities of all the enzymes essential for hepatic gluconeogenesis in dairy cows with induced fatty liver. We aimed to induce severe fatty liver in ten experimental cows by overfeeding them during the dry period while seven control cows were maintained on a restricted diet. To induce a marked negative energy balance, the experimental cows were deprived of feed for 8 h immediately after parturition. In addition, the experimental cows were given a restricted amount of diet during the first 5 d of lactation. Liver samples were collected 1 week before and 1, 2 and 4 weeks after parturition. Before parturition, liver triacylglycerol concentrations did not differ between the two groups. After parturition, the experimental cows developed marked fatty liver as indicated by a higher level of triacylglycerols in the liver compared with the control cows.

Before parturition, all gluconeogenic enzymes in the liver were lower in experimental cows than in control cows. Phosphoenolpyruvate carboxykinase, pyruvate carboxylase and propionyl-CoA carboxylase were significantly lower and fructose 1,6-bisphosphatase and glucose 6-phosphatase tended to be lower in the experimental cows. The activities of two crucial enzymes for gluconeogenesis in ruminants, i.e., phosphoenolpyruvate carboxykinase and propionyl-CoA carboxylase, remained low throughout the sampling period post partum. Activities of pyruvate carboxylase and glucose 6-phosphatase in the experimental cows post partum were upgraded to values similar to those of the control cows. The results showed that the capacity for hepatic gluconeogenesis before parturition was lower in cows with induced fatty liver than in control cows. After parturition, the low activities of crucial gluconeogenic enzymes indicated insufficient production of glucose. It is suggested that the low gluconeogenic capacity leads successively to low blood glucose concentrations, low insulin levels and high rates of mobilization of fatty acid, causing severe hepatic lipidosis.

The effect of abomasal infusion of glucose (120 kJ/d per kg body weight (BW)0·75, 758 mmol/d) on urea production, plasma alanine-N flux rate and the conversion of alanine-N to urea was studied in sheep offered a low-N diet at limited energy intake (500 kJ/d per kg BW0·75), based on hay and grass pellets. Glucose provision reduced urinary N (P = 0·040) and urea (P = 0·009) elimination but this was offset by poorer N digestibility. Urea-N production was significantly reduced (822 v. 619 mmol/d, P = 0·024) by glucose while plasma alanine-N flux rate was elevated (295 v. 342 mmol/d, P = 0·011). The quantity of urea-N derived from alanine tended to be decreased by glucose (127 v. 95 mmol/d) but the fraction of urea production from alanine was unaltered (15 %). Plasma urea and alanine concentrations (plus those of the branched chain amino acids) decreased in response to exogenous glucose, an effect probably related to enhanced anabolic usage of amino acids and lowered urea production.

The effects of diet composition and ration size on the activities of key enzymes involved in intermediary metabolism were studied in the liver of gilthead sea bream (Sparus aurata). High-carbohydrate, low-protein diets stimulated 6-phosphofructo 1-kinase (EC 2.7.1.11), pyruvate kinase (EC 2.7.1.40), glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (EC 1.1.1.44) enzyme activities, while they decreased alanine aminotransferase (EC 2.6.1.2) activity. A high degree of correlation was found between food ration size and the activity of the enzymes 6-phosphofructo 1-kinase, pyruvate kinase, glucose-6-phosphate dehydrogenase (positive correlations) and fructose-1,6-bisphosphatase (EC 3.1.3.11) (negative correlation). These correlations matched well with the high correlation also found between ration size and growth rate in starved fish refed for 22 d. Limited feeding (5 g/kg body weight) for 22 d decreased the activities of the key enzymes for glycolysis and lipogenesis, and alanine aminotransferase activity. The findings presented here indicate a high level of metabolic adaptation to both diet type and ration size. In particular, adaptation of enzyme activities to the consumption of a diet with a high carbohydrate level suggests that a carnivorous fish like Sparus aurata can tolerate partial replacement of protein by carbohydrate in the commercial diets supplied in culture. The relationship between enzyme activities, ration size and fish growth indicates that the enzymes quickly respond to dietary manipulations of cultured fish.

The effects of altering the glucogenic potential of the diet of wether lambs (initial body weight 56·4 kg) offered an energy allowance of 0·25 that required for maintenance on body-weight loss, nitrogen retention and blood and rumen fluid parameters were examined. Whole barley was offered alone or substituted on an isoenergetic basis with 40 or 120 g of a blend of glucose precursors (polyhydric alcohols, propylene glycol, sodium propionate) absorbed on to palm kernel meal (acetona meal), to 36 lambs (no. = 12 per treatment) for 8 weeks. Substituting barley with 40 g acetona meal had a small but non-significant effect in decreasing body-weight loss and plasma aspartate amino transferase activity and in increasing plasma insulin concentration. Increasing the glucogenic potential of the diet by substituting barley with 120 g acetona meal decreased body-weight loss in the early phase of the study, decreased plasma concentrations of non-esterified fatty acids and increased the proportion of propionate in volatile fatty acids in rumen fluid, plasma aspartate amino transferase and gamma-glutamyl transferase activities and plasma insulin concentrations. Increasing the glucogenic potential of the diet did not affect nitrogen retention or plasma glucose concentration. It is concluded that increasing the glucogenic potential of the diet of lambs resulted in an increase in metabolic efficiency at the onset of negative energy balance which was mediated through a change in plasma insulin concentration. The absence of a significant difference in body-weight loss between the diets in the latter part of the study reflects the lack of sustained ketosis.