A new mathematical model for combining growth and energy intake in animals: the case of the growing pig.

A simultaneous model for analysis of net energy intake and growth curves is presented, viewing the animal's responses as a two dimensional outcome. The model is derived from four assumptions: (1) the intake is a quadratic function of metabolic weight; (2) the rate of body energy accretion represents the difference between intake and maintenance; (3) the relationship between body weight and body energy is allometric and (4) animal intrinsic variability affects the outcomes so the intake and growth trajectories are realizations of a stochastic process. Data on cumulated net energy intake and body weight measurements registered from weaning to maturity were available for 13 pigs. The model was fitted separately to 13 datasets. Furthermore, slaughter data obtained from 170 littermates was available for validation of the model. The parameters of the model were estimated by maximum likelihood within a stochastic state space model framework where a transform-both-sides approach was adopted to obtain constant variance. A suitable autocorrelation structure was generated by the stochastic process formulation. The pigs' capacity for intake and growth were quantified by eight parameters: body weight at maximum rate of intake (149-281 kg); maximum rate of intake (25.7-35.7 MJ/day); metabolic body size exponent (fixed: 0.75); the daily maintenance requirement per kg metabolic body size (0.232-0.303 MJ/(day x kg(0.75))); reciprocal scaled energy density (0.192-0.641 kg/MJ(theta(6)) ; a dimensional exponent, theta(6) (0.730-0.867); coefficient for animal intrinsic variability in intake (0.120-0.248 MJ(0.5)) and coefficient for animal intrinsic variability in growth (0.029-0.065 kg(0.5)). Model parameter values for maintenance requirements and body energy gains were in good agreement with those obtained from slaughter data. In conclusion, the model provides biologically relevant parameter values, which cannot be derived by traditional analysis of growth and energy intake data.

A dynamic growth model for prediction of nutrient partitioning and manure production in growing-finishing pigs: Model development and evaluation.

Nutrient loading and air emissions from swine operations raise environmental concerns. The objective of the study was to describe and evaluate a mathematical model (Davis Swine Model) of nutrient partitioning and predict manure excretion and composition on a daily basis. State variables of the model were AA, fatty acids, and a central pool of metabolites that supplied substrate for lipid synthesis and oxidation. The model traced the fate of ingested nutrients and water through digestion and intermediary metabolism into body protein, fat, water, and ash, where body protein and fat represented the body constituent pools. It was assumed that fluxes of metabolites follow saturation kinetics, depending on metabolite concentrations. The main inputs to the model were diet nutrient composition, feed intake, water-to-feed ratio, and initial BW. First, the model was challenged with nutrient partitioning data and then with excretion data. The data had 48 different feeding regimes with contrasting energy and lysine intakes at 2 different stages of growth. The overall observed and predicted mean were 109 and 112 g/d for protein deposition and 132 and 136 g/d for lipid deposition respectively, suggesting minor mean bias. Root mean square prediction error (RMSPE) was used in evaluation of the model for its predictive power. The overall RMSPE was 2.2 and 4.1 g/d for protein and lipid deposition, respectively. The excretion database used for evaluation of the model was constructed from 150 digestibility trials using growing-finishing pig diets that had a wide range of nutrient chemical composition. Nutrient and water excretion were quantified using the principle of mass conservation. The average daily observed and predicted manure production was 3.79 and 3.99 kg/d, respectively, with a RMSPE of 0.49 kg/d. There was a good agreement between observed and predicted mean fecal N output (9.9 and 9.8 g/d, respectively). Similarly, the overall observed and predicted mean urine N output was 21.7 and 21.3 g/d, respectively, suggesting minor mean bias. The RMSPE was 1.9 and 4.1 g/d for fecal and urinary N, respectively. Evaluation of the model showed that the model predicts manure excretion and N content well and can be used to assess environmental mitigation options from swine operations.

Review and an experimental study on the physiological and quantitative aspects of gluconeogenesis in lactating ruminants.

A short review describes quantitative aspects of glucose metabolism in lactating ruminants, including the contribution of different substrates to glucose synthesis as affected by nutritional and hormonal regulation. Gluconeogenesis from propionate (the major gluconeogenic substrate in the fed state) is mainly regulated by feed intake and increased propionate availability and is less affected by insulin and glucagon. Quantitative estimates of amino acid contribution to glucose synthesis are highly variable (from 2 to 40% of glucose flux), but no conclusive data exist by which this large variation can be explained. An experimental model is described by which nutrient exchanges across the liver as well as the contribution of different substrates to glucose synthesis can be quantitated. The novel approach is continuous infusion of differently labelled (3H and 14C) gluconeogenic substrates into multi-catheterized goats. Results are reported from a preliminary experiment using this model. A goat in mid-lactation was fed hay ad libitum, and the experimental treatments were continuous infusion into a mesenteric vein of either sodium propionate or a complete mixture of amino acids. Measurements were nutrient exchanges across portal-drained viscera and liver, whole body glucose flux rate, and hepatic gluconeogenesis from propionate, lactate and glycerol. Glucose synthesis from amino acids was estimated by difference. Net portal appearance and hepatic uptake of propionate and of amino acids were increased by propionate and amino acid infusions, respectively. Glucose flux rate and hepatic glucose output were not affected by treatments. With propionate infusion, the proportions of liver glucose derived from propionate and amino acids were 62 and 19%, respectively. With amino acid infusion, the corresponding figures were 24 and 36%. Glucagon concentration in portal blood plasma was 2.7 times higher on the amino acid than on the propionate treatment. It is concluded that the hepatic metabolism of propionate and glucogenic amino acids into glucose synthesis is regulated to a great extent by the availability of these substrates, and that glucagon may be involved in this regulation. It is further concluded that the described experimental approach is suitable for investigations of nutrient absorption and hepatic gluconeogenesis.

Net portal appearance of volatile fatty acids in sheep intraruminally infused with mixtures of acetate, propionate, isobutyrate, butyrate, and valerate.

The net portal appearance of volatile fatty acids (VFA) was investigated in four ruminally fistulated and multicatheterized sheep. During the experiments, the sheep were fed once every hour for 14 h and intraruminally infused with mixtures of VFA for the 12 h commencing 2 h after the initiation of the hourly feeding protocol. Paired arterial and portal blood samples were obtained hourly during the last 6 h of the experiments. In the control treatment (1), only water was infused intraruminally. In Treatments 2 through 4, the intraruminal infusion rates of propionate (40 mmol/h), isobutyrate (5 mmol/h), and valerate (5 mmol/h) were unchanged. In Treatments 2, 3, and 4, the acetate infusion rate was 100, 60, and 20 mmol/h, respectively, and the butyrate infusion rate was 10, 30, and 50 mmol/h, respectively. Thus, the infusion rate of VFA carbon was constant across Treatments 2 through 4. Portal recovery estimated from the increased net portal appearance in Treatments 2 through 4 compared to the control treatment was 85% for propionate and 60% for isobutyrate, and these recoveries were unaffected by treatment. The portal recovery of butyrate increased (from 21 to 32%) with increasing infusion rate of butyrate and decreasing infusion rate of acetate, as did the portal recovery of valerate (from 14 to 31%). The portal recovery of acetate was 55%, when measured as net portal appearance. Thus, it seems that the capacity for beta-oxidation in ruminal epithelium is limited, which would explain the increasing portal recovery of butyrate and valerate with increasing infusion rate of butyrate, when infusion rate of VFA carbon is unchanged.

Portal-drained visceral metabolism of 3-hydroxybutyrate in sheep.

The present experiment was conducted to study the impact of portal-drained visceral (PDV) metabolism of arterial 3-OH-butyrate on estimates of the portal recovery of intraruminally infused butyrate. Three multicatheterized and rumen-fistulated Leicester ewes were subjected to three intraruminal infusion protocols in a Latin square design: control (C; water), butyrate (B; 20 mmol x h(-1)), and butyrate (20 mmol x h(-1)) + propionate (40 mmol x h(-1)) (BP). During the experiments, the sheep were infused with 1,2,3,4-13C4-D-3-OH-butyrate in a mesenteric vein. Portal recoveries of intraruminally infused butyrate and propionate were obtained by comparing Treatments B and BP, respectively, with Treatment C. The portal net appearance of butyrate and the portal net appearance of butyrate + 3-OH-butyrate accounted for 20 +/- 2% and 48 +/- 14% of intraruminally infused butyrate, respectively. Metabolism by the PDV tissues accounted for 32 to 44% of the whole-body irreversible loss rate of 3-OH-butyrate (12.0 to 24.7 +/- 0.5 mmol x h(-1)). The portal net appearance of butyrate plus the unidirectional PDV output of 3-OH-butyrate accounted for 62 +/- 5% of the intraruminally infused butyrate, and this estimate was comparable to the portal recovery of intraruminally infused propionate (62 +/- 7%). The results from the present study show that the extent of epithelial butyrate oxidation is overestimated and the portal recovery of butyrate carbon underestimated if only portal net appearance rates of butyrate and 3-OH-butyrate are considered.

Metabolism of propionate and 1,2-propanediol absorbed from the washed reticulorumen of lactating cows.

To investigate the metabolism of 1,2-propanediol (PPD) in lactating cows independently of normal rumen microbial metabolism, three ruminally cannulated lactating Holstein cows were subjected to three experimental infusion protocols under washed reticulo-ruminal conditions in a Latin square design. Reticulo-ruminal absorption rates were maintained for 420 min by continuous intraruminal infusion of VFA and PPD. With the control treatment, 1,246 +/- 39 mmol/ h of acetate and 213 +/- 5 mmol/h of butyrate were absorbed from the reticulorumen. With the propionate treatment, 1,148 +/- 39 mmo/h of acetate, 730 +/- 23 mmol/h of propionate and 196 +/- 5 mmol/h of butyrate were absorbed from the reticulorumen. With PPD treatment, 1,264 +/- 39 mmol/h of acetate, 220 +/- 5 mmol/h of butyrate and 721 +/- 17 mmol/h of PPD were absorbed from the reticulorumen. Glucose irreversible loss rate (ILR), as well as the relative enrichment of plasma lactate and alanine, were determined by primed continuous infusion of [U-13C]glucose in a jugular vein. Treatments did not affect (P > 0.10) the plasma concentrations of glucose (4.2 +/- 0.1 mmoVL), alanine (0.14 +/- 0.01 mmol/L), or insulin (80 +/- 25 pmol/L). The plasma concentration of lactate was higher (P < 0.05) with both propionate (0.84 +/- 5 mmol/L) and PPD treatment (0.81 +/- 5 mmol/ L) compared with the control treatment (0.29 +/- 0.5 mmol/L). The plasma concentration of pyruvate was higher (P < 0.05) with the propionate treatment (0.09 +/- 0.01 mmol/L) compared with the control treatment (0.03 +/- 0.01 mmol/L). The plasma concentration of 3-hydroxybutyrate was lower (P < 0.05) with the propionate treatment (0.15 +/- 0.03 mmol/L) compared with the control treatment (0.40 +/- 0.03). With the PPD treatment, the plasma concentrations of pyruvate and 3-hydroxybutyrate were in between the other treatments and tended (P < 0.10) to be different from both. The plasma concentration of PPD increased throughout the infusion period with the PPD treatment and reached a concentration of 4.9 +/- 0.6 mmol/L at 420 min. The ILR of glucose was not affected (P > 0.10) by treatments (441 +/- 35 mmol/h). The relative 13C enrichment of plasma lactate compared with that of glucose decreased (P < 0.05) with the PPD treatment compared with the control treatment (44 to 21 +/- 3%). It was concluded that PPD has a low rate of metabolism in cows without a normal functioning rumen, although about 10% of the absorbed PPD was metabolized into lactate.

A multilevel nonlinear mixed-effects approach to model growth in pigs.

Growth functions have been used to predict market weight of pigs and maximize return over feed costs. This study was undertaken to compare 4 growth functions and methods of analyzing data, particularly one that considers nonlinear repeated measures. Data were collected from an experiment with 40 pigs maintained from birth to maturity and their BW measured weekly or every 2 wk up to 1,007 d. Gompertz, logistic, Bridges, and Lopez functions were fitted to the data and compared using information criteria. For each function, a multilevel nonlinear mixed effects model was employed because it allowed for estimation of all growth profiles simultaneously, and different sources of variation (i.e., sex, pig, and litter effects) were incorporated directly into the parameters. Furthermore, variance in-homogeneity and within-pig correlation were introduced to the functions. Inclusion of a variance of power function and a continuous autoregressive process of first order rendered a substantially improved fit to data for all 4 growth functions. The Lopez function provided the best fit to the data set and was used for characterizing mean growth curves for the 3 sexes (barrows, boars, and gilts). It was estimated that the maximum growth rate occurs at 117, 134, and 96 kg of BW for barrows, boars, and gilts, respectively. Hence, the gilts reached their maximum growth rate at an earlier stage in life compared with boars. Mature size of pigs varied systematically with sex and was estimated to be 466, 537, and 382 kg of BW for the barrows, boars, and gilts, respectively. These estimates are significantly affected by the duration of the experimental period, and it is recommended that future studies looking at estimating the mature size in animals are conducted long enough so that the BW visually stabilizes. Furthermore, studies should consider adding continuous autoregressive process when analyzing nonlinear mixed models with repeated measures.

A multivariate nonlinear mixed effects method for analyzing energy partitioning in growing pigs.

Simultaneous equations have become increasingly popular for describing the effects of nutrition on the utilization of ME for protein (PD) and lipid deposition (LD) in animals. The study developed a multivariate nonlinear mixed effects (MNLME) framework and compared it with an alternative method for estimating parameters in simultaneous equations that described energy metabolism in growing pigs, and then proposed new PD and LD equations. The general statistical framework was implemented in the NLMIXED procedure in SAS. Alternative PD and LD equations were also developed, which assumed that the instantaneous response curve of an animal to varying energy supply followed the law of diminishing returns behavior. The Michaelis-Menten function was adopted to represent a biological relationship in which the affinity constant (k) represented the sensitivity of PD to ME above maintenance. The approach accommodated inclusion of a PD potential (PD(Potential)) concept. This was described by a Gompertz function, which was parameterized in terms of the maximum rate of PD (PD(Max)) and corresponding BW (BW(PDMax)) at that point. Metabolizable energy for LD was equated to the difference between ME intake and the sum of ME used for maintenance and PD. Metabolizable energy designated for PD and LD was used, with efficiencies k(p) and k(f), respectively. The new equations were compared with the van Milgen and Noblet (1999) equations using 2 comprehensive data sets on energy metabolism in growing pigs. The 2 equation sets were evaluated using information criteria, which showed that the new equations performed best for data set II, whereas the reverse was true for the first. For the data set I population, estimates for k(p) and k(f) were 0.57 (SE = 0.05) and 0.84 (SE = 0.03), respectively. Maintenance was quantified as 1.10 (SE = 0.08) MJ/d*kg(0.55). The animal variation in the parameter k(p) was estimated to be 6% CV. The animal variation in PD(Max) and k(f) was estimated to be 9 and 10% of the population estimates, respectively. It was concluded that application of the MNLME framework was superior to the multivariate nonlinear regression model because the MNLME method accounted for correlated errors associated with PD and LD measurements and could also include the random effect of animal. It is recommended that multivariate models used to quantify energy metabolism in growing pigs should account for animal variability and correlated measurement errors.

Post-ruminal or intravenous infusions of carbohydrates or amino acids to dairy cows 1. Early lactation.

The objectives of this study were to compare the effects of post-ruminal and intravenous infusions of wheat starch or glucose (CHO) or a mixture of amino acids (AA) on milk protein yield, nitrogen utilisation, plasma metabolites and mammary extraction rate of dairy cows in early lactation. Eight cow, ruminally fistulated, was assigned to two 4 × 4 Latin squares during 14-day periods, where the last 7 days were for infusions. Infusions were: (1) starch in the abomasum (SP), (2) glucose in the blood (GB), (3) AA in the abomasum (AP), and (4) AA in the blood (AB). The experiment started 54 ± 4 days (mean ± s.e.) post partum (milk yield 33.4 ± 1.7 kg). Daily amounts of nutrients infused were 378, 365, 341, and 333 g for SP, GB, AP and AB, respectively. The cows were fed a basal diet consisting of a concentrate mixture and grass silage (55:45 on dry-matter (DM) basis), and DM intake was 17.2 kg/day. Milk production was affected by site of infusion within substrate, whereas infusion substrates within infusion site (CHO or AA) were of minor importance. Compared with SP infusion, GB infusion increased ( P < 0.05) milk protein yield and concentration by 55 g and 1 g/kg. The AB infusion tended to ( P < 0.10) increase milk yield and ECM and increased ( P < 0.05) protein yield and concentration by 1.8 and 2.2 kg, 83 g and 1.1 g/kg compared with AP infusion, respectively. Nitrogen balance data indicated higher losses of metabolic faecal nitrogen (MFN) by abomasal than by intravenous infusions, and an increased ( P < 0.05) catabolism for AP and AB infusions compared with SP and GB infusions. GB infusion did not increase ( P>0.10) plasma glucose or insulin concentrations above that of SP infusion. Compared with the SP infusion, the GB infusion had minor effect on plasma AA. AP infusion increased ( P < 0.05) plasma non-essential AA (NEAA) concentration compared with AB infusion, whereas infusion site of AA had no effect ( P>0.05) on essential AA (EAA) or branched-chain AA (BCAA). Although a higher milk protein synthesis was observed for AB infusion, the mammary extraction rate was not higher ( P>0.05) than for AP infusion. Across infusion site, AP and AB infusions increased plasma concentration of EAA and BCAA, but compared with GB infusion, the mammary extraction rates tended ( P < 0.10) to be lower. It is concluded that abomasal nutrient infusion increases loss of MFN and that the gastrointestinal metabolism influences the nutrients available for milk synthesis. Our conclusion is that when glucose was infused, AA limited a further milk protein synthesis, but when AA was infused, glucose or energy substrate might have been the limiting factor. Our results verify that glucogenic substrates are limiting when cows are in negative energy balance.

Post-ruminal or intravenous infusions of carbohydrates or amino acids to dairy cows 2. Late lactation.

The objectives of this study were to compare the effects of post-ruminal and intravenous infusions of wheat starch or glucose (CHO) or a mixture of amino acids (AA) on milk protein yield, nitrogen (N) utilisation, plasma metabolites and mammary extraction rate of dairy cows in late lactation. Eight cow, ruminally fistulated, was assigned to two 4 × 4 Latin squares during 14-day periods, where the last 7 days were for infusions. Infusions were: (1) starch in the abomasum (SP), (2) glucose in the blood (GB), (3) AA in the abomasum (AP), and (4) AA in the blood (AB). The experiment started 165 ± 4 days (mean ± s.e.) post partum (milk yield 22.5 ± 1.1 kg) Daily amounts of nutrients infused were 257, 283, 233, and 260 g for SP, GB, AP and AB, respectively. The cows were fed a basal diet consisting of a concentrate mixture and grass silage (55:45 on a dry-matter (DM) basis), where total dry-matter intake (DMI) was 13.3 kg/day. Milk production was affected by site of infusion within substrate, whereas infusion substrates within infusion site (CHO or AA) were of minor importance. Responses to intravenous infusions (GB or AB) were similar to those in early lactation, but more pronounced. Compared with SP infusion, GB infusion increased ( P < 0.05) milk yield, energy-corrected milk (ECM), protein and lactose yield by 1.4 and 0.9 kg, 38 and 59 g, respectively. The AB infusion had 1.4 and 1.3 kg, 51, 52 and 50 g higher ( P < 0.05) milk yield, ECM, protein, fat and lactose yields than the AP infusion, respectively. N balance data indicated higher losses of metabolic faecal nitrogen (MFN) by abomasal than by intravenous infusions, but the catabolism of AA was lower than in early lactation indicated by no difference ( P < 0.05) in urinary N excretion between treatments. Intravenous AA infusion increased plasma glucose and insulin above that of intravenous glucose infusion. The treatment effects on plasma insulin concentrations were higher in late than in early lactation, suggesting a higher sensitivity in late lactation even at similar negative energy balance. Compared with the SP infusion, GB infusion showed lower ( P < 0.05) concentrations of essential AA (EAA) and branched-chain AA (BCAA) resulting in a higher AA utilisation because of a higher milk protein production. AP infusion increased ( P < 0.05) plasma non-essential AA concentration compared with AB infusion, but infusion site of AA had no effect ( P>0.05) on plasma EAA or BCAA. It is concluded that it is the nutrient supply and not the lactation stage per se that is important for the response in milk production. Nevertheless, stage of lactation affects the N metabolism and the response in plasma hormone concentrations even when cows are in negative energy balance in both lactation stages.

Quantitative aspects of protein fractional synthesis rates in ruminants.

Protein fractional synthesis rate (FSR) is a key-factor in the characterisation of ruminant metabolism. Published data from the literature were collected and statistically analysed to isolate the factors influencing FSR. FSR varied largely depending on the tissue considered, over a range from 1 to 20. FSR, with the plasma as the precursor pool for protein synthesis, was halved compared to that of the intracellular pool. The method for supplying the amino acid also significantly affects FSR since the flooding dose technique gave higher FSR estimates than the constant infusion technique. The choice of the labelled amino acid infused influenced FSR. There is a ranking order depending on the tissue or organ. The protein and energy levels of the diets significantly increased FSR, which raises the question of the body nitrogen requirements. Moreover, FSR values were dependent on the physiological status of the animals. To conclude, FSR values should be determined simultaneously with other biological parameters in order to obtain a realistic quantitative estimate of the nitrogen turnover rates during intermediary metabolism.

Quantitative aspects of blood and amino acid flows in cattle.

A quantitative literature review was undertaken on the amino acid fluxes in ruminants and the factors which influence them. Two aspects were considered: blood flow and amino acid uptake by tissues. The statistical relationships indicated that blood flow was influenced by feed intake, metabolizable energy and milk production. The rates of amino acid uptake and release by tissues and organs varied greatly between compartments and between amino acids. The net rates of appearance were the result of a combination of inter- and intratissue phenomena. Simulated balances between supply and requirements revealed the limiting role of certain amino acids, depending on milk production level and growth rate. In conclusion, this approach underlined the technical difficulties involved in obtaining this type of data, and also revealed a lack of data in certain areas. Nevertheless, the observations in this review supported the implementation of the feed units used at the present time, notably for the addition of amino acids.

Absorption and metabolism of short-chain fatty acids in ruminants.

Short-chain fatty acids (SCFA), viz. acetate, propionate and butyrate are quantitatively important substrates in ruminant energy metabolism. In the reviewed literature, 16 44% of ME intake was recovered as portal appearance of SCFA. This is considerably lower than expected when related to the estimated intragastric flux of SCFA. The discrepancy is caused by portal drained viscera metabolism of arterially abundant metabolites e.g., acetate and the metabolism of acetate and butyrate to acetoacetate and D-3-hydroxybutyrate in the absorptive epithelia. Even though considerable variations between experiments on acetate and propionate appearance are found, there seems to be a great deal of evidence that the proportion of gastrointestinally produced acetate and propionate absorbed to the portal blood is 50-75%. The portal recovery of butyrate has been found to be between 10 and 36% dependent on intraruminal infusion rate. It is concluded that major parts of acetate and propionate are directly absorbed to the portal vein. The true absorption rate of acetate can only be estimated by taking the portal drained viscera metabolism of arterial acetate into account. Butyrate is generally found to have a low recovery in the portal vein, but the production of D-3-hydroxybutyrate seems to be underestimated in major parts of the literature. It is therefore necessary to measure portal appearance as well as portal drained viscera metabolism to assess the quantitative as well as the qualitative contribution of SCFA and SCFA metabolites to whole animal metabolism.

Simulation analysis of substrate utilization in the mammary gland of lactating cows.

A kinetic modelling approach was developed and investigated with the aim of predicting the utilization of major substrates in the mammary gland and milk secretion rates in the lactating cow at varying concentrations of substrate in arterial blood. The model includes kinetic equations of transport and metabolism of glucose, acetate, free amino acids and free fatty acids in secretory cells and a phenomenological description of autoregulation of local blood flow, in which an energy criterion of control has been used. The predicted relationships between the rate of milk secretion and glucose levels in the blood are consistent with experimental results. Differential stimulation of alpha-lactalbumin synthesis causes increments in local blood flow and milk secretion rate in the model. The results of the study suggest that there is no simple relationship between the level of substrates in the blood and milk yield and contents of fat and protein in milk. This is because the effect on production of varying patterns of substrate concentrations in the blood is mediated by network interactions at the level of secretory cell metabolism and microcirculation. However, dynamic modelling provides a rational framework for developing such predictive tools.

Portal recovery of short-chain fatty acids infused into the temporarily-isolated and washed reticulo-rumen of sheep.

The present study was undertaken to study the metabolism of short-chain fatty acids (SCFA) by the reticulo-ruminal epithelium and the portal-drained viscera (PDV) under in vivo conditions with no interference from the metabolism of the rumen microbes. The technique of temporary isolation of the reticulo-rumen was applied to wethers implanted with catheters in a mesenteric artery, the hepatic portal vein and the right ruminal vein. Portal blood flow was measured by downstream dilution of p-aminohippuric acid; the PDV uptake of arterial acetate, as well as the whole-body irreversible loss rate (ILR) of acetate, was estimated by [2-(13)C]acetate infusion into the right ruminal vein. The sheep were maintained with a bicarbonate-buffered solution of SCFA in the reticulo-rumen along with continuous intraruminal infusion of SCFA for 4 h. The portal appearance of SCFA of non-reticulo-ruminal origin was estimated before and after the infusion protocol. Of the acetate absorbed by the sheep, 89 (SE 5), 109 (SE 7) and 101 (SE 7)% was recovered as portal net appearance of acetate, portal net appearance of acetate corrected for PDV uptake of arterial acetate and increase in the ILR of acetate respectively. Of the propionate, isobutyrate, butyrate, isovalerate and valerate absorbed by the sheep, 95 (SE 7), 102 (SE 9), 23 (SE 3), 48 (SE 5) and 32 (SE 4)% respectively was recovered as portal net appearance. In contrast to current concepts, the present study showed that the reticulo-ruminal epithelium metabolizes none (or only a small proportion) of the acetate and propionate absorbed from the rumen. This observation could lead to the more efficient use of results obtained with multi-catheterized animals to quantify the net metabolite output of the rumen microbes.