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The effects of high (2.5% of DM) versus normal dietary sodium chloride (NaCl) intake on renal urea-N kinetics and urea-N metabolism were investigated in 9 rumen-cannulated and multicatheterized lactating dairy cows in a crossover design with 21-d periods. It was hypothesized that urinary urea-N excretion would be greater, and BUN concentration lower in response to greater diuresis induced by high NaCl intake. Also, urea-N transport across ruminal and portal-drained viscera (PDV) tissues was hypothesized to be affected by dietary sodium intake. A second experiment was conducted using 8 lactating cows in a crossover design with 14-d periods to test high NaCl (2.5% of DM) versus high KCl (3.2% of DM) intake on milk yield and MUN concentrations. Experiment 1 showed that despite greater diuresis, high NaCl intake had no effect on urinary urea-N excretion or BUN concentration. The high NaCl intake did not affect rumen ammonia concentrations, total rumen VFA concentrations, ruminal venous - arterial concentration differences for ammonia, or ammonia absorption indicating that high NaCl did not adversely affect ruminal fermentation and microbial protein synthesis. High NaCl intake did not affect the total amount of urea-N transport from blood to gut, but ruminal venous - arterial concentration differences for urea-N were lower with high NaCl and ruminal extraction of arterial urea-N was numerically smaller, indicating that the ruminal epithelial urea-N transport was lower with high NaCl. Energy-corrected milk yield was greater with high NaCl (3.2 ± 1.5 kg/d); however, MUN concentrations were not affected by treatment. In experiment 2, ECM was greater with NaCl (1.4 ± 0.31 kg/d) compared with KCl (30.2 and 28.8 ± 0.91 kg ECM/d, respectively). Milk urea-N concentration was lower with KCl, suggesting a urea-N lowering effect in milk not evident with high NaCl intake. In conclusion, the present data show that dietary Na intake of 12 to 13 g/kg DM was followed by greater diuresis but did not affect urea-N excretion or BUN concentration. High NaCl intake did not affect the total amount of urea-N transfer across PDV tissues. Energy-corrected milk yield was greater with high NaCl compared with both control and feeding KCl; however, with KCl, MUN decreased., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

An experiment was conducted to determine if ergot alkaloids affect blood flow to the absorptive surface of the rumen. Steers (n=8) were pair-fed alfalfa cubes and received ground endophyte-infected (Neotyphodium coenophialum) tall fescue (Lolium arundinaceum; E+) seed (0.015 mg ergovaline·kg BW(-1)·d(-1)) or endophyte-free tall fescue (E-) seed via the rumen cannula 2x daily for 7 d at thermoneutral (TN; 22°C) and heat stress (HS; 32°C) conditions. On d 8, the rumen was emptied and rinsed. A buffer containing VFA was incubated in the following sequence: control (CON), 15 μg ergovaline·kg BW(-1) (1×EXT) from a tall fescue seed extract, and 45 μg ergovaline·kg BW(-1) (3×EXT). For each buffer treatment there were two 30-min incubations: a 30-min incubation of a treatment buffer with no sampling followed by an incubation of an identical sampling buffer with the addition of Cr-EDTA and deuterium oxide (D2O). Epithelial blood flow was calculated as ruminal clearance of D2O corrected for influx of physiological water and liquid outflow. Feed intake decreased with dosing E+ seed at HS but not at thermoneutral conditions (TN; P<0.02). Dosing E+ seed decreased serum prolactin (P<0.005) at TN. At HS, prolactin decreased in both groups over the 8-d experiment (P<0.0001), but there was no difference in E+ and E- steers (P=0.33). There was a seed treatment×buffer treatment interaction at TN (P=0.038), indicating that E+ seed treatment decreased reticuloruminal epithelial blood flow at TN during the CON incubation, but the two groups of steers were not different during 1×EXT and 3×EXT (P>0.05). Inclusion of the extract in the buffer caused at least a 50% reduction in epithelial blood flow at TN (P=0.004), but there was no difference between 1×EXT and 3×EXT. There was a seed × buffer treatment interaction at HS (P=0.005), indicating that the reduction of blood flow induced by incubating the extract was larger for steers receiving E- seed than E+ seed. Volatile fatty acid flux was reduced during the 1×EXT and 3×EXT treatments (P<0.01). An additional experiment was conducted to determine the effect of time on blood flow and VFA flux because buffer sequence could not be randomized. Time either increased (P=0.05) or did not affect blood flow (P=0.18) or VFA flux (P>0.80), indicating that observed differences are due to the presence of ergot alkaloids in the rumen. A decrease in VFA absorption could contribute to the signs of fescue toxicosis including depressed growth and performance.

Ruminal absorption of volatile fatty acids (VFA) is quantitatively the most important nutrient flux in cattle. Historically, VFA absorption models have been derived primarily from ruminal variables such as chemical composition of the fluid, volume, and pH. Recently, a mechanistic model incorporated the control of VFA absorption from epithelial surface area of the reticulorumen. In the present study, we hypothesized that ruminal absorption of VFA was controlled through epithelial permeability to VFA and rumen epithelial capillary blood flow. The objective of the study was to construct a model of VFA exchange across the rumen wall that incorporates epithelial blood flow as a driving force for ruminal VFA removal. The bidirectional fluxes between the ruminal and epithelial pool of VFA were assumed mass action driven, given that passive diffusion of nonionized VFA is the dominant transmembrane VFA flux. Parameter estimates were derived by fitting the model to observed data. The model provided reliable unbiased estimates of ruminal VFA absorption and rumen epithelial blood flow. Blood flow was modeled using an equation that considered the effect of butyrate and dietary crude protein intake per kilogram of body weight. The rate constants related to the flux from ruminal fluid to epithelium were in the order isobutyrate < acetate < propionate < butyrate (0.32 ± 0.02, 0.72 ± 0.2, 0.91 ± 0.06, and 0.97 ± 0.02 /h, respectively). The rate constants for fluxes of isobutyrate, acetate, propionate, and butyrate from the rumen epithelium to the ruminal fluid, relative to the pool size of the epithelium, were 4.78, 10.6, 13.4, and 14.3 /h, respectively. Ruminal concentrations of acetate, propionate, butyrate, and isobutyrate were predicted with root mean square prediction errors as percentage of the observed means (RMSPE) of 5.86, 5.75, 11.3, and 4.12, respectively. The epithelial blood flow was predicted with 26.3% RMSPE. Sensitivity analyses indicated that when ruminal butyrate concentration increased from 4.0 to 37.4 mmol/L, blood flow of the epithelium increased 47% and the ruminal disappearance rate of propionate increased 11%. The concentration gradient of propionate between ruminal fluid and epithelium was no more than 3:1 and increased with increasing blood flow. In conclusion, a dynamic model based on rumen epithelial blood flow and bidirectional fluxes of VFA between ruminal fluid and epithelium gave unbiased predictions with low residual error of ruminal VFA absorption under washed rumen conditions. The model indicates that the effect of varying epithelial blood flow on the control of ruminal VFA absorption is related to the concentration gradient of individual VFA between ruminal fluid and epithelial blood. Epithelial blood flow may be an important determinant of ruminal absorption of VFA, a result that has not been evaluated on independent data., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The effects of a 6 versus 24h ruminal urea infusion in lactating dairy cows fed a basal diet deficient in N on ruminal ammonia concentration, arterial urea-N concentration, net portal-drained viscera (PDV) urea-N flux, arterial urea-N extraction across the PDV, and renal urea-N kinetics were investigated. Three Danish Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in major splanchnic blood vessels were randomly allocated to a 3 × 3 Latin square design with 21-d periods. Treatments were ventral ruminal infusion of water for 24h (water INF), 24-h infusion of 15 g of urea/kg of dry matter intake (DMI; 24-h INF), and 6-h infusion of 15 g of urea/kg of DMI (6-h INF). The 6-h INF was initiated 0.5h after the afternoon feeding, and ran until 2230 h. Eight sample sets of arterial, portal, and hepatic blood, ruminal fluid, and urine were obtained at 0.5h before the morning feeding and 0.5, 1.5, 2.5, 3.5, 4.5, 5.5, and 6.5h after feeding (i.e., 9 to 15.5h after the 6h infusion was terminated). A substantial decrease in DMI for 6-h INF compared with 24-h INF and water INF was observed, and it has to be recognized that DMI may have confounding effects. However, the experimental setting plan was met (i.e., to cause changes in the daily pattern of ruminal ammonia and blood urea-N concentrations). The arterial urea-N concentration for 24-h INF and 6-h INF were greater than the arterial urea-N concentration with water INF throughout the sampling window. However, the arterial urea-N concentration for 6-h INF decreased steadily with sampling time reflecting a carryover effect from the ruminal urea infusion. The ruminal ammonia concentration and net portal flux of ammonia for 6-h INF were not different from water INF; hence, no carryover effect on ruminal ammonia concentration was observed. The portal flux of urea-N was not affected by treatment (i.e., even the combination of low ruminal ammonia and high arterial urea-N concentration with 6-h INF was not used by the cow to increase the uptake of urea-N across the PDV). Arterial urea-N extraction across the PDV was increased with water INF especially from 0.5 to 3.5h postprandial relative to the urea infusion treatments, reflecting increased epithelial permeability for urea-N. This indicates that daily ruminal peak of ammonia or blood urea-N concentrations overruled potential signals from low ruminal ammonia concentration observed during the sampling window. In conclusion, dairy cows appear unable to increase transport of urea-N from blood to gut in periods with low ruminal ammonia concentrations, even in a situation with infrequent N supply and apparent carryover effects on blood urea-N. It is speculated that mechanisms responsible for downregulation of epithelial urea-N transport based on daily maximum concentrations of ammonia in the rumen or urea-N in the blood suppresses any short-term signal from low ruminal ammonia during periods with low ruminal N supply., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The effect of reticuloruminal epithelial blood flow on the absorption of propionate as a volatile fatty acid (VFA) marker in 8 lactating Holstein cows was studied under washed rumen conditions. The cows were surgically prepared with ruminal cannulas and permanent catheters in an artery and mesenteric, right ruminal, and hepatic portal veins. The experiment was designed with 2 groups of cows: 4 cows adapted to high crude protein (CP) and 4 to low CP. All cows were subjected to 3 buffers: butyric, ammonia, and control in a randomized replicated 3 × 3 incomplete Latin square design. The buffers (30 kg) were maintained in a temporarily emptied and washed rumen for 40 min. The initial concentration of VFA was 84.2 mmol/L. Butyrate was increased from 4 to 36 mmol/L in butyric buffer by replacement of acetate, and ammonia (NH(3)) was increased from 2.5 to 22.5 mmol/L in ammonia buffer by replacement of NaCl. Increasing amounts of deuterium oxide (D(2)O) were added to the buffers as the order of buffer sequence increased (6, 12, and 18 g of D(2)O). Ruminal clearance of D(2)O was used to estimate epithelial blood flow. To increase accuracy of the epithelial blood flow estimates, data of ruminal liquid marker (Cr-EDTA), and initial and final buffer volumes were fitted to a dynamic simulation model. The model was used to estimate ruminal liquid passages, residual liquid, and water influx (saliva and epithelia water) for each combination of cow and buffer (n=24). Epithelial blood flow increased 49±11% for butyric buffer compared with control. The ruminal disappearance of propionate (marker VFA) was affected by buffer and followed the same pattern as for epithelial blood flow. The correlation between ruminal disappearance of propionate and epithelial blood flow (r=0.56) indicates that the removal of propionate can be limited by epithelial blood flow. The ruminal disappearance of propionate increased 30±12% for the butyric compared with ammonia buffer and 12.5±8% when compared with control. The net portal flux of propionate increased 32±6% in butyric compared with control. In conclusion, rumen epithelial blood flow is positively correlated with ruminal disappearance of propionate and affects the kinetics of ruminal VFA absorption., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The effects of decreasing ruminal urea infusion in lactating dairy cows fed a basal diet deficient in rumen degradable protein on inter-organ urea-N fluxes, epithelial urea-N extraction, and renal urea-N kinetics were investigated. Eight Danish Holstein cows fitted with a ruminal cannula and permanent indwelling catheters in the major splanchnic blood vessels and the gastrosplenic vein were used. The cows were randomly allocated to a triplicate incomplete 3 × 3 Latin square design with 14-d periods. Treatments were continuous ventral ruminal infusion of water, 4.1g of feed urea/kg of dry matter intake, and 8.5 g of feed urea/kg of dry matter intake. Dry matter intake and milk yield decreased linearly with decreasing urea infusion. Arterial blood urea-N and ruminal ammonia concentrations decreased linearly with decreasing urea infusion. In absolute amounts, the urea-N recycling did not increase when urea infusion was decreased. Arterial urea-N extraction across the portal-drained viscera and rumen wall increased linearly with decreasing urea infusion (2.46, 3.65, and 4.32 ± 0.31% and 7.5, 11.5, and 16.9 ± 0.9%, respectively), indicating that cows responded to the changes in N supply. The relative urea-N extraction across the ruminal wall increased compared with the total portal-drained viscera extraction. We observed a postprandial decrease in ruminal extraction of arterial urea-N that might reflect that the activity of the protein, presumably facilitating urea-N transport, is regulated by ruminal ammonia. The urea-N clearance by the kidneys decreased (35, 30, and 25 ± 2L/h) and the urea-N reabsorbed by the kidney increased (42, 51 and 56 ± 3%) with decreasing urea infusion, indicating that the kidneys salvaged urea-N with low-N supply. The urea transporter B mRNA abundance in rumen papillae (papillae harvested at sampling days) was not affected by dietary N supply. The study showed, that rumen wall extraction of arterial urea-N is subjected to both long- and short-term regulation. Extraction increases with decreasing N supply long-term; however, a short-term postprandial decrease in extraction was observed. No association between long-term adaptation of urea-N extraction across the rumen wall and urea transporter B mRNA abundance could be demonstrated., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

To test the hypothesis that dietary N concentrations affect gut epithelial urea transport by modifying the expression of urea transporter B (UT-B) and aquaporins (AQP), the mRNA expression and protein abundance of UT-B and AQP3, AQP7, AQP8, and AQP10 were investigated in ruminal papillae from 9 lactating dairy cows. Ruminal papillae were harvested from cows fed low N (12.9% crude protein) and high N (17.1% crude protein) diets in a crossover design with 21-d periods. The mRNA expression was determined by real-time reverse transcription-PCR and protein abundance by immunoblotting. The mRNA expression of UT-B was not affected by dietary treatment, whereas mRNA expression of AQP3, 7, and 10 were greater in the high N compared with the low N fed cows. Using peptide-derived rabbit antibodies to cow AQP3, 7, and 8, immunoblotting revealed bands of approximately 27, 27, and 24 kDa in ruminal papillae, respectively. A peptide-derived chicken antibody to cow UT-B detected a band of approximately 30 to 32 kDa in ruminal papillae. The abundance of UT-B and AQP3 and 7 were not affected by dietary treatment. In contrast, the abundance of AQP8 was greater in high N compared with low N diets. In conclusion, AQP3, 7, and 8 were found to be expressed in bovine rumen papillae. None of the investigated transcripts or proteins correlated to the increased rumen epithelial urea permeability observed with low dietary N concentration., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Effects of physical changes in consistency of ruminal contents on intraruminal equilibration and net portal fluxes of volatile fatty acids (VFA) in dairy cows were studied. Four Danish Holstein cows (121+/-17 d in milk, 591+/-24 kg of body weight, mean+/-SD) surgically fitted with a ruminal cannula and permanent indwelling catheters in the major splanchnic blood vessels were used. The experimental design was a 4x4 Latin square with a 2x2 factorial design of treatments. Treatments differed in forage (grass hay) particle size (FPS; 3.0 and 30 mm) and feed dry matter (DM) content of the total mixed ration (44.3 and 53.8%). The feed DM did not affect chewing time, ruminal variables, or net portal flux of VFA. However, decreasing the FPS decreased the overall chewing and rumination times by 151+/-55 and 135+/-29 min/d, respectively. No effect of the reduced chewing time was observed on ruminal pH or milk fat percentage. Cows maintained average ventral ruminal pH of 6.65+/-0.02, medial ruminal pH of 5.95+/-0.04, and milk fat of 4.42+/-0.12% with chewing time of 28.0+/-2.1 min/kg of DM when fed short particles. The medial ruminal pool of wet particulate matter was decreased by 10.53+/-2.29 kg with decreasing FPS, thereby decreasing the medial pool of total VFA, acetate, propionate, butyrate, isobutyrate, and isovalerate by 1,143+/-333, 720+/-205, 228+/-69, 140+/-51, 8.0+/-2.3, and 25.2+/-5.6 mmol, respectively. Ventral pool variables were not affected by treatments. Relatively large intraruminal differences of VFA concentrations and pH between the ventral and medial pools were observed, VFA concentrations being largest and pH being the lowest medially. This indicates that the ruminal mat acts as a barrier retaining VFA. The effects of reduced FPS were limited to the VFA pool sizes of the mat, leaving ruminal pH, ruminal VFA concentrations, and net portal flux of VFA unaffected. Consequently reduced FPS affected the intraruminal equilibration of VFA between mat and ventral rumen with an estimated turnover rate of isobutyrate increasing from 66+/-3%/h with long particles to 81+/-3%/h [corrected] with short particles. The estimated ruminal fluid flow and therefore intraruminal VFA transport between medial and ventral phase was not affected by the FPS. In conclusion, the ruminal mat pool of VFA was proportional to the size of the mat and the only detected effects of decreasing FPS were decreasing the mat size and an increasing turnover of the mat pool of VFA., (Copyright (c) 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Ruminal cannulation techniques are frequently used to study fermentation in the ruminant forestomach. Unsatisfactory results with the traditionally applied procedure for cannulation of young calves stimulated the development of a simpler and more robust procedure; this procedure was tested for effects on performance traits and gross anatomy of the gastrointestinal tract compared with a control group not undergoing surgery. Five calves were ruminally cannulated at approximately 10 d of age and 5 matching calves were used as controls. All calves were fed milk replacer and a diet based on clover grass silage and sodium hydroxide-treated wheat. Ruminal fluid was collected from cannulated calves once weekly for 3 consecutive weeks. All calves were euthanized at 43+/-3 d of age. No apparent adverse effects of cannulation were observed. Feed intake, BW gain, and gross anatomy of the gastrointestinal tract were not affected by cannulation. Minimum ruminal pH increased with sampling week, but average ruminal pH, total volatile fatty acids concentration, and volatile fatty acids proportions were not affected by sampling week. In conclusion, the implemented surgical technique was found to have no major effect on apparent animal health and performance traits, and the cannula proved useful for multiple samplings of ruminal contents in young calves., (Copyright 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Two experiments were conducted to evaluate the effects of slow-release urea (SRU) versus feed-grade urea on ruminal metabolite characteristics in steers and DMI, gain, and G:F in growing beef steers. Experiment 1 used 12 ruminally cannulated steers (529 +/- 16 kg of BW) to monitor the behavior of SRU in the ruminal environment. Compared with feed-grade urea, SRU decreased ruminal ammonia concentration (P = 0.02) and tended to increase ruminal urease activity (P = 0.06) without affecting ruminal VFA molar proportions or total concentrations (P > 0.20). After 35 d of feeding, the in situ degradation rate of SRU was not different between animals fed urea or SRU (P = 0.48). Experiment 2 used 180 Angus-cross steers (330 +/- 2.3 kg) fed corn silage-based diets supplemented with urea or SRU for 56 d to evaluate the effects on feed intake, gain, and G:F. The design was a randomized complete block with a 2 x 4 + 1 factorial arrangement of treatments. Treatments included no supplemental urea (control) or urea or SRU at 0.4, 0.8, 1.2, or 1.6% of diet DM. Over the entire 56 d experiment, there were interactions of urea source x concentration for gain (P = 0.04) and G:F (P = 0.01) because SRU reduced ADG and G:F at the 0.4 and 1.6% supplementation concentrations but was equivalent to urea at the 0.8 and 1.2% supplementation concentrations; these effects were due to urea source x concentration interactions for gain (P = 0.06) and G:F (P = 0.05) during d 29 to 56 of the experiment. The SRU reduced DMI during d 29 to 56 (P = 0.01) but not during d 0 to 28, so that over the entire experiment there was no difference in DMI for urea source (P = 0.19). These collective results demonstrate that SRU releases N slowly in the rumen with no apparent adaptation within 35 d. Supplementation of SRU may limit N availability at low (0.4%) concentrations but is equivalent to urea at 0.8 and 1.2% concentrations.

The extensive development of the ruminant forestomach sets apart their N economy from that of nonruminants in a number of respects. Extensive pregastric fermentation alters the profile of protein reaching the small intestine, largely through the transformation of nitrogenous compounds into microbial protein. This process is fueled primarily by carbohydrate fermentation and includes extensive recycling of N between the body and gut lumen pools. Nitrogen recycling occurs via blood and gut lumen exchanges of urea and NH(3), as well as endogenous gut and secretory N entry into the gut lumen, and the subsequent digestion and absorption of microbial and endogenous protein. Factors controlling urea transfer to the gut from blood, including the contributions of urea transporters, remain equivocal. Ammonia produced by microbial degradation of urea and dietary and endogenous AA is utilized by microbial fermentation or absorbed and primarily converted to urea. Therefore, microbial growth and carbohydrate fermentation affect the extent of NH(3) absorption and urea N recycling and excretion. The extensive recycling of N to the rumen represents an evolutionary advantage of the ruminant in terms of absorbable protein supply during periods of dietary protein deficiency, or asynchronous carbohydrate and protein supply, but incurs a cost of greater N intakes, especially in terms of excess N excretion. Efforts to improve the efficiency of N utilization in ruminants by synchronizing fermentable energy and N availability have generally met with limited success with regards to production responses. In contrast, imposing asynchrony through oscillating dietary protein concentration, or infrequent supplementation, surprisingly has not negatively affected production responses unless the frequency of supplementation is less than once every 3 d. In some cases, oscillation of dietary protein concentration has improved N retention compared with animals fed an equal amount of dietary protein on a daily basis. This may reflect benefits of Orn cycle adaptations and sustained recycling of urea to the gut. The microbial symbiosis of the ruminant is inherently adaptable to asynchronous N and energy supply. Recycling of urea to the gut buffers the effect of irregular dietary N supply such that intuitive benefits of rumen synchrony in terms of the efficiency of N utilization are typically not observed in practice.

Four lactating Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in the mesenteric artery, mesenteric vein, hepatic portal vein, and hepatic vein were used in a cross-over design to study the metabolism of propylene glycol (PG). Each cow received 2 treatments: control (no infusion) and infusion of 650 g of PG into the rumen at the time of the morning feeding. Propylene glycol was infused on the day of sampling only. Samples of arterial, portal, and hepatic blood as well as ruminal fluid were obtained at 0.5 h before feeding and at 0.5, 1.5, 2.5, 3.5, 5, 7, 9, and 11 h after feeding. Infusion of PG did not affect ruminal pH or the total concentration of ruminal volatile fatty acids, but did decrease the molar proportion of ruminal acetate. The ruminal concentrations of PG, propanol, and propanal as well as the molar proportion of propionate increased with PG infusion. The plasma concentrations of PG, ethanol, propanol, propanal, glucose, L-lactate, propionate, and insulin increased with PG and the plasma concentrations of acetate and beta-hydroxybutyrate decreased. The net portal flux of PG, propanol, and propanal increased with PG. The hepatic uptake of PG was equivalent to 19% of the intraruminal dose. When cows were dosed with PG, the hepatic extraction of PG was between 0 and 10% depending on the plasma concentration of PG, explaining the slow decrease in arterial PG. The increased net hepatic flux of L-lactate with PG could account for the entire hepatic uptake of PG, which suggests that the primary hepatic pathway for PG is oxidation to l-lactate. The hepatic uptake of propanol increased with PG, but no effects of PG on the net hepatic and net splanchnic flux of glucose were observed. Despite no effect of PG on net portal flux and net hepatic flux of propionate, the net splanchnic flux of propionate increased and the data suggest that propionate produced from hepatic metabolism of propanol is partly released to the blood. The data suggest that PG affects metabolism of the cows by 2 modes of action: 1) increased supply of l-lactate and propionate to gluconeogenesis and 2) insulin resistance of peripheral tissues induced by increased concentrations of PG and propanol as well as a decreased ratio of ketogenic to glucogenic metabolites in arterial blood plasma.

The aim of the present experiment was to test the hypothesis that a barley-based concentrate would induce an acidic ruminal environment in young calves and that increased milk allowance would alleviate this condition. Eight Holstein calves ruminally cannulated at d 7 +/- 1 of age were used to study the effect of variation in barley-based starter concentrate intake induced by 4 different milk allowances (3.10, 4.84, 6.60, and 8.34 kg of milk replacer/d; 123 g of dry matter/kg of milk) on the ruminal environment, blood variables, and fore-stomach development from wk 2 to 5 of age. Twelve ruminal fluid samples were collected during a weekly 24-h sampling in 4 consecutive weeks. Blood samples were collected by venipuncture between 1200 and 1300 h on ruminal sampling days. Rumen papillae development and visceral organ mass were recorded at slaughter. A linear treatment x week effect was observed for concentrate intake, with the calves fed the lowest milk allowance having the fastest increase in concentrate intake whereby these calves reached the same ME intake in wk 5 compared with calves with the highest milk allowance. Effects on ruminal variables were dominated by week of sampling, with minor differences among treatments. Ruminal pH was below 5.5 for 5 to 13 h/d and all calves with concentrate intake above 20 g of dry matter/d were observed to have a daily ruminal pH minimum at pH 5.5 or lower. The ruminal concentration of total volatile fatty acids (VFA) increased from 71 to 133 +/- 9 mmol/L in wk 2 to 5 and was characterized by a relatively high molar proportion of propionate, increasing from 34 to 40 mol/100 mol of VFA in wk 2 to 5. In addition, the presence of ethanol and propanol as well as numerous VFA esters points to a ruminal environment with a relatively high hydrogen pressure. Plasma glucose and insulin responded to the highest milk allowance in wk 2 to 4. Plasma VFA and ketone bodies increased with the lowest milk allowance in wk 4 to 5. At slaughter, empty wet weights of the rumen + reticulum and omasum as well as mass of digesta in these compartments were found to decrease linearly and perirenal fat was found to increase linearly with milk allowance, indicating that the milk allowance changed the body composition of the calves. Lengths of ruminal papillae in the atrium and ventral ruminal sac were not affected by treatment. We concluded that the ruminal environment of young calves fed a barley-based starter concentrate was characterized by a low ruminal pH and high VFA concentration regardless of the milk allowance.

Dairy cows fed silage are subjected to various alcohols and low molecular weight esters. Four lactating Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in the hepatic portal vein, hepatic vein, mesenteric vein, and mesenteric artery were used to study the absorption of alcohols into portal blood and the metabolism of feed alcohols in the rumen and splanchnic tissues. The cows were allocated to 4 experimental treatments in a Latin square design. All treatments were formulated as total mixed rations with the same overall nutrient composition, differing by the source of corn silage. Treatments were a control silage and 3 qualities of problematic corn silage (silage with Fusarium toxin, Penicillium-infected silage, and silage with a high propanol content). Feeding was followed by a decreasing ruminal pH, as well as decreasing molar proportions of ruminal acetate and isobutyrate. The ruminal concentrations of total VFA, ethanol, propanol, 2-butanol, ethyl acetate, propyl acetate, glucose, and L-lactate, and molar proportions of propionate, butyrate, isovalerate, valerate, and caproate increased after feeding. Treatments affected ruminal concentrations of propanol, propyl acetate, and butyrate and a strong correlation was observed between ruminal propyl acetate and the molar proportion of butyrate (r = -0.79). Arterial concentrations of ethanol, propanol, propanal, acetone (sum of acetone and acetoacetate), 3-hydroxybutyrate, L-lactate, glutamate, and glutamine increased, and the arterial concentration of glucose decreased after feeding, but no effects of treatment were observed for arterial variables. The postprandial increase in arterial ethanol was maintained for 5 h. The net portal release of ethanol tended to decrease with the treatment with the lowest ethanol content, and the net splanchnic release of ethanol increased after feeding, but overall, the net splanchnic flux of ethanol was not different from zero, in agreement with the liver being the major organ for alcohol metabolism. The net portal flux and net hepatic flux of propanol were affected by treatment. All dietary ethanol and propanol were accounted for by absorption of the respective alcohol into the portal blood. The hepatic extraction ratios of ethanol and propanol were, on average, 63 to 66%, and no indications of saturation of hepatic alcohol metabolism were observed at any time. We concluded that typical amounts of alcohols in corn silage do not interfere with splanchnic metabolism of any of the measured variables and do not saturate hepatic pathways for alcohol metabolism. However, even low concentrations of alcohols in feed might affect ruminal metabolism and are followed by hours of elevated peripheral blood concentrations of alcohols.

Four steers fitted with a ruminal cannula and chronic indwelling catheters in the mesenteric artery, mesenteric vein, hepatic portal vein, hepatic vein, and the right ruminal vein were used to study VFA absorption from bicarbonate buffers incubated in the washed reticulorumen, and metabolism by splanchnic tissues. Portal and hepatic vein blood flows were determined by infusion of p-aminohippurate into the mesenteric vein. The steers were subjected to four experimental treatments in a Latin square design. The treatments were Control (ruminal bicarbonate buffer with [mmol/kg]: acetate = 72; propionate = 30; isobutyrate = 2.1; butyrate = 12; valerate = 1.2; caproate = 0; and heptanoate = 0); Val (same as control except for valerate = 8 mmol/kg); Cap (same as control except for caproate = 3.5 mmol/kg); and Hep (same as control except for heptanoate = 3 mmol/kg). All buffers were incubated for 90 min in the rumen, and ruminal VFA absorption rates were maintained by continuous intraruminal infusion of VFA. The arterial concentrations of valerate and heptanoate showed a small increase (< or = 1 micromol/L; P < 0.05) with inclusion of the respective acid in the ruminal buffer, but no change (P = 0.57) in arterial concentration of caproate was detected. Valerate increased (P < 0.05) the net portal flux of butyrate and valerate, as well as the net splanchnic flux of propionate, butyrate, and valerate. With Cap and Hep, the net portal flux of caproate and heptanoate accounted for 54 and 45% of ruminal disappearance rates, respectively, indicating that these acids were extensively metabolized by the ruminal epithelium. Caproate was ketogenic both in the ruminal epithelium and in the liver, and Cap increased (P < 0.05) the arterial concentration, ruminal vein minus arterial concentration difference, net hepatic flux, and net splanchnic flux of 3-hydroxybutyrate. The net hepatic flux of glucose decreased (P = 0.02) with Cap and Hep compared with Control and Val; however, no effect (P = 0.14) on the net splanchnic flux of glucose could be detected. We conclude that the strong biological activity of valerate, caproate, and heptanoate warrant increased emphasis on monitoring their ruminal presence and their potential systemic effects on ruminant metabolism.

Four steers fitted with a ruminal cannula and chronic indwelling catheters in the mesenteric artery, mesenteric vein, hepatic portal vein, hepatic vein, and the right ruminal vein were used to study the absorption and metabolism of VFA from bicarbonate buffers incubated in the temporarily emptied and washed reticulorumen. Portal and hepatic vein blood flows were determined by infusion of p-aminohippurate into the mesenteric vein, and portal VFA fluxes were calibrated by infusion of isovalerate into the ruminal vein. The steers were subjected to four experimental treatments in a Latin square design with four periods within 1 d. The treatments were Control (bicarbonate buffer) and VFA buffers containing 4, 12, or 36 mmol butyrate/kg of buffer, respectively. The acetate content of the buffers was decreased with increasing butyrate to balance the acidity. The butyrate absorption from the rumen was 39, 111, and 300 +/- 4 mmol/h for the three VFA buffers, respectively. The ruminal absorption rates of propionate (260 +/- 12 mmol/h), isobutyrate (11.4 +/- 0.7 mmol/h), and valerate (17.3 +/- 0.7 mmol/h) were not affected by VFA buffers. The portal recovery of butyrate and valerate absorbed from the rumen increased (P < 0.01) with increasing butyrate absorption and reached 52 to 54 +/- 4% with the greatest butyrate absorption. The liver responded to the increased butyrate absorption with a decreasing fractional extraction of propionate and butyrate, and with the greatest butyrate absorption, the splanchnic flux was 22 +/- 1% and 18 +/- 1% of the absorbed propionate and butyrate, respectively. The increased propionate and butyrate release to peripheral tissues was followed by increased (P < 0.05) arterial concentrations of propionate (0.08 +/- 0.01 mmol/kg) and butyrate (0.07 +/- 0.01 mmol/kg). Arterial insulin concentration increased (P = 0.01) with incubation of VFA buffers compared with Control and was numerically greatest with the greatest level of butyrate absorption. We conclude that the capacity to metabolize butyrate by the ruminal epithelium and liver is limited. If butyrate absorption exceeds the metabolic capacity, it affects rumen epithelial and hepatic nutrient metabolism and affects the nutrient supply of peripheral tissues.

An observational study was conducted in six Danish dairy herds. A specially designed stomach tube was compared to the rumenocentesis technique as part of the monitoring of rumen pH. In contrast to a previous study, the use of the stomach tube appeared to reduce saliva contamination. However, correlation with the rumenocentesis technique was poor ( r = 0.33; p = 0.019) and a linear model could only partly explain variations between either results. The presence of subclinical rumen acidosis (SRA) was evidenced in one herd only, as judged by results obtained by the rumenocentesis technique. The present study revealed some limitations of the rumenocentesis technique in small or medium-sized herds due to difficulties in selecting sufficient numbers of cows in the respective groups at risk. The finding of two apparently clinical normal cows with rumen pH values below 5.0 leads to the consideration that such fluctuations may be temporary and at least does not give rise to clinical symptoms. However, the long-term effect of such fluctuations is not known. In general, primiparous cows seemed more prone to low ruminal pH values (< 6.0), higher ruminal concentrations of short-chain fatty acids, and possibly to metabolic acidosis, than were multiparous cows. Ruminal propionate was the most precise predictor of rumen pH, whereas milk fat percentage varied greatly between lactational groups. Blood lactate dehydrogenase (LDH), beta-hydroxybutyrate (BHB) and fructosamine as well as urine phosphorus excretion and renal net acid-base excretion (NABE) were related to ruminal acid load, but were not predictive of rumen pH. Monitoring of dairy herds for SRA should be performed routinely and employ several diagnostic tools (rumenocentesis, renal NABE determination) as well as specific knowledge of herd management and feeding routines.

Six steers fitted with a ruminal cannula and chronic indwelling catheters in the mesenteric artery, mesenteric vein, hepatic portal vein, hepatic vein, as well as in the right ruminal vein were used to study metabolism of VFA absorbed from buffers in the emptied and washed reticulorumen. [2-(13)C]Acetate was infused into a jugular vein to study portal-drained visceral (PDV) uptake of arterial acetate, hepatic unidirectional uptake of acetate, and whole-body irreversible loss rate (ILR). Isobutyrate was infused into the right ruminal vein to calibrate VFA fluxes measured in the portal vein. On sampling days, the rumen was emptied and incubated in sequence with a 0-buffer (bicarbonate buffer without VFA), a VFA-buffer plus continuous intraruminal infusion of VFA, and finally another 0-buffer. Ruminal VFA absorption was determined as VFA uptake from the VFA-buffer and metabolic effects determined as the difference between metabolite fluxes with VFA-buffer and 0-buffers. Steady absorption rates of VFA were maintained during VFA-buffer incubations (4 h; 592+/-16, 257+/-5, 127+/-2, 17+/-<1, 20+/-<1 mmol/h, respectively, of acetate, propionate, butyrate, isovalerate, and valerate). The portal flux of acetate corrected for PDV uptake of arterial acetate accounted for 105+/-3% of the acetate absorption from the rumen, and the net portal flux of propionate accounted for 91+/-2% of propionate absorption. Considerably less butyrate (27+/-3%) and valerate (30+/-3%) could be accounted for in the portal vein. The sum of portal VFA and 3-hydroxybutyrate as well as lactate represented 99+/-3% of total VFA acetyl units and 103+/-2% of VFA propionyl units. Estimates are maximum because no accounting was made for lactate derived from glycolysis in the PDV. The net splanchnic flux of VFA, lactate, 3-hydroxybutyrate, and glucose accounted for 64+/-2% of VFA acetyl units and 34+/-5% of VFA propionyl units. Results indicate that there is a low "first-pass" uptake of acetate and propionate in the ruminal epithelium of cattle, whereas butyrate and valerate are extensively metabolized, though seemingly not oxidized to carbon dioxide in the epithelium but repackaged into acetate, 3-hydroxybutyrate, and perhaps other metabolites. When PDV "second-pass" uptake of arterial nutrients is accounted for, PDV fluxes of VFA, lactate, and 3-hydroxybutyrate represent VFA production in the gastrointestinal tract and thereby VFA availability to the ruminant animal.

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.

To investigate the impact of rumen microbial sequestration of VFA carbon on estimates of acetate availability based on intraruminal infusion of [2-(13)C] acetate, three nonlactating or low-yielding dairy cows were continuously intraruminally infused with [2-(13)C]acetate for 26 h. The 13C content of ruminal VFA, duodenal carbon, and fatty acids (FA) and AA isolated from liquid-associated ruminal microbes and duodenal DM was measured by an isotope ratio mass spectrometer interfaced to an elemental analyzer or a gas-liquid chromatograph. The ruminal gross production of acetate was 38 +/- 4 mol/d and could account for about 38% of the DE intake. Of the intraruminally infused 13C in [2-(13)C]acetate, 7.6 +/- 0.9% was recovered at the duodenum. The 13C content of ruminal propionate, butyrate, and valerate increased (P < 0.05) with intraruminal infusion of [2-(13)C]acetate. It was estimated that about 28% of the 13C intraruminally infused in [2-(13)C]acetate could be accounted for by duodenal 13C flow and absorption of non-acetate VFA. A number of FA isolated from liquid-associated ruminal microbes (C6, C12, C14, anteiso C15, and iso C15) were enriched with 13C (P < 0.05) at a level comparable to the enrichment of ruminal butyrate. Any absorption of these FA from the rumen would further contribute to non-acetate 13C uptake. A maximum of 72% of the ruminal gross production of acetate represented acetate absorption from the rumen in the present study. Consequently, previously used models using intraruminal isotope dilution techniques seem not to be appropriate for measuring acetate availability in ruminants. The number of metabolites exchanging carbon with acetate was found to be so high that assessments of the entire range of inter conversions seem to be practically impossible. Portal absorption studies are discussed as an alternative method of estimating VFA availability to the metabolism in ruminants.

The aim of the present work was to estimate volatile fatty acid (VFA) production rate in the rumen of sheep fed two levels of intake using both a tracer (TM; by isotope dilution) and a non-tracer method (NTM; by supplementary infusion) in steady-state conditions. Six wethers received a diet containing 700 g lucerne hay and 300 g ground maize/kg in eight equal meals at 3 h intervals per d. The diet (9.8 MJ metabolizable energy (ME)/kg DM) was offered at 90 % ad libitum consumption (high intake, HI) or 45 % ad libitum consumption (low intake, LI) in a crossover design. Each sheep received five intrarumen VFA solutions infused continuously for 24 h at rates of 250 ml and 165 ml/h for the HI and LI respectively. The first infusion, considered as a control treatment (Con), consisted of a solution of [1-(13)C]propionate (7 mmol/d). The four other solutions were isoenergetic (1.9 MJ ME/kg DM intake) mixtures of unlabelled propionate (C(3)) and butyrate (C(4)) at different levels: 0.90 mol C(4)/kg DM intake; 0.60 mol C(3)/kg DM intake; 0.30 mol C(3)/kg DM intake; 1.35 mol C(3)/kg DM intake. The VFA infusions did not affect rumen fermentation of the basal diet (pH, osmotic pressure, protozoa numbers), and comparable DM digestibility of the diet among the different treatments was observed. Both estimation methods demonstrated a similar increase (1.7-fold) in the rumen VFA production rate of sheep fed at intakes varying between 0.9 to 1.7 times maintenance. Irrespective of the intake level, the rumen production rate of individual VFA was on average 1.5-fold higher when estimated by the TM compared with the NTM. Rumen VFA production rates estimated by the NTM and TM represented 80 % and 120 % ME intake respectively. The difference between NTM and TM estimates seems likely to be caused mainly by overestimation of the VFA production rates by the TM.

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.

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.

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.

Using the technique of vanadate-facilitated [3H]ouabain binding we have developed a simple and reliable assay for measuring the concentration of [3H]ouabain binding sites in small fresh or frozen biopsies of rumen epithelium papillae. In bovine and ovine rumen epithelium obtained from the cranio-ventral rumen sac the concentration of [3H]ouabain binding sites was 1.6-4.9 nmol g dry wt-1 (n = 32) and 3.7-5.2 nmol g dry wt-1 (n = 6), respectively. When incubated in oxygenated Krebs-Ringer bicarbonate buffer fresh biopsies of rumen epithelium maintained a high K+ and low Na+ content for at least 6 h. Na+ loading of the biopsies induced about 20-fold increase of the Na+, K(+)-pump activity based on measurement of ouabain suppressible net [86Rb+] influx. The ouabain suppressible net influx of [86Rb+] measured in Na+ loaded biopsies showed a close correlation to the [3H]ouabain binding capacity (r = 0.80, P < 0.01) and corresponded to 47 +/- 2% (n = 9) of the theoretical maximum flux rate. The ouabain suppressible net influx of K+ and [86Rb+] were linearly related (r = 0.73; P < 0.001). The net Na+ efflux was 1.21 times the net K+ influx. It is concluded that rumen epithelium has a large capacity for active Na+/K+ transport and that there is agreement between the concentration of [3H]ouabain binding sites in the epithelium and the ouabain suppressible rate of net [86Rb+] influx in Na+ loaded biopsies in spite of some uncertainty about the maximum turnover number of the Na+, K(+)-pump in rumen epithelium.