Na transport across a preparation of sheep omasum was studied. All tissues exhibited a serosa-positive short-circuit current (Isc), with a range of 1-4 microeq. h-1. cm-2. A Michaelis-Menten-type kinetic was found between the Na concentration and the Isc (Michaelis-Menten constant for transport of Na = 6.7 mM; maximal transport capacity of Na = 4.16 microeq. h-1. cm-2). Mucosal amiloride (1 mM), phenamil (1 or 10 microM), or serosal aldosterone (1 microM for 6 h) did not change Isc. Removal of divalent cations (Ca and Mg) enhanced Isc considerably from 2.61 +/- 0.24 to a peak value of 11.18 +/- 1.1 microeq. h-1. cm-2. The peak Isc (overshoot) immediately declined to a plateau Isc of approximately 6-7 microeq. h-1. cm-2. Na flux measurements showed a close correlation between changes in Isc and Na transport. Transepithelial studies demonstrated that K, Cs, Rb, and Li are transported, indicating putative nonselective cation channels, which are inhibited by divalent cations (including Ca, Mg, Sr, Ba) and by (trivalent) La. Intracellular microelectrode recordings from the luminal side clearly showed changes of voltage divider ratio when mucosal divalent cations were removed. The obtained data support the assumption of a distinct electrogenic Na transport mechanism in sheep omasum. (+info)
Amino acid flux in ruminal and gastric veins of sheep: effects of ruminal and omasal injections of free amino acids and carnosine.
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The possibility of free amino acid (FAA) and peptide absorption across the ruminant stomach wall was studied in multicatheterized wethers fed every 12 h. During the last third of the feeding cycle, two intraruminal or intraomasal injections of solutions containing increasing amounts of Ser, Gly, Val, Met, Phe, Lys, and carnosine were successively performed. Before injections, a net uptake of each of these FAA was measured in the ruminal and the gastric veins. The ruminal injections produced a linear increase in ruminal FAA concentration. The highest ruminal concentrations (observed with 3 g of FAA and carnosine) ranged between 5 and 14 mM. After ruminal injections, Ser (P < .05), Gly (P < .05), Val (P < .05), Met (P < .10), and Lys (P < .10) uptake decreased and carnosine net release linearly increased (P < .05), suggesting absorption across the ruminal epithelium. Owing to the low net flux generated by high ruminal concentration, the ruminal epithelium permeability to these molecules seemed to be low. After omasal injections, net flux of injected FAA were not modified, suggesting a low permeability of the gastric epithelia to FAA. Carnosine net release linearily increased (P < .05) with increasing level of carnosine injection, indicating the possibility of dipeptide absorption at the gastric level. This study demonstrated in vivo that the stomach epithelia possess the capacity to absorb FAA and small peptides; however, the permeability of these epithelia to these molecules seemed limited. (+info)
Absorption of 2-hydroxy-4-(methylthio)butanoic acid by isolated sheep ruminal and omasal epithelia.
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Alimet (Novus Inter., Inc., St. Louis, MO) feed supplement (an 88% aqueous solution of 2-hydroxy-4-(methylthio) butanoic acid; HMB) is a source of L-Met commonly used in nonruminants and ruminants. The absorption of HMB across ovine omasal and ruminal epithelia was evaluated in this study. Ruminal and omasal epithelia were collected from eight lambs (BW = 67.6 kg +/- 9.1) and mounted in parabiotic chambers that were repeatedly sampled throughout a 60-min incubation. The appearance of HMB (using DL-[5-14C]-HMB as a radiolabeled marker) in serosal buffers increased quadratically (P < .004) with time in both tissues. More (P < .001) HMB appeared in the serosal buffers with omasal than with ruminal epithelia. Both tissues responded similarly, and, after 60 min of incubation, the accumulation of HMB within the tissues increased linearly (P < .001) as substrate concentration (.375, .75, 1.5, 3.0, 6.0, and 12.0 mM) increased in mucosal buffers. As the concentration of HMB in the mucosal buffers increased, there was a quadratic (P < .001) increase in the appearance of HMB in the serosal buffer of the omasal epithelium, indicating some saturation of the system. The increase in serosal appearance of HMB was linear (P < .001) with ruminal tissue. The results indicate that there are probably multiple mechanisms involved in the absorption of HMB. Because saturation was observed in the omasum, it is likely that mediated transport accounts for at least a portion of the absorption of HMB in the omasum. Other mechanisms (e.g., diffusion and(or) paracellular absorption) are responsible for the balance of the absorption. Omasal epithelium appears to have a greater capacity for HMB absorption than ruminal epithelium. The enzymes involved in the conversion of HMB to 2-keto-4-(methylthio)butanoic acid were found in ruminal and omasal epithelia, liver and kidney. These results indicate that HMB can be absorbed across ruminal and omasal epithelium and that HMB can be used as a source of L-methionine. (+info)
Ergot alkaloid transport across ruminant gastric tissues.
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Ergot alkaloids cause fescue toxicosis when livestock graze endophyte-infected tall fescue. It is generally accepted that ergovaline is the toxic component of endophyte-infected tall fescue, but there is no direct evidence to support this hypothesis. The objective of this study was to examine relative and potential transport of ergoline and ergopeptine alkaloids across isolated gastric tissues in vitro. Sheep ruminal and omasal tissues were surgically removed and placed in parabiotic chambers. Equimolar concentrations of lysergic acid, lysergol, ergonovine, ergotamine, and ergocryptine were added to a Kreb's Ringer phosphate (KRP) solution on the mucosal side of the tissue. Tissue was incubated in near-physiological conditions for 240 min. Samples were taken from KRP on the serosal side of the chambers at times 0, 30, 60, 120, 180, and 240 min and analyzed for ergot alkaloids by competitive ELISA. The serosal KRP remaining after incubation was freeze-dried and the alkaloid species quantified by HPLC. The area of ruminal and omasal tissues was measured and the potential transportable alkaloids calculated by multiplying the moles of transported alkaloids per square centimeter of each tissue type by the surface area of the tissue. Studies were conducted to compare alkaloid transport in reticular, ruminal, and omasal tissues and to determine whether transport was active or passive. Ruminal tissue had greater ergot alkaloid transport potential than omasal tissue (85 vs 60 mmol) because of a larger surface area. The ruminal posterior dorsal sac had the greatest potential for alkaloid transport, but the other ruminal tissues were not different from one another. Alkaloid transport was less among reticular tissues than among ruminal tissues. Transport of alkaloids seemed to be an active process. The alkaloids with greatest transport potential were lysergic acid and lysergol. Ergopeptine alkaloids tended to pass across omasal tissues in greater quantities than across ruminal tissues, but their transport was minimal compared to lysergic acid and lysergol. (+info)
Supplementing barley or rapeseed meal to dairy cows fed grass-red clover silage: I. Rumen degradability and microbial flow.
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The present study was conducted to measure the flow of microbial and nonmicrobial N fractions entering the omasal canal of lactating dairy cows fed grass-red clover silage supplemented with barley and rapeseed meal. Four ruminally cannulated Finnish Ayrshire dairy cows were fed, in a 4 x 4 Latin square design, grass-red clover silage alone or supplemented with (on DM basis) 5.1 kg/d of barley, 1.9 kg/d of rape-seed meal or 5.1 kg/d of barley and 1.9 kg/d rapeseed meal. Nonammonia N flow entering the omasal canal was fractionated into microbial and nonmicrobial N using 15N. Microbial N was fractionated into N associated with liquid-associated bacteria, particle-associated bacteria, and protozoa. Supplementation of diets with barley increased microbial N flow entering the omasal canal (P < 0.01) but had no effect on nonmicrobial N flow. Increased microbial N flow was attributed to liquid-associated bacteria and protozoa. Barley had no effect on apparent ruminal N degradability, but increased true ruminal N degradability (P < 0.01). Barley had no effect on urinary N excretion, but increased daily N retention (P = 0.03). Furthermore, barley supplementation decreased ruminal (P = 0.02) and total tract (P < 0.01) NDF digestibility. Supplementation of diets with rapeseed meal increased apparent ruminal N degradability (P < 0.01) and nonmicrobial N flow entering the omasal canal (P < 0.01), but had no effect on true ruminal N degradability. Despite higher N excretion in urine, rapeseed meal improved daily N retention (P < 0.01). Milk yield was increased (P < 0.01) by barley and rapeseed meal supplements, with the responses being additive. Responses attained with barley were primarily due to increased energy supply for ruminal microbes and improvements in energy and protein supply for the animal. However, provision of readily digestible carbohydrates in barley did not improve microbial capture of ruminal ammonia. Benefits associated with rapeseed meal supplementation were explained as an increase in the supply of ruminally undegradable protein. (+info)
Supplementing barley or rapeseed meal to dairy cows fed grass-red clover silage: II. Amino acid profile of microbial fractions.
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Four ruminally cannulated dairy cows were used to examine the effect of diet on the AA composition of rumen bacteria and protozoa, and the flow of microbial and nonmicrobial AA entering the omasal canal. Cows were offered grass-red clover silage alone, or that supplemented with 5.1 kg DM of barley, 1.9 kg DM of rapeseed meal, or 5.1 kg DM of barley and 1.9 kg DM of rapeseed meal according to a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. During the first 10 d of each period, cows had free access to silage and, thereafter intake was restricted to 95% of ad libitum intake. Postruminal digesta flow was assessed using the omasal canal sampling technique in combination with a triple marker method. Liquid- (LAB) and particle- (PAB) associated bacteria were isolated from digesta in the reticulorumen and protozoa from digesta entering the omasal canal. Microbial protein flow was determined using 15N as a microbial marker. Flows of AA entering the omasal canal were similar in cows fed silage diets supplemented with barley or rapeseed meal. However, rapeseed meal increased nonmicrobial AA flow while barley increased the flow of AA associated with LAB and protozoa. Diet had negligible effects on the AA profile of microbial fractions. Comparison of AA profiles across diets indicated differences between LAB and PAB for 10 out of 17 AA measured. Rumen bacteria and protozoa were found to be different for 14 out of 15 AA measured. For grass silage-based diets, energy and protein supplementations appear to alter postruminal AA supply through modifications in the proportionate contribution of microbial and nonmicrobial pools to total protein flow rather than as a direct result of changes in the AA profile of microbial protein. (+info)
Peptide absorption: a review of current concepts and future perspectives.
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Absorption of di- and tripeptides from the gastrointestinal tract is accepted as being an important biological phenomenon. The extent to which peptides are absorbed and the nutritional and metabolic significance of peptide absorption remain unclear. Evidence is strong for the existence of multiple peptide transport systems, including one type that is electrogenic in nature and that requires a protonmotive force and cotransports two H+ for every peptide transported. The rate of absorption of peptides can be responsive to level of dietary intake and level of dietary protein. Peptide absorption seems to be an important physiological process in ruminants, and this process may account for a large portion of absorbed amino acids. An important new observation is that the nonmesenteric portion of the portal-drained viscera of the ruminant is a major site of peptide absorption. These new observations may result in a reshaping of the currently accepted theory concerning protein utilization by ruminants. (+info)
Morphometric and immunohistochemical study of the omasum of red deer during prenatal development.
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The red deer is an important study species because of its value in the national economy and because it provides a wealth of genetic material. To date, there has been little research into the prenatal development of the stomach of ruminants, and none of the red deer. We therefore performed a histological evaluation of the ontogenesis of the omasum in the red deer. Histomorphometric and immunohistochemical analyses were carried out on 50 embryos and fetuses of deer from the initial stages of prenatal life until birth. For test purposes, the animals were divided into five experimental groups: Group I (1.4-3.6 cm crown-rump length, CRL; 30-60 days, 1-25% of gestation); Group II (4.5-7.2 cm CRL; 67-90 days, 25-35% of gestation); Group III (8-19 cm CRL; 97-135 days, 35-50% of gestation); Group IV (21-33 cm CRL; 142-191 days, 50-70% of gestation); and Group V (36-40 cm CRL; 205-235 days, 75-100% of gestation). At 67 embryonic days, the omasum wall was differentiated, and comprised three layers: the epithelial layer, pluripotential blastemic tissue and serosa. The stratification of the epithelial layer was accompanied by changes in its structure, with the appearance of four laminae of different sizes; in order of appearance these were: primary at 67 days, secondary at 90 days, tertiary at 97 days and quaternary at 135 days. At around mid-gestation, lateral evaginations were formed from the stratum basale of the primary and secondary smaller laminae. These were the primitive corneum papillae. From 205 days, the corneum papillae were present in all four sizes of laminae. The histodifferentiation of the lamina propia-submucosa, tunica muscularis and serosa showed patterns of development similar to those reported for the rumen and reticulum of red deer. The omasum of red deer during prenatal life, especially from 67 days of gestation, was shown to be an active structure with full secretory capacity. Its histological development, its secretory capacity (detected by the presence of neutral mucopolysaccharides) and its neuroendocrine nature (detected by the presence of positive non-neuronal enolase cells and the neuropeptides vasoactive intestinal peptide and neuropeptide Y) were parallel to the development of the rumen and the reticulum. However, its prenatal development was later than that of the omasum in sheep, goat and cow. (+info)