Ruminal fatty acid outflow in dry cows fed different sources of linoleic acid: reticulum and omasum as alternative sampling sites to abomasum.

This study was designed to determine the rumen outflow of fatty acids (FA) and biohydrogenation (BH) extent using alternative sampling sites (reticulum and omasum) to abomasum in dry cows fed different sources of FA. Four Holstein non-pregnant dry cows (≥3 parturitions, and 712 ± 125 kg BW), cannulated in the rumen and abomasum, were randomly assigned to a 4 × 4 Latin square design experiment, containing the following treatments: 1) control (CON); 2) soya bean oil (SO), dietary inclusion at 30 g/kg; 3) whole raw soya beans (WS), dietary inclusion at 160 g/kg; and 4) calcium salts of FA (CSFA), dietary inclusion at 32 g/kg. Rumen outflow of nutrients was estimated using the three markers reconstitution system (cobalt-EDTA, ytterbium chloride, and indigestible neutral detergent fibre [NDF]). Diets with FA sources decreased feed intake and increased FA intake. No differences in nutrient intake and digestibility were detected among cows fed diets supplemented with different FA sources. Diets with FA sources reduced the rumen outflow of DM and NDF, hence decreasing their passage rates. In addition, SO diet reduced the ruminal outflow of DM and NDF in comparison with WS and CSFA. Omasal sampling yielded the highest values of rumen outflow of NDF and potentially degradable NDF (pdNDF), whereas the reticular and abomasal samplings yielded intermediate and least values, respectively. The interaction effect between diet and sampling site was observed for rumen outflow of majority FA (except for C16:0, C18:0, and C18:2 trans-10, cis-12) and BH extension of C18:1 cis, C18:2, and C18:3. Calculations derived from abomasal sampling revealed that WS and CSFA diets had lower BH extent of C18:1 cis and C18:2 in comparison with SO, whereas cows fed CSFA had greater BH extent of C18:3 and lower BH extent of C18:1 cis compared to those fed WS. However, the latter results were not similar when calculations were performed based on the reticular and omasal samplings. Thus, there is evidence that neither reticular nor omasal samplings are suitable for estimating rumen outflow of FA in dry cows. In addition, WS and CSFA diets can increase the abomasal flow of polyunsaturated FA in dry cows.

Effect of increasing the proportion of dietary concentrate on gastrointestinal tract measurements and brush border enzyme activity in Holstein steers.

The aim of this study was to determine the time course for adaptation of the reticulo-rumen, omasum, abomasum, and small intestine in response to an abrupt increase in the proportion of grain in the diet. Adaptive responses include tissue and digesta mass, small intestinal length, and brush border enzyme activity in the duodenum, proximal jejunum, and ileum. Twenty-five Holstein steers (213 ± 23 kg; 5 to 7 mo of age) were blocked by body weight, and within block were randomly assigned to 1 of 5 treatments: the control diet (CTRL; 92% chopped grass hay and 8% mineral and vitamin supplement on a dry matter basis) or a moderate grain diet (MGD; 50% chopped grass hay, 42% rolled barley grain, and 8% mineral and vitamin supplement) that was fed for 3 (MGD3), 7 (MGD7), 14 (MGD14), or 21 d (MGD21). Dry matter intake was limited to 2.25% of body weight to ensure that changes in dry matter intake did not confound the results. On the last day of the dietary exposure, steers were slaughtered 2 h after feeding. Reticulo-rumen tissue mass and ruminal epithelium mass in the ventral sac of the rumen were not affected by the MGD. Wet reticulo-ruminal digesta mass decreased from CTRL to MGD7 and then increased, but reticulo-ruminal digesta dry matter mass did not differ between treatments. Omasal mass, omasal tissue mass, and omasum digesta mass decreased linearly with the number of days fed MGD, but abomasal tissue mass tended to increase linearly. Duodenal tissue mass tended to increase linearly, and ileal length increased linearly with the number of days fed MGD. Lactase activity in the proximal jejunum increased linearly and maltase activity in duodenum tended to increase linearly with days fed MGD. Aminopeptidase N activity in the proximal jejunum increased cubically with days fed MGD, and dipeptidylpeptidase IV activity in ileum tended to decrease from CTRL to MGD14 and then tended to increase. Adaptation to a diet with a greater proportion of concentrate involves changes in the mass and length of regions of the gastrointestinal tract and brush border enzyme activity. These changes take place gradually over at least 3 wk.

Distribution of ki-67, alpha smooth muscle actin and vimentin in the reticulum and omasum of Baladi goat

Seven Baladi goats of both sexes (9 to 24 months old) were used to describe the distribution pattern of ki-67, alpha smooth muscle actin (áSMA) and vimentin (VIM) in the reticulum and omasum. This study was carried out using the avidin-biotin immunoperoxidase method. Ki-67 immunostaining was restricted to the basal cells layer in the epithelia of both the reticulum and omasum, suggesting the importance of ki-67 in epithelial cells proliferation and keratin biosynthesis. Immunostaining for áSMA was detected in smooth muscle cells in reticular folds, omasal lamiae and muscularis in both the reticulum and omasum, indicating the critical role of áSMA in muscular motility. The widespread distribution of VIM immunostainings in epithelia, fibroblasts in lamina propria and submucosa, and endothelia of blood vessels supports the importance of VIM as an intermediate filament protein. Detection of VIM in glial cells of enteric plexuses indicates its supportive role in the nervous control of both reticulum and omasum. Overall, this immunohistochemical study revealed non-significant differences in the expression of ki-67, áSMA, and VIM between the reticulum and omasum. This study thus verifies the important roles of ki-67, áSMA and VIM in the structure and function of the reticulum and omasum of Baladi goats

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Contribution of different segments of the gastrointestinal tract to digestion in growing Saanen goats.

This study examined mean retention time (MRT) of particulate and liquid matter in different segments of the gastrointestinal tract (GIT) of growing Saanen goats of different sexes and subjected to different levels of feed restriction. In addition, feeding behavior and total tract digestibility were determined for all animals ahead of slaughter. In total, 54 Saanen goats (18 each of females, castrated males, and intact males) with initial BW 15.3 ± 0.4 kg were used in a 3 × 3 factorial arrangement comprising the 3 sexes and 3 levels of feed restriction (unrestricted/ad libitum, moderate, and severe restriction). Six blocks per sex group, each consisting of 3 goats, were randomly formed and the goats within each block were randomly allocated to 1 of 3 different feed restrictions. The daily amounts of feed offered to animals subjected to moderate and severe feed restriction (approximately 75% and 50% of ad libitum rate, respectively) were determined within block based on the DMI by ad libitum fed goats on the previous day. The MRT of particulate matter was determined either using Yb-labeled diet or indigestible NDF (iNDF) determined in situ as markers. Mean retention time of the liquid phase was determined by Cr-EDTA. Orthogonal polynomial contrasts were used to determine linear and quadratic effect of feed restriction, while the effect of sex was compared by Tukey test. The effects of sex and the interaction between sex and feed restriction were not significant on most of variables evaluated. Eating, ruminating, and total chewing time per g DM and NDF intake increased linearly as feed restriction increased (P ≤ 0.03). Diet digestibility increased quadratically for DM and OM, and linearly for NDF as feed intake decreased (P ≤ 0.03). The MRT of iNDF in the reticulorumen, omasum, abomasum, colon, and total GIT increased linearly with increased feed restriction (P ≤ 0.01). Mean retention time in the cecum varied quadratically, being greatest for animals with moderate feed restriction. The MRT of liquid was quadratically (P ≤ 0.04) affected by feed restriction in the reticulorumen, cecum, and total GIT, with the greatest MRT observed for animals subjected to moderate feed restriction. In conclusion, the level of feed restriction increased the MRT of particulate and liquid matter. The MRT was an important mechanism to increase nutrient supply when animals were subjected to feed restriction, as indicated by increased total tract digestibility.

Digesta sampling sites and marker methods for estimation of ruminal outflow in bulls fed different proportions of corn silage or sugarcane.

The objective of this experiment was to assess ruminal outflow and apparent total-tract digestibility using digesta samples from 3 sites (reticulum, omasum, and abomasum) and 3 marker methods (single marker: indigestible NDF [iNDF; sample without separation]; double marker: iNDF + Co-EDTA [filtered sample]; and triple marker: iNDF + ytterbium [Yb] acetate + Co-EDTA [filtered and centrifuged]) in bulls fed corn silage and sugarcane-based diets. Eight crossbred (Holstein × Zebu) bulls (353 ± 37 kg of BW; 24 ± 1 mo of age) with ruminal and abomasal cannulas were randomly distributed into two 4 × 4 Latin squares that were balanced for residual effects. The following experimental diets were used: 1) 60% corn silage + 40% concentrate, 2) 40% corn silage + 60% concentrate, 3) 60% fresh sugarcane + 40% concentrate, and 4) 40% fresh sugarcane + 60% concentrate. Reticular, omasal, and abomasal digesta samples were collected at 9-h intervals over 3 d. At the end of the experiment, a composite sample was prepared for each bull, and these samples were subsequently assigned to the 3 marker methods. The concentrations of CP, NDF, and iNDF of reticular digesta differed (P < 0.01) from those of the omasum and abomasum. Use of omasal and abomasal samples led to similar estimates of ruminal outflow and ruminal digestibility for DM (P = 0.65), OM (P = 0.68), CP (P = 0.85), and NDF (P = 0.57). In contrast, the ruminal outflow of digesta based on reticular sampling appeared to be underestimated. We recommend sampling from the omasum because sampling from this region is less invasive than sampling from the abomasum. Although we did not observe differences in ruminal NDF digestibility among the different marker methods, we did observe that ruminal digestibility of CP was greater for the single marker method than for the double and triple marker methods; we therefore recommend the use of the double or triple marker method.

Effects of feeding wheat or corn-wheat dried distillers grains with solubles in low- or high-crude protein diets on ruminal function, omasal nutrient flows, urea-N recycling, and performance in cows.

A study was conducted to determine the effects of including either wheat-based (W-DDGS) or corn-wheat blend (B-DDGS) dried distillers grains with solubles as the major protein source in low- or high-crude protein (CP) diets fed to dairy cows on ruminal function, microbial protein synthesis, omasal nutrient flows, urea-N recycling, and milk production. Eight lactating Holstein cows (768.5 ± 57.7 kg of body weight; 109.5 ± 40.0 d in milk) were used in a replicated 4 × 4 Latin square design with 28-d periods (18d of dietary adaptation and 10d of measurements) and a 2 × 2 factorial arrangement of dietary treatments. Four cows in one Latin square were ruminally cannulated for the measurement of ruminal fermentation characteristics, microbial protein synthesis, urea-N recycling kinetics, and omasal nutrient flow. The treatment factors were type of distillers co-product (W-DDGS vs. B-DDGS) and dietary CP content [15.2 vs. 17.3%; dry matter (DM) basis]. The B-DDGS was produced from a mixture of 15% wheat and 85% corn grain. All diets were formulated to contain 10% W-DDGS or B-DDGS on a DM basis. No diet effect was observed on DM intake. Yields of milk, fat, protein, and lactose, and plasma urea-N and milk urea-N concentrations were lower in cows fed the low-CP compared with those fed the high-CP diet. Although feeding B-DDGS tended to reduce ruminal ammonia-N (NH3-N) concentration compared with feeding W-DDGS (9.3 vs. 10.5mg/dL), no differences were observed in plasma urea-N and milk urea-N concentrations. Additionally, dietary inclusion of B-DDGS compared with W-DDGS did not affect rumen-degradable protein supply, omasal flows of total N, microbial nonammonia N (NAN), rumen-undegradable protein, and total NAN, or urea-N recycling kinetics and milk production. However, cows fed the low-CP diet had lower N intake, rumen-degradable protein supply, ruminal NH3-N concentration, and omasal flows of N, microbial NAN, and total NAN compared with those fed the high-CP diet. Feeding the low-CP compared with the high-CP diet also resulted in lower endogenous urea-N production, urea-N recycled to the gastrointestinal tract, and urea-N excretion in urine. In summary, our results indicate that both W-DDGS and B-DDGS can be included as the major protein sources in dairy cow diets without compromising nutrient supply and production performance. However, feeding the low-CP diet lowered omasal flows of microbial protein and metabolizable protein, which, in turn, resulted in lower milk production compared with feeding the high-CP diet.

Effect of dietary phytate on phosphorus digestibility in dairy cows.

The objective was to evaluate the effect of dietary phytate P (Pp) supply on ruminal and postruminal Pp digestion and net disappearance of P from the lower digestive tract of lactating cows. Six ruminally and ileally cannulated crossbred lactating cows were used in 2 incomplete Latin squares with four 21-d periods (17d of diet adaptation, 4d of total collection). Dietary treatments were low Pp, medium Pp, and high Pp, and a high inorganic P (Pi) diet with the same total P content as the highest Pp diet but with P mostly from inorganic sources. The diets contained 0.10, 0.18, 0.29, and 0.11% Pp and 0.43, 0.48, 0.54, and 0.53% total P on a dry matter basis, with cottonseed meal used to increase Pp content. Ytterbium-labeled corn silage and Co-EDTA were used as particulate and liquid phase markers to measure omasal and ileal digesta flow. Omasal and ileal digesta were collected every 6h on d 20 and 21 and rumen contents were collected on d 21. Samples were analyzed for total P (molybdovanadate yellow method), Pi (blue method), and Pp (high performance ion chromatography). Phytate P and total P intake increased linearly with increasing dietary Pp. Ruminal Pp disappearance also increased linearly with dietary Pp but the magnitude of change was small. Small intestinal net disappearance of Pi was not affected by dietary Pp. Phytate P was hydrolyzed in the large intestine but its hydrolysis was not influenced by dietary Pp. Net disappearance of Pi form the large intestine did not vary with dietary Pp. Dry matter digestibility decreased linearly with increasing dietary Pp, as did apparent digestion of P, and fecal P increased linearly. Dry matter digestibility was higher for high Pi than for high Pp, likely due to the effect of cottonseed meal in the latter diet. Replacing a portion of Pp with Pi resulted in decreased P excretion but this effect was confounded with increased fecal dry matter for the high-Pp (high-cottonseed meal) diet. In lactating cows Pp digestibility was not negatively influenced by dietary Pp and fecal P excretion was regulated by dietary total P rather than by form of dietary P.

Effect of silage botanical composition on ruminal biohydrogenation and transfer of fatty acids to milk in dairy cows.

Ruminal biohydrogenation and transfer of fatty acids (FA) to milk were determined for 4 silages with different botanical compositions using 4 multiparous Norwegian Red dairy cows [(mean ± SD) 118 ± 40.9 d in milk, 22.5 ± 2.72 kg of milk/d, 631 ± 3.3 kg of body weight, 3.3 ± 0.40 points on body condition score at the start of the experiment] fitted with rumen cannulas. Treatments consisted of 4 experimental silages: a mix of the first and third cut of organically managed short-term grassland with timothy (Phleum pratense L.) and red clover (Trifolium pratense L.; 2 yr old; ORG-SG); organically managed long-term grassland with a high proportion of unsown species (6 yr old; ORG-LG); conventionally managed ley with perennial ryegrass (Lolium perenne L.; CON-PR); and conventionally managed ley with timothy (CON-TI). The herbages were cut, wilted, and preserved with additive in round bales and fed at 0.90 of ad libitum intake. A barley (Hordeum vulgare L.) concentrate constituted 300 g/kg of dry matter of the total feed offered. A Latin square design (4 × 4) with 3-wk periods and the last week in each period used for sampling was implemented. Omasal flows of FA were measured using Yb acetate, Cr-EDTA, and the indigestible neutral detergent fiber fraction as indigestible markers. The composition of FA was analyzed in feed, omasal digesta, and milk. Compared with ORG-LG, ORG-SG had a higher herbage proportion of red clover (0.36 vs. 0.01) and lower proportions of timothy (0.42 vs. 0.18), smooth meadowgrass (Poa pratensis L.), meadow fescue (Festuca pratensis Huds.), white clover (Trifolium repens L.), dandelion (Taraxacum spp.), and creeping buttercup (Ranunculus repens L.). The silages were well preserved. The concentration of neutral detergent fiber was higher and the concentration of Kjeldahl-N was lower for CON-TI than for the other silages. Silage type had no effect on dry matter intake, but milk yield was lower for CON-TI than for the other silages. Apparent biohydrogenation of C18:3n-3 was lower for ORG-SG (932 g/kg) than for ORG-LG (956 g/kg), CON-PR (959 g/kg), and CON-TI (958 g/kg). Compared with the grass-based silages, ORG-SG and ORG-LG resulted in higher omasal flows of C18:1 trans FA and higher milk fat proportions of C18:1 trans FA and C18:2 cis-9,trans-11. Apparent recovery of C18:3n-3 in milk was higher for ORG-SG (61 g/kg) than for ORG-LG (33 g/kg), CON-PR (34 g/kg), and CON-TI (38 g/kg), and milk fat proportion of C18:3n-3 was higher for ORG-SG than for CON-TI. Milk fat proportions of C16:0 were lower for ORG-SG and ORG-LG compared with those for CON-PR and CON-TI. It was concluded that high proportions of red clover and other dicotyledons in the silages affected ruminal biohydrogenation and increased milk fat proportions of beneficial FA.

Effects of feeding different linseed sources on omasal fatty acid flows and fatty acid profiles of plasma and milk fat in lactating dairy cows.

The aim of this experiment was to study the effects of feeding different linseed sources on omasal fatty acid (FA) flows, and plasma and milk FA profiles in dairy cows. Four ruminally cannulated lactating Holstein-Friesian cows were assigned to 4 dietary treatments in a 4×4 Latin square design. Dietary treatments consisted of supplementing crushed linseed (CL), extruded whole linseed (EL), formaldehyde-treated linseed oil (FL) and linseed oil in combination with marine algae rich in docosahexaenoic acid (DL). Each period in the Latin square design lasted 21 d, with the first 16 d for adaptation. Omasal flow was estimated by the omasal sampling technique using Cr-EDTA, Yb-acetate, and acid detergent lignin as digesta flow markers. The average DM intake was 20.6 ± 2.5 kg/d, C18:3n-3 intake was 341 ± 51 g/d, and milk yield was 32.0 ± 4.6 kg/d. Milk fat yield was lower for the DL treatment (0.96 kg/d) compared with the other linseed treatments (CL, 1.36 kg/d; EL, 1.49 kg/d; FL, 1.54 kg/d). Omasal flow of C18:3n-3 was higher and C18:3n-3 biohydrogenation was lower for the EL treatment (33.8 g/d; 90.9%) compared with the CL (21.8 g/d; 94.0%), FL (15.5 g/d; 95.4%), and DL (4.6 g/d; 98.5%) treatments, whereas whole-tract digestibility of crude fat was lower for the EL treatment (64.8%) compared with the CL (71.3%), FL (78.5%), and DL (80.4%) treatments. The proportion of C18:3n-3 (g/100 g of FA) was higher for the FL treatment compared with the other treatments in plasma triacylglycerols (FL, 3.60; CL, 1.22; EL, 1.35; DL, 1.12) and milk fat (FL, 3.19; CL, 0.87; EL, 0.83; DL, 0.46). Omasal flow and proportion of C18:0 in plasma and milk fat were lower, whereas omasal flow and proportions of biohydrogenation intermediates in plasma and milk fat were higher for the DL treatment compared with the other linseed treatments. The results demonstrate that feeding EL did not result in a higher C18:3n-3 proportion in plasma and milk fat despite the higher omasal C18:3n-3 flow. This was related to the decreased total-tract digestibility of crude fat. Feeding FL resulted in a higher C18:3n-3 proportion in plasma and milk fat, although the omasal C18:3n-3 flow was similar or lower than for the CL and EL treatment, respectively. Feeding DL inhibited biohydrogenation of trans-11,cis-15-C18:2 to C18:0, as indicated by the increased omasal flows and proportions of biohydrogenation intermediates in plasma and milk fat.

Effects of replacing canola meal as the major protein source with wheat dried distillers grains with solubles on ruminal function, microbial protein synthesis, omasal flow, and milk production in cows.

A study was conducted to determine the effects of replacing canola meal (CM) as the major protein source with wheat-based dried distillers grains with solubles (W-DDGS) on ruminal fermentation, microbial protein production, omasal nutrient flow and animal performance. Eight lactating dairy cows were fed in a replicated 4 × 4 Latin square design with 28-d periods (20 d of dietary adaptation and 8 d of measurements). Four cows in one Latin square were ruminally cannulated for measurements of ruminal fermentation characteristics and flow of nutrients at the omasal canal. Cows were fed either a standard barley silage-based total mixed ration containing CM as the major protein supplement (0% W-DDGS, control) or diets formulated to contain 10, 15, and 20% W-DDGS (dry matter basis), with W-DDGS replacing primarily CM. Diets were isonitrogenous (18.9% crude protein) and contained 3.0, 3.2, 3.5, and 3.7% ether extract for 0, 10, 15, and 20% W-DDGS, respectively. Diets contained 50% forage and 50% concentrate. Inclusion of W-DDGS linearly increased dry matter intake (29.5, 31.2, 30.2, and 31.9 kg/d for 0, 10, 15, and 20% W-DDGS, respectively). The addition of W-DDGS in place of CM resulted in a 1.2- to 1.8-kg increase in milk yield (42.9, 44.7, 44.1, and 44.5 kg/d for 0, 10, 15, and 20% W-DDGS); however, a quadratic change in feed efficiency (i.e., milk yield/DM intake) occurred as the dietary level of W-DDGS increased. Treatments did not differ for milk fat, protein, and lactose concentrations; however, quadratic changes were observed in milk yields of fat (1.48, 1.56, 1.62, and 1.55 kg/d for 0, 10, 15, and 20% W-DDGS, respectively), protein (1.44, 1.46, 1.49, and 1.42 kg/d) and lactose (1.96, 2.02, 2.09, and 1.93 kg/d). Ruminal fermentation characteristics did not change except that the inclusion of 20% W-DDGS resulted in a decrease and a tendency for a decrease in molar concentrations of isobutyrate and total volatile fatty acids, respectively. Omasal flow of total bacterial nonammonia N (NAN) and bacterial efficiency (g of total bacterial NAN flow/kg of organic matter truly digested in the rumen) were not different among diets; however, feeding W-DDGS resulted in a quadratic increase in nonammonia nonbacterial N flow at the omasal canal (271, 318, 336, and 311 g/d for 0, 10, 15, and 20% W-DDGS, respectively). These data indicate that W-DDGS can substitute for CM as the major protein source in dairy cow diets without negatively affecting ruminal fermentation, microbial protein production, and omasal nutrient flow, and can potentially increase dry matter intake and milk yield.

Role of tachykinin and neurokinin receptors in the regulation of ovine omasal contractions.

The present study investigated a role of tachykinins (TK) and neurokinin (NK) receptors (NK-R) in the non-cholinergic regulation of omasal contractions in sheep. Semiquantitative reverse transcription (RT)-PCR revealed that both preprotachykinin (PPT)-A and PPT-B mRNA were distributed in the omasal muscle layers and that NK-R type-1 (NK-1R) and type-2 (NK-2R) mRNA were largely expressed in the same tissues. Cumulative application of substance P (SP), neurokinin A (NKA), and neurokinin B (NKB) at 0.03-10µM induced tonic contractions of omasal muscle strips, and the contractile amplitude increased in order of NKB

Role of nitrergic nerves in the regulation of motility of the omasum and abomasum in healthy sheep (Ovis aries).

We examined the physiological role of nitrergic nerves in the regulation of omasal and abomasal motility in conscious healthy sheep and omasal muscle specimens. Nitric oxide (NO)-donor, S-nitroso-acethyl-dl-penicillamine (SNAP, 3-30 nmol/kg per min, i.v.) significantly inhibited omasal electromyographic (EMG) activity, whereas it did not alter EMG activity in the abomasal antrum. However, NO synthase inhibitor, Nomega-nitro-l-arginine-methyl ester (L-NAME, 0.3-3.0 micromol/kg per min, i.v.) did not alter EMG activity of the omasum and abomasum. In the in vitro experiments, SNAP application (6-200 micromol/l) significantly inhibited bethanechol (10 micromol/l)-induced contraction of longitudinal and circular muscles of the omasum. L-NAME application (0.03-3.0 mmol/l) enhanced electric field stimulation-induced contractions of the circular muscles. The results suggest that the omasal muscles are responsive to exogenous NO and that nitrergic nerves innervate the circular muscle layer of the omasum, however, nitrergic nerves are not or scarcely involved the physiological regulation of omasal and possibly abomasal motility in healthy sheep.

Localization of CCK-1R in the omasum and role of CCK in the regulation of omasal contractions in sheep.

The present study examined localization of cholecystokinin receptor (CCK-R) mRNA in the muscle layer of the ovine omasum and role of CCK-R type 1 (CCK-1R) in the regulation of muscle contraction of the omasum. We demonstrated that not only CCK-R type 2 (CCK-2R) mRNA but also CCK-1R mRNA is highly expressed in the muscle layer of the ovine omasum. Application of CCK-8 to muscle strips of the greater curvature of the ovine omasum at 1-100 nM induced tonic contraction in a concentration-dependent manner, and the contractile effect of CCK-8 was inhibited by both CCK-1R antagonist lorglumide (IC(50) 2.7 and 7.9 microM in the longitudinal and circular muscle, respectively) and CCK-2R antagonist PD135,158 (IC(50) 51.4 microM in the longitudinal muscle), indicating that not only CCK-2R but also CCK-1R is functionally expressed in the plasma membrane of smooth muscles in the omasum and mediates action of exogenous CCK. Contractile effect of intravenous infusion of CCK-8 (1-30 pmol/kg/min) on omasal contraction was also confirmed in the in vivo experiments using conscious sheep in the absence and presence of atropine infusion (14.4 nmol/kg/min), and showed that circulating CCK increases omasal electromyographic (EMG) activity at lower plasma concentration than that it inhibits ruminal contractions. Taking account of our previous results in the in vivo study using other CCK-1R antagonist, it is suggested that circulating CCK, even at normal range of plasma concentration, plays a physiological role as a regulator of omasal contractions in sheep and CCK-1R mediates the action of CCK.

Effect of intravenous infusion of proglumide on ruminal motility in conscious sheep (Ovis aries).

The effects of intravenous infusion of proglumide on regular ruminal contractions were examined in conscious sheep using doses that inhibit pancreatic exocrine secretion. After a control period of 20 min, proglumide was infused intravenously for 40 min at a dose of 15, 30 or 60 micromol/kg per min and venous blood was collected. The intravenous infusion of proglumide significantly increased the frequency of ruminal contractions at 15 micromol/kg per min without altering the amplitude, while it significantly decreased the frequency and amplitude of ruminal contractions at 30 and 60 micromol/kg per min in a dose-dependent manner. Proglumide did not increase contractile activity of the omasum, abomasum and duodenum or the plasma concentration of immunoreactive cholecystokinin (CCK). Application of proglumide at 1-30 mmol/L inhibited bethanechol-induced contraction in both longitudinal and circular muscle strips of the dorsal sac of the rumen. These results suggest that proglumide at a low dose acts indirectly on the rumen as a CCK receptor antagonist to increase the frequency of contractions, whereas at higher doses it inhibits cholinergic-induced contraction of the ruminal muscles or acts as an agonist to inhibit contractions in sheep. Hence, proglumide at high doses seems unsuitable for research or therapeutic use as a CCK receptor blockade in sheep.

Effect of varying dietary ratios of alfalfa silage to corn silage on omasal flow and microbial protein synthesis in dairy cows.

Eight ruminally cannulated multiparous Holstein cows that were part of a larger production trial were used to study the effects of varying dietary ratios of alfalfa silage (AS) to corn silage (CS) on omasal flow of nutrients and microbial protein. Cows were blocked by DIM and randomly assigned to 2 replicated 4 x 4 Latin squares (28-d periods). Diets fed contained (dry matter basis): A) 51% AS, 43% rolled high-moisture shelled corn (HMSC), and 3% solvent soybean meal (SSBM); B) 37% AS, 13% CS, 39% HMSC, and 7% SSBM; C) 24% AS, 27% CS, 35% HMSC, and 12% SSBM; or D) 10% AS, 40% CS, 31% HMSC, and 16% SSBM. Crude protein (CP) contents were 17.2, 16.9, 16.6, and 16.2% for diets A, B, C, and D. All 4 diets were high in energy, averaging 49% nonfiber carbohydrates and 24% neutral detergent fiber. Total microbial nonammonia nitrogen flow was lower on diet D (423 g/d) compared with diets A (465 g/d), B (479 g/d), and C (460 g/d). A significant quadratic effect indicated that microbial protein synthesis was maximal at 38% AS. Supply of rumen-degraded protein decreased linearly from 3,068 g/d (diet A) to 2,469 g/d (diet D). Omasal flow of rumen-undegraded protein did not differ among diets and averaged 1,528 g/d. However, when expressed as a percentage of dry matter intake, rumen-undegraded protein increased linearly from 5.59% (diet A) to 6.13% (diet D), probably because CP from SSBM was more resistant to degradation than CP from AS. Essential AA flow was lowest on diet D, and Lys flow tended to be lower on diet D, which may explain the lower milk and protein yields observed on that diet.

Retention of different sizes of electronic identification boluses in the forestomachs of sheep.

Twelve types of electronic identification ruminal boluses of different dimensions were used to obtain a model for predicting their retention in the forestomachs of sheep. Boluses (n = 1,662) were made of ceramic materials, and their dimensions varied in o.d. (9 to 21 mm), length (37 to 68 mm), volume (2.5 to 21.0 mL), and specific gravity (0.85 to 3.91). Each bolus contained a half-duplex, standardized, glass-encapsulated transponder (32 x 3.8 mm). Boluses were administered to sheep (n = 1,497) of different ages by using the appropriate balling guns, and their retention under semiintensive conditions was recorded for at least 2 yr. When a bolus was lost, the sheep was rebolused with a heavier bolus. All sheep wore 2 plastic ear tags: one for the official control of health programs and the other for farm use. To determine the anatomical limit for a bolus passing through the gastrointestinal tract, the size of the reticulo-omasal orifice was measured in 46 adult sheep (male, n = 14; female, n = 32) that died by causes not related to bolus administration during the experiment. No signs of disease or growth alteration were detected in the bolused sheep. Total ear tag losses during the experiment period were 7.5% on average. Bolus retention (5 to 100%) varied according to bolus features and age of the sheep, but it showed a plateau after 18 mo. Inadequately dimensioned boluses were regurgitated or passed through the gastrointestinal tract and were excreted with the feces. The diameter of the reticulo-omasal orifice in adult sheep differed between male and female (23.1 and 21.8 mm, respectively; P < 0.01) and was greater than the o.d. of the retained boluses. Retention rate was predicted from bolus weight and volume by a logistic regression (R2 = 0.997; P < 0.001). When retention data from the literature (59.0 to 100%) were included in the model, the adjustment was slightly lower (R2 = 0.967). As a result, the minimum bolus weight estimated to reach a 99.5% retention rate in sheep varied between 16 and 45 g when volume varied between 3 and 22 mL, for boluses with a specific gravity between 2.0 and 5.2. In conclusion, bolus retention rate in sheep varied dramatically according to their features. For safe and efficient retention of electronic identification boluses in sheep, boluses of small volume and diameter (e.g., < 15 mm) with specific gravity and weight greater than 3.0 and 20 g, respectively, are recommended.

Transport of acetate and sodium in sheep omasum: mutual, but asymmetric interactions.

We have studied the transport of acetate across the isolated epithelium of sheep omasum; no net transport was observed (J(ms) approximately = J(sm)) under Ussing chamber conditions. Low mucosal pH (pH 6.4) significantly enhanced J(ms) acetate and the transport rates of acetate increased linearly and significantly (r2=0.99) with the luminal acetate concentration. The presence of another short chain fatty acid (propionate) did not affect J(ms) acetate significantly. Neither addition of 1 mmol l(-1) DIDS to the mucosal side nor HCO3 replacement caused changes of J(ms) acetate; this does not support the assumption of acetate transport via anion exchange. Addition of 1 mmol l(-1) amiloride to the mucosal side significantly decreased acetate fluxes at high mucosal acetate concentration (100 mmol l(-1)) and low pH (6.4) indicating interaction between acetate uptake in the undissociated form, intracellular release of protons and activation of Na+/H+ exchange (NHE). However, the mutual interaction between Na transport via NHE and acetate transport is asymmetric. Stimulation or inhibition of Na transport via NHE is much more pronounced than the corresponding changes of acetate fluxes. Thus, the obtained results support the conclusion that acetate is transported via simple diffusion and probably predominantly in the protonated form, thereby explaining the positive and mutual interaction between Na transport and short chain fatty acids.

Determination of digesta flow entering the omasal canal of dairy cows using different marker systems.

Four studies were conducted to compare the effect of four indigestible markers (LiCoEDTA, Yb-acetate, Cr-mordanted straw and indigestible neutral-detergent fibre (INDF)) and three marker systems on the flow of digesta entering the omasal canal of lactating dairy cows. Samples of digesta aspirated from the omasal canal were pooled and separated using filtration and high-speed centrifugation into three fractions defined as the liquid phase, small particulate and large particulate matter. Co was primarily associated with the liquid phase, Yb was concentrated in small particulate matter, whilst Cr and INDF were associated with large particles. Digesta flow was calculated based on single markers or using the reconstitution system based on combinations of two (Co + Yb, Co + Cr and Co + INDF) or three markers (Co + Yb + Cr and Co + Yb + INDF). Use of single markers resulted in large differences between estimates of organic matter (OM) flow entering the omasal canal suggesting that samples were not representative of true digesta. Digesta appeared to consist of at least three phases that tended to separate during sampling. OM was concentrated in particulate matter, whilst the liquid phase consisted mainly of volatile fatty acids and inorganic matter. Yb was intimately associated with nitrogenous compounds, whereas Cr and INDF were concentrated in fibrous material. Current data indicated that marker systems based on Yb in combination with Cr or INDF are required for the accurate determination of OM, N and neutral-detergent fibre flow. In cases where the flow of water-soluble nutrients entering the omasal canal is also required, the marker system should also include Co.

Functional anatomy of the omasum in high Arctic Svalbard reindeer (Rangifer tarandus platyrhynchus) and Norwegian reindeer (Rangifer tarandus tarandus).

The structure and fill of the omasum was investigated in summer and in winter in adult female reindeer living on the polar desert and tundra of the high Arctic archipelago of Svalbard and in sub-Arctic mountain habitats in northern Norway. The mean total mass of the omasum in non-lactating adult female Svalbard reindeer was 467 g (0.65 g per 100 g live body mass (BM)) in September and 477 g (1.03 g per 100 g BM) in April. By contrast, the mean mass of the omasum in non-lactating adult reindeer in northern Norway was 534 g (0.83 g per 100 g BM) in September but only 205 g (0.35 g per 100 g BM p < 0.05) in late March, owing to a decrease in both tissue mass and the wet mass of the contents of the organ. The mean absorptive surface of the omasum in Svalbard reindeer was 2300 cm2 in September and 2023 cm2 in April. In Norwegian reindeer, by contrast, the absorptive surface area decreased from 2201 cm2 in September to 1181 cm2 (p < 0.05) in late March. The marked seasonal decline of omasal tissue and contents in Norwegian reindeer probably results from intake of highly digestible forage plants, including lichens, in winter. Svalbard reindeer, a non-migratory sub-species, survive eating poor quality fibrous vascular plants in winter. The absence of any marked seasonal change in the mass, total absorptive surface area or filling of the omasum in Svalbard reindeer in winter despite a substantial decline in body mass presumably reflects their need to maintain maximum absorption of nutrients, including volatile fatty acids, when feeding on such poorly fermentable forage.

Role of nitric oxide in in vitro contractile activity of the third compartment of the stomach in llamas.

OBJECTIVE: To determine the role of nitric oxide and an apamin-sensitive nonadrenergic-noncholinergic inhibitory transmitter in in vitro contractile activity of the third compartment in llamas. SAMPLE POPULATION: Isolated strips of third compartment of the stomach from 5 llamas. PROCEDURE: Strips were mounted in tissue baths containing oxygenated Kreb's buffer solution and connected to a polygraph chart recorder to measure contractile activity. Atropine, guanethidine, and indomethacin were added to tissue baths to inhibit muscarinic receptors, adrenoreceptors, and prostaglandin synthesis. Responses to electrical field stimulation following addition of the nitric oxide antagonist Nwo-nitro-L-arginine methyl ester (L-NAME) and apamin were evaluated. RESULTS: Electrical field stimulation (EFS) resulted in a reduction in the amplitude and frequency of contractile activity, followed by rebound contraction when EFS was stopped. Addition of L-NAME resulted in a significant reduction in inhibition of contractile activity. Addition of apamin also resulted in a significant reduction in inhibitory contractile activity at most stimulation frequencies. The combination of L-NAME and apamin resulted in a significant reduction in inhibition at all frequencies. CONCLUSION: Nitric oxide and a transmitter acting via an apamin-sensitive mechanism appear to be involved in inhibition of contractile activity of the third compartment in llamas. CLINICAL RELEVANCE: Results suggest that nitric oxide plays an important role in mediating contractile activity of the third compartment in llamas. Use of nitric oxide synthase inhibitors may have a role in the therapeutic management of llamas with lesions of the third compartment.