T. Mutsvangwa

Effects of Monensin and Dietary Soybean Oil on Milk Fat Percentage and Milk Fatty Acid Profile in Lactating Dairy Cows

The objective of this study was to investigate the effect of monensin (MN) and dietary soybean oil (SBO) on milk fat percentage and milk fatty acid (FA) profile. The study was conducted as a randomized complete block design with a 2 x 3 factorial treatment arrangement using 72 lactating multiparous Holstein dairy cows (138 +/- 24 d in milk). Treatments were [dry matter (DM) basis] as follows: 1) control total mixed ration (TMR, no MN) with no supplemental SBO; 2) MN-treated TMR (22 g of MN/kg of DM) with no supplemental SBO; 3) control TMR including 1.7% SBO; 4) MN-treated TMR including 1.7% SBO; 5) control TMR including 3.4% SBO; and 6) MN-treated TMR including 3.4% SBO. The TMR (% of DM; corn silage, 31.6%; haylage, 21.2%; hay, 4.2%; high-moisture corn, 18.8%; soy hulls, 3.3%; and protein supplement, 20.9%) was offered ad libitum. The experiment consisted of a 2-wk baseline, a 3-wk adaptation, and a 2-wk collection period. Monensin, SBO, and their interaction linearly reduced milk fat percentage. Cows receiving SBO with no added MN (treatments 3 and 5) had 4.5 and 14.2% decreases in milk fat percentage, respectively. Cows receiving SBO with added MN (treatments 4 and 6) had 16.5 and 35.1% decreases in milk fat percentage, respectively. However, the interaction effect of MN and SBO on fat yield was not significant. Monensin reduced milk fat yield by 6.6%. Soybean oil linearly reduced milk fat yield and protein percentage and linearly increased milk yield and milk protein yield. Monensin and SBO reduced 4% fat-corrected milk and had no effect on DM intake. Monensin interacted with SBO to linearly increase milk fat concentration (g/100 g of FA) of total trans-18:1 in milk fat including trans-6 to 8, trans-9, trans-10, trans-11, trans-12 18:1 and the concentration of total conjugated linoleic acid isomers including cis-9, trans-11 18:2; trans-9, cis-11 18:2; and trans-10, cis-12 18:2. Also, the interaction increased milk concentration of polyunsaturated fatty acids. Monensin and SBO linearly reduced, with no significant interaction, milk concentration (g/100 g of FA) of short- and medium-chain fatty acids (

Influence of Carbohydrate Source on Ruminal Fermentation Characteristics, Performance, and Microbial Protein Synthesis in Dairy Cows

Eight multiparous Holstein cows (676 +/- 57 kg of body weight; 121 +/- 17 d-in-milk) were used in a replicated 4 x 4 Latin square design to determine the effects of 4 sources of carbohydrate on milk yield and composition, ruminal fermentation, and microbial N flow to the duodenum. Four cows in one of the Latin squares were fitted with permanent ruminal cannulae. Diets contained (DM basis) 50% forage in combinations of alfalfa hay and barley silage, and 50% concentrate. The concentrate portion of the diets contained barley, corn, wheat, or oats grain as the primary source of carbohydrate. Intake of DM ranged from 24.0 to 26.2 kg/d, and it tended to be lower in cows fed the wheat-based diet compared with those fed the barley-based diet; consequently, milk yield tended to be lower in cows fed the wheat-based diet compared with those fed the barley-based diet. Cows fed the barley- or wheat-based diets had a lower milk fat content compared with those fed the corn-based diet. Ruminal fermentation characteristics were largely unaffected by the source of dietary carbohydrate, with similar ruminal pH and volatile fatty acid and ammonia concentrations for the first 6 h after the morning feeding. Dietary treatment did not affect total tract apparent digestibility of DM, organic matter, and neutral detergent fiber; however, total tract apparent digestibility of starch in cows fed the oats-based diet was higher compared with those fed the corn-and wheat-based diets. Nitrogen that was used for productive purposes (i.e., N secreted in milk + N apparently retained by the cow) tended to be lower in cows fed the wheat-based diet compared with cows fed the barley-, corn-, or oats-based diets. Urinary purine derivative (PD) excretion was similar in cows fed the barley-, corn-, and wheat-based diets; however, purine derivative excretion was higher in cows fed the barley-based diet compared with those fed the oats-based diet. Consequently, estimated microbial N flow to the duodenum was 49 g/d higher in cows fed the barley-based diet compared with those fed the oats-based diet. Improved production performance with corn and barley diets appeared to be due to greater nutrient absorption than in cows fed oats and wheat diets, rather than improved nutrient utilization efficiency.

Effects of Peripartum Propylene Glycol Supplementation on Nitrogen Metabolism, Body Composition, and Gene Expression for the Major Protein Degradation Pathways in Skeletal Muscle in Dairy Cows

Early-lactating dairy cows mobilize body protein to provide amino acids that are directed toward gluconeogenesis and milk protein synthesis. Propylene glycol (PG) is a precursor of ruminal propionate, and feeding PG has been reported to improve energy supply by increasing blood glucose. Our hypothesis was that feeding PG could spare body protein by providing an alternative source of carbon for gluconeogenesis. The major objectives of this study were 1) to delineate the effects of pre- and postpartum PG supplementation in transition dairy cows on whole-body nitrogen balance, urinary 3-methylhistidine (3-MH) excretion, body composition, and gene expression profiles for the major protein degradation pathways in skeletal muscle; and 2) to characterize the changes in body protein metabolism during the periparturient period. Sixteen pregnant cows (7 primiparous and 9 multiparous) were paired based on expected calving dates and then randomly assigned within each pair to either a basal diet (control) or basal diet plus 600 mL/d of PG. Diets were fed twice daily for ad libitum intake, and PG was fed in equal amounts as a top dress from d -7 to d 45. All measurements were conducted at 3 time intervals starting at d -14 +/- 5, d 15, and d 38 relative to calving. Propylene glycol had no effect on whole-body N balance, urinary 3-MH excretion, or body composition. However, N balance was lower at d 15 and 38, compared with d -14. Urinary excretion of 3-MH was lower at d -14 than at d 15 and 38. Supplemental PG had no effect on body weight (BW) and all components of empty BW. On average, cows fed both diets mobilized 19 kg of body fat and 14 kg of body protein between d -14 and d 38. Supplemental PG had no effect on mRNA abundance in skeletal muscle for m-calpain, and the 14-kDa ubiquitin-carrier protein E2 (14-kDa E2) and proteasome 26S subunit-ATPase components of the ubiquitin-mediated proteolytic pathway; however, PG supplementation downregulated mRNA expression for mu-calpain at d 15, and tended to downregulate mRNA expression for ubiquitin at d 15 and 38. Relative to calving, mRNA abundance for m- and mu-calpain, ubiquitin, and 14-kDa E2 were greater at d 15 compared with d -14 and d 38. In summary, these results indicate that transitional effects on whole-body metabolism and gene expression for the Ca(2+)-dependent and ubiquitin-mediated proteolytic pathways in skeletal muscle were more pronounced than those elicited by PG supplementation.

Supplemental butyrate does not enhance the absorptive or barrier functions of the isolated ovine ruminal epithelia1

Our objective was to determine if increasing the ruminal butyrate concentration would improve the selective permeability of ruminal epithelia. Suffolk wether lambs (n = 18) with an initial BW of 47.4 ±1.4 kg were housed in individual pens (1.5 × 1.5 m) with rubber mats on the floor. Lambs were blocked by initial BW into 6 blocks and, within block, were randomly assigned to either the control (CON) or 1 of 2 butyrate supplementation amounts (i.e., 1.25% or 2.50% butyrate as a proportion of DMI). With the exception of butyrate supplementation, all lambs were fed a common diet (90% concentrate and 10% barley silage). After a 14-d feeding period, lambs were killed, and ruminal epithelia from the ventral sac were mounted in Ussing chambers. To facilitate the Ussing chamber measurements, only 1 lamb was killed on an individual day. Thus, the starting date was staggered so that all lambs were exposed to the same experimental protocol. In Ussing chambers, epithelia were incubated using separate mucosal (pH 6.2) and serosal (pH 7.4) bathing solutions. Then 1-14C-butyrate (74 kBq/10 mL) was added to the mucosal side and was used to measure the mucosal-to-serosal flux (J(ms-butyrate)) in 2 consecutive 60-min flux periods with simultaneous measurement of transepithelial conductance (G(t)). During the first (challenge) flux period, the mucosal buffer solution was either acidified to pH 5.2 (ACID) or used as a control (pH 6.2; SHAM). Buffer solutions bathing the epithelia were replaced before the second flux period (recovery). Total ruminal short-chain fatty acid and butyrate concentrations were greater (P = 0.001) in lambs fed 2.50% compared with those fed 0% or 1.25% butyrate. The J(ms-butyrate) was less for lambs fed 1.25% and 2.50% butyrate [3.00 and 3.12 μmol/(cm2·h), respectively] than for CON [3.91 μmol/(cm2· h)]. However, no difference (P = 0.13)was observed for G(t). An ex vivo treatment × flux period interaction was detected (P = 0.003) for J(ms-butyrate), where no differences were present between ACID and SHAM during the challenge period, but the Jms-butyrate was less for ACID than for SHAM during recovery. These results indicate that large increases in the ruminal butyrate concentration decrease the selective permeability of the isolated ruminal epithelia.

Effects of partial ruminal defaunation on urea-nitrogen recycling, nitrogen metabolism, and microbial nitrogen supply in growing lambs fed low or high dietary crude protein concentrations

Urea-nitrogen recycling to the gastrointestinal tract (GIT), N metabolism, and urea transporter-B (UT-B) mRNA abundance in ruminal epithelium were evaluated in partially defaunated (PDFAUN) and faunated (FAUN) growing lambs fed 2 levels (10%, low, or 15%, high) of dietary CP (DM basis). Four Suffolk ram lambs (43.9 +/- 1.4 kg initial BW) were used in a 4 x 4 Latin square design with 27-d periods. Sunflower oil was fed (6%; DM basis) as an anti-protozoal agent. Nitrogen balance was measured from d 22 to 26, with concurrent measurement of urea-N kinetics using continuous intrajugular infusions of [(15)N(15)N]-urea. Feeding sunflower oil decreased (P < 0.01) total ruminal protozoa by 88%, and this was associated with a decrease (P < 0.01) in ruminal ammonia-N concentrations. Endogenous production of urea-N (UER; 26.1 vs. 34.6 g/d) and urea-N loss in urine (UUE; 10.1 vs. 15.7 g/d) were less (P < 0.01), and urea-N entering the GIT (GER; 16.0 vs. 18.9 g/d) tended to be less (P = 0.06) in PDFAUN as compared with FAUN lambs. However, as a proportion of UER, GER was greater (P < 0.01) and the proportion of recycled urea-N that was utilized for anabolism (i.e., UUA) tended to be greater (P = 0.09) in PDFAUN lambs. Partial defaunation increased (P < 0.01) microbial N supply. The UER, GER, and UUE were greater (P < 0.01) in lambs fed the high diet. However, as a proportion of UER, GER and its anabolic use were greater (P < 0.01) in lambs fed the low diet. The expression of UT-B mRNA in PDFAUN lambs was numerically greater (by 20%; P = 0.15) compared with FAUN lambs. In summary, results indicate that part of the mechanism for improved N utilization in defaunated ruminants is an increase in the proportion of endogenous urea-N output that is recycled to the GIT, thus potentially providing additional N for microbial growth.

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.

Effects of partial replacement of dietary starch from barley or corn with lactose on ruminal function, short-chain fatty acid absorption, nitrogen utilization, and production performance of dairy cows.

In cows fed diets based on corn-alfalfa silage, replacing starch with sugar improves milk production. Although the rate of ruminal fermentation of sugar is more rapid than that of starch, evidence has been found that feeding sugar as a partial replacement for starch does not negatively affect ruminal pH despite increasing diet fermentability. The mechanism(s) for this desirable response are unknown. Our objective was to determine the effects of replacing barley or corn starch with lactose (as dried whey permeate; DWP) on ruminal function, short-chain fatty acid (SCFA) absorption, and nitrogen (N) utilization in dairy cows. Eight lactating cows were used in a replicated 4 × 4 Latin square design with 28-d periods and source of starch (barley vs. corn) and level of DWP (0 vs. 6%, DM basis) as treatment factors. Four cows in 1 Latin square were ruminally cannulated for the measurement of ruminal function, SCFA absorption, and N utilization. Dry matter intake and milk and milk component yields did not differ with diet. The dietary addition of DWP tended to increase ruminal butyrate concentration (13.6 vs. 12.2 mmol/L), and increased the Cl(-)-competitive absorption rates for acetate and propionate. There was no sugar effect on minimum ruminal pH, and the duration and area when ruminal pH was below 5.8. Minimum ruminal pH tended to be lower in cows fed barley compared with those fed corn (5.47 vs. 5.61). The duration when ruminal pH was below pH 5.8 tended to be shorter (186 vs. 235 min/d), whereas the area (pH × min/d) that pH was below 5.8 was smaller (47 vs. 111) on the corn than barley diets. Cows fed the high- compared with the low-sugar diet had lower ruminal NH3-N concentration. Feeding the high-sugar diet tended to increase apparent total-tract digestibility of dry matter and organic matters and increased apparent total-tract digestibility of fat. Apparent total-tract digestibility of N tended to be greater in cows fed barley compared with those fed corn, whereas apparent total-tract digestibility of acid-digestible fiber was greater in cows fed corn compared with those fed barley. In conclusion, partially replacing dietary corn or barley starch with sugar upregulated ruminal acetate and propionate absorption, suggesting that the mechanisms for the attenuation of ruminal acidosis when sugar is fed is partly mediated via increased SCFA absorption.

Short communication: Interrelationship between butyrate and glucose supply on butyrate and glucose oxidation by ruminal epithelial preparations.

The aim of this study was to determine whether dietary Na-butyrate supplementation affects butyrate and glucose oxidation by ruminal epithelial preparations and whether this effect can be acutely modulated by substrate (glucose and butyrate) supply. Eighteen Suffolk wether lambs (6 lambs/treatment) were blocked by body weight and, within block, randomly assigned to the control treatment (CON) or to diets containing differing Na-butyrate inclusion rates (1.58 or 3.16%) equating to 1.25 (B1.25), and 2.50% (B2.50) butyrate on a dry matter basis, respectively. All lambs received their diet for a period of 14 d. After dietary adaptation, lambs were killed and the ruminal epithelium was harvested from the ventral sac, minced finely, and used for in vitro incubations. Incubation medium contained either a constant concentration of glucose (4 mM) with increasing butyrate concentrations (0, 5, 15, 25, or 40 mM) or a constant butyrate concentration (15 mM) with increasing glucose concentrations (0, 1, 2, 4, or 8 mM) to allow for the evaluation of whether acute changes in the concentration of metabolic substrates affect the oxidation of glucose and butyrate. We observed no interactions between the in vivo and in vitro treatments. Increasing dietary butyrate supplementation linearly decreased glucose oxidation by ruminal epithelial preparations, but had no effect on butyrate oxidation. Increasing butyrate concentration in vitro decreased (cubic effect) glucose oxidation when butyrate concentration ranged between 5 and 15 mM; however, glucose oxidation was increased with a butyrate concentration of 40 mM. Butyrate oxidation decreased (cubic effect) as glucose concentration increased from 1 to 4 mM; however, butyrate oxidation increased when glucose was included at 8mM. The results of this study demonstrate that dietary butyrate supplementation can decrease glucose oxidation by the ruminal epithelium, but the relative supply of glucose and butyrate has a pronounced effect on substrate oxidation.

Serosal-to-mucosal urea flux across the isolated ruminal epithelium is mediated via urea transporter-B and aquaporins when Holstein calves are abruptly changed to a moderately fermentable diet

Urea transport (UT-B) proteins are known to facilitate urea movement across the ruminal epithelium; however, other mechanisms may be involved as well because inhibiting UT-B does not completely abolish urea transport. Of the aquaporins (AQP), which are a family of membrane-spanning proteins that are predominantly involved in the movement of water, AQP-3, AQP-7, and AQP-10 are also permeable to urea, but it is not clear if they contribute to urea transport across the ruminal epithelium. The objectives of this study were to determine (1) the functional roles of AQP and UT-B in the serosal-to-mucosal urea flux (Jsm-urea) across rumen epithelium; and (2) whether functional adaptation occurs in response to increased diet fermentability. Twenty-five Holstein steer calves (n=5) were assigned to a control diet (CON; 91.5% hay and 8.5% vitamin and mineral supplement) or a medium grain diet (MGD; 41.5% barley grain, 50% hay, and 8.5% vitamin and mineral) that was fed for 3, 7, 14, or 21 d. Calves were killed and ruminal epithelium was collected for mounting in Ussing chambers under short-circuit conditions and for analysis of mRNA abundance of UT-B and AQP-3, AQP-7, and AQP-10. To mimic physiologic conditions, the mucosal buffer (pH 6.2) contained no urea, whereas the serosal buffer (pH 7.4) contained 1 mM urea. The fluxes of (14)C-urea (Jsm-urea; 26 kBq/10 mL) and (3)H-mannitol (Jsm-mannitol; 37 kBq/10 mL) were measured, with Jsm-mannitol being used as an indicator of paracellular or hydrophilic movement. Serosal addition of phloretin (1 mM) was used to inhibit UT-B-mediated urea transport, whereas NiCl2 (1 mM) was used to inhibit AQP-mediated urea transport. Across treatments, the addition of phloretin or NiCl2 reduced the Jsm-urea from 116.5 to 54.0 and 89.5 nmol/(cm(2) × h), respectively. When both inhibitors were added simultaneously, Jsm-urea was further reduced to 36.8 nmol/(cm(2) × h). Phloretin-sensitive and NiCl2-sensitive Jsm-urea were not affected by diet. The Jsm-urea tended to increase linearly as the duration of adaptation to MGD increased, with the lowest Jsm-urea being observed in animals fed CON [107.7 nmol/(cm(2) × h)] and the highest for those fed the MGD for 21 d [144.2 nmol/(cm(2) × h)]. Phloretin-insensitive Jsm-urea tended to increase linearly as the duration of adaptation to MGD increased, whereas NiCl2-insensitive Jsm-urea tended to be affected by diet. Gene transcript abundance for AQP-3 and UT-B in ruminal epithelium increased linearly as the duration of MGD adaptation increased. For AQP-7 and AQP-10, gene transcript abundance in animals that were fed the MGD was greater compared with that of CON animals. These results demonstrate that both AQP and UT-B play significant functional roles in urea transport, and they may play a role in urea transport during dietary adaptation to fermentable carbohydrates.

Effects of dietary crude protein and rumen-degradable protein concentrations on urea recycling, nitrogen balance, omasal nutrient flow, and milk production in dairy cows

The objective of this study was to determine how interactions between dietary crude protein (CP) and rumen-degradable protein (RDP) concentrations alter urea-nitrogen recycling, nitrogen (N) balance, omasal nutrient flow, and milk production in lactating Holstein cows. Eight multiparous Holstein cows (711±21kg of body weight; 91±17d in milk at the start of the experiment) were used in a replicated 4×4 Latin square design with a 2×2 factorial arrangement of dietary treatments and 29-d experimental periods. Four cows in one Latin square were fitted with ruminal cannulas to allow ruminal and omasal sampling. The dietary treatment factors were CP (14.9 vs. 17.5%; dry matter basis) and RDP (63 vs. 69% of CP) contents. Dietary RDP concentration was manipulated by including unprocessed or micronized canola meal. Diet adaptation (d 1-20) was followed by 8d (d 21-29) of sample and data collection. Continuous intrajugular infusions of [(15)N(15)N]-urea (220mg/d) were conducted for 4d (d 25-29) with concurrent total collections of urine and feces to estimate N balance and whole-body urea kinetics. Proportions of [(15)N(15)N]- and [(14)N(15)N]-urea in urinary urea, and (15)N enrichment in feces were used to calculate urea kinetics. For the low-CP diets, cows fed the high-RDP diet had a greater DM intake compared with those fed the low-RDP diet, but the opposite trend was observed for cows fed the high-CP diets. Dietary treatment had no effect on milk yield. Milk composition and milk component yields were largely unaffected by dietary treatment; however, on the low-CP diets, milk fat yield was greater for cows fed the low-RDP diet compared with those fed the high-RDP diet, but it was unaffected by RDP concentration on the high-CP diets. On the high-CP diets, milk urea nitrogen concentration was greater in cows fed the high-RDP diet compared with those fed the low-RDP diet, but it was unaffected by RDP concentration on the low-CP diets. Ruminal NH3-N concentration tended to be greater in cows fed the high-CP diet compared with those fed the low-CP diet, and it was greater in cows fed the high-RDP diet as compared with those fed the low-RDP diet. Nitrogen intake and both total N and urea-N excretion in urine were greater for cows fed the high-CP diet compared with those fed the low-CP diet. However, N balance and urinary excretion of purine derivatives were unaffected by dietary treatment. Urea-N entry rate (UER) was greater in cows fed the high-CP diet compared with those fed the low-CP diet; however, UER was unaffected by dietary RDP concentration. The proportion of urea-N recycled to the gastrointestinal tract (as a percentage of UER) was greater in cows fed the low-CP diet compared with those fed the high-CP diet. In summary, reducing dietary CP concentration decreased urinary N excretion but had no effect on milk yield, thus resulting in an overall improvement in milk N efficiency.