Effects of canola meal inclusion rate in starter mixtures for Holstein heifer calves on dry matter intake, average daily gain, ruminal fermentation, plasma metabolites, and total-tract digestibility.

The objective of this study was to determine the effects of the canola meal (CM) inclusion rate in pelleted starter mixtures for Holstein heifer calves on dry matter intake, average daily gain, ruminal fermentation, plasma metabolites, and total-tract digestibility. Fifty Holstein heifer calves were blocked by birth date and body weight and, within block, randomly assigned to 1 of 5 pelleted starter treatments with 0, 15, 30, 45, or 60% of the crude protein supplied by CM instead of soybean meal (SBM). Pellets were formulated to be similar in crude protein (24.3%), starch (26.6%), and neutral detergent fiber (17.8%) and were provided to calves starting on d 8 of age, with starter intake measured daily. From 8.0 ± 0.0 (mean ± standard deviation) d of age through d 35.3 ± 2.4, calves were fed milk replacer at 15% of body weight, offered in 3 equal feedings at 0600, 1500, and 2100 h. After that, a gradual 21-d step-down weaning process was imposed, where no further milk replacer was provided starting on d 57.0 ± 0.0. Data for milk replacer and starter intake were calculated to determine weekly averages. On d 62.2 ± 0.8 of age, blood was collected every 4 h and analyzed for glucose, ß-hydroxybutyrate, insulin, and urea concentrations. From d 66.2 ± 0.8 of age and extending for 3 d, fecal samples were collected every 12 h with a 3-h daily offset, to estimate fecal nutrient output and to determine apparent total-tract digestibility. Additionally, ruminal fluid (d 70.2 ± 0.8 of age) was sampled at 1300 h through an esophageal tube connected to a vacuum pump. The pH of ruminal fluid was measured, and ruminal fluid was analyzed to determine short-chain fatty acid and ammonia concentrations. Data were analyzed with fixed effect of treatment and random effect of block. Polynomial contrasts were calculated to assess linear, quadratic, and cubic effects with repeated measures statement for variables analyzed over time. Starter intake, average daily gain, body weight, and feed efficiency did not differ among treatments. Crude protein and ether extract digestibility were affected in a cubic manner, where CP was greatest for CM0, CM30, and CM45, and ether extract digestibility was least for CM15 and CM60. The molar proportion of acetate responded cubically, but the proportions of propionate and butyrate did not differ among treatments. Ruminal ammonia and plasma urea concentrations were not affected by CM inclusion rate. In conclusion, CM can replace up to 60% of the CP provided from SBM without affecting starter intake and growth of calves.

Canola meal or soybean meal as protein source and the effect of microencapsulated sodium butyrate supplementation in calf starter mixture. I. Performance, digestibility, and selected blood variables.

Two studies were conducted to assess the effect of protein source and microencapsulated sodium butyrate (MSB) inclusion in pelleted starter mixtures on growth performance, gain to feed (G:F) ratio, nutrient digestibility, and selected blood metabolites in calves. In study 1, 28 Holstein bull calves (8.7 ± 0.8 d of age and 43.0 ± 4.4 kg; mean ± SD) were allocated to 1 of 4 treatments in a 2 × 2 factorial arrangement and fed a pelleted starter mixture containing canola meal (CM, 35% as fed) or soybean meal (SM, 24% as fed) as the main source of protein, with or without supplemental MSB (0.3% as fed). Starter mixtures were formulated to be similar for crude protein, Lys, and Met, and were fed ad libitum. Calves were weaned after 42 d of milk replacer feeding (51.7 ± 0.8 d of age) and observed for another 21 d. Furthermore, selected blood metabolites were measured on d 21, 42, and 63 of the study, and nutrient digestibility was measured after weaning. In study 2, 60 Holstein heifer calves (9.1 ± 0.8 d of age and 43.2 ± 4.2 kg) were assigned to the same treatments as in study 1. The calves were weaned after 49 d of milk replacer feeding (59.1 ± 0.8 d of age) and observed for an additional 14 d. Milk replacer and starter mixture intake and fecal score were recorded daily, whereas body weight (BW) was recorded weekly. In study 1, calves fed starter mixtures containing CM had or tended to have lesser preweaning starter intake, weaning average daily gain (ADG), weaning and overall G:F ratio, and postweaning total-tract dry matter digestibility, as opposed to those fed starter mixtures with SM. However, these differences did not affect overall starter intake, overall ADG, or final BW. Supplementation with MSB only tended to increase the preweaning starter mixture intake. In study 2, heifer calves that were fed starter mixtures with CM had greater cumulative starter intake after weaning, but the protein source in the starter mixture had no effect on ADG, BW, or G:F ratio. Inclusion of MSB in starter mixtures for calves tended to decrease postweaning starter mixture intake. In conclusion, use of CM or SM as the main source of protein in starter mixture resulted in similar growth performance of bull and heifer calves; however, CM use in starter mixtures reduced starter intake, ADG, and G:F ratio at least at some points of rearing. Supplementation of MSB had minor effects on the growth performance of calves.

Canola meal or soybean meal as protein source and the effect of microencapsulated sodium butyrate supplementation in calf starter mixture. II. Development of the gastrointestinal tract.

The aim of this study was to assess the effect of protein source, either soybean meal (SM) or canola meal (CM), and microencapsulated sodium butyrate (MSB) supplementation in a pelleted starter mixture on the development of the gastrointestinal tract (GIT) in dairy calves. Twenty-eight bull calves (8.7 ± 0.8 d of age and 43.0 ± 4.4 kg; mean ± SD) were assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement: CM as a main source of protein without or with MSB or SM without or with MSB. Calves were fed starters ad libitum and exposed to a gradual weaning program, with weaning taking place on 51.7 ± 0.8 d of age. Calves were observed for an additional 3 wk after weaning and slaughtered on d 72.1 ± 0.9 of age, after which the GIT was dissected. Morphometric measurements were recorded, and samples for determination of ruminal fermentation, histology, gene expression, and brush border enzyme activities were collected. Canola meal use in the starter mixture increased abomasal tissue weight, jejunal tissue weight and length, and mRNA expression of SLC16A4 (formerly known as MCT4) and FFAR2 (GPR43) in the ruminal epithelium, and decreased ruminal ammonia and mRNA expression of SLC15A2 (PEPT2) and SLC6A14 (ATB0+) in the proximal small intestine and ileum, respectively. However, MSB inclusion in the starter mixture decreased ruminal papillae length, ruminal epithelial surface, and ruminal epithelium dry weight, while increasing mRNA expression of SLC16A1 (MCT1) in ruminal epithelia. Reduced ruminal surface area associated with MSB supplementation was the most apparent when MSB was combined with CM in the starter mixture. Additionally, MSB supplementation decreased the thickness of omasal epithelium, omasal epithelium living strata, and stratum corneum, and increased duodenal and ileal aminopeptidase A enzymatic activity and ileal aminopeptidase N enzymatic activity. Overall, CM might increase growth of the GIT of calves, particularly of the small intestine, but may negatively affect intestinal epithelium function and peptide and AA absorption. Supplementation of MSB has a negative effect on the ruminal and omasal epithelium development, particularly when combined in a starter mixture with CM.

Effect of heat-treated canola meal and glycerol inclusion on performance and gastrointestinal development of Holstein calves.

The objectives of this study were to assess the effect of using heat-treated canola meal (CM) and glycerol inclusion in starter mixtures on starter intake, growth, and gastrointestinal tract development in Holstein bull calves. In the first study, a protocol for the heat treatment of CM was evaluated by comparing commercial CM that was exposed to 0, 100, 110, or 120°C of heat treatment for 10 min. Following heat treatment, in situ crude protein (CP) ruminal degradability and estimated intestinal CP digestibility were assessed. It was observed that the degradable fractions of dry matter and CP in CM decreased linearly with increasing temperature of heat treatment. The estimated intestinal CP digestibility was greatest when CM was heated to 110°C. In the second study, 28 bull calves were used in a randomized complete block design. Calves were fed pelleted starters containing CM or CM that was heat-treated to 110°C for 10 min. Diets also contained 0 or 5% glycerol on a dry matter basis. The study lasted 51 d, ending on the first day of weaning. Starter intake, average daily gain (ADG), ruminal short-chain fatty acid concentrations, morphology of the rumen and small intestine, gene expression (MCT1, GPR41, GPR43, UTB, AQP3, PEPT1, PEPT2, ATB0+, and EAAC1) in the ruminal, jejunal, and ileal epithelium, and brush border enzyme activities in the duodenum, jejunum, and ileum were investigated. Few interactions between heat-treated CM and glycerol inclusion were observed. Feeding heat-treated CM did not affect starter intake. However, feeding heat-treated CM to calves tended to reduce ADG and decreased the weight of ruminal and jejunal tissue. Heat treatment did not affect gene expression or brush border enzyme activities in the small intestine. Glycerol inclusion tended to increase cumulative starter intake and increased cumulative body weight gain. Use of glycerol reduced ruminal pH and increased the concentration of ruminal short-chain fatty acids. Additionally, glycerol inclusion increased abomasal, duodenal, jejunal, and cecal digesta weights and tended to increase the weight of the jejunal tissue. Glycerol supplementation tended to downregulate the expression of MCT1 in the ruminal epithelium, and upregulated the expression of MCT1 in the epithelium of proximal jejunum. In conclusion, heat treatment of CM may negatively affect calf growth and gastrointestinal tract development. Glycerol inclusion may increase starter intake, ADG, ruminal fermentation, and intestinal development in calves when CM is used as a main source of protein in pelleted starter mixture.

The effect of docosahexaenoic acid-rich algae supplementation in milk replacer on performance and selected immune system functions in calves.

The aim of this study was to determine the effect of docosahexaenoic acid-rich algae (DHA-RA) supplementation in milk replacer (MR) on performance, selected cytokine expression in lymphocytes, and blood immunoglobulin concentration in newborn dairy calves. Forty female Holstein-Friesian calves (8.6 ± 0.8 d old and 41.1 ± 4.3 kg; mean ± standard deviation) were blocked by date of birth and allocated into 4 experimental groups (10 animals/group): (1) not supplemented with DHA-RA, (2) supplemented with 9 g of DHA-RA/d in MR, (3) supplemented with 18 g of DHA-RA/d in MR, and (4) supplemented with 27 g of DHA-RA/d in MR. Milk replacer was fed in an amount equal to 900 g of MR powder/d (as fed), 2 times a d, for 49 d. Starter mixture (SM) was fed ad libitum beginning on d 15 of the study. Each calf was in the study over a period of 49 d. The MR and SM intake and fecal score were recorded daily and body weight was recorded weekly. Blood samples were collected before the morning feeding, at the beginning of the study, every consecutive week, and at the end of the study for morphology and smear analysis, serum immunoglobulin level (IgG, IgA, and IgM), and lymphocyte isolation. The mRNA isolated from lymphocytes was checked for TNFα, IL-1ß, IL-6, and p65 expression. Average daily gain between d 1 to 14 of the study increased quadratically with increasing dose of DHA-RA. However, average daily gain between d 15 to 49 of the study tended to linearly decrease and over the whole study linearly decreased with increasing dose of DHA-RA. The MR intake decreased linearly between d 1 to 14 of the study and over the whole study, and mean SM intake decreased quadratically with increasing dose of DHA-RA. Feed efficiency increased quadratically and fecal score decreased quadratically during the first 14 d of the study. Increasing dose of DHA-RA led to cubic changes in feed efficiency and fecal score between d 15 and 49 of the study. Overall, over the whole study period a tendency was observed for lower fecal score for the DHA-RA supplemented groups. Interleukin-1ß mRNA expression decreased linearly, whereas the mRNA expression of p65 and TNFα as well as serum IgG concentration tended to decrease linearly with increasing dose of supplemental DHA-RA. No effect of group was found on IgA and IgM serum level and the majority of blood parameters. Altogether, treatment worsened production variables but seemed to have a beneficial effect on the immune system of calves.

Invited review: Use of butyrate to promote gastrointestinal tract development in calves.

Promotion of microbial butyrate production in the reticulorumen is a widely used method for enhancing forestomach development in calves. Additional acceleration of gastrointestinal tract (GIT) development, both the forestomach and lower parts of the GIT (e.g., abomasum, intestine, and also pancreas), can be obtained by dietary butyrate supplementation. For this purpose, different sources (e.g., butyrate salts or butyrins), forms (e.g., protected or unprotected), methods (e.g., in liquid feed or solid feed), and periods (e.g., before or after weaning) of butyrate administration can be used. The aim of this paper was to summarize the knowledge in the field of butyrate supplementation in feeds for newborn calves in practical situations, and to suggest directions of future studies. It has been repeatedly shown that supplementation of unprotected salts of butyrate (primarily sodium salt) in milk replacer (MR) stimulates the rumen, small intestine, and pancreas development in calves, with a supplementation level equating to 0.3% of dry matter being sufficient to exert the desired effect on both GIT development and growth performance. On the other hand, the effect of unprotected butyrins and protected forms of butyrate supplementation in MR has not been extensively investigated, and few studies have documented the effect of butyrate addition into whole milk (WM), with those available focusing mainly on the growth performance of animals. Protected butyrate supplementation at a low level (0.3% of protected product in DM) in solid feed was shown to have a potential to enhance GIT development and performance of calves fed MR during the preweaning period. Justification of this form of butyrate supplementation in solid feed when calves are fed WM or after weaning needs to be documented. After weaning, inclusion of unprotected butyrate salts in solid feed was shown to increase solid feed intake, but the effect on GIT development and function has not been determined in detail, and optimal levels of supplementation are also difficult to recommend based on available reports. Future studies should focus on comparing different sources (e.g., salts vs. esters), forms (e.g., protected vs. unprotected), and doses of supplemental butyrate in liquid feeds and solid feeds and their effect not only on the development of rumen, abomasum, and small intestine but also the omasum and large intestine. Furthermore, the most effective source, form, and dose of supplemental butyrate in solid feed depending on the liquid feed program (e.g., MR or WM), stage of rearing (e.g., pre- or postweaning), and solid composition (e.g., lack or presence of forage in the diet) need to be determined.

Age-related changes in mRNA expression of selected surface receptors in lymphocytes of dairy calves.

The aim of this study was to determine age-related changes in the mRNA expression of four clusters of differentiation (CD: e.g. CD5, CD21, CD22 and CD23) in lymphocytes of calves. Blood samples were collected from 10 Holstein heifers on day 2, 22 and 56 of life and used for lymphocyte isolation. Subsequently, the mRNA was isolated from lymphocytes and the relative expression of CD5, CD21, CD22 and CD23 was investigated using quantitative real-time PCR with GAPDH as a reference gene. CD5, CD21 and CD23 mRNA expression increased linearly (p ≤ 0.04) with calf age, whereas CD22 mRNA expression did not change in the investigated period (p > 0.05). Age related changes in CD5, CD21 and CD23 mRNA expression suggest their importance in the process of lymphocyte maturation in calves.

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.

Short communication: Effect of canola meal use as a protein source in a starter mixture on feeding behavior and performance of calves during the weaning transition.

The aim of this study was to determine the effect of canola meal use as a protein source in a starter mixture (SM) on feeding behavior and performance of calves during weaning transition. A total of 36 female Holstein calves of a mean age 14.9±1.6 d and body weight 40.1±4.2 kg (mean ± SD) were allocated to 1 of 3 treatments differing in the main source of protein for the SM (12 calves per treatment): (1) soybean meal (TSBM); (2) soybean meal and canola meal (TSBM/TCM); and (3) canola meal (TCM). The SM was offered for ad libitum consumption beginning on the first day of the study, whereas milk replacer (MR) was fed in amounts equal to 900 g (as fed) per day from d 1 to 35 and 450 g/d from d 36 to 42 of the study. Calves were completely weaned on d 43 of the study (57.9±1.6 d of age; mean ± SD), and their performance was monitored for an additional 2 wk. Calf body weight was recorded weekly, and MR and SM intake and fecal fluidity were recorded daily. Feeding behavior of calves during weaning transition, including frequency (no./d), time (min/d), and rate (g/min) of eating the SM as well as frequency and time of drinking water, was monitored on 6 calves per treatment for 2 consecutive days before MR step-down (d 34-35), at MR step-down (d 41-42), and after weaning (d 48-49 of study). Starter mixture intake tended to be higher for TSBM calves as compared with TSBM/TCM calves from d 1 to 35 of the study but was not different between TSBM and TCM calves and was not different between treatments in the whole study period. Calves from TCM treatment had reduced average daily gain (ADG) and feed efficiency (g of ADG/kg of dry matter intake) and a higher fecal score in the period from d 1 to 35 of the study and had lower feed efficiency and tended to have lower ADG in the whole study period as compared with TSBM calves. Average daily gain and feed efficiency did not differ between TSBM and TSBM/TCM calves. Frequency of eating the SM and drinking water as well as time and rate of eating the SM and time of drinking water did not differ between treatments. It is concluded that presence of canola meal in a SM does not affect feeding behavior and performance of calves during weaning transition but has a negative effect on ADG, feed efficiency, and number of days with diarrhea during the preweaning phase of rearing.

Short communication: Effect of inclusion rate of microencapsulated sodium butyrate in starter mixture for dairy calves.

The aim of this study was to determine the effect of different inclusion rates of microencapsulated sodium butyrate (M-SB) in the starter mixture (SM) on performance of dairy calves. Forty female Holstein calves with a mean (± SD) age of 12.8 (± 1.5) d were allocated to 1 of 4 treatments (10 calves/treatment) and fed SM without (M-SB-0) or with 0.3% (M-SB-0.3), 0.6% (M-SB-0.6), or 0.9% (M-SB-0.9) of M-SB (as fed) during a 49-d period of milk replacer feeding. The milk replacer was fed at 670 g/d divided into 2 equal meals. Starter mixture with or without M-SB was offered for ad libitum consumption beginning on the first day of the trial. Body weight of calves was recorded weekly, whereas intakes of milk replacer and SM and fecal fluidity were recorded daily. Intake of SM decreased linearly with increasing M-SB inclusion rate. Average daily gain decreased and body weight gain tended to decrease linearly with increasing amounts of M-SB in SM, but feed efficiency was not affected. Fecal score and number of days with diarrhea increased cubically with increasing M-SB inclusion rate in SM. Under the conditions of the current study, supplementation of SM with M-SB had a negative effect on performance of calves.

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.

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.

Effect of microencapsulated sodium butyrate in the close-up diet on performance of dairy cows in the early lactation period.

Two trials were conducted to determine the effect of sodium butyrate microencapsulated within triglyceride matrix (Na-butyrate) in the close-up period on performance of dairy cows and rumen papillae development. In trial 1, 26 Holstein-Friesian cows were randomly allocated to 2 groups (13 cows/group) and fed prepartum a total mixed ration (TMR) without or with 300g of Na-butyrate/d from 30 d before expecting calving to parturition. After calving, the same lactational TMR without Na-butyrate was offered to both treatments. Dry matter intake and milk yield were monitored daily to 60 d in milk, and body condition of cows was scored on d 30, 21, and 4 before parturition and d 14, 31, and 60 after parturition. On d 15, 10, and 5 before parturition blood samples were collected from 6 cows randomly chosen from each group and analyzed for plasma ß-hydroxybutyrate and nonesterified fatty acids concentrations. No differences in dry matter (DM) intake, milk yield, body condition score, or plasma ß-hydroxybutyrate and nonesterified fatty acids concentrations was observed between treatments; however, in the last 5 d before parturition the cows receiving Na-butyrate ate 1.7kg of DM/d more, on average, as compared with control cows. In trial 2, 12 Holstein-Friesian growing bulls (404±48; body weight ± SD) were used to determine the effect of Na-butyrate inclusion in the diet on rumen papillae development. Bulls were randomly allocated to 2 groups (6 bulls/group) and fed TMR without or with 2% (on a dry matter basis) of Na-butyrate for 21 d. At the end of the study, bulls were killed and rumen fluid and rumen tissue samples from dorsal and ventral sac of the rumen were collected. No effect of Na-butyrate supplementation on BW of bulls and DMI during the trial period was observed. Sodium butyrate supplementation increased total short-chain fatty acid concentration in the rumen but had no effect on rumen pH, molar proportions of short-chain fatty acids, and NH3-N concentration. In dorsal sac of the rumen, papillae length and papillae cross-section surface area were increased as a result of Na-butyrate supplementation, whereas in the ventral sac a reverse effect was observed (significant treatment × location in the rumen interaction). Both in the dorsal and ventral sac of the rumen, dietary Na-butyrate increased rumen muscle layer thickness. Altogether, results of this study suggest that Na-butyrate supplementation in the close-up diet may have a potential to enhance rumen papillae growth and rumen adaptation to postpartum diet but lactation performance was not affected under conditions of the current study.

Differences in monocarboxylic acid transporter type 1 expression in rumen epithelium of newborn calves due to age and milk or milk replacer feeding.

The aim of the present study was to investigate whether besides age and solid feed intake, monocarboxylic acid transporter type 1 (MCT1) expression in the rumen epithelium of calves is affected by liquid feed type [whole milk (WM) or milk replacer (MR)]. Thirty bull calves at the mean age of 5 days were randomly allocated to five experimental groups (six calves/group). Six calves were slaughtered immediately after allocation to the trial (5 days of life), eighteen calves were fed MR and slaughtered at week intervals (on 12, 19, 26 days of life respectively), and six calves were fed WM and slaughtered at the 26 days of life. MCT1 protein abundance and the MCT1 mRNA level were investigated in the dorsal and ventral sack of the rumen. Solid feed intake and short-chain fatty acids (SCFA) concentration in the rumen fluid increased linearly with calves' age. The amount of the MCT1 protein and mRNA in the dorsal sac of rumen as well as the amount of MCT1 protein in the cranial ventral sac of rumen also increased linearly with calves' age. Calves fed WM had greater solid feed intake in the last week of the study as compared to calves fed MR, but SCFA concentration in the rumen fluid was not different. MCT1 mRNA expression in the cranial dorsal sac of rumen and protein MCT1 expression in both dorsal and ventral cranial sack of the rumen were higher in calves fed WM as compared to calves fed MR. This study confirmed age-dependent changes of MCT1 expression in the rumen epithelium of newborn calves and showed that its expression might be affected by liquid feed type.

Effect of method of delivery of sodium butyrate on maturation of the small intestine in newborn calves.

The effect of sodium butyrate (SB) supplementation in milk replacer (MR), starter mixture (SM), or both on small intestine maturation in newborn calves was investigated. Twenty-eight male calves with a mean age of 5 (± 1) d were randomly allocated into 1 of 4 groups (7 animals per group) and fed (1) MR and SM, without SB (MR(-) and SM(-), respectively; MR(-)/SM(-)); (2) MR(-) and SM supplemented with SB encapsulated within triglyceride matrix (SM(+), 0.6% as fed; MR(-)/SM(+)); (3) MR supplemented with crystalline SB (MR(+), 0.3% as fed) and SM(-) (MR(+)/SM(-)); or (4) MR(+) and SM(+) (MR(+)/SM(+)). The MR was offered in amounts equal to 10% of initial body weight of the calf. The SM was blended with whole corn grain (50/50; wt/wt) and offered ad libitum as a starter diet. Calves were slaughtered at 26 d (± 1) of age and small intestine development was investigated. Treatment with MR(+) decreased villus height in the proximal jejunum and decreased villus height, crypt depth, and tunica mucosa thickness in the middle jejunum, whereas treatment with SM(+) tended to increase small intestine weight and crypt depth in the proximal jejunum, and increased villus height in the distal jejunum. In the duodenum, crypt depth and tunica mucosa thickness were greater for the MR(-)/SM(+) group compared with MR(-)/SM(-), MR(+)/SM(-), and MR(+)/SM(+) groups. In the ileum, crypt depth was less for MR(-)/SM(+) compared with MR(-)/SM(-). Supplementation with SB in both MR and SM enhanced cell proliferation and decreased apoptosis in the middle jejunum mucosa. Regarding brush border enzyme activities, addition of SB to MR increased lactase activity in the middle jejunum and maltase activity in the distal jejunum, and tended to increase lactase activity in the distal jejunum, aminopeptidase A activity in the middle jejunum and ileum, and aminopeptidase N activity in the ileum. In contrast, SM(+) increased dipeptidylpeptidase IV activity in the distal jejunum and tended to increase aminopeptidase N in the distal jejunum. In conclusion, both MR(+) and SM(+) affected small intestine development in newborn calves. This effect depended on the method of SB delivery but MR(+) generally had a more pronounced effect. No synergistic effect of SB supplementation into MR and SM was found.

Effects of duration of moderate increases in grain feeding on endotoxins in the digestive tract and acute phase proteins in peripheral blood of yearling calves.

Effects of duration of grain feeding on the concentration of endotoxic lipopolysaccharide (LPS) in digesta throughout the digestive tract and on acute phase proteins and LPS in peripheral blood were determined in Holstein yearling calves. Twenty-five Holstein yearling steer calves received either a forage-based diet containing 92% hay and 8% of a mineral and vitamin pellet on a dry matter basis (CON) or a moderate-grain diet, obtained by replacing 41.5% of the hay in the forage-based diet with barley grain, for 3 (MG3), 7 (MG7), 14 (MG14), or 21 d (MG21) before slaughter. Immediately before slaughter, blood samples were collected from the jugular vein. Immediately after slaughter, digesta samples were collected from the rumen, jejunum, ileum, cecum, colon, and rectum. Rumen liquid digesta, digesta from the intestines, and peripheral blood plasma were analyzed for LPS. Peripheral blood plasma and serum were analyzed for the acute phase proteins amyloid A, haptoglobin, and LPS-binding protein. Feeding the grain diet increased the LPS concentration in rumen fluid linearly from 15,488 endotoxin units (EU)/mL for CON to 70,146 EU/mL for MG7. Concentrations of LPS in rumen fluid in MG14 and MG21 were 61,944 and 56,234 EU/mL, respectively, and did not differ. The LPS concentrations in jejunal digesta were much lower than that in digesta elsewhere in the digestive tract, which suggests that ruminal LPS is broken down in the abomasum or proximal jejunum. The concentration of digesta LPS in the ileum was higher than that of digesta elsewhere in the intestines and similar to that in rumen fluid. The duration of grain feeding increased the LPS concentration in digesta in the ileum and cecum and tended to increase that in the colon cubically. Concentrations of LPS in this part of the digestive tract were highest in the MG3 and MG21 groups. The highest concentrations of LPS in digesta in the cecum, colon, and rectum were 3.7, 3.8, and 5.6 times higher than that in CON, respectively. Grain feeding and the increase in LPS in digesta were not accompanied by an acute phase response or a detectable concentration of LPS in peripheral blood. The absence of LPS in peripheral blood and the lack of increase in acute phase proteins indicated that the grain feeding protocol used in the current study and the accompanying changes in LPS concentrations of the digesta did not result in systemic inflammation.

Effect of butyrate sources in a high-concentrate diet on rumen structure and function in growing rams.

Dietary butyrate is considered to have mostly positive impacts on the ruminal epithelium. However, its supplementation in a high-concentrate diet may not be justified as excessive ruminal butyrate may negatively affect the rumen. Furthermore, butyrate impact on the rumen may depend on its source. Thirty-two Swiniarka growing rams (30.6 ± 2.5 kg; 11-14 months of age) were used to investigate the effect of a high-concentrate diet and sodium butyrate (SB) or tributyrin (TB) supplementation in a high-concentrate diet on the rumen structure and selected functions. The rams were allocated to four treatments and fed diets with: (1) low concentrate inclusion (22.5% of diet DM; L); (2) high concentrate inclusion (60% of diet DM; H); (3) H with SB (3.2% of diet DM; H+SB); and (4) H with TB (2.93% of diet DM; H+TB). The preplanned contrasts were used for treatment comparisons (L vs H treatments (H, H+SB, and H+TB), H vs H+SB, and H vs H+TB). The BW, BW gain and DM intake did not differ between treatments. In the atrium ruminis, epithelium thickness did not differ between the L and H treatments (P = 0.46), tended to be higher for H+SB than for H (P = 0.09) but did not differ between H+TB and H (P = 0.61). The expression of downregulated in adenoma was higher for L than for H treatments (P = 0.03) but was not affected by SB or TB supplementation (P ≥ 0.26). In the ventral rumen, the mucosa surface and epithelium thickness were lower for L than for H treatments (P < 0.01), were or tended to be higher for H+SB than for H (P ≤ 0.06) but did not differ between H+TB and H (P ≥ 0.26). The expression of monocarboxylate transporter 1 was lower for L than for H treatments (P = 0.02) but was not affected by SB or TB supplementation (P ≥ 0.28). The expression of putative anion transporter-1 and downregulated in adenoma did not differ between the L and H treatments (P ≥ 0.76); however, expression of the former tended to be higher and the latter tended to be lower for H+SB than for H (P ≤ 0.09), whereas no differences were observed between H+TB and H (P ≥ 0.14). In summary, SB supplementation, but not TB supplementation, in a high-concentrate diet stimulated ruminal epithelium growth and affected short-chain fatty acid transporters expression in the ruminal epithelium.

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.

Effect of increased intake of concentrates and sodium butyrate supplementation on reticulorumen macroanatomy and reticulorumen fermentation in growing rams.

Increased ruminal butyrate production is considered to have mostly positive impacts on rumen macro- and microanatomy and its functions. However, excessive ruminal butyrate production may also affect the rumen negatively. Forty-two growing rams were allocated into six treatments and fed a diet with low (22.5% of diet DM; LOW) or high (60% of diet DM; HIGH) inclusion of concentrates in combination with no, low (1.6% of diet DM) or high (3.2% of diet DM) sodium butyrate (SB) supplementation to obtain low or high reticuloruminal (RR) pH with different concentrations of butyrate. Both absolute (L/day) and relative (% of BW) water intake increased linearly with increasing dose of SB (P ≤ 0.02). The RR fluid pH was lower for HIGH compared to LOW treatments (P < 0.01) but was not affected by SB supplementation (P = 0.35). Total short-chain fatty acid concentration, propionate and valerate concentrations in the RR fluid were higher for HIGH compared to LOW treatments (P ≤ 0.01), but were not affected by SB supplementation (P ≥ 0.22). Reticuloruminal butyrate was higher for HIGH compared to LOW treatments and increased linearly with increasing dose of SB (P < 0.01). High concentrate inclusion in the diet (P < 0.01) decreased and SB supplementation tended to (P = 0.10) decrease fibrolytic activity in the RR. Increasing doses of SB linearly decreased acetate, isovalerate and NH3-N concentrations in RR fluid, and RR digesta DM weight (g DM/kg BW; P ≤ 0.02). Relative RR and rumen tissue weights (g/kg BW) were higher for LOW compared to HIGH (P ≤ 0.03) treatments but were not affected by SB inclusion in the diet (P ≥ 0.35). Also, there was no impact of concentrates or SB inclusion in the diet on ruminal epithelium DM weight (mg/cm2), either in the ventral or dorsal sac of the rumen (P ≥ 0.14). Under conditions of the current study, SB supplementation in the diet decreased RR digesta DM concentration and weight, acetate, isovalerate and NH3-N concentration in the RR fluid, and tended to reduce fibrolytic activity in the RR. At least part of this response could be due to increased intake of water, and consequently passage of digesta from the RR to lower regions of the gastrointestinal tract.

Effect of increased intake of concentrates and sodium butyrate supplementation on ruminal epithelium structure and function in growing rams.

Increased ruminal butyrate production is considered to have a positive impact on rumen epithelium growth and function. However, excessive ruminal butyrate production may affect the rumen negatively, particularly when the rumen is already challenged with low pH. The aim of this study was to determine the effect of the inclusion of concentrates in the diet and sodium butyrate (SB) supplementation on ruminal epithelium growth and function in growing rams. Forty-two rams (27.8 ± 7.3 kg; 9-14 months of age) were allocated into six treatments and fed a diet with low (22.5% of diet DM; LOW) or high (60% of diet DM; HIGH) inclusion of concentrates in combination with no (SB0), 1.6% (SB1.6) or 3.2% (SB3.2) of diet DM inclusion of SB. There was no impact of the investigated factors on papilla dimensions and mucosa surface area, either in the atrium ruminis or ventral rumen (P ≥ 0.11). Stratum corneum thickness was higher for HIGH compared to LOW treatments (P ≤ 0.04), independently of the location in the rumen. In the atrium ruminis, the epithelium and living strata thickness quadratically increased due to SB supplementation for LOW treatments but quadratically decreased for HIGH treatments (concentrate inclusion × butyrate supplementation interaction; P ≤ 0.03); conversely, in the ventral sac of the rumen, a thicker epithelium was observed due to both increased concentrate inclusion in the diet and SB supplementation (P < 0.01) but living strata thickness was increased only by SB supplementation (linear effect; P < 0.01). The epithelium damage index in the ventral sac of the rumen was higher for LOW compared to HIGH treatments (P = 0.02). Increased inclusion of concentrates in the diet increased mRNA expression of monocarboxylate transporter 1 in both the epithelium of the atrium ruminis and ventral rumen, occludin in the epithelium of the atrium ruminis and downregulated in adenoma in the epithelium of the ventral rumen (P ≤ 0.02). Protein expression of claudin-4 in the epithelium of the ventral rumen was the highest for the HIGH/SB1.6 and HIGH/SB3.2 treatments (significant effect of interaction between main effects; P < 0.01). Under the conditions of the current study, increased intake of concentrates had mostly positive effects on ruminal epithelium in growing rams, and the same was observed for the effect of SB supplementation. However, the effect of SB supplementation was at least partially affected by the inclusion of concentrates in the diet.