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.

Relationship between maximum eye temperature and plasma cortisol concentration in racehorses during intensive training.

The aim of the study was to determine the utility of maximum eye temperature measured by infrared thermography (IRT) as a stress indicator compared with plasma cortisol concentration in Thoroughbred and Arabian racehorses. The study included thirty racehorses undergoing standard training for racing. Measurements of maximum eye temperature and blood collection for plasma cortisol concentration were carried out before training (BT), and within 5 (5AT) and 120 minutes (120AT) after the end of the each training session in three repetitions, with a monthly interval. Both parameters were elevated at 5AT compared to BT (p⟨0.001). Compared to BT, at 120AT the maximum eye temperature remained elevated (p⟨0.001) and plasma cortisol concentration decreased (p⟨0.001). The study indicated significant weak correlations (r=0.220; p⟨0.001) between both measurements at all time points. The results support the use of IRT technique to monitor the response of horses to stress, potentially improving animal management and welfare.

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.

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.

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.

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 butyrate infusion into the rumen on butyrate flow to the duodenum, selected gene expression in the duodenum epithelium, and nutrient digestion in sheep.

The aim of the study was to determine the effect of butyrate infusion into the rumen on butyrate flow to the duodenum, expression of short-chain fatty acid (SCFA) transporters (monocarboxylate transporter-1, -2, and -4) and receptors (G protein coupled receptor-41 and -43) in the duodenal epithelium and nutrient digestion in sheep. Eight wethers (39.0 ± 3.00 kg; mean ± SD) with ruminal and T-shape duodenal cannulas were allocated to 4 × 4 replicated Latin square design with each experimental period lasting for 21 d (12 d of adaptation and 9 d for data and sample collection). Experimental treatments were: 1) distilled water infusion into the rumen (CONT); 2) 15 g/d of butyric acid infusion into the rumen (BUT15); 3) 30 g/d of butyric acid infusion into the rumen (BUT30); and 4) 45 g/d of butyric acid infusion into the rumen (BUT45). The daily dose of butyrate was infused into the rumen via the rumen cannula, with 200 mL of solution of butyric acid and distilled water, at a constant rate (0.1389 mL/min) throughout the day using a peristaltic pump. Correspondingly, 200 mL/d of distilled water was infused into the rumen of CONT. The wethers were fed daily 900 g of chopped meadow hay and 200 g of concentrate in two equal meals at 0600 and 1800 h. Butyrate infusion into the rumen did not affect total SCFA concentration in the rumen fluid ( > 0.11). Molar proportion of butyrate in total SCFA linearly increased, and molar proportion of acetate and isovalerate linearly decreased ( ≤ 0.02) with an increasing amount of butyrate infused into the rumen. The molar proportion of butyrate in total SCFA in the duodenal digesta linearly increased ( < 0.01), and butyrate flow to duodenum tended to linearly increase ( = 0.06) with an increasing dose of exogenous butyrate delivered to the rumen. Butyrate infusion into the rumen did not affect ( ≥ 0.14) the mRNA expression of monocarboxylate transporter-2 and -4 and G protein coupled receptor-43 in the duodenal epithelium. The G protein coupled receptor-41 and monocarboxylate transporter-1 mRNA expression in the duodenal epithelium was very low in many of the analyzed samples. Digestibility of organic matter, neutral detergent fiber, and acid detergent fiber in the stomach (forestomach and abomasum) decreased for BUT15 and BUT30 and then increased for BUT45 (quadratic, ≤ 0.04); however, neither digestibility in the intestine nor total tract digestibility differed between treatments ( ≥ 0.10).

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.

Insulin-like growth factors 1 and 2 (IGF-1 and IGF-2) mRNA levels in relation to the gastrointestinal tract (GIT) development in newborn calves.

The study was aimed at determining the age-depended changes in insulin-like growth factors 1 and 2 (IGF-1 and IGF-2) mRNA levels in relation to the gastrointestinal tract (GIT) development in newborn calves. Twenty four male calves (5-day-old) were randomly allocated to 4 groups which were slaughtered at day 5, 12, 19 and 26 of their life, respectively. Tissue samples from the rumen, abomasum, duodenum, jejunum and liver were taken for analysis, and the level of IGF-1 and IGF-2 mRNA was determined using RT semi-quantitative PCR method. Both IGF-1 and IGF-2 mRNA level was the biggest on 5 day of life, significantly decreased up to day 12-19 in most of analyzed GIT parts, and started to increase thereafter. In the rumen the reduction in the amount of IGFs transcripts was associated with the reduction in papillae length and tunica muscularis thickness. The abomasum weight and tunica mucosa thickness increased from the 5 day up to 19 day of life, whereas abomasal IGF-1 mRNA level decreased together with calf's age up to 19 day of life, and the level of IGF-2 mRNA did not change. The reduction in IGFs mRNA level after 5 day of life in the duodenum (IGF-1 and IGF-2) and in the jejunum (IGF-1) was associated with reduction in villi length (duodenum and jejunum), and the increase of crypt depth (duodenum).

Effect of sodium butyrate supplementation in milk replacer and starter diet on rumen development in calves.

Rumen development is an important factor determining early solid feed intake and performance in cattle. A popular trend towards early weaning of newborn dairy calves necessitated looking for ways of accelerating the gastrointestinal tract (GIT) development. The present study aimed to determine the effect of sodium butyrate (NaB) supplementation in milk replacer and starter diet on rumen development in rearing calves. Fourteen bull calves (5-day-old) were randomly allocated to two groups: Control (C) and NaB. The later received 0.3 % NaB in milk replacer and starter diet. Animals were in experiment up to age of 26 days. Addition of NaB to milk replacer and starter diet had no effect on daily growth rate, but reduced the weight loss observed in C calves in first 11 days of age. Additionally, the NaB calves weighed more at the end of the study and tended to have higher growth rate in the whole trial period (P<0.15). The NaB calves showed a tendency toward higher reticulorumen weight (P=0.13) and higher reticulorumen weight expressed as a percent of whole stomach weight (P=0.02) as compared to control. Histometry analysis indicated larger rumen papillae length and width (P<0.01) in NaB group, and no change in muscle layer thickness, as compared to control. Plasma glucagon-like peptide-2 relative increase was higher in NaB group than in C group, and may be involved in rumen development. In conclusion, supplementation of the diet (milk replacer and starter diet) with NaB may enhance rumen development in neonatal calves.