Effects of weaning age and pace on blood metabolites, cortisol concentration, and mRNA abundance of inflammation-related genes in gastrointestinal, adipose, and liver tissue of Holstein dairy calves.

The objective of this study was to evaluate the effects of weaning age and pace on blood metabolites, cortisol concentration, and mRNA abundance of inflammation-related genes in Holstein dairy calves. A total of 70 1-d-old calves (38.8 ± 4.4 kg BW ± SD), blocked by sex and birth BW, were randomly assigned to a 2 × 2 factorial arrangement of treatments. The first factor was weaning age, which was either early (6 wk) or late (8 wk). The second factor was weaning pace, which was either abrupt (4 steps down over 3 d; the initial milk replacer was 7.6 L, which was reduced by 1.9 L in each step-down) or gradual (7 steps down over 14 d; the initial milk replacer was 7.6 L, which was reduced by 1.09 L in each step-down), generating early-abrupt (EA), early-gradual (EG), late-abrupt (LA), and late-gradual (LG) treatments. All treatments had 10 female and 8 male calves, except EA that had 1 fewer male calf. Milk replacer (24% CP, 17% fat) was bottle fed, up to 1,200 g/d, twice daily (0600 h and 1800 h). The EA and EG treatment calves received 46.2 kg of milk replacer, and the LA and LG treatment calves received 63 kg of milk replacer. The study had 2 cohorts (2020, n = 40; 2021, n = 31), and each cohort included all treatments. Blood was collected from the jugular vein at 0900 h at 3 and 7 d of age, and a day before starting and a day after weaning completion. Male calves were humanely killed a day after weaning. Rumen, jejunum, large intestine, liver, omental adipose and perirenal adipose tissues were sampled to determine the mRNA abundance of inflammation-related genes. Weaning pace, age, pace × age, birth BW, and sex were included as fixed and cohort was included as random effects in the model. Blood metabolites and cortisol were analyzed as repeated measures, and sampling day, pace × sampling day, and age × sampling day were also included as additional fixed effects. Significance was noted at P ≤ 0.05 and tendencies when 0.05

Effect of pH and lipopolysaccharide on tight junction regulators and inflammatory markers in intestinal cells as an experimental model of weaning transition in dairy calves.

Acidic conditions combined with the presence of lipopolysaccharide (LPS) may increase the permeability of gastrointestinal epithelium. Feeding starch-rich starter to dairy calves is associated with ruminal acidosis and decreases the pH of other segments of the gastrointestinal tract, and that affects the tight junction regulator. The objective was to evaluate the effect of the combination of different pH (7.4 vs. 6.0) and LPS concentrations (0, 0.5, 10 ng/mL) in intestinal cells on tight junction regulators, inflammatory markers, and permeability. The human colon carcinoma Caco-2 cell line was used with the main treatment of pH and LPS in a 2 × 3 factorial arrangement. The pH was acidic (pH 6.0) or physiologic (pH 7.4), whereas LPS was 0, 0.5, or 10 ng/mL. After cells reached 70%-80% of confluence, the media were replaced with each respective treatment medium. Cells were treated for 3 h for mRNA abundance analysis, 3 and 6 h for protein abundance determination, and 3, 6, 12, and 24 h for permeability determination. Protein abundance of the myosin light-chain kinase (MYLK) and toll-like receptor 4 (TLR4) were measured by western blot. The mRNA abundance of IL-8, MYLK, peroxisome proliferator activated receptor gamma, and nuclear factor kappa B (Nfkb1) was determined by real-time, quantitative PCR. Paracellular permeability was determined with Lucifer yellow after 21 d of incubation. Cell culture was performed in biological triplicate; each biological replicate for real-time, quantitative PCR had 2 technical replicates, and for protein abundance and permeability assay had one technical replicate. The MIXED procedure of SAS (SAS Institute Inc.) was used with LPS, pH, and pH × LPS as fixed effects. Significance was declared at P ≤ 0.05 and tendencies when 0.05 < P ≤ 0.10. Increasing LPS doses did not affect the protein abundance of MYLK and TLR4, nor mRNA abundance of MYLK and PPRG. The LPS tended to increase mRNA abundance of IL-8 while pH × LPS interactively increased mRNA abundance of Nfkb1, where mRNA abundance of Nfkb1 was lower in cells exposed to pH 6.0 when combined with 0 and 10 ng/mL of LPS. Contrary to expectations, LPS did not affect the permeability of Caco-2 cells. The mRNA abundance of MYLK was greater at pH 6.0 versus pH 7.4. Also, protein abundance of TLR4 was lower at pH 6.0 than pH 7.4, and it decreased when exposure increased to 6 h. In addition, mRNA abundance of IL-8 was lower at pH 6.0 versus pH 7.4. Permeability was greater at pH 6.0 versus 7.4 after 6, 12, and 24 h of treatment. In summary, the effect of LPS and its interaction with pH showed less impact than expected on dependent variables measured, which might be attributed to the adopted clinically achievable LPS doses likely not being high enough to draw a strong response as observed in the literature. On the other hand, pH was far more relevant, modulating mRNA abundance of inflammatory markers, tight junction regulators, and permeability in in vitro colon cell models.

Effects of weaning strategies on health, hematology, and productivity in Holstein dairy calves.

Weaning strategies in dairy calves vary considerably, though the effect on animal health is unclear. This study examined the effects of calf weaning age (6 vs. 8 wk) and pace (abrupt vs. gradual) on performance, blood, and health parameters in dairy calves. The experiment consisted of a 2 × 2 factorial arrangement of treatments, where the factors included weaning age (early vs. late) and weaning pace (abrupt vs. gradual). Holstein calves (n = 72), blocked by sex and birth weight, were randomly assigned to one of 4 treatments (n = 18 per treatment): early-abrupt (EA), early-gradual (EG), late-abrupt (LA), and late-gradual (LG). Milk replacer (24% crude protein, 17% fat; up to 1,200 g/d) was fed twice daily; water, calf starter (18% crude protein), and chopped alfalfa hay were fed ad libitum. Daily intakes of milk replacer, calf starter, and forage were recorded from birth until end of weaning. Body weight, selected health measures, blood hematology, and fecal scores were obtained 1 d preweaning and 1 d postweaning. Calves were orally bolused with a rumen pH logger for the last 3 d of the weaning transition and rumen pH was measured continuously. Data were analyzed with age, pace, age × pace interaction, birthweight, and sex as fixed effects, and starting date as a random effect. Greater age at weaning increased respiration, whereas gradual-weaned calves had lower respiration rate. Heart rate was lower in gradual than in abrupt weaned groups. Fecal score had a marginal increase in late-weaned groups and significantly increased in gradually weaned groups. No difference was detected in body core temperature by age, pace, or interaction. During the weaning transition, average daily gain was lower in LA than EA and gradually weaned groups had an increased average daily gain. Change in grain intake, but not forage intake, was greater in gradually weaned groups. Mean rumen pH marginally increased from EG to LG and from LA to LG. No difference was detected among treatments in red or white blood cell counts, and hemoglobin. Procalcitonin was marginally highest in the LA group, while blood hematocrit increased in abruptly weaned groups. Overall, calf health is affected by both age and pace of weaning, though the health parameters affected by age and pace differ.

Effects of Saccharomyces cerevisiae boulardii (CNCM I-1079) on feed intake, blood parameters, and production during early lactation.

The periparturient period is a metabolically demanding time for dairy animals because of increased nutrient requirements for milk yield. The objective of this study was to investigate the effect of feeding Saccharomyces cerevisiae boulardii (CNCM I-1079), a commercial active dry yeast (ADY), in dairy cows on productive and metabolic measures during the periparturient period. Primiparous (n = 33) and multiparous (n = 35) cows were fed a close-up total mixed ration (TMR) before calving and a lactation TMR postpartum. Three weeks before expected calving time, animals were blocked by parity and body weight and then randomly assigned to either control group (control; n = 34) or treatment (ADY; n = 34). All animals were housed in a tie-stall barn with individual feed bunks; the ADY animals received supplementary Saccharomyces cerevisiae boulardii (CNCM I-1079), top dressed daily at a predicted dosage of 1.0 × 1010 cfu (12.5 g) per head. Blood samples were collected weekly along with milk yield and milk composition data; feed intake data were collected daily. Serum samples were analyzed for glucose, nonesterified fatty acid, ß-hydroxybutyrate, haptoglobin (Hp), and the cytokines tumor necrosis factor-α, IL-6, and IL-18. Colostrum samples collected within the first 6 to 10 h were analyzed for somatic cell score and IgG, IgA, and IgM concentrations. Data were analyzed using PROC GLIMMIX in SAS with time as a repeated measure; model included time, parity, treatment, and their interactions. The ADY groups had greater milk yield (39.0 ± 2.4 vs. 36.7 ± 2.3 kg/d) and tended to produce more energy-corrected milk with better feed efficiency. There was no difference in plasma glucose, serum nonesterified fatty acid, serum ß-hydroxybutyrate, Hp, IL-6, or IL-18 due to ADY treatment. The tumor necrosis factor-α increased in ADY-supplemented animals (1.17 ± 0.69 vs. 4.96 ± 7.7 ng/mL), though week, parity, and their interactions had no effect. Serum amyloid A tended to increase in ADY-supplemented animals when compared to control animals and was additionally affected by week and parity; there were no significant interactions. No difference in colostrum IgG, IgA, and IgM was observed between treatments. Supplementing transition cow TMR with ADY (CNCM I-1079) improved milk production and tended to improve efficiency in early lactation; markers of inflammation were also influenced by ADY treatment, though the immunological effect was inconsistent.

Effects of milk replacer allowances and levels of starch in pelleted starter on nutrient digestibility, whole gastrointestinal tract fermentation, and pH around weaning.

The objectives of this study were to examine the effects of pelleted starter diets differing in starch and neutral detergent fiber (NDF) content when fed differing levels of milk replacer (MR) on nutrient digestibility, whole gastrointestinal tract fermentation, pH, and inflammatory markers in dairy calves around weaning. Calves were randomly assigned to 1 of 4 dietary treatments (n = 12 per treatment) in a 2 × 2 factorial design based on daily MR allowance and amount of starch in pelleted starter (SPS): 0.691 kg of MR per day [dry matter (DM) basis] with starter containing low or high starch (12.0% and 35.6% starch on DM basis, respectively), and 1.382 kg of MR per day (DM) with starter containing low or high starch. All calves were housed in individual pens with straw bedding until wk 5 when bedding was covered. Calves were fed MR twice daily (0700 and 1700 h) containing 24.5% crude protein (DM) and 19.8% fat (DM), and had access to pelleted starter (increased by 50 g/d if there were no refusals before weaning and then 200 g/d during and after weaning) and water starting on d 1. Calves arrived between 1 and 3 d of age and were enrolled into an 8-wk study, with calves undergoing step-down weaning during wk 7. Starting on d 35, an indwelling pH logger was inserted orally to monitor rumen pH until calves were dissected at the end of the study in wk 8. Higher SPS calves showed an increase in rumen pH magnitude (1.46 ± 0.07) compared with low SPS calves (1.16 ± 0.07), a decrease in rumen pH in wk 8 (high SPS: 5.37 ± 0.12; low SPS: 5.57 ± 0.12), and a decrease in haptoglobin in wk 8 (high SPS: 0.24 ± 0.06 g/L; low SPS: 0.49 ± 0.06 g/L). The majority of differences came from increased starter intake in general, which suggests that with completely pelleted starters the differences in starch and NDF do not elicit drastic changes in fermentation, subsequent end products, and any resulting inflammation in calves around weaning.

Performance and visceral tissue growth and development of Holstein calves fed differing milk replacer allowances and starch concentrations in pelleted starter.

The objectives of this study were to investigate how milk replacer (MR) allowance and differing concentrations of starch and neutral detergent fiber in starter alters visceral tissue and overall growth of the calf. Calves were randomly assigned to 1 of 4 dietary treatments (n = 12 per treatment) arranged in a 2 × 2 factorial based on daily MR allowance (MRA) and amount of starch in pelleted starter (SPS) as follows: 0.691 kg of MR/d [dry matter (DM) basis] with starter containing low or high starch (12.0% and 35.6% starch, respectively) and 1.382 kg of MR/day (DM) with starter containing low or high starch. All calves were housed in individual pens with straw bedding until wk 5 when bedding was covered to minimize intake. Calves were fed MR twice daily (0700 and 1700 h) containing 24.5% crude protein (DM) and 19.8% fat (DM), and had access to pelleted starter (increased by 50 g/d if there were no refusals before weaning, and then 200 g/d during and after weaning) and water starting on d 1. Calves arrived between 1 and 3 d of age and were enrolled into an 8-wk study, with calves undergoing step-down weaning during wk 7. Intakes were measured daily, and body weight (BW) and blood samples were recorded and collected weekly. Calves were dissected in wk 8 for visceral tissue measurements. Overall, there was increased MR DM intake for the high- (0.90 ± 0.01 kg/d; ± SE) compared with the low-MRA (0.54 ± 0.01 kg/d) calves, whereas starter DM intake increased in low- (0.47 ± 0.05 kg/d) compared with high-MRA (0.20 ± 0.05 kg/d) calves, which was driven by increases in wk 6, 7, and 8. High-MRA calves had increased BW during wk 2, 3, 4, 5, 6, and 7. The difference in BW disappeared by wk 8, with overall average daily gain having a tendency to be increased in high (0.57 ± 0.04 kg/d) compared with low-MRA (0.50 ± 0.04 kg/d) calves, whereas average daily gain was increased in high-MRA calves during wk 2 and 3 and increased in low-MRA calves during wk 7 and 8. There were several differences throughout visceral tissue measurements, but most notably, an increase in rumen mass (i.e., full, empty, and digesta weights) in low- compared with high-MRA calves, as well as in low- compared with high-SPS calves was observed. The length, width, and 2-dimensional area of rumen papillae were also increased in low- (area: 0.88 ± 0.03 mm2) compared with high-MRA (0.46 ± 0.03 mm2) calves. The majority of differences were attributed to increased MR allowance, which contributed to reduced pelleted starter intake by more than 50% and reduced rumen development, whereas differences in starch intake from the completely pelleted starter had minimal effects on overall growth and tissue measurements.

Production performance and nitrogen metabolism in dairy cows fed supplemental blends of rumen undegradable protein and rumen-protected amino acids in low- compared with high-protein diets containing corn distillers grains.

Feeding corn dried distillers grains with solubles (DDGS) in low crude protein (CP) diets could limit N waste in lactating cows. However, it also could possibly reduce metabolizable AA supply, especially Lys, and compromise milk production. Therefore, the objective of this study was to evaluate the effects of feeding supplemental blends of rumen undegradable protein (RUP) and rumen-protected (RP) AA in a low compared with high CP diet containing corn DDGS on milk production and selected measures of N utilization. Six multiparous Holstein cows (619.3 ± 49.8 kg of body weight; 26.8 ± 6.2 d in milk) were subjected to a split-plot, 3 × 3 Latin square design (21-d periods) with dietary CP content [low (14.6%; LP) or high (16.6%; HP)] as the whole-plot factor, and blend of RUP and RP-AA [control (CON), no supplement; blend A (0.11 kg/cow per d); or blend B (0.45 kg/cow per d)] as the sub-plot factor. All diets contained 10% corn DDGS; blends of RUP and RP-AA were top-dressed during morning feeding. There was no dietary CP content × supplemental blend interaction for all measured variables. Nutrient (dry matter, organic matter, neutral detergent fiber, acid detergent fiber, and CP), milk and milk component yields, and feed and apparent N efficiency did not differ for cows fed the low- compared with the high-protein diet. However, apparent total-tract CP digestibility, blood and milk urea-N concentrations, and urinary excretion (g/d) of N and urea-N were lower for cows fed the low-protein compared with the high-protein diet. There was no supplemental blend effect on nutrient intake and apparent total-tract digestibility, and milk and milk component yields. Except for a tendency for total urinary purine derivative excretion and microbial N flow to be lower for cows fed blend B compared with CON but not blend A, there was no supplemental blend effect on measures of N utilization. Both dietary CP content and supplemental blend of RUP and RP-AA had a marginal effect on the plasma free AA profile. In summary, reducing dietary CP content in diets containing corn DDGS had no effect on lactation performance, possibly accounting for the lack of a positive response following the provision of supplemental blends of RUP and RP-AA. However, reducing dietary CP content resulted in a decrease in blood and milk urea-N concentrations, and urinary excretion of N and urea-N, suggestive of an improvement in the efficiency of N use.

Effects of supplemental butyrate and weaning on rumen fermentation in Holstein calves.

The objectives of this study were to determine the effects of the weaning transition and supplemental rumen-protected butyrate on subacute ruminal acidosis, feed intake, and growth parameters. Holstein bull calves (n = 36; age = 10.7 ± 4.1 d; ± standard deviation) were assigned to 1 of 4 treatment groups: 2 preweaning groups, animals fed milk replacer only (PRE-M) and those fed milk replacer, calf starter, and hay (PRE-S); and 2 postweaning groups, animals fed milk replacer, calf starter, and hay without supplemental rumen-protected butyrate (POST-S) or with supplemental rumen-protected butyrate at a rate of 1% wt/wt during the 2-wk weaning transition (POST-B). Milk replacer was provided at 1,200 g/d; starter, water, and hay were provided ad libitum. Weaning took place over 14 d by reducing milk replacer provision to 900 g/d in wk 7, 600 g/d in wk 8, and 0 g/d in wk 9. Rumen pH was measured continuously for 7 d during wk 6 for PRE-S and PRE-M and during wk 9 for POST-S and POST-B. After rumen pH was measured for 7 d, calves were euthanized, and rumen fluid was sampled and analyzed for volatile fatty acid (VFA) profile. Individual feed intake was recorded daily, whereas, weekly, body weights were recorded, and blood samples were collected. Compared with PRE-M, PRE-S calves tended to have a greater total VFA concentration (35.60 ± 11.4 vs. 11.90 ± 11.8 mM) but mean rumen pH was unaffected (6.25 ± 0.22 vs. 6.17 ± 0.21, respectively). Between PRE-S (wk 6) and POST-S (wk 9), calf starter intake increased (250 ± 219 vs. 2,239 ± 219 g/d), total VFA concentrations increased (35.6 ± 11.4 vs. 154.4 ± 11.8 mM), but mean rumen pH was unaffected (6.25 ± 0.22 vs. 6.40 ± 0.22, respectively). Compared with POST-S, POST-B calves had greater starter intake in wk 7, 8, and 9, but POST-B tended to have lower total VFA concentration (131.0 ± 11.8 vs. 154.4 ± 11.8 mM) and lower mean ruminal pH (5.83 ± 0.21 vs. 6.40 ± 0.22). In conclusion, the weaning transition does not appear to affect rumen pH and VFA profile, but supplementing rumen-protected butyrate during the weaning transition increased starter intake and average daily gain. Further, these data suggest that the ability of the rumen to manage rumen pH changes fundamentally postweaning. Why weaned calves with lower rumen pH can achieve higher calf starter intakes is unclear; these data suggest the effect of rumen pH on feed intake differs between calves and cows.

Effect of butyrate on passive transfer of immunity in dairy calves.

The objectives of this study were to determine the effects of supplemental butyrate on (1) Ig production in dams and (2) Ig absorption in their calves. Twenty dry dams fed a close-up total mixed ration were assigned to either a control treatment (CTRL-D) or a butyrate treatment where the close-up total mixed ration was supplemented with butyrate at 1% of dry matter intake (wt/wt; BUT-D). At calving, calves were assigned to 1 of 2 treatments: a control group fed colostrum replacer only (CTRL-C) and a butyrate group fed colostrum replacer with supplemental butyrate at 2.5% (wt/vol; BUT-C). Serum IgG, glucose, and ß-hydroxybutyrate were measured weekly in both dams and calves. Additionally, calves were weighed weekly to determine average daily gain. In dams, serum IgG concentration was not different between CTRL-D and BUT-D (1,785 ± 117 vs. 1,736 ± 137 mg/dL, respectively), nor was there a change in Ig levels in the colostrum between control and butyrate groups. Serum total protein did not differ between CTRL-D and BUT-D dams. Dam dry matter intake did not differ between CTRL-D and BUT-D but did decrease 1 wk before parturition. Compared with CTRL-C calves, BUT-C calves had significantly decreased serum IgG concentration at 24 h (2,110 ± 124 vs. 1,400 ± 115 mg/dL), wk 1 (1,397 ± 121 vs. 866 ± 115 mg/dL), and wk 2 (1,310 ± 121 vs. 797 ± 115 mg/dL). Additionally, apparent efficiency of absorption was lower for the BUT-C group compared with the CTRL-C group (35.3 ± 2.1 vs. 25.9 ± 2.0). Differences in serum Ig concentrations between the CTRL-C and BUT-C groups did not affect average daily gain (0.59 ± 0.05 vs. 0.48 ± 0.05 kg/d, respectively), serum glucose concentrations, or serum ß-hydroxybutyrate concentrations. These data demonstrate that butyrate inclusion in colostrum negatively affects IgG absorption in newborn calves, whereas calf body weight gains were unaffected.

Short communication: Effects of meloxicam administration on protein metabolism and growth performance in transported Jersey calves.

Our objective was to investigate the effects of administering the nonsteroidal anti-inflammatory drug meloxicam (MEL) before transport on various indicators of protein metabolism and growth performance over the first 96 h after transport in Jersey calves. Calves (age ± SD; 2 ± 1 d) sourced from a commercial farm were randomly administered, at 1 mg/kg of body weight, either meloxicam (MEL; n = 11) or a whey protein placebo (CON; n = 10) orally before transport to a calf facility (669 km; 8.5-h road trip). Calves were weighed and rectal temperature was recorded before departure (0 h), on arrival (8.5 h), and 96 h after arrival. Blood was collected at the same time as calves were weighed, and samples were analyzed for total protein (0-h sample), cortisol (0- and 8.5-h samples), haptoglobin (0- and 96-h samples), and amino acids, 3-methylhistidine, and urea-N (96 h). Milk replacer (MR) intake was recorded on arrival and over the next 4 d. Serum total protein concentration did not differ for CON and MEL calves. Plasma cortisol concentration was similar across treatments at 0 h; however, it was lower for CON than for MEL calves at 8.5 h. Although serum haptoglobin concentration tended to be greater for CON than MEL calves 96 h after transport, 3-methylhistidine and plasma urea-N concentrations did not differ across treatments. Plasma Asp, Asn, Glu, Lys, Met, Ser, and Trp were greater and plasma Arg, Gly, Pro, and Thr concentrations tended to be greater at 96 h after arrival for MEL compared with CON calves. Intake of MR and average daily gain were higher in MEL than in CON calves. In summary, although it had no effect on 3-methylhistidine or urea-N concentrations, administration of MEL before transport tended to reduce haptoglobin concentration, altered the amino acid profile, and was beneficial in preventing a decrease in MR intake and average daily gain in Jersey calves.

Effects of subacute ruminal acidosis and low feed intake on short-chain fatty acid transporters and flux pathways in Holstein steers.

The objective of this study was to investigate the role of protein-mediated transport pathways for short-chain fatty acid flux across the ruminal epithelium, using subacute ruminal acidosis (SARA) and feed restriction as models. Twenty-one Holstein steers (216.8 ± 31.4 kg BW) were individually housed and fed a total mixed ration (TMR) with a 50:50 forage:concentrate ad libitum for 5 d. After the 5 d diet adjustment period, calves were assigned 1 of 3 treatments: control (CTRL) calves were fed the TMR ad libitum on d 1, subacute ruminal acidosis calves were given 25% of their ad libitum DMI on d 1 and then given a barley grain challenge at 30% of ad libitum DMI on d2 (ACID) calves were given 25% of their ad libitum DMI on d 1 and then given a barley grain challenge at 30% of ad libitum DMI on d 2, and feed restriction (FR) calves were given 25% of their ad libitum DMI for 5 d. Reticuloruminal pH was continuously measured during the entire study. At the end of the study, rumen tissue was harvested and acetate and butyrate flux were measured. Selective inhibitors were used to differentiate total flux (TOTAL), protein-mediated flux (PMF), and passive diffusion flux (PDF). The duration that rumen pH was <5.6 was greater in ACID calves compared with CTRL and FR calves (57 ± 90 vs. 519.71 ± 90 vs. 30 ± 90 min/d for CTRL, ACID, and FR, respectively; < 0.01). Total acetate flux was greater in FR than in CTRL (630.6 ± 38.9 vs. 421.1 ± 41.4 nmol/cm × h, respectively; < 0.01), but no difference was observed between CTRL and ACID (421.1 ± 41.4 vs. 455.4 ± 38.9 nmol/cm × h, respectively). Also, total butyrate flux was greater in FR than in CTRL (1,241.9 ± 94.8 vs. 625.5 ± 86.3 nmol/cm × h, respectively; < 0.01), but no difference was detected between CTRL and ACID (625.5 ± 86.3 vs. 716.7 ± 81.0 nmol/cm × h, respectively). For butyrate flux, PMF was greater for FR than for CTRL (479.21 ± 103.9 vs. 99.9 ± 86.3 nmol/cm × h, respectively; < 0.01), but no difference was observed between the CTRL and ACID treatments (99.9 ± 86.3 vs. 90.2 ± 81.0 nmol/cm × h, respectively). Immunofluorescence analysis showed an increase in monocarboxylate cotransporter isoform 1 abundance in the FR treatment compared with the ACID treatment (9,250 ± 1,648 vs. 4,187 ± 1,537 arbitrary units, respectively; = 0.03) but not compared with the CTRL treatment (9,250 ± 1,648 vs. 7,241 ± 1,648 arbitrary units, respectively; = 0.15). These data identify a short-term adaptive response of the ruminal epithelium to dietary changes that involves PMF and PDF.

The periparturient period is associated with structural and transcriptomic adaptations of rumen papillae in dairy cattle.

The structural and functional adaption of the rumen epithelium during the transition period is largely undescribed. To characterize the adaptation of the rumen epithelium during transition, multiparous dairy cattle (n=12) fitted with rumen fistulas and fed a low-energy dry cow diet (1.37 Mcal/kg, net energy for lactation) were transitioned abruptly to a high-energy lactating cow diet (1.68 Mcal/kg, net energy for lactation) immediately after parturition. Rumen papillae were biopsied at -3, +1, and +6 wk relative to calving. The histology of morphology of the rumen papillae was evaluated under the light microscope and electron microscope, and mRNA profiling was performed using an Affymetrix GeneChip Bovine Gene 1.0 ST Array (Affymetrix, Santa Clara, CA). Data preprocessing was conducted using the robust multi-array average method, and detection of significant genes was conducted using ANOVA. Also, the Benjamini-Hochberg false discovery rate of 0.1 was applied. Microscopic examination of rumen papillae revealed an increase in epithelial desquamation during early lactation as sloughing scores increased from 1.7 ± 0.2 at -3 wk to 4.1 ± 0.3 and 3.4 ± 0.2 at +1 and + 6 wk, respectively. A total of 1,011 (-3 vs. +1 wk) and 729 (-3 vs. +6 wk) differentially expressed genes were identified (false discovery rate of 0.10, P<10(-3), fold-change ± 1.2 cut-off). A group of differentially expressed genes involved in desmosome assembly (DSG1, CDSN), epidermal growth factor signaling (EGFR, EREG), transforming growth factor ß signaling (TGFB1), and the insulin-like growth factor-axis (GHR, IGFBP2, IGFBP3, CTGF) was also validated using PCR. Gene network analysis found that EGFR, GHR, and TGFB1 were focal points of the top pathways, thereby supporting the importance of these regulatory genes to the adaptive response of rumen papillae in early lactation. The microscopic and transcriptomic changes in this study provide insight into the mechanisms responsible for the rapid rate of cellular and molecular adaptations of rumen papillae tissue during the transition period in dairy cattle. In conclusion, the experimental data support the hypothesis that rumen papillae adapt in early lactation by altering their gene expression patterns and, thus, their epithelial structure.

Active dry Saccharomyces cerevisiae can alleviate the effect of subacute ruminal acidosis in lactating dairy cows.

The objective of the study was to determine the effect of active dry Saccharomyces cerevisiae (ADSC) supplementation on dry matter intake, milk yield, milk components, ruminal pH, and microbial community during a dietary regimen that leads to subacute ruminal acidosis (SARA). Sixteen multiparous, rumen-cannulated lactating Holstein cows were randomly assigned to 1 of 2 dietary treatments that included ADSC (Biomate; AB Vista, Marlborough, UK; 8 × 10(10) cfu/head per day) or control. During wk 1 to 6, all cows received a high-forage (HF) diet (77:23, forage:concentrate). Cows were then abruptly switched during wk 7 to a high-grain (HG) diet (49:51, forage:concentrate) and remained on the HG until the end of wk 10. Feed intake and milk yields were recorded daily. Ruminal pH was recorded continuously using an indwelling system for 1 to 2 d per week during the pre-experimental phase, and wk 6, 7, and 10. Ruminal digesta samples were collected at the end of the experiment and analyzed for relative change in microbial communities using real-time quantitative PCR. Cows were considered to have SARA if the duration below pH 5.6 was ≥300 min/d. Ruminal pH during wk 6 (HF plateau) was not different across treatments (15 ± 46 min/d at pH <5.6). The dietary regimen successfully induced SARA during wk 7 (transition from HF to HG diet), and ruminal pH (551 ± 46 min/d at pH <5.6) was not different across treatments. However, cows receiving ADSC had an improved ruminal pH (122 ± 57 vs. 321 ± 53 min/d at pH <5.6) during wk 10 (HG plateau) compared with control. Additionally, cows receiving ADSC had a better dry matter intake (23.3 ± 0.66 vs. 21.6 ± 0.61 kg/d) and 4% fat-corrected milk yield (29.6 ± 1.2 vs. 26.5 ± 1.2 kg/d) than control cows during the HG phase (wk 8 to 10). During HG feeding, cows receiving ADSC had greater total volatile fatty acid and propionate concentrations (175 ± 7.5 vs. 154 ± 7.5 and 117 ± 6.1 vs. 94 ± 5.7 mM for ADSC and control, respectively) and lower acetate:propionate ratio (0.26 ± 0.5 vs. 0.36 ± 0.05 for ADSC and control, respectively). Microbial analyses conducted on samples collected during wk 10 showed that cows supplemented with S. cerevisiae had a 9-fold, 2-fold, 6-fold, 1.3-fold, and 8-fold increase in S. cerevisiae, Fibrobacter succinogenes, Anaerovibrio lipolytica, Ruminococcus albus, and anaerobic fungi, respectively, which suggested an increase in cellulolytic microbes within the rumen. Cows supplemented with ADSC had 2.2-fold reduction in Prevotella albensis, which is a gram-negative bacterium predominant during SARA. Prevotella spp. are suggested to be an important source of lipopolysaccharide responsible for inflammation within the rumen. Cows supplemented with ADSC had a 2.3-fold increase in Streptococcus bovis and a 12-fold reduction in Megasphaera elsdenii. The reduction in M. elsdenii may reflect lower concentration of lactic acid within the rumen for ADSC cows. In conclusion, ADSC supplementation to dairy cows was demonstrated to alleviate the condition of SARA caused by abrupt dietary changes from HF to HG, and can potentially improve rumen function, as indicated by greater numbers of cellulolytic microorganisms within the rumen.

Effects of starch content of calf starter on growth and rumen pH in Holstein calves during the weaning transition.

The objective of this study was to evaluate the effects of substituting high fiber byproducts for dry ground corn in calf starter on growth and rumen pH during the weaning transition. Holstein bull calves were raised on an intensified nursing program using milk replacer containing 26% CP and 18% fat. Calves were fed a texturized calf starter containing either dry ground corn at 18.8% of dry matter (DM; CRN), beet pulp replacing dry ground corn at 10.2% dietary DM (BP), or triticale dried distillers grains with solubles replacing dry ground corn and high-protein feedstuffs at 18.6% of dietary DM (DDGS) in the pellet; treatment calf starters differed only in the pellet portion. Starch concentrations of CRN, BP, and DDGS were 35.3, 33.4, and 31.4%, respectively. After a calf consumed 2.50 kg of starter for 3 consecutive days, a small ruminant rumen pH data logger was inserted orally and rumen pH was measured continuously for 4d. Calves were then killed and rumen fluid was sampled to determine volatile fatty acid profile. No difference was found in overall average daily gain or growth rates of hip height, withers height, and heart girth. During the weaning transition, rate of increase in calf starter intake was greater for calves fed DDGS compared with those fed CRN (87.7 vs. 77.5 g/d), but lower for calves fed BP compared with CRN (68.1 vs. 77.5 g/d). The area under pH 5.8 (470 vs. 295 min × pH/d) or pH 5.2 (72.7 vs. 16.4 min × pH/d) was greater for calves fed DDGS than those fed CRN. Rumen pH profile was not affected by BP treatment compared with CRN, but calves fed BP tended to have greater water intake than those fed CRN (6.6 vs. 5.8 L/d). Volatile fatty acid profile was not affected by treatment with the exception of molar proportion of butyrate, which tended to be lower for calves fed BP compared with those fed CRN (15.0 vs. 16.6%). Hay intake was positively correlated to mean rumen pH for calves used in this study (r=0.48). Decreasing dietary starch concentration did not mitigate rumen acidosis in calves during weaning transition, and low rumen pH did not adversely affect growth during the weaning transition.

Effects of feeding a calf starter on molecular adaptations in the ruminal epithelium and liver of Holstein dairy calves.

The objective of this study was to elucidate the effect of feeding a calf starter on the volatile fatty acid (VFA) profile in the rumen and on expression of genes involved in epithelial intracellular pH regulation, butyrate metabolism, and hepatic urea cycle during the weaning transition. Twenty Holstein bull calves were fed either milk replacer and hay (MR) or milk replacer, hay, and a commercial texturized calf starter (MR+S) in a randomized complete block design. All calves were fed 750 g/d of milk replacer as the basal diet. Calves on the MR+S treatment were also fed starter ad libitum, and the energy intake of calves within blocks was maintained by supplementing the MR group with extra milk replacer that was equivalent to the energy intake from calf starter. Calves were killed 3 d after they consumed 680 g/d of calf starter for 3 consecutive days. Calves fed MR+S had higher VFA concentrations in the rumen (99.1±8.1 vs. 64.6±8.6 mM) and a higher molar proportion of butyrate (15.6±1.7 vs. 7.9±1.9%) than calves fed MR. Relative abundance of mRNA for monocarboxylate transporter isoform 1 was higher (1.45 vs. 0.53), and that of Na(+)/H(+) exchanger isoform 3 (0.37 vs. 0.82) and 3-hydroxy-3-methylglutaryl synthase isoform 1 (0.40 vs. 0.94) lower for the MR+S treatment compared with the MR treatment. In the liver, relative mRNA abundances of argininosuccinate synthetase isoform 1 (2.67 vs. 1.56), argininosuccinate lyase (1.44 vs. 0.99), and arginase isoform 1 (3.21 vs. 1.74) were greater for MR+S than for MR calves. Calf starter consumption appeared to increase fermentation in the rumen and affected expression of genes involved in cholesterol synthesis and intracellular pH regulation in ruminal epithelium, and those involved in urea cycle in the liver.

Short communication: Effect of calf starter on rumen pH of Holstein dairy calves at weaning.

The objective of this study was to evaluate the effect of feeding calf starter on rumen pH of dairy calves during weaning transition. Twenty Holstein bull calves were paired into 10 blocks by starting date of the study and body weight, and fed either milk replacer and hay (MR) or MR, hay, and a commercial texturized calf starter (MR+S) in a randomized complete block design. All calves were fed 750 g/d of milk replacer as the basal diet. Calves on MR+S treatment were also fed a calf starter ad libitum to maintain similar energy intake between calves within blocks, and MR calves were fed additional milk replacer that was equivalent to energy from calf starter intake. When MR+S calves consumed a calf starter at 680 g/d for 3 consecutive d, rumen pH of a MR+S calf and his MR counterpart was measured continuously for 3 d using a small ruminant rumen pH measurement system. Treatment did not affect minimum pH, mean pH, maximum pH, standard deviation of mean pH, and duration or area under pH 5.8, indicating that calf starter consumption did not appear to affect rumen pH. However, hay intake was negatively correlated to area under pH 5.8, with a breakpoint at 0.080 kg/d intake, suggesting hay intake might play an important role in mitigating ruminal acidosis in dairy calves during weaning transition.