Ruminal bacterial community shifts in grain-, sugar-, and histidine-challenged dairy heifers

Ruminal bacterial community composition (BCC) and its associations with ruminal fermentation measures were studied in dairy heifers challenged with combinations of grain, fructose, and histidine in a partial factorial study. Holstein-Friesian heifers (n = 30) were randomly allocated to 5 triticale grain-based treatment groups: (1) control (no grain), (2) grain [fed at a dry matter intake (DMI) of 1.2% of body weight (BW)], (3) grain (0.8% of BW DMI) + fructose (0.4% of BW DMI), (4) grain (1.2% of BW DMI) + histidine (6 g/head), and (5) grain (0.8% of BW DMI) + fructose (0.4% of BW DMI) + histidine (6 g/head). Ruminal fluid was collected using a stomach tube 5, 115, and 215 min after consumption of the rations and bacterial 16S ribosomal DNA sequence data was analyzed to characterize bacteria. Large variation among heifers and distinct BCC were evident in a between-group constrained principal components analysis. Bacterial composition in the fructose-fed heifers was positively related to total lactate and butyrate concentrations. Bacterial composition was positively associated with ruminal ammonia, valerate, and histamine concentrations in the grain-fed heifers. The predominant phyla were the Firmicutes (57.6% of total recovered sequences), Bacteroidetes (32.0%), and candidate phylum TM7 (4.0%). Prevotella was the dominant genus. In general, grain or histidine or their interactions with time had minimal effects on the relative abundance of bacterial phyla and families. Fructose increased and decreased the relative abundance of the Firmicutes and Proteobacteria phyla over time, respectively, and decreased the abundance of the Prevotellaceae family over time. The relative abundance of the Streptococcaceae and Veillonellaceae families was increased in the fructose-fed heifers and these heifers over time. A total of 31 operational taxonomic units differed among treatment groups in the 3.6 h sampling period, Streptococcus bovis was observed in fructose fed animals. The TM7 candidate phylum had an increased abundance of sequence reads by over 2.5 fold due to the introduction of histidine into the diet. Rapid changes in BCC can occur in a short period after a single substrate challenge and the nature of these changes may influence ruminal acidosis risk and differ from those in cattle exposed to substrate challenges over a longer time period.     

Effects of grain, fructose, and histidine on ruminal pH and fermentation products during an induced subacute acidosis protocol

The effects of grain, fructose, and histidine on ruminal pH and fermentation products were studied in dairy cattle during an induced subacute acidosis protocol. Thirty Holstein heifers were randomly allocated to 5 treatment groups: (1) control (no grain); (2) grain [fed at a crushed triticale dry matter intake (DMI) of 1.2% of body weight (BW)]; (3) grain (0.8% of BW DMI)+fructose (0.4% of BW DMI); (4) grain (1.2% of BW DMI)+histidine (6 g/head); and (5) grain (0.8% of BW DMI)+fructose (0.4% of BW DMI)+histidine (6 g/head) in a partial factorial arrangement. Heifers were fed 1 kg of grain daily with ad libitum access to ryegrass silage and alfalfa hay for 10 d. Feed was withheld for 14 h before challenge day, on which heifers were fed 200 g of alfalfa hay and then the treatment diets immediately thereafter. Rumen samples were collected 5 min after diet ingestion, 60 min later, and at 3 subsequent 50-min intervals. Grain decreased ruminal pH and increased ammonia, total volatile fatty acid (VFA), acetate, butyrate, propionate, and valerate concentrations compared with controls. The addition of grain had no effect on ruminal D- and L-lactate concentrations. Fructose markedly decreased ruminal pH and markedly increased D- and L-lactate concentrations. Fructose increased total VFA and butyrate and decreased valerate concentrations. Although histidine did not have a marked effect on ruminal fermentation, increased concentrations of histamine were observed following feeding. This study demonstrates that the substitution of some grain for fructose can lower ruminal pH and increase VFA and lactate concentrations, warranting further investigation into the role of sugars on the risk of acidosis in dairy cattle.     

Effects of feed additives on rumen and blood profiles during a starch and fructose challenge

We evaluated the effect of feed additives on the risk of ruminal acidosis in Holstein heifers (n = 40) fed starch and fructose in a challenge study. Heifers were randomly allocated to feed additive groups (n = 8 heifers/group): (1) control (no additives); (2) virginiamycin (VM); (3) monensin + tylosin (MT); (4) monensin + live yeast (MLY); and (5) sodium bicarbonate + magnesium oxide (BUF). Heifers were fed 2.5% of body weight (BW) dry matter intake (DMI) per day of a total mixed ration (62:38 forage:concentrate) and feed additives for a 20-d adaptation period. Fructose (0.1% of BW/d) was included for the last 10 d of the adaptation period. On d 21, heifers were fed to target a DMI of 1.0% of BW of wheat, fructose at 0.2% of BW, and their feed additives. Rumen fluid samples obtained by stomach tube and blood samples were collected weekly as well as during a 3.6-h period on challenge day (d 21). Virginiamycin and BUF groups maintained a consistently high DMI across the 20-d adaptation period. The MLY heifers had low DMI of the challenge ration. Average daily gain and feed conversion ratio were not affected by feed additives. All rumen and plasma measures changed weekly over adaptation and over the challenge sampling period with the exception of rumen total lactate and histamine concentrations, plasma oxidative stress index, and ceruloplasmin. Substantial within- and between-group variation was observed in rumen and plasma profiles at challenge sampling. No significant group changes were observed in rumen total volatile fatty acids, propionate, acetate-to-propionate ratio, isobutyrate, caproate, isovalerate, total lactate, d- and l-lactate, and pH measures on challenge day. Acetate concentration was increased in the BUF and control groups on challenge day. Butyrate concentration was lower in the MLY and MT groups compared with other groups at challenge. Valerate concentrations were lowest in the control, VM, and BUF groups and lactate concentrations were numerically lower in the MLY, VM, and BUF groups. Total lactate concentrations were >10 mM for each group throughout the challenge. Ammonia concentrations were lower in the MLY and MT groups. Histamine concentrations were decreased in MLY and increased in the VM and BUF groups. Plasma oxidative stress measures were not influenced by feed additives weekly or on challenge day, except for an increase in biological antioxidant potential in the control, VM, and MT groups on challenge day. Despite the large within-animal variation, all feed additives modified rumen function and may influence the risk of acidosis by different mechanisms; however, none stabilized the rumen in all heifers.     

Feeding high proportions of barley grain stimulates an inflammatory response in dairy cows

The objective of this study was to evaluate effects of feeding increasing proportions of barley grain on acute phase response in lactating dairy cows. Eight cannulated primiparous (60 to 140 d in milk) Holstein dairy cows were assigned to 4 diets in a 4 × 4 Latin square experimental design. The experimental period lasted for 21 d, with 11 d of adaptation and 10 d of measurements. Cows were fed the following diets: 1) no barley grain in the diet, 2) 15% barley grain, 3) 30% barley grain, and 4) 45% barley grain, as well as barley and alfalfa silage and alfalfa hay at 85, 70, 55, and 40% [dry matter (DM) basis]. All cows were supplemented with a 15% concentrate mix. Blood and rumen fluid samples were collected on d 1, 3, 5, 7, and 10 of the measurement period, and pH and endotoxin content were measured in rumen samples. Concentrations of serum amyloid A, lipopolysaccharide-binding protein, haptoglobin, and C-reactive protein in plasma were measured by ELISA. Feeding high proportions of barley grain at 0, 15, 30, and 45% of DM was associated with lower feed intake (32.6, 32.9, 27.34, and 25.18 kg/d ± 1.30, respectively), lower ruminal pH (6.8, 6.7, 6.7, and 6.5 ± 0.03, respectively), and higher DM intake (13.33, 15.28, 14.68, and 16.04 ± 0.63 kg/d, respectively) and milk production (27.2, 28.2, 29.0, and 31.0 ± 1.2 kg/d, respectively). Ruminal endotoxin increased in cows receiving 30 and 45% barley grain (5,021, and 8,870 ± 393 ng/mL, respectively) compared with those fed no grain or 15% barley grain (654 and 790 ± 393 ng/mL, respectively). Plasma concentrations of serum amyloid A, lipopolysaccharide-binding protein, and C-reactive protein increased in cows given higher (30 and 45%) proportions of grain. Plasma haptoglobin was not affected by treatments. In conclusion, feeding dairy cows high proportions (30 and 45% DM basis) of barley grain was associated with lower feed intake and rumen pH, increased endotoxin in the rumen fluid, and stimulation of an inflammatory response.     

Effects of dietary energy and protein density on plasma concentrations of leptin and metabolic hormones in dairy heifers

The hormonal and metabolic signals that communicate the level of body energy reserves to the reproductive-mammary axis remain undefined in dairy cattle; consequently, our hypothesis was that leptin may fulfill this role. Our objectives were to determine the effects of diets differing in energy and protein density on dry matter intake (DMI), growth traits [body weight (BW), body condition score (BCS), back-fat (BF) thickness], and temporal changes in plasma concentrations of leptin, insulin, growth hormone (GH), insulin-like growth factor-1 (IGF-1), glucose, and nonesterified fatty acids (NEFA) in dairy heifers during the pre- and postpubertal periods. In period 1, heifers were randomly allotted (n = 10/diet) at 103 kg of BW to diets for a predicted average daily gain of 1.10 (high, H), 0.80 (medium, M), or 0.50 kg/d (low, L). Five heifers in each of the H and L groups were further studied during period 2, either at 12 mo of age (HA, LA) or at 330 kg of BW (HW, LW). The data provide evidence that 1) DMI (18%), BW (17%), and BF (5%) together explained 40% of the variation in plasma leptin concentrations (r² = 0.396); 2) unlike the acute postprandial increase in plasma insulin as a result of increased nutrient density (H 1.42 ± 0.09, M 1.02 ± 0.09, L 0.68 ± 0.11 ng/mL), plasma leptin concentrations did not respond acutely with a distinct postprandial profile; 3) although plasma leptin concentrations increased with age, leptin at puberty did not differ among treatment groups (H 5.63 ± 2.48, M 4.28 ± 0.55, L 4.12 ± 0.72 ng/mL) and there was no evidence of an abrupt transition in prepubertal plasma leptin concentrations; 4) plasma leptin concentrations may not be a critical trigger for puberty in rapidly growing heifers, but are apparently essential for puberty in heifers with normal or restricted growth rates; and 5) plasma concentrations of insulin (H 0.59 ± 0.07, M 0.43 ± 0.09, L 0.30 ± 0.09 ng/mL), IGF-1 (H 151.08 ± 16.47, L 82.51 ± 17.47 ng/mL), and glucose (H 81.35 ± 3.39, M 73.59 ± 2.34, L 68.25 ± 3.39 mg/dL) reflected nutrient density, whereas GH (H 1.82 ± 0.23, L 5.87 ± 0.45 ng/mL) and NEFA (H 209.54 ± 50.83, L 234.93 ± 48.97 μM) were inversely related to the plane of nutrition. Collectively, these data suggest that plasma concentrations of leptin may play a role in long-term regulation of energy reserves and puberty in growing Holstein heifers.     

Effects of prepartum 2,4-thiazolidinedione on metabolism and performance in transition dairy cows

Thiazolidinediones (TZD) are potent synthetic ligands for peroxisome proliferator-activated receptor-γ that have been shown previously to reduce plasma nonesterified fatty acids and increase peripartal dry matter intake (DMI) in dairy cows. Data from Holstein cows (n = 36) entering their second or greater lactation were used to determine whether late prepartum administration of TZD would affect periparturient metabolism, milk production, and ovarian activity. Cows were administered 0, 2.0, or 4.0 mg of TZD/kg of BW by intrajugular infusion once daily from 21 d before expected parturition until parturition. Plasma samples were collected daily from 22 d before expected parturition through 21 d postpartum and twice weekly from wk 4 through 9 postpartum. In response to increasing TZD dosage, plasma nonesterified fatty acid concentrations decreased linearly during the postpartum period (d 0 to +21: 348, 331, 268 ± 31 μEq/L, respectively). Plasma concentrations of glucose were highest in cows administered 4.0 mg of TZD/kg of BW during the peripartum and postpartum periods (d −7 to +7: 57.9, 57.8, 61.1 ± 0.8 mg/dL and d 0 to +21: 51.6, 49.3, 54.7 ± 1.1 mg/dL, respectively). Plasma concentrations of β-hydroxybutyrate were increased during the peripartum period by TZD administration (9.6, 9.9, 10.2 ± 0.3 mg/dL) but were not affected during the postpartum period. Plasma insulin was not affected by treatment during any time period. Postpartum liver triglyceride content was decreased linearly (11.0, 10.4, 4.2 ± 1.6%) and glycogen content was increased linearly (2.16, 2.38, 2.79 ± 0.19%) by prepartum TZD administration. Prepartum TZD administration linearly increased DMI during the peripartum period (d −7 to +7: 16.1, 17.2, 17.3 ± 0.5 kg/d). Cows administered TZD prepartum maintained higher postpartum body condition scores than control cows (wk 1 through 9: 2.77, 2.89, 3.02 ± 0.05). There was no effect of prepartum TZD on milk yield; however, yields of 3.5% fat-corrected milk (52.2, 54.6, 48.0 ± 1.6 kg/d) and most other milk components were decreased in cows that received 4.0 mg of TZD/kg of BW prepartum. Prepartum TZD administration linearly decreased the number of days to first ovulation (29.3, 28.3, 19.0 ± 3.6 d). These results suggest that prepartum administration of TZD improves metabolic health and DMI of periparturient dairy cows and may decrease reliance on body fat reserves during early lactation.     

Phlorizin induces lipolysis and alters meal patterns in both early- and late-lactation dairy cows

Phlorizin is known to increase whole-body glucose demand, but it has also stimulated lipolysis in past studies in ruminants. Increased lipolysis complicates studies of dry matter intake (DMI) regulation by hepatic oxidation by providing the liver with additional oxidative substrate. Therefore, to assess whether increased glucose demand selectively increases DMI for cows in negative energy balance, phlorizin was administered to early- and late-lactation cows. Six Holstein cows in early lactation (19 ± 6 DIM, 50.0 ± 1.8 kg/d of milk, mean ± SD) and 6 Holstein cows in late lactation (228 ± 18 DIM, 30.6 ± 1.9 kg/d of milk) were randomly assigned to treatment sequence in a crossover design. Periods were 14 d with 7-d adaptation periods and 7 d of treatment. Phlorizin (4 g/d) and propylene glycol (carrier and control) were administered subcutaneously every 6 h throughout the treatment periods. Feeding behavior and DMI data were collected for the final 4 d of each treatment period; blood samples and total urine output were collected on d 4 of each treatment period. Phlorizin caused urinary loss of glucose at 333 g/d in early-lactation cows and 532 g/d in late-lactation cows. Phlorizin increased plasma nonesterified fatty acid concentrations similarly in early- and late-lactation cows, but did not significantly alter plasma insulin concentrations. Treatment with phlorizin tended to decrease meal size, but also decreased intermeal interval, resulting in no effect on DMI. The effects of phlorizin on lipolysis, feeding behavior, and DMI are not dependent on relative energy balance.     

Rumen digestion and nutritional efficiency of dairy heifers limit-fed a high forage ration to four levels of dry matter intake

Eight rumen-cannulated Holstein dairy heifers [beginning body weight (BW) 340 (±5) kg and age 14.5 (±1) mo] were fed a high forage diet at 4 levels of intake. Diets were composed of grass silage, grass hay, and corn silage as the forage components and offered at 1.25, 1.50, 1.75, and 2.00% of BW to heifers in a replicated Latin square design. Diets were incubated in situ in heifers receiving all 4 levels of feed. Blood and rumen were sampled at 2-h intervals for 24 h, rumen contents were emptied, and total fecal and urine collection was made. Dietary intake increased in proportion to feed offered until dry matter intake (DMI) was 1.92% BW, after which a statistically determined plateau was evident due to greater refusals when feed was offered at 2.00% BW. In situ degradation of feed was not affected by intake level, which, combined with the greater turnover rate of rumen contents, leads to the inference that rate of passage was increased with increasing intake. Rumen pH decreased and rumen volatile fatty acid concentration and microbial protein flow to the small intestine (estimated using urinary purine derivative excretion) increased as intake increased. Manure excretion increased as DMI increased at a rate 2.54 times greater than increases in DMI; this increase was entirely due to greater excretion of wet feces because urine excretion did not change with intake level. Nitrogen digestibility decreased and N retention increased linearly as the level of feed offered increased. Efficiency of N retention was minimized when feed was offered at 1.25% BW; all levels of feed offered above this level resulted in equivalent efficiencies. From the results of this experiment it can be concluded that when dairy heifers are limit-fed a high forage diet, the efficiency of nutrient utilization is increased as intake decreases, but reducing DMI below 1.50% BW reduced efficiency.     

Variation in plasma concentrations of insulin-like growth factor-I in pasture-fed Holstein cows

The objective of this 5-wk study was to determine dietary effects on plasma concentrations of insulin-like growth factor-I (IGF-I), as well as milk production and milk components in pasture-fed dairy cows. Thirty-two Holstein cows 4 to 5 wk postpartum were randomly assigned to 4 dietary subgroups. Feed was provided twice daily ad libitum at 0900 and 1600 h composed of fresh-cut pasture, meadow hay, and pelleted cereal grain to achieve differing levels of DMI and ME density (LL: 16.6 kg of DMI and 174 MJ of ME; HL: 17.3 kg of DMI and 181.1 MJ of ME; LH: 15.4 kg of DMI and 183.1 MJ of ME; HH: 17.9 kg of DMI and 213.3 MJ of ME, with the first letter indicating DMI and the second ME, and with H indicating high and L indicating low, respectively). The first day cows were placed on their diets was designated d 0. Concentrations of IGF-I were measured in frozen-thawed samples of plasma using a verified ELISA. Dietary treatment had affected plasma concentrations of IGF-I by d 7 with cows on high ME diets having greater IGF-I concentrations at d 14 (83.7 vs. 45.6 ng/mL) than cows on the low ME diets. The level of DMI had less effect on plasma concentrations of IGF-I at d 14 (72.2 vs. 57.1 ng/mL). Dietary treatment effects on these concentrations had stabilized by d 21. Day-to-day variation in mean plasma concentrations of IGF-I within each dietary treatment was low during an intensive period of daily sampling for 14 d (from d 22 to 35). Within-cow day-to-day variation was also low compared with that among cows within the same dietary group and was associated with a high repeatability in the day-to-day concentration of IGF-I in individual cows. Intraclass correlation coefficients for IGF-I ranged from 0.56 (± 0.14) to 0.88 (± 0.06) with a combined (pooled) value for the 4 subgroups of 0.77 (± 0.05). The ME and DMI effects (H vs. L) at d 35 were 79.3 vs. 41.4 and 62.0 vs. 55.7 ng/mL, respectively. Although the ME and DMI differences also affected milk yield and compositional parameters, the effects were not as proportionately great as those measured for IGF-I. Altering the ME or DMI components of the pasture-based diets produced changes in plasma IGF-I concentrations that did not become stabilized for 3 wk, but were then highly repeatable for individual cows within each dietary group. Both observations have relevance to interpreting data related to plasma concentrations of IGF-I in lactating Holstein cows.     

Evaluation of net energy expenditures of dairy cows according to body weight changes over a full lactation

Equations that predict daily dry matter intake (DMI) of a lactating cow could be evaluated by comparing the predicted accumulation of energy in body weight (BW) over the course of lactation with the observed BW evolution. However, to do so requires that first the energy balance calculations from observed DMI are evaluated. The purpose of the work reported here was to determine the degree of deviation of predicted from observed BW, according to net energy for lactation (NE_L) balance calculated from weekly observations of DMI, BW, and fat-corrected milk production in 21 sets of full-lactation data, and to determine an appropriate correction of the NE_L bias for subsequent DMI prediction evaluations. When the National Research Council maintenance equation 0.08 × BW(kg)^0.75 was used in energy balance calculation, BW was overpredicted with an increasing difference between the cumulative predicted BW and observed BW as lactation progressed. Placing all the error of BW prediction into maintenance energy expenditures resulted in a best-fit equation of 0.096 ± 0.003 Mcal/kg of BW^0.75. A time-dependent equation was also developed, in which weekly maintenance expenditures were determined as the NE_L expenditure to yield a zero NE_L balance and could be described by a second-order polynomial equation related to week of lactation (WOL) where maintenance NE_L = [−0.0227(± 0.0098) × WOL² + 1.352(± 0.456) × WOL + 78.09(± 4.92) Mcal/kg of BW^0.75] × 10⁻³. Average maintenance energy expenditure at the onset of lactation was approximately 0.08 Mcal/kg of BW^0.75, and this value increased to a plateau at wk 15 of lactation of approximately 0.098 Mcal/kg of BW^0.75. Standard deviations between data sets of weekly maintenance parameter estimates throughout lactation were large but consistent at approximately 25% of the mean. Revision of the maintenance energy expenditure estimate substantially improved BW prediction by the energy balance model. On average, the 0.096 Mcal of NE_L/kg of BW^0.75 equation resulted in the best BW predictions, although substantial variation existed around this value.     

Perturbations of plasma metabolites correlated with the rise of rumen endotoxin in dairy cows fed diets rich in easily degradable carbohydrates

Feeding dairy cows diets high in easily degradable carbohydrates increases the incidence of rumen and systemic metabolic disorders; however, the triggering factor is not well understood. In this study, dairy cows were fed 4 different amounts of barley grain-based concentrate at 15, 30, 45, and 60% (dry matter basis) of a total mixed ration to determine whether alterations in the rumen environment would be associated with perturbations of the plasma profile of selected metabolites. In addition, associations among free rumen endotoxin and several plasma metabolites were determined. The study was a replicated 4 × 4 Latin square design with 8 rumen-cannulated lactating dairy cows (60 ± 15 d in milk). Multiple rumen fluid and blood plasma samples were collected and analyzed for pH and rumen fluid endotoxin and for concentrations of glucose, insulin, cholesterol, β-hydroxybutyrate (BHBA), nonesterified fatty acids (NEFA), and lactate in the plasma. Rumen pH decreased below 6.0, from 8 to 12 h after the morning feeding, with the augmentation of the proportion of concentrate in the diet of ≥30%. Feeding diets with >30% concentrate resulted in a rise of free endotoxin in the rumen fluid (8.87 ± 0.39 μg/mL). Inclusion of 60% concentrate in the total mixed ration was associated with enhanced concentrations of glucose (64.5 ± 1.0 mg/dL) and lactate (540.9 ± 36.5 μmol/L) and lowered cholesterol (265.5 ± 13.7 mg/dL), BHBA (449.1 ± 47.4 μmol/L), and NEFA (138.8 ± 19.1 μEq/L) in the blood plasma. The regression analysis revealed that greater concentrations of plasma lactate and lower concentrations of cholesterol, BHBA, and NEFA were related to the rise of rumen endotoxin. Interestingly, 93% of the increase in the plasma lactate was explained by the rise of rumen endotoxin. Moreover, the analysis revealed inverse relationships of rumen endotoxin with plasma cholesterol (R² = 0.47), BHBA (R² = 0.37), and NEFA (R² = 0.50) and a biphasic response of plasma insulin (R² = 0.58). Taken together, feeding dairy cows diets rich in rumen-degradable carbohydrates and low in fiber led to lower rumen pH and a large accumulation of rumen endotoxin; the latter was correlated with perturbations of plasma metabolites allied to carbohydrate and lipid metabolism.     

Effects of intravenous glucose infusion and nutritional balance on serum concentrations of nonesterified fatty acids, glucose, insulin, and progesterone in nonlactating dairy cows

The objective of this study was to evaluate serum concentrations of nonesterified fatty acids, glucose, insulin, and progesterone in nonlactating dairy cows according to nutritional balance and glucose infusion. Ten nonlactating, ovariectomized Gir × Holstein cows were stratified by body weight (BW) and body condition score (BCS) on d −28 of the study, and randomly assigned to 1) negative nutrient balance (NB) or 2) positive nutrient balance (PB). From d −28 to d 0, cows were allocated according to nutritional treatment (5 cows/treatment) into 2 low-quality pastures with reduced forage availability. However, PB cows individually received, on average, 3 kg/cow per day (as-fed) of a concentrate during the study. All cows had an intravaginal progesterone releasing device inserted on d −14, which remained in cows until the end of the study. Cow BW and BCS were assessed again on d 0. On d 0, cows within nutritional treatment were randomly assigned to receive, in a crossover design containing 2 periods of 24 h each, 1) intravenous glucose infusion (GLU; 0.5 g of glucose/kg of BW, as a 5% glucose solution administered, on average, at 32 mL/min over a 3-h period), or 2) intravenous saline infusion (SAL; 0.9% solution infused on average at 32 mL/min over a 3-h period). Prior to the beginning of each period, all cows were fasted for 12 h. Blood samples were collected, relative to the beginning of the infusion, at −12 and −11.5 h (beginning of fasting), and at −0.5, 0, 0.5, 1, 2, 3, 4, 5, and 6 h. Following the last blood collection of period 1, cows received (PB) or not (NB) concentrate and were returned to their respective pastures. Changes in BCS and BW were greater in NB cows compared with PB cows (−0.60 and −0.25 ± 0.090 for BCS, respectively; −22.4 and 1.2 ± 6.58 kg for BW, respectively). Cows receiving GLUC had greater glucose concentrations from 0.5 to 3 h relative to infusion compared with SAL cows. Insulin concentrations were greater in PB cows assigned to GLUC compared with SAL cohorts at 0.5 and 3 h following infusion, whereas NB cows assigned to GLUC had greater insulin concentrations compared with SAL cohorts at 0.5, 1, 2, and 3 h. Progesterone concentrations were greater in PB cows assigned to GLUC at 2, 3, and 4 h following infusion compared with SAL cohorts. In conclusion, the effects of glucose infusion on serum concentrations of insulin and progesterone in nonlactating dairy cows were dependent on cow nutritional status.     

Selection of barley grain affects ruminal fermentation, starch digestibility, and productivity of lactating dairy cows

The objective of this study was to evaluate the effects of 2 lots of barley grain cultivars differing in expected ruminal starch degradation on dry matter (DM) intake, ruminal fermentation, ruminal and total tract digestibility, and milk production of dairy cows when provided at 2 concentrations in the diet. Four primiparous ruminally cannulated (123 ± 69 d in milk; mean ± SD) and 4 multiparous ruminally and duodenally cannulated (46 ± 14 d in milk) cows were used in a 4 × 4 Latin Square design with a 2 × 2 factorial arrangement of treatments with 16-d periods. Primiparous and multiparous cows were assigned to different squares. Treatments were 2 dietary starch concentrations (30 vs. 23% of dietary DM) and 2 lots of barley grain cultivars (Xena vs. Dillon) differing in expected ruminal starch degradation. Xena had higher starch concentration (58.7 vs. 50.0%) and greater in vitro 6-h starch digestibility (78.0 vs. 73.5%) compared with Dillon. All experimental diets were formulated to supply 18.3% crude protein and 20.0% forage neutral detergent fiber. Dry matter intake and milk yield were not affected by treatment. Milk fat concentration (3.55 vs. 3.29%) was greater for cows fed Dillon compared with Xena, but was not affected by dietary starch concentration. Ruminal starch digestion was greater for cows fed high-starch diets compared with those fed low-starch diets (4.55 vs. 2.49 kg/d), and tended to be greater for cows fed Xena compared with those fed Dillon (3.85 vs. 3.19 kg/d). Ruminal acetate concentration was lower, and propionate concentration was greater, for cows fed Xena or high-starch diets compared with cows fed Dillon or low-starch diets, respectively. Furthermore, cows fed Xena or high-starch diets had longer duration that ruminal pH was below 5.8 (6.6 vs. 4.0 and 6.4 vs. 4.2 h/d) and greater total tract starch digestibility (94.3 vs. 93.0 and 94.3 vs. 93.0%) compared with cows fed Dillon or low-starch diets, respectively. These results demonstrate that selection of barley grain can affect milk fat production and rumen fermentation to an extent at least as great as changes in dietary starch concentration.     

Short communication: Effect of a stable pen management strategy for precalving cows on dry matter intake, plasma nonesterified fatty acid levels, and milk production

During the close-up transition period, dairy cows are at risk for negative energy balance due to increasing energy demands and decreasing feed intake. This can result in postparturient health problems and decreased milk production after calving. Cows are frequently regrouped during this period, which can negatively affect feeding and resting behavior. The hypothesis was that housing in a stable pen during the close-up transition period should result in a more settled environment resulting in fewer displacements from the feed bunk, which would result in improved feed intake, energy balance [lower nonesterified fatty acid (NEFA) concentrations], and lactation performance. This study addresses precalving pen grouping strategies, which have the potential to affect feed intake and energy balance. A randomized complete block design with pen as the experimental unit was used to compare a stable (S) housing strategy (cows with similar calving dates added to a precalving pen at once) to the more traditional dynamic (D) housing strategy (cows added up to 2 times per week to a precalving pen). Twice-weekly blood samples were collected for NEFA analysis and cow interactions within the pen were observed. Dry matter intake (DMI), milk production, and postparturient health problems were recorded. Mean DMI for the duration of the 28 d of the study was not different (S: 25.5 ± 1.6 vs. D: 25.7 ± 1.0 kg/d), and when examined over time relative to calving, no treatment by time interaction was observed. Concentrations of NEFA were not different when cows initially entered the pens (S: 0.21 ± 0.10 vs. D: 0.18 ± 0.04 mEq/L) and remained not different for the time intervals closer to calving (d −9 to −14: S: 0.28 ± 0.09 vs. D: 0.21 ± 0.04; d −3 to −6: S 0.36 ± 0.10, D 0.32 ± 0.05 mEq/L). Pen grouping strategy did not affect DMI, plasma NEFA concentrations, or milk production.     

Evaluation of equations to predict dry matter intake of dairy heifers

Daily pen dry matter intakes (DMI, n = 9,275) were collected over a 28-mo period at the University of Wisconsin's Integrated Dairy Research Facility. Heifers were housed in pens containing 8 Holstein or Holstein × Jersey crossbred heifers/pen. Heifer diets were formulated to energy and protein requirement twice monthly, with feed intake, dietary nutrient density, and ambient temperature recorded daily. Heifers were weighed at 60-d intervals, and mean pen body weights (BW) were estimated for each day between the weigh dates using the interval average daily gain as a regression coefficient. Prediction of heifer DMI was evaluated using the equations of NRC (2001), Quigley et al. (1986), or alternative random effects mixed models or nonlinear exponential models. The effects of breed, BW, temperature and neutral detergent fiber deviation (NDFdv) were considered as independent variables. Holstein and crossbred heifer DMI was predicted with reasonable precision [standard error (SE) < 0.86 kg/d], by the NRC (2001) or Quigley et al. (1986) equations, but heifer DMI was over- or underpredicted for heifers >500 kg, respectively. Improved heifer DMI prediction equations were achieved with exponential models. For Holsteins (SE = 0.71 kg/d), the prediction equation was: DMI (kg/d) = 15.79 × [1 - e^{(−0.00210 × BW)}] − 0.0820 × NDFdv, where NDFdv = (dietary neutral detergent fiber as a % of dry matter) - {22.07 + [0.08714 × BW] - [0.00007383 × (BW)²]}. For crossbred heifers (SE = 0.60 kg/d), the prediction equation was: DMI (kg/d) = 13.48 × [1 - e^{(−0.00271 × BW)}] - 0.0824 × NDFdv where NDFdv = (dietary neutral detergent fiber as a % of dry matter) - {23.11 + [0.07968 × BW] - [0.00006252 × (BW)²]}. Alternative exponential DMI model equations when dietary neutral detergent fiber is unknown were also developed. The Holstein DMI equation (SE = 0.73 kg/d) was: DMI (kg/d) = 15.36 × [1 - e^{(−0.00220 × BW)}], and the crossbred DMI equation (SE = 0.81 kg/d) was: DMI (kg/d) = 12.91 × [1 - e^{(−0.00295 × BW)}].     

Effects of a supplemental yeast culture on heat-stressed lactating Holstein cows

Multiparous, lactating Holstein cows (n = 23; 120 ± 30 d in milk, 690 ± 67 kg of body weight) housed in climatic chambers were randomly assigned to 1 of 2 dietary treatments: a diet containing a novel yeast culture formulation (YC) for heat stress (n = 12, 10 g/d) or a control diet (n = 11). The trial length was 28 d and consisted of a 7-d thermal neutral period (TN; 18°C, 20% humidity) followed by 21 d of heat stress (HS; cyclical daily temperatures ranging from 29.4 to 37.8°C and 20% humidity). Cows were individually fed a total mixed ration consisting primarily of alfalfa hay and steam-flaked corn. During TN, the YC feeding had no effect on production variables or most body temperature indices. During HS, all body temperature indices increased and YC had no effect on rump surface temperature, respiration rate, or sweating rates. Cows fed YC had lower rectal temperatures at 1200 and 1800 h (40.29 vs. 40.02°C and 40.35 vs. 40.12 ± 0.07°C, respectively) compared with control-fed cows. Cows fed both diets lost body weight (42 kg) during HS, but there were no differences between diets. Control-fed cows had increased dry matter intake (DMI) and milk yield (19.1 vs. 17.9 ± 0.5 kg/d and 32.15 vs. 29.15 ± 0.02 kg/d, respectively) compared with YC-fed cows, but intake and milk production were similar between diets when evaluated on a body weight basis. Heat stress progressively decreased DMI (29%) and milk yield, with milk production reaching a nadir (33%) in the third week. Heat stress decreased milk protein (7%) and lactose (5%) levels, but did not alter milk fat content. Heat-stressed cows were in calculated negative energy balance (−1.91 ± 0.70 Mcal/d) and this was unaffected by diet. Independent of diet, HS decreased plasma glucose (11%), but neither diet nor HS altered basal nonesterified fatty acid levels. Heat stress increased plasma urea N concentrations (11.5 vs. 14.8 ± 0.4 mg/dL). Despite YC-fed cows having slightly reduced body temperatures indices, feeding YC did not prevent the negative effects of HS.     

The effect of calcium-naloxone treatment on blood calcium, beta-endorphin, and acetylcholine in milk fever

Milk fever is a postpartum syndrome of cows characterized by acute hypocalcemia, which reduces the release of acetylcholine (ACH), inducing flaccid paralysis and recumbency. Our aim was to evaluate the effect of calcium (Ca²⁺) combined with naloxone (Nx, an opioid antagonist; Ca²⁺-Nx) on plasma concentrations of ACH, ß-endorphin (ßE), and Ca²⁺ just before treatment (T0) and at 15, 30, and 90 min after treatment (T15, T30, and T90, respectively). Thirty cows were divided into 3 groups of 10 cows each. In group A1, cows affected by milk fever were treated (i.v.) with a combination of 0.2 mL/kg of body weight (BW) of Ca²⁺ borogluconate (20%) and 0.01 mg/kg of BW of Nx hydrochloride dihydrate. In group A2, cows affected by milk fever were treated (i.v.) with 2 mL/kg of BW of Ca²⁺ borogluconate (20%). In group C, healthy cows were treated (i.v.) with a combination of 0.2 mL/kg of BW of Ca²⁺ borogluconate (20%) and 0.01 mg/kg of BW of Nx hydrochloride dihydrate. Cows underwent treatments within 24 h of calving. Blood samples were collected at T0 and at T15, T30, and T90 for quantitative determination of ACH, ßE, and Ca²⁺. The cows in groups A1 and A2 recovered within a mean of 20 ± 10 min, although 4 cows in group A2 underwent a relapse. Blood Ca²⁺ concentrations in group C increased slightly at T30 and at T90 (T30: 8.8 ± 0.6 mg/dL; T90: 8.7 ± 0.6 mg/dL) after treatment, whereas the response in groups affected by milk fever was similar, even though Ca²⁺ concentrations showed a sharp increase (A1: 8.9 ± 0.8 mg/dL; A2: 6.0 ± 0.7 mg/dL), particularly at T15 in group A1. Concentrations of ßE showed a similar pattern in groups A1 and C, with an increase at T15 (A1: 8.2 ± 1.0 ng/mL; C: 2.7 ± 0.4 ng/mL) and a subsequent decrease until T90 (A1: 1.4 ± 0.3 ng/mL; C: 1.4 ± 0.4 ng/mL), whereas ßE remained constant throughout in group A2. Concentrations of ACH in group A1 decreased significantly between T0 and T15, T30, and T90 (T0: 7.2 ± 1.1 nmol/L; T15: 4.2 ± 1.2 nmol/L; T30: 2.9 ± 0.8 nmol/L; T90: 3.1 ± 0.3 nmol/L), whereas in group A2, it did not change. In group C, concentrations of ACH decreased at T15 and increased again at T30 (T15: 1.1 ± 0.3 nmol/L; T30: 3.2 ± 0.7 nmol/L). Our results suggest that administration of Ca²⁺-Nx, which restored the physiological Ca²⁺ concentrations, might have an effect on nicotinic receptors by restoring the normal neuromuscular transmission at the motor endplate.     

Effects of dietary cation-anion difference on intake, milk yield, and blood components of the early lactation cow

Early lactation Holsteins cows (15 primiparous and 18 multiparous) were offered rations with dietary cation-anion difference, calculated as mEq (Na + K − Cl − S)/100 g of feed dry matter (DCAD:S), of 20, 35, or 50 mEq from d 0 (calving) to 42 d postpartum (August 20, 2000 to January 9, 2001) to determine the effects of increasing DCAD:S on dry matter intake (DMI), milk yield, and blood metabolites. For DCAD:S of 20, 35, and 50, DMI was 3.30, 3.38, 2.96 kg/100 kg of body weight (BW); milk yield was 25.5, 24.2, and 22.4 kg/d, respectively. No differences were observed for concentration or yield of milk fat or milk protein. Serum Ca, P, Mg, Na, K, Cl, cation-anion difference, insulin, and glucose did not differ with DCAD. Serum HCO₃⁻ was 26.07, 25.88, and 27.64 mEq/L for 20, 35, and 50 DCAD:S. Serum Ca, Mg, Na, and K concentrations were greater for primiparous cows (9.52 mg/dL, 2.35 mg/dL, 140.03 mEq/L, 4.66 mEq/L, respectively) than for multiparous cows (9.27 mg/dL, 2.12 mg/dL, 137.63 mEq/L, 4.46 mEq/ L, respectively). A DCAD:S between 23 and 33 mEq/ 100 g of dry matter (DM) appears to be adequate during cool weather for the milk yield that occurred in the present study based on DMI (kg/100 kg of BW), whereas DCAD:S of 50 mEq/100 g of DM may be excessive and could be too alkaline or unpalatable, resulting in decreased DMI (kg/100 kg of BW).     

Short communication: Prediction of intake in dairy cows under tropical conditions

A meta-analysis was conducted to develop a model for predicting dry matter intake (DMI) in dairy cows under the tropical conditions of Brazil and to assess its adequacy compared with 5 currently available DMI prediction models: Agricultural and Food Research Council (AFRC); National Research Council (NRC); Cornell Net Carbohydrate and Protein System (CNCPS; version 6); and 2 other Brazilian models. The data set was created using 457 observations (n = 1,655 cows) from 100 studies, and it was randomly divided into 2 subsets for statistical analysis. The first subset was used to develop a DMI prediction equation (60 studies; 309 treatment means) and the second subset was used to assess the adequacy of DMI predictive models (40 studies; 148 treatment means). The DMI prediction model proposed in the current study was developed using a nonlinear mixed model analysis after reparameterizing the NRC equation but including study as a random effect in the model. Body weight (mean = 540 ± 57.6 kg), 4% fat-corrected milk (mean = 21.3 ± 7.7 kg/d), and days in milk (mean = 110 ± 62 d) were used as independent variables in the model. The adequacy of the DMI prediction models was evaluated based on coefficient of determination, mean square prediction error (MSPE), root MSPE (RMSPE), and concordance correlation coefficient (CCC). The observed DMI obtained from the data set used to evaluate the prediction models averaged 17.6 ± 3.2 kg/d. The following model was proposed: DMI (kg/d) = [0.4762 (±0.0358) × 4% fat-corrected milk + 0.07219 (±0.00605) × body weight^0.75] × (1 – e^{−0.03202 (±0.00615) × [days in milk + 24.9576 (±5.909)]}). This model explained 93.0% of the variation in DMI, predicting it with the lowest mean bias (0.11 kg/d) and RMSPE (4.9% of the observed DMI) and the highest precision [correlation coefficient estimate (ρ) = 0.97] and accuracy [bias correction factor (C_b) = 0.99]. The NRC model prediction equation explained 92.0% of the variation in DMI and had the second lowest mean bias (0.42 kg/d) and RMSPE (5.8% of the observed DMI), as well as the second highest precision (ρ = 0.94) and accuracy (C_b = 0.98). The CNCPS and AFRC DMI prediction models explained 93.0 and 85.0% of the variation in DMI but underpredicted DMI by 1.8 and 1.4 kg/d, respectively. These 2 models (CNCPS and AFRC) resulted, respectively, in RMSPE of 11.3 and 10.7% of the observed DMI, with moderate to high precision (ρ = 0.81 and 0.82) and accuracy (C_b = 0.84 and 0.89). The remaining 2 models resulted in the poorest results, underpredicting DMI by 2.3 and 1.9 kg/d, with RMSPE of 22.8 and 14.9% of the observed DMI and moderate to low precision (ρ = 0.49 and 0.76) and accuracy (C_b = 0.81 and 0.86). The new model derived from the current meta-analytical approach provided the best accuracy and precision for predicting DMI in lactating dairy cows under Brazilian conditions.     

Effects of feeding polyphenols from pomegranate extract on health, growth, nutrient digestion, and immunocompetence of calves

Objectives were to determine effects of feeding pomegranate extract (POMx) rich in polyphenols on performance, health, nutrient digestion, and immunocompetence of calves in the first 70 d of age. Holstein calves (n = 67), at 2 ± 1 d of age (d 0 = birth day) were randomly assigned to 0 (control), 5 (POMx5), or 10 g/d (POMx10) of pomegranate extract containing 16.9% gallic acid equivalent (GAE) to result in intakes of 0, 850 and 1,700 mg of GAE/d or an average of approximately 0, 15, and 30 mg of GAE/kg of body weight (BW) per day. All calves received colostrum during the first 24 h, pasteurized milk thereafter until 61 d of age, and grain was fed ad libitum for the first 70 d of age. Calves were housed in individual hutches, and grain intake, attitude and fecal scores, incidence and duration of health disorders, and treatments for health problems were evaluated daily. Body weight was measured on 2 consecutive days at 2, 30, and 70 d of age and averaged for each measurement. Concentrations of glucose and 3-hydroxybutyrate were measured in plasma. Nutrient digestion was measured using total fecal collection during a 3-d period. Neutrophil phagocytic and killing activities and antibody response to immunization with ovalbumin were measured. Peripheral blood mononuclear cells were cultured and cytokine production measured. Feeding POMx had no effect on intake or BW gain in the first 30 d of age, but after 30 d of age, both grain dry matter intake and BW gain decreased with increasing addition of POMx, which resulted in calves that were 1.8 and 4.3 kg lighter at 70 d of age for POMx5 and POMx10, respectively, compared with controls. Feeding POMx did not influence dry matter, organic matter, or starch digestibility, but it reduced crude protein and fat digestion. Plasma concentrations of glucose and 3-hydroxybutyrate were similar among treatments throughout the first 70 d of age. Measures of calf health such as fecal and attitude scores, risk of fever, and rectal temperature were not altered by treatments. Similarly, neutrophil phagocytic and killing activities did not differ among treatments. On the contrary, feeding POMx increased synthesis of interferon-γ and interleukin-4 by peripheral blood mononuclear cells and improved total immunoglobulin G responses to ovalbumin vaccination. These results suggest that feeding POMx top-dressed onto the grain suppresses intake of grain and digestibility of fat and protein, likely because of the high tannin content. Nevertheless, polyphenols from POMx enhanced mitogen-induced cytokine production and response to vaccination, which might benefit immune competence of calves and potentially health. Additional studies are warranted to minimize the effect of POMx on intake and digestibility and to better understand the mechanisms by which polyphenols improve immune response of calves.