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ABSTRACT: Increasing nutrient supply to dairy calves has well known benefits; however, the effects of milk replacer (MR) composition when supplied in higher amounts are not fully understood, particularly in the first weeks of life. To better understand the metabolism of macronutrient supply in young calves (21 d old), we investigated diurnal patterns of heat production and substrate oxidation in young calves fed MR with an incremental supply of fat, lactose, or protein. Thirty-two groups of 3 mixed-sex Holstein-Friesian newborn calves (3.4 ± 1.6 d of age), were randomly assigned to one of 4 dietary treatments and studied for 21 d. Diets consisted of a basal MR (23.3% CP, 21.2% EE, and 48.8% lactose of DM) fed at 550 kJ/kg BW0.85 per day (CON; n = 24), or the basal MR incrementally supplied with 126 kJ of digestible energy/BW0.85 per day as milk fat (+FAT; n = 23), lactose (+LAC; n = 24), or milk protein (+PRO; n = 23). Calves were fed MR in 2 daily meals and had ad libitum access to water, but were not supplied with any calf starter nor forage. After 2 weeks of adaptation to the diets, groups of 3 calves were placed for 1 wk in an open-circuit respiration chamber for nitrogen and energy balance measurements (lasting 7 d). On d 3, glucose oxidation kinetics was estimated by using [U-13C]glucose. Measurements included total heat production (total energy [HP], activity [Hact] expenditure, resting metabolic rate [RMR]), respiration quotient (RQ), carbohydrate (COX) and fat oxidation (FOX) in 10 min. intervals and averaging these values per hour over days. Incremental supply of lactose and fat increased body fat deposition, with observed patterns in RMR indicating that this increase occurred primarily after the meals. Specifically, the average daily RMR was highest in the +PRO group and lowest in the CON treatment. The HP was higher in the +PRO group and throughout the day, hourly means of HP were higher in this treatment mainly caused by an increase in Hact. The recovery of 13CO2 from oral pulse-dosed [U-13C]glucose was high (77%), and not significantly different between treatments, indicating that ingested lactose was oxidized to a similar extent across treatments. Increasing lactose supply in young calves increased fat retention by reduction in fatty oxidation. Calves fed a MR with additional protein or fat raised RMR persistently throughout the day, whereas the extra lactose supply only affects RMR after the meal. Dietary glucose was almost completely oxidized (77% based on (13C) glucose measurement) regardless of nutrient supplementation. Extra protein supply increased HP and FOX compared with similar intakes of fat and lactose. Fasting heat production (FHP) of young, group-housed calves is comparable to literature values and unaffected by energy intake. Overall, these findings deepen our understanding of how different nutrients affect metabolic processes, fat retention, and energy expenditure in young dairy calves.

ABSTRACT: Milk replacers (MR) for calves contain alternative fat sources as substitute for milk fat. This substitution leads to differences in fat properties, such as the fatty acid profile and the triglyceride structure. This study evaluated how fat composition in MR affects gastrointestinal health, blood redox parameters, and postprandial metabolism in calves fed twice daily. Forty-five individually housed male Holstein-Friesian calves (2.3 ± 0.85 d of age) were assigned to 1 of 15 blocks based on the age and the day of arrival. Within each block, calves were randomly assigned to 1 of 3 experimental diets and received their respective diet from arrival until 35 d after arrival. The 3 experimental diets (n = 15 per treatment group) consisted of an MR with a blend of vegetable fats containing rapeseed and coconut (VG), an MR with only animal fats from lard and dairy cream (AN), and an MR containing a mixture of animal and vegetable fats including lard and coconut (MX). The fatty acid profile of each MR was formulated to resemble that of bovine milk fat while using only 2 fat sources. All MR were isoenergetic, with 30% fat (% DM), 24% crude protein, and 36% lactose. Chopped straw and water were available ad libitum from arrival onward but no starter feed was provided. Daily milk allowances were 6.0 L from d 1 to 5, 7.0 L from d 6 to 9, and 8.0 L from d 10 to 35, divided into 2 equal meals and prepared at 135 g/L (13.5% solids). Fecal appearance was scored daily; calves were weighed and blood was drawn on arrival and weekly thereafter. Urine and feces were collected over a 24-h period at wk 3 and 5 to determine apparent total-tract digestibility and assess gastrointestinal permeability using indigestible markers. Postprandial metabolism was evaluated at wk 4 by sequential blood sampling over 7.5 h, and the abomasal emptying rate was determined by acetaminophen appearance in blood. Fat composition in MR did not affect growth, MR intake, gastrointestinal permeability, nor nutrient digestibility. The percentage of calves with abnormal fecal scores was lower at wk 2 after arrival in calves fed VG than MX, whereas AN did not differ from the other treatments. Calves fed AN and MX had higher thiobarbituric acid reactive substances measured in serum than VG, whereas plasma ferric-reducing ability was greater in calves fed MX than VG. Postprandial acetaminophen concentrations did not differ across treatment groups, but the area under the curve was smaller in calves fed VG than in the other 2 treatments, which is indicative of a slower abomasal emptying. Postprandial serum triglyceride concentration was greater in calves fed AN than VG, whereas MX did not differ from the other treatments. Based on these outcomes, all 3 fat blends can be considered suitable for inclusion in MR for calves.

ABSTRACT: Fat composition in milk replacers (MR) for calves differs from bovine milk fat in multiple ways. The aim of the study was to investigate the impact of different approaches of formulating fat in MR on growth, ad libitum intakes of MR and solid feeds, as well as blood metabolites in dairy calves. Upon 24 to 96 h after birth, 63 calves were acquired from dairy farms and incorporated into the study. Calves were blocked based on arrival day and randomly assigned within each block to one of 3 treatments differing in MR fat composition (n = 21 per group): VG was based on vegetable fats including 80% rapeseed and 20% coconut fats; AN was formulated with animal fats including 65% lard and 35% dairy cream; and MX with a mixture of 80% lard and 20% coconut fats. All 3 MR contained 30% fat, 24% crude protein, and 36% lactose and were formulated to have a fatty acid profile resembling that of milk fat. From arrival onward (3.1 ± 0.84 d of age; means ± standard deviation), calves were group housed and were offered an ad libitum supply of MR at 135 g/L (13.5% solids). Weaning was gradual and induced between wk 7 and 10, after which calves were fed only solid feeds. Starter feed, chopped straw, and water were offered ad libitum throughout the study. Calves were weighed, and blood was collected weekly until d 84 after arrival. Preweaning average daily gain was greater in calves fed AN (915 g/d) than other treatments (783 g/d), whereas no differences were detected in the weaning and postweaning phases. Preweaning MR intake was greater in calves fed AN than MX from wk 2 to 6 and was also higher in calves fed AN than VG in wk 5 and 6. Consistently, the number of rewarded visits during the ad libitum phase was greater in calves fed AN than MX, whereas VG showed no differences. This led to a higher preweaning total metabolizable energy intake in calves fed AN than in calves fed VG and MX. Serum cholesterol was higher, and serum albumin was lower in calves fed VG than other treatments. The proportion of high-density lipoprotein cholesterol in total plasma cholesterol was lower and that of low-density lipoprotein (LDL) cholesterol was higher in calves fed VG compared with other treatments. Overall, the fatty acid profile of plasma largely mirrored the MR fat composition during the preweaning period. Feeding AN enhanced MR intake and improved preweaning growth compared with other treatments. Feeding VG resulted in a marked increase in plasma cholesterol, particularly in the form of LDL cholesterol, which could be linked to an excessive intake of polyunsaturated fatty acids. These findings underscore the importance of formulating the fat content of MR to be similar to bovine milk fat.

ABSTRACT: Quantifying the water and mineral losses in feces is essential to determine the optimal composition of oral rehydration solutions (ORS) for diarrheic animals. In a randomized complete block design, this study evaluated water, mineral, and blood acid-base balance of calves with naturally occurring diarrhea receiving ORS or a placebo. On d 0, 45 calves (age: 18 ± 3.2 d; mean ± SD) were selected based on the presence of visual signs of diarrhea, such as dirty tail or wet feces, along with clinical symptoms evaluated by measuring the skin turgor and the degree of enophthalmos. On d 1, calves were divided into blocks of 3 animals based on blood base excess (BE) measured at 0900 h, and within each block, calves were randomly assigned to 1 of 3 treatments (15 calves per treatment) including (1) a hypertonic ORS (HYPER; Na+ = 110 mmol/L; 370 mOsm/kg; strong ion difference [SID] = 60 mEq/L), (2) a hypotonic ORS with low Na+ (HYPO; Na+ = 77 mmol/L; 278 mOsm/kg; SID = 71 mEq/L), and (3) a placebo consisting of lukewarm water with 5 g/L of whey powder (CON). Milk replacer (MR) was fed through teat buckets twice daily at 0630 h and 1700 h in 2 equally sized meals of 2.5 L from d 1 to 3 and of 3.0 L on d 4 and 5. Treatments consisting of 2.0 L lukewarm solutions were administered between milk meals from d 1 to 3 at 1200 h and 2030 h through teat buckets. Refusals of MR and treatments were recorded daily, and blood samples were collected from the jugular vein once daily at arrival in the afternoon of d 0 and at 0900 h from d 1 to 5 after arrival. Urine and feces were collected quantitatively over a 48-h period from 1200 h on d 1 to 1200 h on d 3, and a representative sample of each 24-h period was stored. In addition, the volume of extracellular fluid was evaluated on d 2 by postprandial sampling over a 4-h period relative to the injection of sodium thiosulfate at 1300 h. Total daily fluid intake (MR, treatment, and water) from d 1 to 3 was greater in HYPER (LSM ± SEM; 8.9 ± 0.36 L/d) and HYPO (7.8 ± 0.34 L/d) than in CON (6.6 ± 0.34 L/d). This resulted in a greater water balance (water intake − fluid output in urine and feces) in calves receiving ORS (59.6 ± 6.28 g/kg BW per 24 h vs. 39.6 ± 6.08 g/kg BW per 24 h). Fecal Na+ losses were greater in HYPER than in the other treatments (81 ± 12.0 mg/kg BW per 24 h vs. 24 ± 11.8 mg/kg BW per 24 h). Blood pH was higher in HYPO (7.41 ± 0.016) than CON (7.35 ± 0.016) over the 5 monitoring days, whereas HYPER (7.37 ± 0.017) did not differ with other treatments. In this experimental model, diarrheic calves were likely unable to absorb the high Na+ load from HYPER, resulting in greater Na+ losses in feces, which might have impaired the alkalinizing capacity of HYPER. In contrast, HYPO significantly sustained blood acid-base balance compared with CON, whereas HYPER did not. This suggests that low tonicity ORS with a high SID are more suitable for diarrheic calves.

ABSTRACT: Significant differences exist in the composition of current milk replacers (MR) and bovine whole milk. This study investigated how the macronutrient profile of 3 different MR formulations containing varying amounts of fat, lactose, and protein, and a whole milk powder (WP), affect postprandial metabolism and gut permeability in male Holstein calves. Sixty-four calves (45.4 ± 4.19 kg [mean ± SD] and 1.8 ± 0.62 d of age) were blocked in order of arrival to the facility and within each block, calves were randomly assigned to 1 of 4 treatments. Treatments included a high-fat MR (HF: 25.0% dry matter [DM] fat, 22.5% protein, 38.6% lactose; n = 14), a high-lactose MR (HL: 44.6% lactose, 22.5% protein, 18.0% fat; n = 17), a high-protein MR (HP: 26.0% protein, 18.0% fat, 41.5% lactose; n = 17), and WP (26.0% fat, 24.5% protein, 38.0% lactose; n = 16). Calves were fed 3.0 L (135 g/L) 3 times daily at 0600, 1200, and 1800 h with a teat bucket. Milk intake was recorded daily for the first 28 d after arrival, and blood sampling and body weight measurements were performed at arrival and on d 7, 14, 21, and 27. Gut permeability was estimated from fractional urinary excretion of indigestible markers (Cr-EDTA, lactulose, and d-mannitol) administered as a single dose on d 21 instead of the morning milk meal. Digestibility was determined simultaneously from a total collection of feces over 24 h. Postprandial dynamics were measured on d 28 by sequential blood sampling over 7.5 h. Dry matter intake of MR over 28 d was slightly greater in calves fed HL and HP than in WP. Recovery of Cr-EDTA and d-mannitol over a 24-h urine collection was greater in calves fed WP and HP than HL calves. Apparent total-tract digestibility of crude ash, protein, and fat did not differ among treatments; however, DM digestibility was lower in calves fed WP than in other treatment groups. In addition, abomasal emptying, as indicated by the area under the curve (AUC) for acetaminophen, was slower in calves fed WP than in calves fed HF and HL. The AUC for postprandial plasma glucose was lower in calves fed HL than WP and HF and lower in calves fed HP than WP. The AUC for postprandial serum insulin was greater in calves fed HP than WP and HF, whereas calves fed HL did not differ from the other treatments. Postprandial triglycerides were greater in calves fed WP, and postprandial adiponectin was higher in calves fed HL than other treatments. The high content of lactose and protein in MR had a major effect on postprandial metabolism. This raises the possibility of optimizing MR formulations to maintain metabolic homeostasis and influence development.

ABSTRACT: Inclusion of urea in dairy cattle diets is often limited by negative effects of high levels of feed urea on dry matter intake (DMI) and efficiency of rumen N utilization. We hypothesized that supplying urea postruminally would mitigate these limitations and allow greater inclusion of urea in dairy cattle diets. Four rumen-fistulated Holstein-Friesian dairy cows (7 ± 2.1 lactations, 110 ± 30.8 d in milk; mean ± standard deviation) were randomly assigned to a 4 × 4 Latin square design to examine DMI, milk production and composition, digestibility, rumen fermentation, N balance, and plasma constituents in response to 4 levels of urea continuously infused into the abomasum (0, 163, 325, and 488 g/d). Urea doses were targeted to linearly increase the crude protein (CP) content of total DMI (diet plus infusion) by 0%, 2%, 4%, and 6% and equated to 0%, 0.7%, 1.4%, and 2.1% of expected DMI, respectively. Each 28-d infusion period consisted of a 7-d dose step-up period, 14 d of adaptation, and a 7-d measurement period. The diet was fed ad libitum as a total mixed ration [10.9% CP, 42.5% corn silage, 3.5% grass hay, 3.5% wheat straw, and 50.5% concentrate (dry matter basis)] and was formulated to meet 100%, 82%, and 53% of net energy, metabolizable protein, and rumen-degradable protein requirements, respectively. Linear, quadratic, and cubic effects of urea dose were assessed using polynomial regression assuming the fixed effect of treatment and random effects of period and cow. Dry matter intake and energy-corrected milk yield responded quadratically to urea dose, and milk urea content increased linearly with increasing urea dose. Apparent total-tract digestibility of CP increased linearly with increasing urea dose and ruminal NH3-N concentration responded quadratically to urea dose. Mean total VFA concentration was not affected by urea dose. The proportion of N intake excreted in feces decreased linearly and that excreted in urine increased linearly in response to increasing urea dose. The proportion of N intake excreted in milk increased linearly with increasing urea dose. Urinary urea excretion increased linearly with increasing urea dose. Microbial N flow responded cubically to urea dose, but the efficiency of microbial protein synthesis was not affected. Plasma urea concentration increased linearly with increasing urea dose. Regression analysis estimated that when supplemented on top of a low-CP diet, 179 g/d of postruminal urea would maximize DMI at 23.4 kg/d, corresponding to a dietary urea inclusion level of 0.8% of DMI, which is in line with the current recommendations for urea inclusion in dairy cattle diets. Overall, these results indicate that postruminal delivery of urea does not mitigate DMI depression as urea dose increases.

ABSTRACT: The composition of milk replacer (MR) for calves greatly differs from that of bovine whole milk, which may affect gastrointestinal development of young calves. In this light, the objective of the current study was to compare gastrointestinal tract structure and function in response to feeding liquid diets having a same macronutrient profile (e.g., fat, lactose, protein) in calves in the first month of life. Eighteen male Holstein calves (46.6 ± 5.12 kg; 1.4 ± 0.50 d of age at arrival; mean ± standard deviation) were housed individually. Upon arrival, calves were blocked based on age and arrival day, and, within a block, calves were randomly assigned to either a whole milk powder (WP; 26% fat, DM basis, n = 9) or a MR high in fat (25% fat, n = 9) fed 3.0 L 3 times daily (9 L total per day) at 135 g/L through teat buckets. On d 21, gut permeability was assessed with indigestible permeability markers [chromium (Cr)-EDTA, lactulose, and d-mannitol]. On d 32 after arrival, calves were slaughtered. The weight of the total forestomach without contents was greater in WP-fed calves. Furthermore, duodenum and ileum weights were similar between treatment groups, but jejunum and total small intestine weights were greater in WP-fed calves. The surface area of the duodenum and ileum did not differ between treatment groups, but the surface area of the proximal jejunum was greater in calves fed WP. Urinary lactulose and Cr-EDTA recoveries were greater in calves fed WP in the first 6 h post marker administration. Tight junction protein gene expression in the proximal jejunum or ileum did not differ between treatments. The free fatty acid and phospholipid fatty acid profiles in the proximal jejunum and ileum differed between treatments and generally reflected the fatty acid profile of each liquid diet. Feeding WP or MR altered gut permeability and fatty acid composition of the gastrointestinal tract and further investigation are needed to understand the biological relevance of the observed differences.

ABSTRACT: During weaning, withdrawal of milk replacer is not directly compensated for by an increase in solid feed intake. Therefore, greater fat inclusion in the starter might mitigate this temporary dietary energy decline. However, fat inclusion in solid feeds may generally limit rumen fermentability and development. To address these potentially conflicting outcomes, we conducted 2 experiments to evaluate the effect of supplementing a high-fat extruded pellet mixed with a calf starter on feed intake, performance, and nutrient digestibility in calves. In experiment 1, 60 Holstein bull calves were blocked by serum IgG (2,449 ± 176 mg/dL) and date of arrival (2.5 ± 0.5 d of age). Within each block, calves were randomly assigned to 1 of 3 treatments: a standard control calf starter (CON; 3.1% fat) and mixtures of CON with 10% inclusion of 1 of 2 different high-fat extruded pellets containing 85% of either hydrogenated free palm fatty acids (PFA, 7.1% fat) or hydrogenated rapeseed triglycerides (RFT, 6.7% fat). Calves were offered milk replacer up to 920 g/d until 42 d of age, followed by a gradual weaning period of 7 d. Calves had ad libitum access to the starter diets, straw, and water. No differences were observed between CON, PFA, and RFT calves on body weight (BW) or average daily gain (ADG) until 49 d of age. From weaning (50 d) until 112 d, PFA calves had a greater BW and ADG than RFT and CON animals. Moreover, PFA calves had the highest intakes of starter, straw, calculated metabolizable energy, and crude protein after weaning. Overall, no differences were present in blood β-hydroxybutyrate and glucose concentrations between treatments; however, calves in the RFT treatment had a higher concentration of insulin-like growth factor-1. In experiment 2, 24 Holstein bull calves at 3 mo of age were assigned to 1 of 8 blocks based on arrival BW and age. Within each block, calves were randomly assigned to 1 of the 3 treatments previously described for experiment 1. Calves on the RFT treatment had the lowest total-tract apparent dry matter and fat digestibility, potentially explaining the differences in performance observed between PFA and RFT calves. Inclusion of the PFA pellet at 10% with a calf starter improved BW, solid feed, and energy intake after weaning. However, these benefits were conditioned by fat source and its digestibility.

ABSTRACT: Milk replacers (MR) for calves usually contain more lactose and less fat than bovine whole milk (WM). There are insufficient data to determine whether these MR formulations are optimal for calves fed at high planes of nutrition. Thus, the effect of 3 MR formulations and a WM powder were evaluated on growth, feeding behavior, and blood metabolites in 96 male Holstein calves fed ad libitum and with 45.5 ± 4.30 kg (mean ± standard deviation) BW at arrival. Calves were blocked based on arrival sequence, and randomly assigned within block to one of the 4 treatments (n = 24 calves/group): a high-fat MR (25.0% fat, dry matter basis; 22.5% protein, 38.6% lactose; 21.3 MJ/kg; HF), a high lactose MR (44.6% lactose, 22.5% protein, 18.0% fat; 19.7 MJ/kg; HL), a high protein MR (26.0% protein, 18.0% fat, 41.5% lactose; 20.0 MJ/kg; HP), and a WM powder (26.0% fat; 24.5% protein, 38.0% lactose; 21.6 MJ/kg; WP). In the first 2 wk after arrival, calves were individually housed and were fed 3.0 L of their respective liquid feed 3 times daily at 135 g/L. They were then moved to group housing and fed ad libitum until d 42 after arrival. Weaning was gradual and took place between d 43 and 70 after arrival; thereafter, calves were fed solids only. Concentrates, chopped straw, and water were available ad libitum throughout the study. Body weight was measured, and blood was collected at arrival and then weekly thereafter from wk 1 to 12. Weight gain and height were greater in HL than WP calves. In the preweaning phase, HL and HP-fed calves consumed more milk than WP, and HL-fed calves consumed more milk than HF calves. In wk 10, starter feed intakes were lower in HF calves than in the other groups. In the preweaning phase, ME intakes were the same for all treatments. This suggests that milk intakes were regulated by the energy density of the milk supplied. The percentage of calves requiring therapeutic interventions related to diarrhea was greater in WP-fed calves (29%) than HF and HL calves (4%), whereas HP (13%) did not differ with other groups. This was coupled with lower blood acid–base, blood gas, and blood sodium in WP than in MR-fed calves. Calves fed HF had greater serum nonesterified fatty acids compared with other groups, and greater serum amyloid A compared with WP and HL calves. Among the serum parameters, insulin-like growth factor-1 and lactate dehydrogenase correlated positively with ME intake and average daily gain. The high lactose and protein intakes in HL and HP calves led to greater insulin-like growth factor-1 concentrations than in WP-fed calves. Although growth differences were limited among MR groups, the metabolic profile largely differed and these differences require further investigation.

The Kempen system is a dairy feeding system in which diet is provided in the form of a compound feed (CF) and hay offered ad libitum. Ad libitum access to CF and hay allows cows in this system to achieve a high DM intake (DMI). Out of physiological concerns, the voluntary hay intake could be increased and the consumption pattern of CF could be manipulated to maintain proper rumen functioning and health. This study investigated the effects of an artificial hay aroma and CF formulation on feed intake pattern, rumen function and milk production in mid- to late-lactating dairy cows. Twenty Holstein–Friesian cows were assigned to four treatments in a 4 × 4 Latin square design. Diet consisted of CF and grass hay (GH), fed separately, and both offered ad libitum, although CF supply was restricted in maximum meal size and speed of supply by an electronic system. Treatments were the combination of two CF formulations – high in starch (CHS) and fibre (CHF); and two GH – untreated (UGH) and the same hay treated with an artificial aroma (TGH). Meal criteria were determined using three-population Gaussian–Gaussian–Weibull density functions. No GH × CF interaction effects on feed intake pattern characteristics were found. Total DMI and CF intake, but not GH intake, were greater (P < 0.01) in TGH treatment, and feed intake was not affected by type of CF. Total visits to feeders per day, visits to the GH feeder, visits to the CF feeder and CF eating time (all P < 0.01) were significantly greater in cows fed with TGH. Meal frequency, meal size and meal duration were unaffected by treatments. Cows fed CHF had a greater milk fat (P = 0.02), milk urea content (P < 0.01) and a greater milk fat yield (P < 0.01). Cows fed TGH had a greater milk lactose content and lactose yield (P < 0.05), and milk urea content (P < 0.01). Cows fed TGH had smaller molar proportions of acetic acid and greater molar proportions of propionic acid compared with UGH. In conclusion, treatment of GH with an artificial aroma increased CF intake and total DMI, but did not affect hay intake. Additionally, GH treatment increased the frequency of visits to both feeders, and affected rumen volatile fatty acid profile. Type of CF did not affect meal patterns, ruminal pH, nor fermentation profiles.

When supplementing lamb diets with vitamin E, an equivalence factor of 1.36 is used to discriminate between RRR-α-tocopheryl acetate and all-rac-α-tocopheryl acetate. However, more recent studies suggest a need for new equivalence factors for livestock animals. The current study aimed to determine the effect of RRR- and all-rac-α-tocopheryl acetate supplementation on α-tocopherol deposition in lamb tissues. A total of 108 Rasa Aragonesa breed lambs were fed increasing amounts of all-rac-α-tocopheryl acetate (0.25, 0.5, 1.0 and 2.0 g/kg compound feed) or RRR-α-tocopheryl acetate (0.125, 0.25, 0.5 and 1.0 g/kg compound feed) by adding them to a basal diet that contained 0.025 g/kg feed of all-rac-α-tocopheryl acetate as part of the standard vitamin and mineral mixture. The diets were fed for the last 14 days before slaughtering at 25.8±1.67 kg BW. Within 20 min after slaughter samples of muscle, heart, liver, brain and spleen were frozen at −20°C until α-tocopherol analysis. Increased supplementation of either vitamin E sources led to a significant increase (P < 0.001) in α-tocopherol concentration in all tissues studied. The tissue with the highest α-tocopherol concentration was the liver followed by spleen, heart and muscle. At similar supplementation levels (0.25, 0.50 and 1.0 g/kg compound feed), α-tocopherol content in the selected tissues was not affected by α-tocopherol source. However, the ratios between RRR- and all-rac-α-tocopheryl acetate increased with the increasing α-tocopherol supplementation (at 0.25 and 1.0 g/kg compound feed), from 1.06 to 1.16 in muscle, 1.07 to 1.15 in heart, 0.91 to 0.94 in liver and 0.98 to 1.10 in spleen. The highest relative proportion of Ʃ2S (sum of SSS-, SSR-, SRS- and SRR-α-tocopherol)-configured stereoisomers was found in the liver of lambs supplemented with all-rac-α-tocopheryl acetate accounting for up to 35 to 39% of the total α-tocopherol retained, whereas the proportion of Ʃ2S-configured stereoisomers in the other tissues accounted for

The increasing availability of automated milk dispensers on dairy farms facilitates ad libitum milk supply but weaning calves from high milk allowances is challenging. This study evaluated effects of gradual weaning methods on starter intake, growth, selected blood parameters and weaning distress in ad libitum fed dairy calves during weaning and early post-weaning periods. Thirty-six male Holstein (n = 30) or crossbred (n = 6) calves were individually housed from days 2 to 14 of age and had ad libitum access to milk replacer (MR) from teat buckets. From days 15 to 84 of age, calves were grouped and had ad libitum access to MR, starter, straw and water from automated feeders. At day 35, calves were blocked (age and breed), and randomly assigned to a weaning method: (1) linear fixed (LIN), MR supply was stepped down to 6 l/day on day 36, and linearly reduced between days 36 to 63 from 6 to 2 l/day. (2) Step-down (STEP), MR supply was stepped down to 6 l/day from days 36 to 48, 4 l/day from days 49 to 56 and 2 l/day from days 57 to 63. (3) Dynamic (DYN), at day 36, MR supply was reduced for each individual calf to 75% of the average voluntary consumption between day 29 and 35, then maintained for 9 days, reduced to 50% for 10 days, and to 25% for 9 days. The DYN calves received more MR during weaning than LIN calves, whereas STEP calves had intermediate MR intake. Starter intake was not affected by weaning method. The DYN calves (1.33±0.08 kg/day) grew faster and were heavier than STEP calves (1.10±0.08 kg/day) during post-weaning period, whereas no difference was observed between LIN calves (1.23±0.08 kg/day) and others. At days 70 and 84, concentrations of β-hydroxybutyric acid were higher in LIN calves compared to STEP and DYN calves. Hair cortisol concentrations were not affected by weaning method. During the gradual weaning process CP intake seemed to recovered earlier than metabolizable energy (ME) intake in all treatments, suggesting that ME rather than CP could be the first limiting factor for growth during weaning. These results highlight the post-weaning benefits of DYN and LIN weaning methods when compared with more abrupt step-down strategies.

Male Holstein calves commonly receive minimal quantities of milk replacer (MR) to speed up weaning and reduce costs. Studies with Holstein female calves show that early life feed restriction affects energy metabolism later in life. Aiming to test this hypothesis, 120 Holstein bull calves (48.4 ± 2.2 kg of BW and 20 ± 3.2 d of age) housed in 24 pens were blocked and randomized to two treatments: A low calf MR allowance (LP) (two daily doses of 2 l each, 582 g/d of DM), or a high MR allowance (HP) (two daily meals of 4 l each, 1164 g/d of DM). Calves were weaned at day 49 of the study and slaughtered at 32.8 ± 0.5 weeks of age. Throughout the study, animals had ad libitum access to a common compound feed, straw, and water. Twenty-four animals were randomly selected for an intravenous glucose tolerance test (IVGTT). The IVGTT was performed at week 6 and 12 of the study and consisted of an intravenous glucose infusion and sequential blood sampling up to 90 min after glucose infusions. Calves were heavier for HP until week 12, after which the difference disappeared. By design, the MR intake was higher in the HP group resulting in a higher energy intake and a higher average daily gain in the preweaning phase. Blood glucose curves were not different at week 6, but at week 12, 5 min after the infusion, glucose was higher in HP calves. Insulin curves were not different at week 6. Nevertheless, in week 12, a higher insulin concentration was observed for HP 5, 10, 15, 20, 30, 35, and 45 min’ postinfusion, indicating a higher requirement of insulin to control glycemia. Differences between HP and LP calves were also observed for the quantitative insulin sensitivity check index, maximum insulin concentration, and insulin delta at week 12. Blood glucose reached maximum concentration within 5–10 min of the IVGTT test, and the concentration was, on average, 8.58 and 10.80 mmol/l at weeks 6 and 12, respectively. Insulin reached maximum concentration within 10–15 min of the IVGTT, and concentrations were, on average, 33.32 and 32.61 μUI/ml at weeks 6 and 12, respectively. Doubling MR supply improved animal growth up to weaning, but these differences disappeared by the end of the feeding period. Despite similar responses to glucose infusions preweaning, higher milk supply seemed to decrease insulin sensitivity after weaning.

The recent trend in the dairy industry towards elevated planes of milk feeding of young calves requires reconsideration of calf milk replacer (CMR) formulations. The fat:lactose ratio in CMR is typically lower than that of whole milk and effects of increasing fat inclusion at the expense of lactose in CMR on nutrient metabolism and gut function of rearing calves are not sufficiently understood. Therefore, the current study aimed to determine the effect of increasing replacement of lactose by fat on growth performance, nutrient digestibility and metabolism, and glucose/insulin sensitivity. A total of 40 male calves (1.7 ± 0.10 days of age, 46.7 ± 0.76 kg BW) were blocked based on arrival date and randomly assigned to one of four treatments containing differing levels of fat and lactose (18F: 18.8% and 47.6%; 22F: 22.3% and 42.8%; 26F: 26.0% and 38.6%; 30F: 30.1% and 33.8%, fat and lactose DM, respectively). Calves were individually housed for the duration of the 11 week study and received their CMR (150 g/L) twice a day (0700 and 1600 h) from a teat bucket. The CMR feeding schedule consisted of 6 L/d from d 2 to 14, 7 L/d from d 15 to 56, and then 4 L/d during gradual weaning from 56 to 63 days. Pelleted starter, chopped straw, and water were available ad libitum throughout the study. Measurements included feed intake, growth, nutrient digestibility, fecal composition, and blood parameters. A glucose tolerance test (GTT) was performed between 28 and 32 days of age. By design, metabolizable energy intake from CMR increased linearly with fat level, but this did not result in a difference in BW, ADG, or concentrate intake. Fecal composition remained unaffected by treatment except for higher fat content in 22F compared to 26F. Also, plasma non-esterified fatty acids and total bilirubin differences were limited to 22F having the highest values whereas 26F had the lowest values. Regarding the GTT, total area under the curve (AUC) for glucose was highest in 22F and 26F and lowest in 18F whereas the AUC between 30 and 60 min for glucose was highest in 26F and lowest in 18F and 22F. Overall, altering the lactose:fat ratio in CMR did not affect growth performance while having minor effects on nutrient metabolism, but future investigation should focus on physiological consequences and effects in adult life to understand biological impact of fat and lactose levels in CMR for rearing calves.

Skeletal muscle is vital in maintaining metabolic homeostasis and adapting to the physiological needs of pregnancy and lactation. Despite advancements in understanding metabolic changes in dairy cows around calving and early lactation, there are still gaps in our knowledge, especially concerning muscle metabolism and the changes associated with drying off. This study aimed to characterize the skeletal muscle metabolome in the context of the dietary and metabolic changes occurring during the transition from the cessation of lactation to the resumption of lactation in dairy cows. Twelve Holstein dairy cows housed in tie stalls were dried off 6 weeks (wk) before the expected calving date. Cows were individually fed ad libitum total mixed rations composed of grass silage, corn silage, and concentrate during lactation and of corn silage, barley straw, and concentrate during the dry period. The metabolome was characterized in skeletal muscle samples (M. longissimus dorsi) collected on wk -7 (9 d before dry-off), -5 (6 d after dry-off), and wk -1, and 1 relative to calving. The targeted metabolomics approach was conducted using the MxP Quant 500 kit (Biocrates Life Sciences AG) with liquid chromatography, flow injection, and electrospray ionization triple quadrupole mass spectrometry. Statistical analysis on the muscle metabolite data was performed using MetaboAnalyst 5.0, which allowed us to conduct various multivariate analyses such as principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), informative heat map generation, and hierarchical clustering. The statistical analysis revealed a clear separation between pregnancy (wk -7, -5, and -1) and post-calving (wk 1). Starting 5 wk before calving and continuing through the first wk thereafter, the concentration of 3-methylhistidine (3-MH) in the muscle increased. This coincided with an increase in the concentrations of 11 AA (Phe, His, Tyr, Trp, Arg, Asn, Leu, Ile, Gly, Ser, and Thr) in the first wk after calving, whereas Gln decreased. l-arginine pathway metabolites (homoarginine, ornithine, citrulline, and asymmetric dimethylarginine), betaine, and sarcosine followed a similar pattern, increasing from wk -7 to -5, but decreasing from wk -1 to 1. The transition from pregnancy to lactation was associated with an increase in concentrations of the long-chain acylcarnitine species C16, C16:1, C18, and C18:1 in the muscle, whereas the concentrations of phosphatidylcholine and sphingomyelin in the muscle remained stable. The significant changes observed in the metabolome mainly concerned the AA and AA-related metabolites, indicating muscle protein breakdown in the first wk after calving. The metabolites produced by the L-Arg pathway might contribute to regulating skeletal muscle mass and function in periparturient dairy cows. The elevated concentrations of long-chain acylcarnitine species in the muscle in the first wk after calving suggest incomplete fatty acid oxidation, likely due to insufficient metabolic adaptation in response to the fatty acid load around the time of calving., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Increasing nutrient supply to dairy calves has well known benefits; however, the effects of milk replacer (MR) composition when supplied in higher amounts are not fully understood, particularly in the first weeks of life. To better understand the metabolism of macronutrient supply in young calves (21 d old), we investigated diurnal patterns of heat production and substrate oxidation in young calves fed MR with an incremental supply of fat, lactose, or protein. Thirty-two groups of 3 mixed-sex Holstein-Friesian newborn calves (3.4 ± 1.6 d of age), were randomly assigned to one of 4 dietary treatments and studied for 21 d. Diets consisted of a basal MR (23.3% CP, 21.2% EE, and 48.8% lactose of DM) fed at 550 kJ/kg BW 0.85 per day (CON; n = 24), or the basal MR incrementally supplied with 126 kJ of digestible energy/BW 0.85 per day as milk fat (+FAT; n = 23), lactose (+LAC; n = 24), or milk protein (+PRO; n = 23). Calves were fed MR in 2 daily meals and had ad libitum access to water, but were not supplied with any calf starter nor forage. After 2 weeks of adaptation to the diets, groups of 3 calves were placed for 1 wk in an open-circuit respiration chamber for nitrogen and energy balance measurements (lasting 7 d). On d 3, glucose oxidation kinetics was estimated by using [U- 13 C]glucose. Measurements included total heat production (total energy [HP], activity [H act ] expenditure, resting metabolic rate [RMR]), respiration quotient (RQ), carbohydrate (COX) and fat oxidation (FOX) in 10 min. intervals and averaging these values per hour over days. Incremental supply of lactose and fat increased body fat deposition, with observed patterns in RMR indicating that this increase occurred primarily after the meals. Specifically, the average daily RMR was highest in the +PRO group and lowest in the CON treatment. The HP was higher in the +PRO group and throughout the day, hourly means of HP were higher in this treatment mainly caused by an increase in H act . The recovery of 13 CO 2 from oral pulse-dosed [U- 13 C]glucose was high (77%), and not significantly different between treatments, indicating that ingested lactose was oxidized to a similar extent across treatments. Increasing lactose supply in young calves increased fat retention by reduction in fatty oxidation. Calves fed a MR with additional protein or fat raised RMR persistently throughout the day, whereas the extra lactose supply only affects RMR after the meal. Dietary glucose was almost completely oxidized (77% based on ( 13 C) glucose measurement) regardless of nutrient supplementation. Extra protein supply increased HP and FOX compared with similar intakes of fat and lactose. Fasting heat production (FHP) of young, group-housed calves is comparable to literature values and unaffected by energy intake. Overall, these findings deepen our understanding of how different nutrients affect metabolic processes, fat retention, and energy expenditure in young dairy calves., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

We expected mitigation of the hypophagic effects of urea (U) with a coated urea (CU) product that aimed to partially shift urea supply to the post-ruminal gastrointestinal tract. Ruminal release and post-ruminal digestibility of CU was evaluated in vitro, followed by a randomised complete block experiment (54 Holstein-Friesian cows; 177 ± 72 days in milk). Soybean meal (SBM) was partially (PR) or fully (FR) replaced on an isonitrogenous basis by beet pulp and U or CU. Urea sources were included at 12 (U-PR, CU-PR) and 19 (U-FR, CU-FR) g/kg dietary dry matter (DM). Hypophagic effects were similar for U-PR and CU-PR (-11% vs. -7%), and for U-FR and CU-FR (-13% vs. -12%) compared with SBM (average 25.8 kg DM intake/d). Compared with SBM, U-PR and CU-PR reduced yields of milk (-8%) and protein (-12%), U-PR reduced yield of fat (-9%) and fat- and protein-corrected-milk (FPCM; -9%), and CU-PR tended to reduce FPCM yield (-5%). Compared with SBM, U-FR and CU-FR respectively reduced yields of milk (-21%, -22%), protein (-25%, -26%), fat (both -14%), lactose (-20%, -21%), and FPCM (-17%, -19%), and lowered N (-15%, -12%) and feed (-8%, trend, -9%) efficiency. Human-edible protein efficiency approximately doubled with U-PR and CU-PR and approximately tripled with U-FR and CU-FR compared with SBM. Milk composition and plasma urea concentration were similar between U and CU, except for a trend for a greater plasma urea concentration with U-PR compared with CU-PR. Dry matter intake patterns differed for CU-PR compared with U-PR and for CU-FR compared with U-FR, suggesting effects of urea release rate or location on feeding behaviour. Overall, replacing SBM with U or CU reduced DM intake and milk production and affected nutrient efficiencies. Coated urea influenced DM intake pattern but did not affect total DM intake or milk production compared with U., (© 2024 The Author(s). Journal of Animal Physiology and Animal Nutrition published by Wiley‐VCH GmbH.)

Recently reviewed development objectives and feeding practices in young dairy calves require an adaptation of nutrient recommendations set for milk replacer (MR) composition. Nutrient requirements of calves younger than 21 d of age, and those of calves fed with high levels of MR are insufficiently quantified. The efficiency at which macronutrients are utilized, particularly protein, substantially diminishes with age, and little data exists for the first weeks of life. In addition, in older preruminant and ruminant calves, protein and energy can be simultaneously limiting for protein gain. Whether this also applies to calves in the first weeks of life is unknown. Therefore, this study aimed to quantify the responses in protein and fat gain to incremental supply of protein, fat, or lactose to MR in very young calves. Thirty-two groups of 3 mixed-sex Holstein-Friesian newborn calves (3.4 ± 1.6 d of age), were randomly assigned to 1 of 4 dietary treatments applied for 19 d: a basal MR (23.3% crude protein, 21.2% crude fat, and 48.8% lactose, percentages of dry matter), provided at 550 kJ/kg of metabolic body weight (BW 0.85 ) per day (CON; n = 24), or the basal MR incrementally supplied with 126 kJ of digestible energy per BW 0.85 per day as milk fat (+FAT; n = 23), lactose (+LAC; n = 24), or milk protein (+PRO; n = 23). Calves were fed MR in 2 daily meals and had ad libitum access to water, but did not have access to calf starter nor any other solid feed. After 2 wk of adaptation to their respective diets, groups of calves were placed for 1 wk in an open-circuit respiration chamber for nitrogen and energy balance measurements (5 d). The incremental nutrient efficiencies indicate what percentage of extra intake of nutrients is retained. In this study, we observed that with every 100-g increase in protein intake, 52% was converted into protein deposition, and 44% contributed to heat production. Similarly, a 100-g increase in fat intake resulted in 67% being stored as fat, 22% being released as heat, and only 5% being retained as protein. Likewise, a 100-g increment in lactose intake led to 49% being stored as fat, with 38% being released as heat. Additional protein intake was not deposited as fat; extra energy intake (fat and additional lactose) increased postabsorptive N efficiency in young calves., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

This study aimed to investigate the metabolic changes in the livers of dairy cows from 1 wk before dry off to 1 wk after calving. Twelve high-yielding Holstein cows were included in a longitudinal study and housed in a tiestall barn. The cows were dried off at 6 wk before the expected calving date (dry period length = 42 d). During the entire lactation, the cows were milked twice daily at 0600 and 1700 h. Liver biopsies were taken from each cow at 4 different times: wk -7 (before drying off), -5 (after drying off), -1 and +1 relative to calving. A targeted metabolomics approach was performed by liquid chromatography and flow injection with electrospray ionization triple quadrupole mass spectrometry using the MxP Quant 500 kit (Biocrates Life Sciences AG). A total of 185 metabolites in the liver were used for the final data analysis. Principal component analysis revealed a clear separation by days of sampling, indicating a notable shift in metabolic phenotype from late lactation to the dry period and further changes after calving. Changes were observed in several classes of compounds, including AA and biogenic amines. In particular, the changes in acylcarnitines (AcylCN), phosphatidylcholines (PC), sphingomyelins (SM), and bile acids (BA) indicated extensive remodeling of the hepatic lipidome. The changes in AcylCN concentrations in early lactation suggest incomplete fatty acid oxidation in the liver, possibly indicating mitochondrial dysfunction or enzymatic imbalance. In addition, the changes in PC and SM species in early lactation indicate altered cell membrane composition, which may affect cell signaling and functionality. In addition, changes in BA concentrations and profiles indicate dynamic adaptations in BA synthesis, as well as lipid digestion and absorption during the observation period. In particular, principal component analysis showed an overlapping distribution of liver metabolites in primiparous and multiparous cows, indicating no significant difference between these groups. In addition, Volcano plots showed similar liver metabolism between primiparous and multiparous cows, with no significant fold changes (>1.5) in any metabolite at significant P-values (false discovery rate <0.05). These results provide valuable insight into the physiological ranges of liver metabolites during dry period and calving in healthy dairy cows and should contribute to the design and interpretation of future metabolite-based studies of the transition dairy cow., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Milk replacers (MR) for calves contain alternative fat sources as substitute for milk fat. This substitution leads to differences in fat properties, such as the fatty acid profile and the triglyceride structure. This study evaluated how fat composition in MR affects gastrointestinal health, blood redox parameters, and postprandial metabolism in calves fed twice daily. Forty-five individually housed male Holstein-Friesian calves (2.3 ± 0.85 d of age) were assigned to 1 of 15 blocks based on the age and the day of arrival. Within each block, calves were randomly assigned to 1 of 3 experimental diets and received their respective diet from arrival until 35 d after arrival. The 3 experimental diets (n = 15 per treatment group) consisted of an MR with a blend of vegetable fats containing rapeseed and coconut (VG), an MR with only animal fats from lard and dairy cream (AN), and an MR containing a mixture of animal and vegetable fats including lard and coconut (MX). The fatty acid profile of each MR was formulated to resemble that of bovine milk fat while using only 2 fat sources. All MR were isoenergetic, with 30% fat (% DM), 24% crude protein, and 36% lactose. Chopped straw and water were available ad libitum from arrival onward but no starter feed was provided. Daily milk allowances were 6.0 L from d 1 to 5, 7.0 L from d 6 to 9, and 8.0 L from d 10 to 35, divided into 2 equal meals and prepared at 135 g/L (13.5% solids). Fecal appearance was scored daily; calves were weighed and blood was drawn on arrival and weekly thereafter. Urine and feces were collected over a 24-h period at wk 3 and 5 to determine apparent total-tract digestibility and assess gastrointestinal permeability using indigestible markers. Postprandial metabolism was evaluated at wk 4 by sequential blood sampling over 7.5 h, and the abomasal emptying rate was determined by acetaminophen appearance in blood. Fat composition in MR did not affect growth, MR intake, gastrointestinal permeability, nor nutrient digestibility. The percentage of calves with abnormal fecal scores was lower at wk 2 after arrival in calves fed VG than MX, whereas AN did not differ from the other treatments. Calves fed AN and MX had higher thiobarbituric acid reactive substances measured in serum than VG, whereas plasma ferric-reducing ability was greater in calves fed MX than VG. Postprandial acetaminophen concentrations did not differ across treatment groups, but the area under the curve was smaller in calves fed VG than in the other 2 treatments, which is indicative of a slower abomasal emptying. Postprandial serum triglyceride concentration was greater in calves fed AN than VG, whereas MX did not differ from the other treatments. Based on these outcomes, all 3 fat blends can be considered suitable for inclusion in MR for calves., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Fat composition in milk replacers (MR) for calves differs from bovine milk fat in multiple ways. The aim of the study was to investigate the impact of different approaches of formulating fat in MR on growth, ad libitum intakes of MR and solid feeds, as well as blood metabolites in dairy calves. Upon 24 to 96 h after birth, 63 calves were acquired from dairy farms and incorporated into the study. Calves were blocked based on arrival day and randomly assigned within each block to one of 3 treatments differing in MR fat composition (n = 21 per group): VG was based on vegetable fats including 80% rapeseed and 20% coconut fats; AN was formulated with animal fats including 65% lard and 35% dairy cream; and MX with a mixture of 80% lard and 20% coconut fats. All 3 MR contained 30% fat, 24% crude protein, and 36% lactose and were formulated to have a fatty acid profile resembling that of milk fat. From arrival onward (3.1 ± 0.84 d of age; means ± standard deviation), calves were group housed and were offered an ad libitum supply of MR at 135 g/L (13.5% solids). Weaning was gradual and induced between wk 7 and 10, after which calves were fed only solid feeds. Starter feed, chopped straw, and water were offered ad libitum throughout the study. Calves were weighed, and blood was collected weekly until d 84 after arrival. Preweaning average daily gain was greater in calves fed AN (915 g/d) than other treatments (783 g/d), whereas no differences were detected in the weaning and postweaning phases. Preweaning MR intake was greater in calves fed AN than MX from wk 2 to 6 and was also higher in calves fed AN than VG in wk 5 and 6. Consistently, the number of rewarded visits during the ad libitum phase was greater in calves fed AN than MX, whereas VG showed no differences. This led to a higher preweaning total metabolizable energy intake in calves fed AN than in calves fed VG and MX. Serum cholesterol was higher, and serum albumin was lower in calves fed VG than other treatments. The proportion of high-density lipoprotein cholesterol in total plasma cholesterol was lower and that of low-density lipoprotein (LDL) cholesterol was higher in calves fed VG compared with other treatments. Overall, the fatty acid profile of plasma largely mirrored the MR fat composition during the preweaning period. Feeding AN enhanced MR intake and improved preweaning growth compared with other treatments. Feeding VG resulted in a marked increase in plasma cholesterol, particularly in the form of LDL cholesterol, which could be linked to an excessive intake of polyunsaturated fatty acids. These findings underscore the importance of formulating the fat content of MR to be similar to bovine milk fat., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this pilot study was to generate data to support the development of an experimental model of hindgut acidosis to further understand its systemic consequences independently of rumen acidosis. Four ruminally fistulated multiparous Holstein cows (213 ± 11 d in milk) were subjected to 2 consecutive experimental periods (P1 and P2), separated by a 3-d washout. Experimental periods were 96 h long from the baseline to the final measurements but expanded over 5 calendar days (d 0-4). Abomasal infusions of saline and corn starch (2.8 kg/d) were performed for the first 72 h (d 0-3) of P1 and P2, respectively. Final measurements were performed 24 h after the end of the infusions (d 4). Each cow was used as its own control by comparing P2 to P1. Postruminal-intestinal permeability was assessed by Cr appearance in blood after a pulse dose administration of Cr-EDTA into the abomasum on d 2 (48 h after infusion initiation) of each period. Starch infusion during P2 was associated with a milk protein yield increase (3.3%) and a decrease in milk urea nitrogen (11%). Fecal dry matter increased (8.8%), and starch content tended to increase (∼2 fold) during P2. There was a period-by-day interaction for fecal pH as it decreased during starch infusion (1.3 pH points) but remained constant during P1. Although fecal lactate was not detectable during P1, it consistently increased during starch infusion. Fecal alkaline phosphatase activity also increased (∼17 fold) in association with starch infusion. Two hours after Cr-EDTA administration, blood Cr concentration was higher during starch infusion, resulting in a tendency for a treatment-by-hour interaction. Furthermore, blood d-lactate increased (∼2.5 fold), serum Cu decreased (18%), and blood urea nitrogen, cholesterol, and Ca tended to decrease (9.4%, 1.2%, and 2.4%, respectively), relative to P1. The current results suggest that hindgut acidosis was successfully induced by postruminal starch infusion, leading to gut damage and increased intestinal permeability. However, indications of systemic inflammation were not observed. The herein described preliminary results will require confirmation in a properly powered study., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this study was to characterize changes in the serum metabolome and various indicators of oxidative balance in dairy cows starting 2 wk before dry-off and continuing until wk 16 of lactation. Twelve Holstein dairy cows (body weight 745 ± 71 kg, body condition score 3.43 ± 0.66; mean ± SD) were housed in a tiestall barn from 10 wk before to 16 wk after parturition. Cows were dried off 6 wk before the expected calving date (mean dry period length = 42 d). From 8 wk before calving to 16 wk after calving, blood samples were taken weekly to study redox metabolism by determining antioxidant capacity, measured as the ferric-reducing ability of plasma, reactive oxidative metabolites, oxidative stress index, oxidative damage of lipids, measured as thiobarbituric acid reactive substances, and glutathione peroxidase activity. According to these results, dairy cows had the lowest serum antioxidant capacity and greater levels of oxidative stress during the dry-off period and the early postpartum period. For metabolomics, a subset of serum samples including wk -7 (before dry-off), -5 (after dry-off), -1, 1, 5, 10, and 15 relative to calving were used. A targeted metabolomics approach was performed using liquid chromatography and flow injection with electrospray ionization triple quadrupole mass spectrometry using the MxP Quant 500 kit (Biocrates Life Sciences AG). A total of 240 metabolites in serum were used in the final data analysis. Principal component analysis revealed a clear separation by days of sampling, indicating a remarkable shift in metabolic phenotype between the dry period and late and early lactation. Changes in many non-lipid metabolites associated with one-carbon metabolism, the tricarboxylic acid cycle, the urea cycle, and AA catabolism were observed in the study, with changes in AA serum concentrations likely related to factors such as energy and nitrogen balance, digestive efficiency, and changing diets. The study confirmed an extensive remodeling of the serum lipidome in peripartum dairy cows, highlighting the importance of changes in acylcarnitine (acylCN), phosphatidylcholines (PC), and triacylglycerols (TG), as they play a crucial role in lipid metabolism. Results showed that short-chain acylCN increased after dry-off and decreased thereafter, whereas lipid-derived acylCN increased around parturition, suggesting that more fatty acids could enter mitochondria. Phospholipids and sphingolipids in serum showed changes during lactation. In particular, concentrations of sphingomyelins, PC, and lysoPC decreased around calving but increased in mid- and late lactation. In contrast, concentrations of TG remained consistently low after parturition. The serum concentrations of bile acids fluctuated during the dry period and lactation, with glycocholic acid, cholic acid, glycodeoxycholic acid, and taurocholic acid showing the greatest concentrations. These changes are likely due to the interplay of diet, liver function, and the ability of the gut microbiota to convert primary to secondary bile acids. Overall, these descriptive results may aid in hypothesis generation and in the design and interpretation of future metabolite-based studies in dairy cows. Furthermore, they contribute to our understanding of the physiological ranges in serum metabolites relative to the lactation cycle of the dairy cow., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Significant differences exist in the composition of current milk replacers (MR) and bovine whole milk. This study investigated how the macronutrient profile of 3 different MR formulations containing varying amounts of fat, lactose, and protein, and a whole milk powder (WP), affect postprandial metabolism and gut permeability in male Holstein calves. Sixty-four calves (45.4 ± 4.19 kg [mean ± SD] and 1.8 ± 0.62 d of age) were blocked in order of arrival to the facility and within each block, calves were randomly assigned to 1 of 4 treatments. Treatments included a high-fat MR (HF: 25.0% dry matter [DM] fat, 22.5% protein, 38.6% lactose; n = 14), a high-lactose MR (HL: 44.6% lactose, 22.5% protein, 18.0% fat; n = 17), a high-protein MR (HP: 26.0% protein, 18.0% fat, 41.5% lactose; n = 17), and WP (26.0% fat, 24.5% protein, 38.0% lactose; n = 16). Calves were fed 3.0 L (135 g/L) 3 times daily at 0600, 1200, and 1800 h with a teat bucket. Milk intake was recorded daily for the first 28 d after arrival, and blood sampling and body weight measurements were performed at arrival and on d 7, 14, 21, and 27. Gut permeability was estimated from fractional urinary excretion of indigestible markers (Cr-EDTA, lactulose, and d-mannitol) administered as a single dose on d 21 instead of the morning milk meal. Digestibility was determined simultaneously from a total collection of feces over 24 h. Postprandial dynamics were measured on d 28 by sequential blood sampling over 7.5 h. Dry matter intake of MR over 28 d was slightly greater in calves fed HL and HP than in WP. Recovery of Cr-EDTA and d-mannitol over a 24-h urine collection was greater in calves fed WP and HP than HL calves. Apparent total-tract digestibility of crude ash, protein, and fat did not differ among treatments; however, DM digestibility was lower in calves fed WP than in other treatment groups. In addition, abomasal emptying, as indicated by the area under the curve (AUC) for acetaminophen, was slower in calves fed WP than in calves fed HF and HL. The AUC for postprandial plasma glucose was lower in calves fed HL than WP and HF and lower in calves fed HP than WP. The AUC for postprandial serum insulin was greater in calves fed HP than WP and HF, whereas calves fed HL did not differ from the other treatments. Postprandial triglycerides were greater in calves fed WP, and postprandial adiponectin was higher in calves fed HL than other treatments. The high content of lactose and protein in MR had a major effect on postprandial metabolism. This raises the possibility of optimizing MR formulations to maintain metabolic homeostasis and influence development., (© 2024, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Quantifying the water and mineral losses in feces is essential to determine the optimal composition of oral rehydration solutions (ORS) for diarrheic animals. In a randomized complete block design, this study evaluated water, mineral, and blood acid-base balance of calves with naturally occurring diarrhea receiving ORS or a placebo. On d 0, 45 calves (age: 18 ± 3.2 d; mean ± SD) were selected based on the presence of visual signs of diarrhea, such as dirty tail or wet feces, along with clinical symptoms evaluated by measuring the skin turgor and the degree of enophthalmos. On d 1, calves were divided into blocks of 3 animals based on blood base excess (BE) measured at 0900 h, and within each block, calves were randomly assigned to 1 of 3 treatments (15 calves per treatment) including (1) a hypertonic ORS (HYPER; Na + = 110 mmol/L; 370 mOsm/kg; strong ion difference [SID] = 60 mEq/L), (2) a hypotonic ORS with low Na + (HYPO; Na + = 77 mmol/L; 278 mOsm/kg; SID = 71 mEq/L), and (3) a placebo consisting of lukewarm water with 5 g/L of whey powder (CON). Milk replacer (MR) was fed through teat buckets twice daily at 0630 h and 1700 h in 2 equally sized meals of 2.5 L from d 1 to 3 and of 3.0 L on d 4 and 5. Treatments consisting of 2.0 L lukewarm solutions were administered between milk meals from d 1 to 3 at 1200 h and 2030 h through teat buckets. Refusals of MR and treatments were recorded daily, and blood samples were collected from the jugular vein once daily at arrival in the afternoon of d 0 and at 0900 h from d 1 to 5 after arrival. Urine and feces were collected quantitatively over a 48-h period from 1200 h on d 1 to 1200 h on d 3, and a representative sample of each 24-h period was stored. In addition, the volume of extracellular fluid was evaluated on d 2 by postprandial sampling over a 4-h period relative to the injection of sodium thiosulfate at 1300 h. Total daily fluid intake (MR, treatment, and water) from d 1 to 3 was greater in HYPER (LSM ± SEM; 8.9 ± 0.36 L/d) and HYPO (7.8 ± 0.34 L/d) than in CON (6.6 ± 0.34 L/d). This resulted in a greater water balance (water intake - fluid output in urine and feces) in calves receiving ORS (59.6 ± 6.28 g/kg BW per 24 h vs. 39.6 ± 6.08 g/kg BW per 24 h). Fecal Na + losses were greater in HYPER than in the other treatments (81 ± 12.0 mg/kg BW per 24 h vs. 24 ± 11.8 mg/kg BW per 24 h). Blood pH was higher in HYPO (7.41 ± 0.016) than CON (7.35 ± 0.016) over the 5 monitoring days, whereas HYPER (7.37 ± 0.017) did not differ with other treatments. In this experimental model, diarrheic calves were likely unable to absorb the high Na + load from HYPER, resulting in greater Na + losses in feces, which might have impaired the alkalinizing capacity of HYPER. In contrast, HYPO significantly sustained blood acid-base balance compared with CON, whereas HYPER did not. This suggests that low tonicity ORS with a high SID are more suitable for diarrheic calves., (© 2024, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Inclusion of urea in dairy cattle diets is often limited by negative effects of high levels of feed urea on dry matter intake (DMI) and efficiency of rumen N utilization. We hypothesized that supplying urea postruminally would mitigate these limitations and allow greater inclusion of urea in dairy cattle diets. Four rumen-fistulated Holstein-Friesian dairy cows (7 ± 2.1 lactations, 110 ± 30.8 d in milk; mean ± standard deviation) were randomly assigned to a 4 × 4 Latin square design to examine DMI, milk production and composition, digestibility, rumen fermentation, N balance, and plasma constituents in response to 4 levels of urea continuously infused into the abomasum (0, 163, 325, and 488 g/d). Urea doses were targeted to linearly increase the crude protein (CP) content of total DMI (diet plus infusion) by 0%, 2%, 4%, and 6% and equated to 0%, 0.7%, 1.4%, and 2.1% of expected DMI, respectively. Each 28-d infusion period consisted of a 7-d dose step-up period, 14 d of adaptation, and a 7-d measurement period. The diet was fed ad libitum as a total mixed ration [10.9% CP, 42.5% corn silage, 3.5% grass hay, 3.5% wheat straw, and 50.5% concentrate (dry matter basis)] and was formulated to meet 100%, 82%, and 53% of net energy, metabolizable protein, and rumen-degradable protein requirements, respectively. Linear, quadratic, and cubic effects of urea dose were assessed using polynomial regression assuming the fixed effect of treatment and random effects of period and cow. Dry matter intake and energy-corrected milk yield responded quadratically to urea dose, and milk urea content increased linearly with increasing urea dose. Apparent total-tract digestibility of CP increased linearly with increasing urea dose and ruminal NH 3 -N concentration responded quadratically to urea dose. Mean total VFA concentration was not affected by urea dose. The proportion of N intake excreted in feces decreased linearly and that excreted in urine increased linearly in response to increasing urea dose. The proportion of N intake excreted in milk increased linearly with increasing urea dose. Urinary urea excretion increased linearly with increasing urea dose. Microbial N flow responded cubically to urea dose, but the efficiency of microbial protein synthesis was not affected. Plasma urea concentration increased linearly with increasing urea dose. Regression analysis estimated that when supplemented on top of a low-CP diet, 179 g/d of postruminal urea would maximize DMI at 23.4 kg/d, corresponding to a dietary urea inclusion level of 0.8% of DMI, which is in line with the current recommendations for urea inclusion in dairy cattle diets. Overall, these results indicate that postruminal delivery of urea does not mitigate DMI depression as urea dose increases., (© 2023, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

We aimed to evaluate the effects of post-ruminal supply of urea (PRU) on nutritional status, and liver metabolism of pregnant beef cows during late gestation. Twenty-four Brahman dams, pregnant from a single sire, and weighing 545 kg ± 23 kg were confined into individual pens at 174 ± 23 d of gestation, and randomly assigned into one of two dietary treatments up to 270 d of gestation: Control (CON, n = 12), consisting of a basal diet supplemented with conventional urea, where the cows were fed with diets containing 13.5 g conventional urea per kg dry matter; and PRU (PRU, n = 12), consisting of a basal diet supplemented with a urea coated to extensively prevent ruminal degradation while being intestinally digestible, where the cows were fed with diets containing 14,8 g urea protected from ruminal degradation per kg dry matter. Post-ruminal supply of urea reduced the urine levels of 3-methylhistidine (P = 0.02). There were no differences between treatments for dry matter intake (DMI; P = 0.76), total digestible nutrient (TDN) intake (P = 0.30), and in the body composition variables, such as, subcutaneous fat thickness (SFT; P = 0.72), and rib eye area (REA; P = 0.85). In addition, there were no differences between treatments for serum levels of glucose (P = 0.87), and serum levels of glucogenic (P = 0.28), ketogenic (P = 0.72), glucogenic, and ketogenic (P = 0.45) amino acids, neither for urea in urine (P = 0.51) as well as urea serum (P = 0.30). One the other hand, enriched pathways were differentiated related to carbohydrate digestion, and absorption, glycolysis, pyruvate metabolism, oxidative phosphorylation, pentose phosphate pathway, and biosynthesis of amino acids of the exclusively expressed proteins in PRU cows. Shifting urea supply from the rumen to post-ruminal compartments decreases muscle catabolism in cows during late gestation. Our findings indicate that post-ruminal urea supplementation for beef cows at late gestation may improve the energy metabolism to support maternal demands. In addition, the post-ruminal urea release seems to be able to trigger pathways to counterbalance the oxidative stress associated to the increase liver metabolic rate., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Santos et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Prebiotic compounds may be supplemented in the diet to improve animal health and performance in a variety of ways. In dairy cattle, the transition from pregnancy through parturition and lactation represents a critical life stage with many concurrent stressors. The objectives of this study were to evaluate responses to the provision of a hindgut-targeted prebiotic compound (calcium gluconate; HFCG) when supplemented prepartum and/or postpartum in a 2 × 2 factorial design. One hundred and sixty-four multiparous Holstein cattle were enrolled and followed from approximately 21 d prior to calving until 100 d of lactation. Treatments were administered as a pelleted compound feed offered in the rotary milking parlor once daily prepartum and thrice daily postpartum. Information pertaining to milk production and body weight were automatically recorded by the milking equipment, and information pertaining to reproductive and health performance was recorded by farm staff. Cattle that received HFCG prepartum were confirmed pregnant approximately 21 d earlier ( P  = 0.024). Cattle that received HFCG both pre- and postpartum had 9% to 10% higher yields of milk protein, fat, and energy-corrected milk ( P  ≤ 0.037) from weeks 4 to 9 of lactation relative to those that received HFCG exclusively prepartum. Conversely, cattle that received HFCG exclusively postpartum had 9% to 10% higher yields of milk protein, fat, and energy-corrected milk ( P  ≤ 0.037) from weeks 9 to 14 of lactation relative to those that received exclusively the negative control in both periods. The mechanism underlying these responses remains unclear, however, we hypothesize that these responses are due to localized reductions in inflammation in the gut and/or signaling to extragastrointestinal tissues altering energy partitioning and balance., Competing Interests: All authors are employed by Trouw Nutrition, which has a commercial interest in the product being investigated., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science.)

The ratio of n-6 to n-3 fatty acid (FA) is between 2 and 10 times higher in milk replacer (MR) than in whole milk, which may promote inflammation and compromise the integrity of the intestinal epithelium. To evaluate how decreasing the n-6:n-3 FA ratio of MR affects gastrointestinal (GIT) permeability and inflammatory status, 30 dairy calves (2.8 ± 1.06 d of age; mean ± standard deviation) were randomly assigned to be fed an MR with an n-6:n-3 FA ratio of 40:1 (CON; 29.3% crude fat of DM; n = 15) or 6.5:1 (n-3; 29.1% crude fat of DM; n = 15). Calves were fed 7.0 L/d in 2 meals. Calves were weighed and fecal consistency was analyzed weekly. On d 22, calves were administered Cr-EDTA, lactulose, and d-mannitol to assess GIT permeability. Blood and total urine were sequentially collected for 6 and 24 h, respectively, and analyzed for marker content. Whole blood collected 4 h after the meal was subjected to an ex vivo lipopolysaccharide (LPS) challenge to evaluate cytokine secretion from blood cells. Calves were euthanized on d 25 for collection of intestinal tissue samples. Tissue samples were processed to assess FA composition by gas chromatography, histomorphology by bright-field microscopy, and gene expression of tight junction proteins, lipid metabolism enzymes, and immune molecules by real-time quantitative PCR. Data were analyzed using PROC GLIMMIX in SAS (version 9.4, SAS Institute Inc.). Growth performance and fecal consistency were unaffected. Calves fed MR with a lower ratio of n-6 to n-3 FA had 2-fold higher n-3 FA contents and 2-fold lower ratios of n-6 to n-3 FA in proximal jejunum and ileum tissues. Total urinary recovery (0-24 h relative to marker administration) and plasma concentrations of the markers were unaffected. Expression of TJP1 tended to be higher in proximal jejunum tissue and lower in ileum tissue of n-3 calves. The expression of TLR4 and TNFA tended to be higher and CD14 was higher in ileum tissue of n-3 calves. Plasma concentrations of interleukin-4 were decreased in response to the ex vivo LPS challenge in n-3 calves. Histomorphology and GIT permeability were largely unaffected by treatment. Furthermore, the inclusion of linseed and algae oil may promote inflammation, as suggested by greater concentrations of the acute-phase proteins haptoglobin and serum amyloid A postprandially, demonstrating that fat sources should be evaluated for their suitability for MR formulations. Understanding how MR composition affects dairy calf health may improve nutritional strategies on farm., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The composition of milk replacer (MR) for calves greatly differs from that of bovine whole milk, which may affect gastrointestinal development of young calves. In this light, the objective of the current study was to compare gastrointestinal tract structure and function in response to feeding liquid diets having a same macronutrient profile (e.g., fat, lactose, protein) in calves in the first month of life. Eighteen male Holstein calves (46.6 ± 5.12 kg; 1.4 ± 0.50 d of age at arrival; mean ± standard deviation) were housed individually. Upon arrival, calves were blocked based on age and arrival day, and, within a block, calves were randomly assigned to either a whole milk powder (WP; 26% fat, DM basis, n = 9) or a MR high in fat (25% fat, n = 9) fed 3.0 L 3 times daily (9 L total per day) at 135 g/L through teat buckets. On d 21, gut permeability was assessed with indigestible permeability markers [chromium (Cr)-EDTA, lactulose, and d-mannitol]. On d 32 after arrival, calves were slaughtered. The weight of the total forestomach without contents was greater in WP-fed calves. Furthermore, duodenum and ileum weights were similar between treatment groups, but jejunum and total small intestine weights were greater in WP-fed calves. The surface area of the duodenum and ileum did not differ between treatment groups, but the surface area of the proximal jejunum was greater in calves fed WP. Urinary lactulose and Cr-EDTA recoveries were greater in calves fed WP in the first 6 h post marker administration. Tight junction protein gene expression in the proximal jejunum or ileum did not differ between treatments. The free fatty acid and phospholipid fatty acid profiles in the proximal jejunum and ileum differed between treatments and generally reflected the fatty acid profile of each liquid diet. Feeding WP or MR altered gut permeability and fatty acid composition of the gastrointestinal tract and further investigation are needed to understand the biological relevance of the observed differences., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Background: Trace metals are supplemented in cattle to prevent nutrient deficiencies. Levels supplemented to mitigate worst-case basal supply and availability scenarios can, however, result in trace metal intakes far above the nutritional requirements of dairy cows with high feed intakes., Objectives: We evaluated Zn, Mn, and Cu balance in dairy cows from late lactation through the subsequent mid-lactation, a period of 24 wk characterized by large changes in dry matter intake., Methods: Twelve Holstein dairy cows were housed in a tie-stall from 10 wk before to 16 wk after parturition and fed 1 unique lactation diet when lactating and a dry cow diet otherwise. After 2 wk of adaptation to the facility and diet, Zn, Mn, and Cu balances were determined at weekly intervals, by calculating the difference between total intakes and complete fecal, urinary, and milk outputs, with the latter 3 fluxes quantified over a 48-h period. Repeated measure mixed models were used to evaluate the effects on trace mineral balances over time., Results: The Mn and Cu balances of cows were not significantly different from 0 mg/d between 8 wk prepartum and calving (P ≥ 0.54), when dietary intake was the lowest of the period evaluated. However, when dietary intake was highest, between wk 6 and 16 postpartum, positive Mn and Cu balances were observed (80 and 20 mg/d, respectively, P ≤ 0.05). Cows were in positive Zn balance throughout the study except during the first 3 wk after calving during which the Zn balance was negative., Conclusions: Large adaptations occur in trace metal homeostasis in transition cows in response to changes in dietary intake. High dry matter intakes, associated with high milk production of dairy cows, combined with current Zn, Mn, and Cu supplementation practices may exceed regulatory homeostatic mechanisms resulting in potential body accumulation of Zn, Mn, and Cu., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Various studies with growing ruminants report increases in nitrogen use efficiency (NUE) when feeding oscillating (OS) dietary CP, whereas limited research with lactating dairy cows demonstrates a lack of improvement in NUE when feeding OS diets. We hypothesised that a total mixed ration (TMR) delivering OS CP (48-h phases of 134 and 171 g CP/kg DM, respectively) compared to a static CP TMR (ST; 152 g CP/kg DM) would result in similar or increased urinary purine derivative excretion (as a marker of microbial protein synthesis (MPS)) and greater urinary nitrogen excretion in lactating dairy cows. Responses in intake, production, apparent total tract digestibility (ATTD), nutrient balance, and estimated MPS were evaluated using faecal and urine collection in 12 multiparous cows (172 ± 39 d in milk) in a randomised complete block design, where total urinary output was estimated indirectly. All measurements were taken during d 8 (at 1700) to d 16 (at 1700) of the 16-d study that followed a 28-d period in which cows already received their respective treatments. Dry matter intake, yields of milk, protein, fat, lactose, and fat- and protein-corrected milk were similar for ST and OS. Milk composition, BW, and body condition score also did not differ between treatments, except for a tendency for increased milk urea concentration with OS (13.7 vs 12.4 mg/dL). Feed efficiency, NUE and ATTD of organic matter, NDF, CP and gross energy did not differ, but ATTD of crude fat (658 vs 627 g/kg) and starch (980 vs 975 g/kg) increased, and ATTD of DM (702 vs 691 g/kg) tended to increase with OS. Milk energy as a proportion of digested energy tended to decrease with OS (34.6 vs 37.1%), but other energy metabolism variables were not affected by treatment. Estimated urinary nitrogen excretion increased (165 vs 144 g/d), estimated urinary nitrogen as a proportion of nitrogen intake tended to increase (25.3 vs 22.7%), and milk nitrogen as a proportion of digested nitrogen decreased (47.3 vs 51.8%) in response to OS. Estimated urinary excretion of creatinine (184 vs 165 mmol/d), uric acid (29 vs 20 mmol/d) and urea (3.1 vs 2.5 mol/d) increased, but other nitrogen metabolism parameters were not affected by OS. Overall, oscillating dietary CP content did not affect lactational performance, milk NUE, or estimated MPS. However, ATTD of some nutrients increased, postabsorptive energy use for milk synthesis tended to decrease, and estimated urinary nitrogen losses increased with OS., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

Situations in which cattle are feed-deprived over extensive periods of time are common in the context of transport and is an animal welfare concern which may also compromise growth and carcass yield grade. This study investigated how the main components of an oral rehydration solution would affect BW loss and blood parameters in feed-deprived bulls. Three dose-response experiments were performed with 24 bulls each (n = 6 per treatment) to study the effect of mineral concentration in Study I (0, 100, 200 and 300 mOsm/kg), the K + to Na + ratio in Study II (25:75, 40:60, 60:40 and 75:25), and glycerol concentration in Study III (0%, 1%, 2% and 4% of the final solution). The blocking factor was initial BW and treatments were randomly assigned within each block. Measurements included fluid intakes, BW, and blood parameters at 0, 24 and 48 h relative to the start of feed deprivation. In Study I, increasing mineral concentration in solution linearly decreased BW losses at 48 h. At 24 and 48 h, serum urea linearly decreased with increasing mineral concentration. At 48 h, blood K + and Na + linearly increased, resulting in increased blood osmolarity. Additionally, at 24 h feed deprivation, blood pH linearly increased with increasing osmolality. In Study II, BW losses decreased with increasing K + to Na + ratio at 24 h, but not at 48 h. No effect of the K + to Na + ratio was found on blood parameters, apart from a trend for a linear decrease of blood osmolarity at 48 h. In Study III, serum urea tended to linearly decrease with increasing glycerol inclusion at 24 h, while blood glucose linearly increased with glycerol inclusion at 24 and 48 h. These combined results indicated that a solution with an osmolality of 200 mOsm/kg and a high K + to Na + ratio would effectively mitigate BW losses and maintain blood acid-base balance, whereas high glycerol inclusion sustained blood glucose in feed-deprived bulls., (© 2022 Nutreco Nederland B.V. Journal of Animal Physiology and Animal Nutrition published by Wiley-VCH GmbH.)

The objective of this study was to characterize total-tract nutrient digestibility, energy balance, and N balance in the critical dietary and metabolic transitions of the lactation cycle. Twelve dairy cows were housed in tiestalls from 10 wk before to 16 wk after parturition. After 2 wk of adaptation to the facility and diet, digestibility of organic matter (OM), neutral detergent fiber (NDF), starch, and N were measured, and energy and N balances determined at weekly intervals by total collection of feces, urine, and milk over 48 h. Cows were individually fed ad libitum a grass silage- and corn silage-based total mixed ration during lactation and a corn silage- and barley straw-based total mixed ration during the dry period. Effects of stage of lactation were evaluated by clustering week in 5 groups: late lactation (wk -8 to -7), dry period (wk -6 to -1), and 3 early lactation periods (wk 1 to 5, wk 6 to 10, and wk 11 to 16). In lactation, apparent total-tract digestibility of OM, NDF, and starch was lowest in the first 5 wk of lactation. From wk 2 to 16 after parturition, apparent nutrient digestibility of all nutrients increased linearly, but with a negative quadratic component for dry matter, OM, and NDF, to levels comparable to those reported in last 2 wk of the previous lactation. However, differences in digestibility across lactation stage were moderate, illustrated by the difference between OM digestibility in late lactation (last 2 wk, 74.8%) and early lactation (first 5 wk, 72.5%). Cows were in negative energy balance for the first 8 wk after calving, and in negative N balance for the first 4 wk after calving. Based on energy and N balance, we predicted that 36.5 kg of body fat and 3.5 kg of body protein were gained in the last 8 wk before calving, and that 47.5 kg of body fat and 7.6 kg of body protein were mobilized in the first weeks of lactation. These predicted changes in body mass, both the gain before calving and loss after calving, were greater by 37% and 10%, respectively, than fluctuations in measured body weight (corrected for predicted gut fill and fetus weights). At wk 1 and 2 postpartum, body N loss corresponded to 25 and 29%, respectively, of total N excretion in milk, and body energy loss corresponded to 64% and 44%, respectively, of the energy exported to milk, illustrating the important contribution of N and energy from body stores to milk production in early lactation. Metabolic N efficiency, measured as total N output (milk and body) over digestible N input (from diet and body), averaged 54.4% in the last 2 wk of lactation, increased to 65.9% 2 wk after calving, and decreased linearly as lactation advanced to 61.9% by wk 16. Short (48 h) but weekly repetition of total collection of feces and urine appears to be a suitable approach to evaluate temporal changes in nutrient digestibility, energy balance, and N balance across lactation and the dry period., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this study was to determine the effect of dietary energy source (fat vs. carbohydrate) in calf milk replacer (MR) on growth performance parameters and feed intake in rearing calves. In a randomized complete block design, 68 Holstein calves [40 females and 28 males; (mean ± SD) body weight (BW): 43.7 ± 1.43 kg] were assigned to 17 blocks of 4 calves based on birth date and parity of the dam. Within each block, calves were randomly assigned to 1 of 2 treatments: a high-lactose MR (HL; 17% fat; 44% lactose; n = 34), or a high-fat MR (HF; 23% fat; 37% lactose; n = 34). Lactose was exchanged for fat on a weight per weight basis, resulting in a 6% difference in metabolizable energy density per kilogram of MR. The feeding plan started with 6 L/d for 7 d, then 8 L/d for 35 d, 6 L/d for 7 d, and finally, 4 L/d for 7 d. Milk replacer allowances were offered in 2 meals per day at 140 g/L. Measurements included daily MR, starter and straw intakes, weekly BW, and blood metabolites, including nonesterified fatty acids (NEFA) and glucose, on wk 4, 6, 8, and 10. Increasing fat at the expense of lactose did not affect MR intake or solid feed intake during the preweaning and weaning periods. However, HF calves tended to consume more solid feed than HL calves during the postweaning period (2.63 ± 0.08 vs. 2.52 ± 0.08 kg/d). Additionally, average daily gain (HF = 0.78 ± 0.02, HL = 0.77 ± 0.02 kg/d) and final BW (HF = 98.8 ± 1.53, HL = 97.7 ± 1.57 kg) were not affected by MR composition. Nevertheless, NEFA concentration was higher in HF calves than in HL calves (0.21 ± 0.01 vs. 0.17 ± 0.01 mmol/L), and glucose concentration was higher in HF calves (6.52 ± 0.23 vs. 5.86 ± 0.23 mmol/L). Under the conditions of this study, HF calves consumed similar amounts of solid feed and grew comparably to the HL calves; however, the isonitrogenous replacement of lactose by fat had evident metabolic effects, such as increased blood NEFA and glucose concentrations., (© 2022.)

During weaning, withdrawal of milk replacer is not directly compensated for by an increase in solid feed intake. Therefore, greater fat inclusion in the starter might mitigate this temporary dietary energy decline. However, fat inclusion in solid feeds may generally limit rumen fermentability and development. To address these potentially conflicting outcomes, we conducted 2 experiments to evaluate the effect of supplementing a high-fat extruded pellet mixed with a calf starter on feed intake, performance, and nutrient digestibility in calves. In experiment 1, 60 Holstein bull calves were blocked by serum IgG (2,449 ± 176 mg/dL) and date of arrival (2.5 ± 0.5 d of age). Within each block, calves were randomly assigned to 1 of 3 treatments: a standard control calf starter (CON; 3.1% fat) and mixtures of CON with 10% inclusion of 1 of 2 different high-fat extruded pellets containing 85% of either hydrogenated free palm fatty acids (PFA, 7.1% fat) or hydrogenated rapeseed triglycerides (RFT, 6.7% fat). Calves were offered milk replacer up to 920 g/d until 42 d of age, followed by a gradual weaning period of 7 d. Calves had ad libitum access to the starter diets, straw, and water. No differences were observed between CON, PFA, and RFT calves on body weight (BW) or average daily gain (ADG) until 49 d of age. From weaning (50 d) until 112 d, PFA calves had a greater BW and ADG than RFT and CON animals. Moreover, PFA calves had the highest intakes of starter, straw, calculated metabolizable energy, and crude protein after weaning. Overall, no differences were present in blood β-hydroxybutyrate and glucose concentrations between treatments; however, calves in the RFT treatment had a higher concentration of insulin-like growth factor-1. In experiment 2, 24 Holstein bull calves at 3 mo of age were assigned to 1 of 8 blocks based on arrival BW and age. Within each block, calves were randomly assigned to 1 of the 3 treatments previously described for experiment 1. Calves on the RFT treatment had the lowest total-tract apparent dry matter and fat digestibility, potentially explaining the differences in performance observed between PFA and RFT calves. Inclusion of the PFA pellet at 10% with a calf starter improved BW, solid feed, and energy intake after weaning. However, these benefits were conditioned by fat source and its digestibility., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Ruminants have evolved with the capability to recycle endogenous urea to the gastrointestinal tract (GIT). Ruminal ammonia derived from urea recycling makes a net contribution to digestible N flow if it is used to synthesise microbial protein. The dynamics of urea recycling and its quantitative importance to the N economy of ruminants are affected by dietary and physiological factors. In general, the transfer of endogenous urea to the GIT is related positively to blood urea concentration and rumen-fermentable energy supply and negatively to ruminal ammonia concentration. After consumption of a meal rich in rumen-degradable N, ruminal ammonia concentrations peak and can exceed the rate of carbohydrate fermentation, resulting in inefficient ammonia capture by microbes. These periods are characterised by greater ruminal ammonia efflux and reduced urea influx. A low ruminal ammonia concentration over time can stimulate recycling of endogenous urea-N to the rumen and its capture into microbial protein and reduce N excretion. Shifting protein digestion to the postruminal GIT can reduce ruminal ammonia concentration and increase plasma urea concentration, conditions that should promote greater reliance on urea recycling to meet N requirements of the rumen. Their ability to use non-protein N, of dietary or endogenous origin, to synthesise metabolisable protein and subsequently meat and milk contributes positively to the human-edible protein efficiency of ruminants. Dietary urea is rapidly degraded to ammonia in the rumen, and high rates of ammonia absorption across the rumen wall when a urea-rich meal is consumed can lead to hypophagic and toxic effects associated with urea feeding. Non-protein N absorbed in the postruminal GIT can contribute substantially to net urea and ammonia uptake into the portal vein, which reflects the potential for targeted urea release in postruminal sections of the GIT. In this review, we suggest that the regulation of urea recycling to the rumen is a critical step towards improved efficiency of ruminal N utilisation. We describe an approach by which postruminal urea supplementation, as an alternative to its ruminal application, may allow a slow and steady return of N to the rumen, avoid peaks in ammonia concentration associated with feeding, confer a greater and more efficient microbial synthesis, and improve fibre digestion compared with conventional urea supplementation., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)