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A 2 × 2 factorial arrangement of treatments was used to investigate the interactive effects of dietary macronutrient composition [high-starch, low-fat, low-fiber (HsLFF) than low-starch, high-fat, high-fiber (LsHFF)] and N source [soybean meal (SBM) or partially replaced by slow-release urea (SRU)] on lactation performance, rumen fermentation, N utilization efficiency, nutrient digestibility, blood metabolites, and feeding behavior in cows. A replicated 4 × 4 Latin square design was used involving 12 multiparous Holstein cows (milk yield of 40.5 ± 5.6 kg/d, BW of 590 ± 20 kg; 81 ± 12 d in milk). The HsLFF diet contained 300 g/kg starch, 31.3 g/kg fat, and 301 g/kg NDF, without straw or additional fat. In contrast, the LsHFF diet contained 195.5 g/kg starch, 60.8 g/kg fat, and 367.5 g/kg NDF, enriched with wheat straw (100 g/kg), and additional fat (34 g/kg). The diets were formulated to be iso-nitrogenous and isocaloric. Cows fed the HsLFF diet had greater DM intake, digestibility of DM and CP, milk yield and milk protein %, but lower intakes of NDF, and physically effective NDF, and milk fat % than cows fed the LsHFF diet. Replacing SBM with SRU significantly increased milk solids yield without affecting other lactation performance or BW. Cows fed the LsHFF diet had higher ruminal pH and branched-chain VFA proportions but lower total VFA concentrations compared with the HsLFF diet, while those on the LsHFF-SRU diet had the highest ruminal ammonia levels. Compared with the HsLFF diet, cows fed LsHFF had lower NE intake, milk energy output and energy requirement for maintenance, although energy balances were similar among groups. The HsLFF diet improved N utilization, resulting in higher N content in milk and lower N excretion in feces. Blood metabolite studies showed significant interactions between the main factors, particularly for blood glucose and creatinine, with the lowest levels in cows fed the LsHFF-SRU and LsHFF-SBM diets. In addition, alanine aminotransferase levels were higher in cows fed the LsHFF diet than in cows fed the HsLFF diet. This could indicate early-stage liver stress due to the metabolic imbalance caused by a high-fat, low-starch diet, which can alter energy metabolism. Cows on the HsLFF-SRU diet had the highest glucose levels, indicating possible changes in carbohydrate metabolism or a higher metabolic rate. The concentration of BUN increased steadily after feeding in the LsHFF diet and peaked after 4 h in the LsHFF-SRU diet, with no difference between N sources in the HsLFF diet. Partial replacement of SRU with SBM had no effect on BUN. The interaction between the main factors had a significant effect on MUN content, which was highest in LsHFF-SRU and lowest in HsLFF-SRU, with no difference between the N sources in the HsLFF diets. Overall, while diets with reduced starch and increased fiber and fat compromised lactation performance, partially substituting SRU with SBM helped maintain milk production and milk nitrogen efficiency. However, the LsHFF-SRU diet was less efficient in N utilization, as shown by higher levels of ruminal ammonia, BUN and MUN., (©, 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 investigated the long-term effects of feeding 5-d transition milk (TRANS) compared with milk replacer (MR) on parameters, including intake, growth, feeding behavior and oxidative stress. Fifty Holstein calves (30 females and 20 males) were fed either 12 L/day TRANS or MR for the first 5 d after an initial colostrum feeding of 3.5 L. Thereafter, all calves were fed with 12 L of MR/d (140 g/L) and were gradually weaned starting in wk 8 until wk 14. Throughout the 14 wk the calves had unrestricted access to concentrate (up to 9.8 kg/calf/day), hay, and water. After weaning all heifers were fed a total mixed ration for young cows. Oxidative status was assessed in blood samples from birth to first insemination. Parameters assessed included the ferric reducing ability of plasma (FRAP) for antioxidant capacity and the concentration of reactive oxygen metabolites by the dROM (detection of reactive oxygen metabolites) assay. In addition, the activity of glutathione peroxidase (GSH-Px), oxidative damage in the form of lipid peroxidation as thiobarbituric acid reactive substances (TBARS) and as advanced oxidation protein products (AOPP) were measured. An oxidative stress index (OSi) was calculated: dROM/FRAP x 100. Total protein (TP) concentration was also quantified via the Bradford assay. The only significant difference in feeding behavior between the 2 treatment groups was a higher concentrate intake by the TRANS calves during the weaning phase. Body weight and ADG did not differ significantly between the TRANS and MR groups. TRANS calves showed a trend for fewer cases of health disorders. Markers of oxidative status, including TBARS, AOPP, GSHPx, FRAP and ROM, showed no treatment effects but varied significantly over time. Of note, the oxidative stress index as ratio between pro- and antioxidants in both groups peaked during weaning and then returned to baseline, suggesting an effective response to this transition phase Overall, the results indicate that feeding TRANS during the first 5 d of life had no long-term effect on the parameters studied as compared with MR feeding under the present rearing conditions. These results provide insight into the changes of oxidative status with age and confirm that the relatively high milk feeding level, slow and late weaning enables calves to adapt well to solely solid feed., (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/).)

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/).)

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/).)

In a previously established animal model, 38 multiparous Holstein cows were assigned to 2 groups fed different diets to achieve either a normal (NBCS) or high (HBCS) body condition score (BCS) and backfat thickness (BFT) until dry-off at -49 d before calving (NBCS: BCS <3.5 [3.02 ± 0.24) and BFT <1.2 cm [0.92 ± 0.21]; HBCS: BCS >3.75 [3.82 ± 0.33] and BFT >1.4 cm [2.36 ± 0.35], mean ± SD). The groups were also stratified for comparable milk yields (NBCS: 10,361 ± 302 kg; HBCS: 10,315 ± 437 kg; mean ± SD). The cows were then fed the same diet during the dry period and subsequent lactation, maintaining the differences in BFT and BCS throughout the study. Using the serum metabolomics data, we created a classification model that identified different metabotypes. Machine learning classifiers revealed a distinct cluster labeled HBCS-PN (HBCS predicted normal BCS) among over-conditioned cows. These cows showed higher feed intake and better energy balance than the HBCS-PH (high BCS predicted high BCS) group, while milk yield was similar. The aim of this study was to investigate the changes in the hepatic transcriptome of cows differing in serum-metabotype postpartum. We performed hepatic transcriptome analysis in cows from 3 metabolic clusters: HBCS-PH (n = 8), HBCS-PN (n = 6), and normal BCS predicted normal BCS (NBCS-PN, n = 8) on d 21 (±2) postpartum. Liver tissue from cows expressed a total of 13,118 genes aligned with the bovine genome. A total of 48 differentially expressed genes (DEG; false discovery rate ≤0.1 and fold-change >1.5) were found between NBCS-PN and HBCS-PH cows, whereas 24 DEG (14 downregulated and 10 upregulated) were found between HBCS-PN and HBCS-PH cows. The downregulated DEG (n = 31) in NBCS-PN cows compared with HBCS-PH cows are involved in biosynthetic processes such as lipid, lipoprotein, and cholesterol synthesis (e.g., APOA1, MKX, RPL3L, CANT1, CHPF, FUT1, ZNF696), cell organization, biogenesis, and localization (e.g., SLC12A8, APOA1, BRME1, RPL3L, STAG3, FBXW5, TMEM120A, SLC16A5, FGF21), catabolic processes (e.g., BREH1, MIOX, APOBEC2, FBXW5, NUDT16), and response to external stimuli (e.g., APOA1, FGF21, TMEM120A, FNDC4), whereas upregulated DEG (n = 17) are related to signal transduction and cell motility (e.g., RASSF2, ASPN, SGK1, KIF7, ZEB2, MAOA, ACKR4, TCAF1), suggesting altered metabolic adaptations during lactation. Our results showed 24 DEG between HBCS-PN and HBCS-PH in the liver. The expression of SLC12A8, SLC16A5, FBXW5, OSGIN1, LAMA3, KDELR3, OR4X17, and INHBE, which are responsible for regulating cellular processes was downregulated in HBCS-PN cows compared with HBCS-PH cows. In particular, the downregulation of SLC12A8 and SLC16A5 expression in HBCS-PN cows indicates lower metabolic load and reduced need for NAD + biosynthesis to support mitochondrial respiratory processes. The upregulation of MAOA, ACKR4, KIF27, SFRP1, and CAV2 in the liver of HBCS-PN cows may indicate adaptive mechanisms to maintain normal liver function in response to increased metabolic demands from over-conditioning. These molecular differences underscore the existence of distinct metabolic types in cows and provide evidence for the role of the liver in shaping different metabolic patterns., (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/).)

To investigate the effects of supplemental monensin administration on the metabolic responses of dairy cows, a systematic review and dose-response meta-analysis were conducted. Initially, 604 studies were identified through comprehensive database searches, including Google Scholar, Scopus, Science Direct, and PubMed, using key words related to dairy cows, monensin, and metabolic outcomes. After a 2-stage screening process, 51 articles with a total of 60 experiments were selected for meta-analysis based on criteria such as study implementation date between 2001 and 2022, presence of a control group that did not receive monensin supplementation, reporting of at least 1 outcome variable, and presentation of means and corresponding errors. The meta-analysis used the 1-stage random-effects method, and sensitivity analyses were performed to assess the robustness of the results. The results showed that the administration of monensin at a dosage of 19 to 26 mg/kg was inversely related to methane emissions and that the administration of monensin at a dosage of 18 to 50 mg/kg resulted in a significant decrease in dry matter intake. Administration of monensin at doses of 13 to 28 and 15 to 24 mg/kg also resulted in a significant decrease in ruminal acetate proportion and an increase in propionate proportion, respectively, with no effects on ruminal butyrate, NH 3 , or pH levels. We found no effects on blood parameters or nitrogen retention, but a significant negative correlation was observed between monensin supplementation and fecal nitrogen excretion. Based on the analysis of all variables evaluated, the optimal dose range of monensin was estimated to be 19 to 24 mg/kg., (© 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/).)

The current study was conducted to examine the effect of l-carnitine (LC) supplementation on telomere length and mitochondrial DNA copy number (mtDNAcn) per cell in mid-lactation cows challenged by lipopolysaccharide (LPS) in blood and liver. The mRNA abundance of 31 genes related to inflammation, oxidative stress, and the corresponding stress response mechanisms, the mitochondrial quality control and the protein import system, as well as the phosphatidylinositol 3-kinase/protein kinase B pathway, were assessed using microfluidics integrated fluidic circuit chips (96.96 dynamic arrays). In addition to comparing the responses in cows with or without LC, our objectives were to characterize the oxidative and inflammatory status by assessing the circulating concentration of lactoferrin (Lf), haptoglobin (Hp), fibrinogen, derivates of reactive oxygen metabolites (dROM), and arylesterase activity (AEA), and to extend the measurement of Lf and Hp to milk. Pluriparous Holstein cows were assigned to either a control group (CON, n = 26) or an LC-supplemented group (CAR; 25 g LC/cow per day; d 42 ante partum to d 126 postpartum (PP), n = 27). On d 111 PP, each cow was injected intravenously with LPS (Escherichia coli O111:B4, 0.5 µg/kg). The mRNA abundance was examined in liver biopsies of d -11 and +1 relative to LPS administration. Plasma and milk samples were frequently collected before and after the challenge. After LPS administration, circulating plasma fibrinogen and serum dROM concentrations increased, whereas AEA decreased. Moreover, serum P4 initially increased by 3 h after LPS administration and declined thereafter irrespective of grouping. The Lf concentrations increased in both groups after LPS administration, with the CAR group showing greater concentrations in serum and milk than the CON group. After LPS administration, telomere length in blood increased, whereas mtDNAcn per cell decreased; however, both remained unaffected in liver. For mitochondrial protein import genes, the hepatic mRNA abundance of the translocase of the mitochondrial inner membrane (TIM)-17B was increased in CAR cows. Moreover, TIM23 increased in both groups after LPS administration. Regarding the mRNA abundance of genes related to stress response mechanisms, 7 out of 14 genes showed group × time interactions, indicating a (local) protective effect due to the dietary LC supplementation against oxidative stress in mid-lactating dairy cows. For mtDNAcn and telomere length, the effects of the LPS-induced inflammation were more pronounced than the dietary supplementation of LC. Dietary LC supplementation affected the response to LPS primarily by altering mitochondrial dynamics. Regarding mRNA abundance of genes related to the mitochondrial protein import system, the inner mitochondrial membrane translocase (TIM complex) seemed to be more sensitive to dietary LC than the outer mitochondrial membrane translocase (TOM complex)., (© 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/).)

Colostrum contains nutrients, immunoglobulins, and various bioactive compounds such as microRNA (miRNA). Less is known about the temporal changes in miRNA profiles in ruminant milk samples during the first week postpartum. In this study, we characterized and compared the profiles of miRNA in the small extracellular vesicles (sEV) isolated from colostrum (CM, collected immediately after parturition, n = 8) and transition milk (TM, collected 7 d postpartum, n = 8) from eight 1-yr-old Guanzhong dairy goats with a milk yield of approximately 500 kg/year. A total of 192 unique sEV-associated miRNA (transcripts per million >1 at least 4 samples in either CM or TM) were identified in all samples. There were 29 miRNA uniquely identified in the TM samples while no miRNA was uniquely identified in the CM samples. The abundance of the top 10 miRNA accounted for 82.4% ± 4.0% (± SD) of the total abundance, with let-7 families (e.g., let-7a/b/c-5p) being predominant in all samples. The top 10 miRNA were predicted to target 1,008 unique genes that may regulate pathways such as focal adhesion, TGF-β signaling, and axon guidance. The expression patterns of EV miRNA were similar between the 2 sample groups, although the abundance of let-7c-5p and miR-30a-3p was higher, whereas that of let-7i-5p and miR-103-3p was lower in CM than in TM. In conclusion, the core miRNAome identified in the samples from CM and TM may play an important role in cell proliferation, bone homeostasis, and neuronal network formation in newborn goat kids. The lack of differential miRNA expression between the CM and TM samples may be due to a relatively short sampling interval in which diet composition, intake and health status of ewes, and environment were relatively stable., (© 2023.)

Transition milk (TRM) is a rich source of bioactive components that promotes intestinal development and growth, and reduces the susceptibility to diarrhoea in calves. The objective of this study was to characterise the effects of replacing pasteurised waste milk (none-saleable milk containing antibiotic and/or drug residues) with pasteurised TRM for 3 wk on blood metabolites of dairy calves at 21 d of age. A total of 84 healthy newborn female Holstein calves was blocked by birth order and assigned randomly to four treatment groups with partial replacement of pasteurised waste milk by TRM (second milking after parturition) at 0 (0 L/day TRM + 6 L/day milk), 0.5 (0.5 L/day TRM + 5.5 L/day milk), 1 (1 L/day TRM + 5 L/day milk), or 2 L (2 L/day TRM + 4 L/day milk) for a 21-day period. Serum metabolome was determined by liquid chromatography with tandem mass spectrometry-based metabolomics analysis on a subset of 26 randomly selected individuals from calves fed pasteurised waste milk (CON, 6 L/d milk; n = 13) or TRM (2 L/d TRM + 4 L/d milk; n = 13) at 21 d of age. The identified metabolites (194 out of 265) were categorised according to chemical class and the number of metabolites per class in the serum, amongst which glycerophospholipids 16% (n = 43), fatty acyls 7% (n = 19), organic acids 7% (n = 18), organic heterocyclic compounds 5% (n = 13), benzenoids 5% (n = 12), sphingolipids 5% (n = 12), organic oxygen compounds 4% (n = 11), and nucleic acids 3% (n = 9), were the predominant types. Significant differences in metabolites were determined by the volcano plot. Applying the volcano plot, only two metabolites (ceramide and phosphatidylserine) were significantly different between CON and TRM. Overall, our results suggested that prolonged TRM feeding for 3 wk had little effect on the serum metabolome of the dairy calves. We speculate that the potential effects of feeding TRM for 3 wk compared with waste milk were spatially limited to affect the composition of the local gut microbial community and the growth or function of the intestinal epithelium, not allowing detection of the likely effects in the serum through a metabolomic approach., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

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/).)

Stress reduces milk and milk components synthesis and increases maintenance requirements of cows. The major stress-related alterations involve enhanced secretion of glucocorticoids and increased sympathetic nervous system activity, which results in biochemical and physiologic changes. In dairy cows exposed to social (ie housing conditions, overstocking, regrouping, feed delivery), physiological (ie initiation of lactation and parturition), or physical (ie heat or cold stress) stressors, responses involve alterations in energy balance and nutrient partitioning. The capacity of the animal to synthesize milk fat largely depends on the availability of substrates for lipid synthesis from the diet, ruminal fermentation or adipose tissue stores, all of which can be altered under stress conditions. Indeed, milk fat concentration is particularly responsive to diet and environment modifications, where a wide range of nutritional and non-nutritional factors influence milk fat output. Milk fat synthesis is an energy demanding process, and extremely sensitive to stress factors during lactation and the involvement of multiple organs. Recent studies examining social, physical, and physiological stressors have provided important insights into how differences in milk yield and milk components may be associated with biological responses to stress factors in dairy cows. This review focuses primarily on the role of stress sources and indicators to which the dairy cow is exposed in regulating milk fat synthesis. We will review the role of nutritional and non-nutritional factors on milk fat synthesis in dairy cows under stress conditions., Competing Interests: Declaration of Competing Interest There are no conflicts of interest for any of the authors., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Bull-fattening diets in Europe and most developed countries around the world have traditionally been based on corn silage, starch-rich, and high-energy/ high-protein supplemental feeds. The impact of climate change on crop yields feed availability, and price volatility, requires new and adapted feeding strategies, including for fattening bulls. Therefore, the objective of this study was to compare the growth performance and economic impact of a representative, conventional corn silage-based (CONVL) total mixed ration, and a dry (DRY) total mixed ration (TMR) fed to Simmental bulls. For nine months (272 days), 24 bulls (215 ± 10 kg BW) were randomly assigned to one of two TMR feeding groups (n = 12 per group). The DRY-TMR was primarily characterised by the nutrient fibre source, exclusively based on straw and other by-products. The diets were formulated and balanced based on the Cornell Net Carbohydrate and Protein System. After 272 days of fattening, bulls were slaughtered. Feed intake, average daily gain (ADG)/DM intake (DMI) ratio, and nutrient intake were affected by treatment, time, and their interaction (P < 0.05). The treatment affected neither acid detergent lignin intake nor starch intake. Compared with CONVL bulls, animals fed DRY-TMR consumed more non-fibre carbohydrates and rumen undegradable neutral detergent fibre, showing lesser dry and fresh matter intake and less metabolisable energy and physically effective neutral detergent fibre intake. Despite differences in nutrient intake (P < 0.05), particle size distribution between the two diets and growth performance were not different (P = 0.45). Simmental bulls in both treatment groups reached target weight in a shorter time due to high ADG of 1.87 kg (DRY-TMR) and 1.84 kg (CONVL). Both treatments achieved a positive profit margin (598 ± 28 €/bull). While total income per bull and dressing percentage did not differ between treatments, the substantially higher feed costs (P < 0.01) of the DRY-TMR resulted in a higher (P = 0.04) income over feed cost in favour of the CONVL treatment group. Despite the higher feed cost of DRY compared with CONVL diets, the better ADG/DMI ratio (P < 0.01) of DRY-TMR contributed to lower absolute feed quantity requirements during the fattening period. Due to the positive profit margin and high ADG results, DRY-TMR solutions for fattening bulls based on straw and by-products can be considered a promising alternative feeding strategy., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

The purpose of this article is to review body condition scoring and the role of body fat reserves in relation to insulin sensitivity and metabolic phenotyping. This article summarizes body condition scoring assessment methods and the differences between subcutaneous and visceral fat depots in dairy cows. The mass of subcutaneous and visceral adipose tissue (AT) changes significantly during the transition period; however, metabolism and intensity of lipolysis differ between subcutaneous and visceral AT depots of dairy cows. The majority of studies on AT have focused on subcutaneous AT, and few have explored visceral AT using noninvasive methods. In this systematic review, we summarize the relationship between body fat reserves and insulin sensitivity and integrate omics research (e.g., metabolomics, proteomics, lipidomics) for metabolic phenotyping of cows, particularly overconditioned cows. Several studies have shown that AT insulin resistance develops during the prepartum period, especially in overconditioned cows. We discuss the role of AT lipolysis, fatty acid oxidation, mitochondrial function, acylcarnitines, and lipid insulin antagonists, including ceramide and glycerophospholipids, in cows with different body condition scoring. Nonoptimal body conditions (under- or overconditioned cows) exhibit marked abnormalities in metabolic and endocrine function. Overall, reducing the number of cows with nonoptimal body conditions in herds seems to be the most practical solution to improve profitability, and dairy farmers should adjust their management practices accordingly., (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/).)

Mobilization of body reserves including fat, protein, and glycogen is necessary to overcome phases of negative nutrient balance typical for high-yielding dairy cows during the periparturient period. Skeletal muscle, the largest internal organ in mammals, plays a crucial role in maintaining metabolic homeostasis. However, unlike in liver and adipose tissue, the metabolic and regulatory role of skeletal muscle in the adaptation of dairy cows to the physiological needs of pregnancy and lactation has not been studied extensively. The functional integrity and quality of skeletal muscle are maintained through a constant turnover of protein, resulting from both protein breakdown and protein synthesis. Thus, muscle protein breakdown (MPB) and synthesis are intimately connected and tightly controlled to ensure proper protein homeostasis. Understanding the regulation of MPB, the catabolic component of muscle turnover, and its assessment are therefore important considerations to provide information about the timing and extent of tissue mobilization in periparturient dairy cows. Based on animal models and human studies, it is now evident that MPB occurs via the integration of 3 main systems: autophagy-lysosomal, calpain Ca 2+ -dependent cysteine proteases, and the ubiquitin-proteasome system. These 3 main systems are interconnected and do not work separately, and the regulation is complex. The ubiquitin-proteasomal system is the most well-known cellular proteolytic system and plays a fundamental role in muscle physiology. Complete degradation of a protein often requires a combination of the systems, depending on the physiological situation. Determination of MPB in dairy cows is technically challenging, resulting in a relative dearth of information. The methods for assessing MPB can be divided into either direct or indirect measurements, both having their strengths and limitations. Available information on the direct measures of MPB primarily comes from stable isotopic tracer methods and those of indirect measurements from assessing expression and activity measures of the components of the 3 MPB systems in muscle biopsy samples. Other indirect approaches (i.e., potential indicators of MPB), including ultrasound imaging and measuring metabolites from muscle degradation (i.e., 3-methylhistidine and creatinine), seem to be applicable methods and can provide useful information about the extent and timing of MPB. This review presents our current understanding, including methodological considerations, of the process of MPB in periparturient dairy cows., (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 the current study was to develop a predictive model for calf disease detection in the preweaning period using data from automated milk feeders (AMF). A deep convolutional neural network (CNN) architecture for the detection of respiratory disease and diarrhea in dairy calves was developed. German Holstein calves were fed milk replacer either ad libitum (up to 25 L/d; n = 32) or restrictively (6 L/d; n = 32) via AMF from 10 ± 3 d of life on. Concentrate, hay, and water were freely available. Calf health parameters were scored daily. The AMF measured milk replacer (MR) intake, number of rewarded visits, number of unrewarded visits, and drinking speed. A calf was considered sick if its fecal score was 3 or 4 and its respiratory score was 2 or 3. Only data from AMF up to 47 d of age were included in the analysis. This cut in the data was made to avoid data from the weaning period. Data were split in 80:20 ratios for training and testing data sets according to the Pareto principle. A minimum sensitivity of 80% was considered an appropriate requirement for the prediction models. Considering all calves in group housing, cross-validation of the test data set showed a sensitivity of 83% and a specificity of 79%, with a positive predictive value and a negative predictive value of 37 and 97%, respectively. The area under the curve of the receiver operating characteristic for the deep CNN model was 0.81 for all group-housed calves. The CNN model yielded sensitivity and specificity of 83 and 71%, respectively (for ad libitum-fed calves), and 82 and 87%, respectively (for restricted-fed calves), with good area under the curve-receiver operating characteristic (0.77 to 0.87), indicating that the CNN models can predict calf disease in both groups with different MR allowances. The permutation feature importance was measured by the decrease in model accuracy, and features (behaviors) were summarized in descending order of their relative importance to the CNN model. Drinking speed and MR intake were the main factors to predict calf disease in calves fed ad libitum. The number of unrewarded visits to the milk feeder and MR intake were the main factors to predict calf disease in restricted-fed calves. Despite the relatively small sample size, the results provide strong evidence that daily feeding behavior data from AMF can be used to identify calves at risk for disease. In conclusion, despite a very good testing performance of the CNN model, the relatively low daily prevalence of calf disease in the present study resulted in a high proportion of false-positive alarms., (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/).)

Telomeres cap the ends of eukaryotic chromosomes, and the telomere length (TL) is related to cellular age. The mitochondrial DNA copy number (mtDNAcn) reflects the abundance of mitochondria in a cell. In addition to generating energy, mitochondria are also the main producers of reactive oxygen species, which in turn can accelerate TL attrition and impair mitochondrial function. Nutrition in early life could influence mtDNAcn and TL in later life. In the present study, we investigated the effects of feeding different levels of milk replacer (MR) on TL shortening and energetic status by examining mtDNAcn of heifers during their first year of life. In this study, whole blood samples were obtained from German Holstein heifer calves 36 to 48 h after birth (wk 1) and at wk 12 and wk 16 of life (n = 37), as well as from 31 calves when reaching 1 yr of age. Calves were fed either a high level of MR (14% solids) at 10 L/d (1.4 kg of MR/d; n = 18) or a restrictive low level at 5.7 L/d (0.8 kg of MR/d; n = 19) until linear weaning in wk 13 to 14 of life. Additional whole blood samples were taken from their respective dams 36 to 48 h after calving. Relative TL (qT) and mtDNAcn in cells from whole blood were measured by multiplex quantitative PCR. The greatest qT values were observed in neonates (36-48 h after birth), with decreasing qT values thereafter. Delta qT values were calculated as ΔqT = qT (first year of life) - initial qT (36-48 h after birth). We found no effect of the feeding regimen on qT values, but qT decreased with age. The mtDNAcn was lowest in neonates, increased until wk 12 of life, and then remained at a constant level until after weaning (wk 16). After the first year of life, mtDNAcn was decreased and returned to levels comparable to those of the neonatal stage. No differences in mtDNAcn were detectable between feeding groups within each time point. When comparing the values of qT and mtDNAcn between the calves and their dams after calving (36-48 h after birth and after calving), greater values were observed in calves than in dams. Delta qT values were negative in all but 2 calves (on the restricted diet), indicating that the change in TL with age was not uniform among individual animals, whereas no difference in mean ΔqT values occurred between the feeding groups. Additional analyses of the correlation between qT, mtDNAcn, and various indicators of oxidative status from birth until wk 16 of life did not indicate major interactions between oxidative status, qT and mtDNAcn. The results of this study support an age-dependent decrease of TL in calves independent of the MR feeding level and show the dynamic changes of mtDNAcn in early life., (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/).)

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., (© 2022, 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/).)

This study examined the effects of frequency of feeding high-solids milk and milk replacer (MMR) blend combined with dry matter (DM) content of starter feed on the performance of dairy calves. Forty-eight Holstein calves (40.2 ± 3.33 kg of body weight) were randomly assigned to 1 of 4 treatments in a completely randomized design. Based on a combination of 2 MMR feeding frequencies (once versus twice daily) and 2 starter DM levels (91.1 and 50% DM), 4 treatments were designed as (1) once daily + dry starter feed (OD), (2) once daily + wet starter feed (OW), (3) twice daily + dry starter feed (TD), and (4) twice daily + wet starter feed (TW). The amount of milk replacer powder added to whole milk was adjusted at each feeding frequency to ensure that calves received similar solids from the MMR blend via the twice-daily (total solids = 18.6%) or once-daily (total solids = 31.0%) programs. Starter intake in the preweaning period did not differ between treatments (average 0.39 kg of DM/d), but the postweaning starter intake was greater in calves fed the wet starter (OW and TW) than in those fed the dry starter (OD and TD). Calves receiving the high-solids MMR blend twice versus once daily grew faster before weaning, which was reflected by a greater weaning weight in the TD and TW groups (88.9 kg) than in the OD and OW groups (83.2 kg). The daily growth rate after weaning did not differ between treatments and averaged 1.15 kg/d. Gain-to-feed ratio during the preweaning period was greater in calves from TD and TW (mean 0.67) than in calves from the OD or OW groups (mean 0.58). Postweaning gain-to-feed ratio was not different among treatments and averaged 0.38. Days to first diarrhea and duration of diarrhea (4.66 ± 0.38), as well as days that animals were treated with antibiotics, remained unaffected with treatments. Although preweaning body weight gain was lower with once-daily milk feeding, calves were twice as heavy at weaning as at birth in both milk feeding programs regardless of starter moisture content, which may qualify the once-daily milk feeding program as a cost-effective management strategy in commercial calf-rearing operations. Feeding solutions with a high solids content (above 15%) may be associated with an increased risk of abomasal bloat and should be used with caution., (© 2022.)

The objectives of this study were (1) to characterize the interindividual variation in the relationship between antepartum (ap) backfat thickness (BFT) and subsequent BFT loss during early lactation in a large dairy herd using cluster analysis; (2) to compare the serum concentrations of metabolites (nonesterified fatty acids, β-hydroxybutyrate), metabolic hormones (leptin and adiponectin), and an inflammatory marker (haptoglobin) among the respective clusters; and (3) to compare lactation performance and uterine health status in the different clusters. An additional objective was (4) to investigate differences in these serum variables and in milk yield of overconditioned (OC) cows that differed in the extent of BFT loss. Using data from a large study of 1,709 multiparous Holstein cows, we first selected those animals from which serum samples and BFT results (mm) were available at d 25 (±10) ap and d 31 (±3 d) postpartum (pp). The remaining 713 cows (parity of 2 to 7) were then subjected to cluster analysis: different approaches based on the BFT of the cows were performed. K-means (unsupervised machine learning algorithm) clustering based on BFT-ap alone identified 5 clusters: lean (5-8 mm BFT, n = 50), normal (9-12 mm, n = 206), slightly fat (SF; 13-16 mm, n = 203), just fat (JF; 16-22 mm, n = 193), and very fat (VF; 23-43 mm, n = 61). Clustering by difference between BFT-ap and BFT-pp (ΔBFT) also revealed 5 clusters: extreme loss (17-23 mm ΔBFT, n = 16), moderate loss (9-15 mm, n = 119), little loss (4-8 mm, n = 326), no loss (0-3 mm, n = 203), and gain (-8 to -1 mm, n = 51). Based on the blood variables measured, our results confirm that cows with greater BFT losses had higher lipid mobilization and ketogenesis than cows with less BFT loss. The serum variables of cows that gained BFT did not differ from normal cows. Milk yield was affected by the BFT-ap cluster, but not by the ΔBFT cluster. Cows categorized as VF had lesser milk yield than other clusters. We further compared the OC cows that had little or no BFT loss (i.e., 2% of VF, 12% of JF, and 31% of SF, OC-no loss, n = 85) with the OC cows that lost BFT (OC-loss, n = 135). Both NEFA and BHB pp concentrations and milk yield were greater in OC-loss cows compared with the OC-no loss cows. The serum concentration of leptin ap was greater in OC-loss than in the OC-no loss cows. Overall, OC cows lost more BFT than normal or lean cows. However, those OC cows with a smaller loss of BFT produced less milk than OC cows with greater losses., (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/).)

Forty-eight newborn Holstein dairy calves [40 ± 3.4 (SD) of kg of body weight (BW); 24 females and 24 males] were used in a completely randomized design to investigate the effects of a mixture of phytobiotic-rich herbal extract (Immunofin, IMPE) incorporated into milk on performance, ruminal fermentation, bacterial population, and serum biochemical metabolites during the preweaning period. Calves had free access to calf starter and clean water from d 6 until weaning. The treatments were the control (CON; without additive) and IMPE at 4, 8, and 12 mL/d. The treatments had no significant effect on total dry matter intake, weight gain, and BW at weaning. The incidence of diarrhea was lower in calves fed 8 mL of IMPE/d compared with CON. At weaning, body measurements (except for front leg circumference) were not affected by IMPE treatment. Relative to the CON group, front leg circumference was significantly decreased by IMPE supplementation. Serum IgG concentration was not significantly increased by IMPE supplementation compared with the CON group. Triglyceride concentration decreased in calves receiving 4, 8, and 12 mL/d of IMPE compared with the CON groups. In contrast to the CON group, serum albumin and total serum protein concentrations increased with IMPE supplementation. Calves receiving 4 mL/d of IMPE had a greater abundance of total bacteria, Ruminococcus albus, Ruminococcus flavefaciens, and Fibrobacter succinogenes compared with the other treatments. Molar proportions of acetate increased in calves fed IMPE (at 12 mL/d) compared with calves fed CON. Ruminal N-NH 3 concentrations decreased linearly with the increase in IMPE supplementation. The results of the present study suggest that the addition of IMPE to milk may improve some health and immunity conditions, blood metabolite concentrations, and increase the abundance of some cellulolytic bacteria in the rumen of Holstein dairy calves. The use of IMPE may be an alternative to feeding antibiotics at subtherapeutic concentrations to improve calf health and immunity status., (© 2022, 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 dietary supplementation with magnesium oxide and calcium-magnesium dolomite on milk fat synthesis and milk fatty acid profile or persistency in milk fat synthesis after their cessation in dairy cows under milk fat depression conditions. Twenty-four multiparous dairy cows in early lactation (mean ± standard deviation; 112 ± 14 d in milk) were used in a randomized complete block design. Milk fat depression was induced in all cows for 10 d by feeding a diet containing 35.2% starch, 28.7% neutral detergent fiber, and 4.8% total fatty acid (dry matter). The experiment was conducted in 2 periods. During the Mg-supplementation period (d 1-20), cows were randomly assigned to (1) the milk fat depression diet used during the induction phase (control; n = 8), (2) the control diet plus 0.4% magnesium oxide (MG; n = 8), or (3) the control diet plus 0.8% calcium-magnesium dolomite (CMC; n = 8). Compared with the control group, feeding the magnesium-supplemented diets increased milk fat concentration and yield by 12% within 4 d. During the 20-d Mg-supplementation period, both the MG and CMC diets increased milk fat concentration and yield, as well as 3.5% fat-corrected milk and energy-corrected milk yield, without affecting dry matter intake, milk yield, and milk protein and lactose concentrations. In the Mg-cessation period (d 21-30), all cows received the control diet, which resulted in a greater milk fat concentration and yield in the cows that had already received the MG and CMC diets in the Mg-supplementation period. Whereas, milk fat concentration and yield remained high after discontinuation of the magnesium-containing alkalizer until d 27. The difference in milk fat synthesis was associated with lower trans-10 C18:1 (-22%) and higher trans-11 C18:1 (+12.5%) concentrations in milk during the Mg-supplementation period. Furthermore, it was evident that within 2 d of supplementation, the trans-10:trans-11 ratio was lower in MG and CMC cows compared with cows receiving the control. This suggested that the effect of magnesium-based alkalizers on milk fat synthesis was mediated via a shift in ruminal biohydrogenation of cis-9,cis-12 C18:2 in the rumen. In conclusion, abrupt addition of magnesium oxide and calcium-magnesium dolomite increased milk fat synthesis, which persisted for 7 d after cessation of magnesium-based alkalizers. A similar ability to recover milk fat synthesis and normal fatty acid biohydrogenation pathways was observed for magnesium oxide and calcium-magnesium dolomite., (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 investigate the effects of milk allowances equal to 526 g/d as moderate (MOD) versus 790 g/d of milk dry matter as high (HI), and starter diets containing 18% or 23% crude protein (CP), on growth performance, blood metabolites, and purine derivative (PD) excretion in the urine of dairy calves. A total of 52 female Holstein dairy calves (40.8 kg of body weight) were randomly assigned to the experimental diets. The treatments were (1) moderate milk and 18% CP starter diet (MOD-18CP); (2) MOD and 23% CP starter diet (MOD-23CP); (3) high milk and 18% CP starter diet (HI-18CP); and (4) HI and 23% CP starter diet (HI-23CP). Calves had free access to a starter feed and water and were weaned on d 53 but remained in the study until d 73. Urine samples were collected during the preweaning period (for 6 consecutive days between d 35 and 40) and postweaning period (for 6 consecutive days between d 65 and 70) to investigate urinary excretion of PD. Starter feed intake, β-hydroxybutyrate (BHB), and blood urea concentrations were reduced; however, average daily gain (ADG) and blood glucose levels increased in calves fed HI before weaning compared with MOD. During the preweaning period, high milk feeding increased total urinary PD excretion but decreased it after weaning. The 23CP diet resulted in higher feed intake and ADG before weaning and higher excretion of allantoin and total excretion of PD compared with the 18CP diet. The HI-23CP treatment resulted in the greatest withers and hip heights at weaning and final measurement, as well as the highest preweaning blood insulin concentrations. In terms of rumen development, MOD-23CP showed the greatest benefits based on starter intake, blood BHB concentration, and urinary excretion of PD. Based on the higher urinary excretion of PD found in HI-fed calves before weaning, it is possible that milk feeding overestimates estimated microbial yield. The results suggest that feeding starters with a higher proportion of CP may help maintain a more balanced ratio of CP to ME during high milk feeding, to avoid protein deficiency due to low starter intake. When calves are fed a high milk allowance, urine excretion of PD may be misinterpreted as a measure of estimated microbial growth and rumen development; this should be considered during calculations of estimated microbial yield in milk-fed calves., (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/).)

This study aimed at characterizing the effects of dietary l-carnitine supplementation on hepatic fatty acid (FA) metabolism during inflammation in mid-lactating cows. Fifty-three pluriparous Holstein dairy cows were randomly assigned to either a control (CON, n = 26) or an l-carnitine supplemented (CAR; n = 27) group. The CAR cows received 125 g of a rumen-protected l-carnitine product per cow per day (corresponding to 25 g of l-carnitine/cow per day) from d 42 antepartum (AP) until the end of the trial on d 126 postpartum (PP). Aside from the supplementation, the same basal diets were fed in the dry period and during lactation to all cows. In mid lactation, each cow was immune-challenged by a single intravenous injection of 0.5 μg of LPS/kg of BW at d 111 PP. Blood samples were collected before and after LPS administration. The mRNA abundance of in total 39 genes related to FA metabolism was assessed in liver biopsies taken at d -11, 1, and 14 relative to LPS (d 111 PP) and also on d 42 AP as an individual covariate using microfluidics integrated fluidic circuit chips (96.96 dynamic arrays). In addition to the concentrations of 3 selected proteins related to FA metabolism, acetyl-CoA carboxylase α (ACACA), 5' AMP-activated protein kinase (AMPK), and solute carrier family 25 member 20 (SLC25A20) were assessed by a capillary Western blot method in liver biopsies from d -11 and 1 relative to LPS from 11 cows each of CAR and CON. On d -11 relative to LPS, differences between the mRNA abundance in CON and CAR were limited to acyl-CoA dehydrogenase (ACAD) very-long-chain (ACADVL) with greater mRNA abundance in the CAR than in the CON group. The liver fat content decreased from d -11 to d 1 relative to the LPS injection and remained at the lower level until d 14 in both groups. One day after the LPS challenge, lower mRNA abundance of carnitine palmitoyltransferase 1 (CPT1), CPT2, ACADVL, ACAD short-chain (ACADS), and solute carrier family 22 member 5 (SLC22A5) were observed in the CAR group as compared with the CON group. However, the mRNA abundance of protein kinase AMP-activated noncatalytic subunit gamma 1 (PRKAG1), ACAD medium-chain (ACADM), ACACA, and FA binding protein 1 (FABP1) were greater in the CAR group than in the CON group on d 1 relative to LPS. Two weeks after the LPS challenge, differences between the groups were no longer detectable. The altered mRNA abundance before and 1 d after LPS pointed to increased transport of FA into hepatic mitochondria during systemic inflammation in both groups. The protein abundance of AMPK was lower in CAR than in CON before the LPS administration. The protein abundance of SLC25A20 was neither changing with time nor treatment and the ACACA protein abundance was only affected by time. In conclusion, l-carnitine supplementation temporally altered the hepatic mRNA abundance of some genes related to mitochondrial biogenesis and very-low-density lipoprotein export in response to an inflammatory challenge, but with largely lacking effects before and 2 wk after LPS., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Postnatal metabolism depends on maturation of key metabolic pathways around birth. In this regard, endogenous glucose production is impaired in calves born preterm. Concerning protein metabolism, the rates of protein turnover are greater during the neonatal period than at any other period of postnatal life. The mammalian target of rapamycin (mTOR) and the ubiquitin-proteasome system (UPS) are considered as the major regulators of cellular protein turnover. The objectives of this study were to investigate (1) the changes in plasma AA profiles, (2) the mRNA abundance of mTOR signaling and UPS-related genes in skeletal muscle, and (3) the mRNA abundance of branched-chain AA (BCAA) catabolic enzymes in skeletal muscle and adipose tissue in neonatal calves with different degree of maturation during the transition to extrauterine life. Calves (n = 7/treatment) were born either preterm (PT; delivered by cesarean section 9 d before term) or at term (T; spontaneous vaginal delivery) and were left unfed for 1 d. Calves in treatment TC were also spontaneously born but were fed colostrum and transition milk for 4 d. Blood samples were collected from all calves at birth and at 24 h of life. Additional blood samples were taken 2 h after feeding (26 h of life) for PT and T calves, and on d 4 of life for TC, to determine plasma glucose, urea, and AA. Tissue samples from 3 muscles [M. longissimus dorsi (MLD), M. semitendinosus (MST), and M. masseter (MM)], and kidney fat were collected following euthanasia at 26 h after birth (PT, T) or on d 4 of life (TC) at 2 h after feeding. The concentrations of the majority of plasma AA (Ala, Gln, Asn, Cit, Lys, Orn, Thr, and Tyr), nonessential AA, and total AA were greater during the first 24 h and also before and 2 h after feeding in PT than in T. The ratio of plasma BCAA to the aromatic AA (Tyr and Phe) was greatest in TC, followed by T, and least in PT. The mRNA abundance of mTOR and ribosomal protein S6 kinase 1 (S6K1) in MLD and MM was greater in PT and T than in TC. The mRNA abundance of muscle-specific ligases FBXO32 (F-box only protein 32) in the 3 different skeletal muscles and TRIM63 (tripartite motif containing 63) in MLD was greater in PT and T than in TC; in MM, TRIM63 mRNA was greatest in PT. The mRNA for BCKDHA and BCKDHB (the α and β polypeptide of branched-chain α-keto acid dehydrogenase) in kidney fat was elevated in PT and T compared with TC, suggesting a possible enhancement of BCAA oxidation as energy source to cover the energetic and nutritional postnatal demands in PT and T in a starved state. The increased abundances of mTOR-associated signaling factors and muscle-specific ligase mRNA indicate a greater rate of protein turnover in muscles of PT and T in a starved state. Elevated plasma concentrations of several AA may result from enhanced muscle proteolysis and impaired conversion to glucose in the liver of PT calves., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This observational study determined the lipidome of cow milk during subclinical intramammary infection (IMI) by non-aureus staphylococci (NAS), also defined as coagulase-negative staphylococci, using an untargeted approach. Among the pathogens causing bovine IMI, NAS have become the most frequently isolated bacteria from milk samples. Although the application of system biology approaches to mastitis has provided pivotal information by investigating the transcriptome, proteome, peptidome, and metabolome, the milk lipidome during mammary gland inflammation remains undisclosed. To cover this gap, we determined the milk lipidome of 17 dairy cows with IMI caused by NAS (NAS-IMI), and we compared the results with those of healthy quarter milk from 11 cows. The lipidome was determined following a liquid chromatography-quadrupole time-of-flight mass spectrometry approach. Sixteen subclasses of lipids were identified in both groups of animals. From 2,556 measured lipids, the abundance of 597 changed more than 10-fold in quarter milk with NAS-IMI compared with healthy quarters. The results demonstrate the influence of NAS-IMI on the milk lipidome, implying significant changes in lipid species belonging to the family of triacylglycerols and sphingomyelins, and contribute to the understanding of inflammatory processes in the bovine udder, highlighting potential novel biomarkers for improving mastitis diagnostics., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study investigated the effects of feeding finely ground starter diets containing either 18 or 22% crude protein (CP) content [dry matter (DM) basis] and high or low ratios of rumen-undegradable protein to rumen-degradable protein (RUP:RDP) on growth performance, nutrient digestibility, ruminal fermentation, blood metabolites, and urinary purine derivatives in dairy calves. A total of 48 three-day-old female Holstein dairy calves with 40.2 ± 2.5 kg of initial body weight (BW) were randomly assigned in a complete randomized block design to a 2 × 2 factorial arrangement of treatments (12 calves/treatment). Treatments were as follows: (1) finely ground starter diet (mean particle size = 0.69 mm) with 18% CP and low RUP:RDP ratio [low ratio (LR) = 26:74; 18CP-LR]; (2) finely ground starter diet with 18% CP and high RUP:RDP ratio [high ratio (HR) = 35:65; 18CP-HR]; (3) finely ground starter diet with 22% CP and low RUP:RDP ratio (22CP-LR); (4) finely ground starter diet with 22% CP and high RUP:RDP ratio (22CP-HR) on DM bases. Blocking was based on the day of treatment assignment, and treatments were randomly assigned within each block. Calves received 4 L of milk daily from d 3 to 10, 7 L/d from d 11 to 40, 4 L/d from d 41 to 49, and 2.5 L/d from d 50 to 53, and then all calves were weaned but remained in the experiment until d 83 of age. The results showed that overall average daily gain (ADG), weaning BW, and feed efficiency (FE) were greater in 22% CP treatments than in 18% CP. Increasing the starter CP content from 18 to 22% of DM did not influence overall starter feed intake, milk intake, total dry matter intake (DMI), postweaning ADG, and FE of calves. No effect of RUP:RDP ratio was observed for starter feed intake, milk intake, total DMI, preweaning ADG, FE, and grams of CP per megacalorie of metabolizable energy. The RUP intake and postweaning ADG were greater for calves fed the HR diets than for those fed the LR diets. The digestibility of neutral detergent fiber was greater, and the digestibility of OM tended to be greater, and the ruminal concentrations of total short-chain fatty acids (SCFA), acetate proportion, and acetate-to-propionate ratio were greater in 22% CP than in 18% CP. A 2-way interaction between starter protein content and time was observed for total ruminal SCFA, acetate proportion, and acetate-to-propionate ratio, indicating that starter CP concentration had more effect on ruminal parameters. Preweaning urinary purine derivatives, preweaning microbial protein synthesis, and postweaning urinary nitrogen were greater for calves fed the 22CP diets than for those fed the 18CP diets but were not affected by the different RUP:RDP ratios. The concentrations of blood glucose and insulin were greater in 22% CP than in 18% CP diets. The blood insulin concentration was greater when calves received the HR diets compared with the LR diets. Therefore, we conclude that greater starter protein content can have beneficial effects on growth performance, probably through increased microbial protein synthesized and preweaning blood insulin concentration; however, a greater RUP:RDP ratio showed marginal effects on growth performance during the postweaning period., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

In the dairy cow, late gestation and early lactation are characterized by a complexity of metabolic processes required for the homeorhetic adaptation to the needs of fetal growth and milk production. Skeletal muscle plays an important role in this adaptation. The objective of this study was to characterize the metabolome in skeletal muscle (semitendinosus muscle) and in serum of dairy cows in the context of the physiological changes occurring in early lactation and to test the effects of dietary supplementation (from d 1 in milk onwards) with conjugated linoleic acids (sCLA; 100 g/d; supplying 7.6 g of cis-9,trans-11 CLA and 7.6 g of trans-10,cis-12 CLA per cow/d; n = 11) compared with control fat-supplemented cows (CTR; n = 10). The metabolome was characterized in skeletal muscle samples collected on d 21 and 70 after calving in conjunction with their serum counterpart using a targeted metabolomics approach (AbsoluteIDQ p180 kit; Biocrates Life Sciences AG, Innsbruck, Austria). Thereby 188 metabolites from 6 different compound classes (acylcarnitines, amino acids, biogenic amines, glycerophospholipids, sphingolipids, and hexoses) were quantified in both sample types. In both groups, dry matter intake increased after calving. It was lower in sCLA than in CTR on d 21, which resulted in reduced calculated net energy and metabolizable protein balances. On d 21, the concentrations of dopamine, Ala, and hexoses in the skeletal muscle were higher in sCLA than in CTR. On d 21, the changed metabolites in serum were mainly long-chain (>C24) diacyl phosphatidylcholine PC (PC-aa) and acyl-alkyl phosphatidylcholine (PC-ae), along with lysophosphatidylcholine acyl (lysoPC-a) C26:1 that were all lower in sCLA than in CTR. Supplementation with CLA affected the muscle concentrations of 22 metabolites on d 70 including 10 long-chain (>C22) sphingomyelin (SM), hydroxysphingomyelin [SM(OH)], PC-aa, and PC-ae along with 9 long-chain (>C16) lysoPC-a and 3 metabolites related to amino acids (spermine, citrulline, and Asp). On d 70, the concentrations of lysoPC-a C18:2 and C26:0 in serum were higher in the sCLA cows than in the CTR cows. Regardless of treatment, the concentrations of Ile, Leu, Phe, Lys, His, Met, Trp, and hydroxybutyrylcarnitine (C4-OH) decreased, whereas those of ornithine, Gln, and trans-4-hydroxyproline (t4-OH-Pro) increased from d 21 to 70 in muscle. The significantly changed metabolites in serum with time of lactation were 28 long-chain (>C30) PC-ae and PC-aa, 7 long-chain (>C16) SM and SM(OH), along with lysoPC-a C20:3 that were all increased. In conclusion, in addition to other significantly changed metabolites, CLA supplementation mainly led to changes in muscle and serum concentrations of glycerophospholipids and sphingolipids that might reflect the phospholipid compositional changes in muscle. The metabolome changes observed in sCLA on d 21 seem to be, at least in part, due to the lower DMI in these cows. The changes in the muscle concentrations of AA from d 21 to 70, which coincided with the steady energy and MP balances, might reflect a shift of protein synthesis/degradation balance toward synthesis., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Dairy calves may benefit from extending the duration of feeding transition milk (TM; the subsequent two to six milkings after parturition) to enhance performance and health during early life. The objective of this study was to assess the effect of replacing pasteurized waste milk (non-saleable milk containing antibiotic and/or drug residues) with pasteurized TM for 3 weeks on the growth performance and health of dairy calves. A total of 84 healthy newborn female Holstein calves were blocked by birth order and assigned randomly to 4 treatment groups with partial replacement of pasteurized waste milk by TM (second milkings after parturition) at 0 (0 l/day TM + 6 l/day milk), 0.5 (0.5 l/day TM + 5.5 l/day milk), 1 (1 l/day TM + 5 l/day milk), or 2 l (2 l/day TM + 4 l/day milk) for a 21-day period. From day 22 onward, all calves were fed individually with 6 l/day pasteurized waste milk. Calves were weaned on day 60 and monitored until day 90 of the study. Liquid feed DM intake (DMI) was increased with increasing levels of TM (P = 0.001). Starter feed DMI and total DMI (liquid feed DMI + starter feed DMI) were not affected by the treatment effect. Calves were fed 2 l/day TM gained more BW compared with those in the control group during the postweaning and overall periods. The average daily gain tended (P = 0.06) to be higher in calves fed 2 l/day TM compared with calves fed 0 (+ 65 g/day), 0.5 (+ 53 g/day), or 1 (+ 76 g/day) l/day TM during the preweaning period. Daily weight gain was also higher in calves fed 2 l/day TM compared with calves in the control group during the postweaning (+ 137 g/day; P = 0.04) and overall (+89 g/day; P < 0.01) periods, respectively. Calves fed 2 l/day TM had a higher feed efficiency compared with calves in the control group during all studied periods. The calves fed TM2 had a lower chance of having diarrhea compared with other treatment groups. Duration but not the frequency of diarrhea was lower in calves fed TM2 vs TM0 (2.5 vs 4.2days; P = 0.03). In general, partial replacement of waste milk with TM (2 l/day) may be recommended to feed dairy calves at an early stage of life to support a higher growth rate and health benefits., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

This study investigated the effects of feeding dairy calves starter diets containing 19% or 22% crude protein (CP) content on a dry matter basis and either supplemented or not with soybean oil (SBO, 0 vs. 3%, dry matter basis) on growth performance, digestibility, urinary nitrogen, and purine derivatives (PD) excretion. A total of 48 female Holstein dairy calves (mean 39.8 kg of body weight) were randomly distributed to experimental diets in a 2 × 2 factorial arrangement of treatments. The 4 dietary treatments were (1) starter diet without SBO supplement and 19% CP (NSBO-19CP), (2) starter diet without SBO supplement and 22% CP (NSBO-22CP), (3) starter diet with 3% SBO and 19% CP (SBO-19CP), and (4) starter diet with 3% SBO and 22% CP (SBO-22CP). Milk feeding value was similarly based on a constant protocol across experimental treatments and calves had ad libitum access to water and starter diets throughout the study. All calves were weaned on d 63 of age and remained in the study until d 83 of age. Calves supplemented with SBO had lower starter feed intake and average daily gain (ADG) and lower feed efficiency (FE) but had a higher fecal score indicating a higher likelihood of diarrhea occurrence compared with unsupplemented calves. Wither heights, digestibilities of organic matter, CP, and neutral detergent fiber were decreased, and ruminal volatile fatty acids tended to be reduced, and the molar proportion of ruminal butyrate (preweaning) and acetate (postweaning) reduced by supplemental SBO. The urinary allantoin and total PD excretion were reduced; however, urinary nitrogen excretion was increased when calves were supplemented with SBO. The CP amount did not affect starter feed intake, FE, or diarrhea occurrence rate, whereas the 22CP diets increased neutral detergent fiber digestibility, improved ADG (tendency), and increased allantoin and urinary PD excretion compared with the 19CP diets. The starter feed intake, ADG, FE, diarrhea occurrence rate, nutrient digestibility, and ruminal fermentation were not affected by the interaction between starter SBO and CP level; however, hip height and total PD in calves that received the SBO-22CP diets were higher than those fed the SBO-19CP diets. In conclusion, based on our experimental conditions, supplemental SBO could not be recommended for dairy calves. Furthermore, our findings indicate that SBO has negative effects on performance more attributed to reducing starter intake, digestibility, and ruminal volatile fatty acid concentration rather than because of a limitation of starter metabolizable protein supply and intestinal amino acid availability. Therefore, our results indicate that feeding the higher starter CP content is not a viable strategy to compensate for the negative effects of SBO supplementation on the growth performance of dairy calves., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to differentiate the effects of acute heat stress (HS) from those of decreased dry matter intake (DMI) during the prepartum period on metabolism, colostrum, and subsequent production of dairy cows. Holstein dairy cows (n = 30) with similar parity and body weight were randomly assigned to 1 of 3 treatments on 45 d before calving: (1) cooled (CL, n = 10) conditions with ad libitum feed intake, (2) HS conditions with ad libitum feed intake (n = 10), and (3) pair-fed cooled (CLPF, n = 10) with reduced DMI similar to the HS group while housed under cooled conditions. The reduction in the amount of feed offered to the CLPF cows was calculated daily as the percentage decrease from the average DMI of HS cows relative to the CL cows. For CLPF and CL cows, barns provided shade, sprinklers, and fans, whereas the HS cows were provided only with shade. Cows in all groups received individually the same total mixed ration. Cows were dried off 60 d before the expected calving. Cows in the HS group and, by design, the CLPF cows had reduced DMI (~20%) during the experiment. Heat stress decreased gestation length, first colostrum yield, and calf birth weight compared with CL and CLPF cows. Milk yield decreased 21% (5 kg) in the HS and 8% (2 kg) in CLPF cows, indicating that reduced feed intake during late gestation accounted for 60% of the total reduced milk yield. The CLPF cows exhibited an elevated NEFA concentration compared with the CL and HS cows. The HS cows had a greater mRNA abundance of HSP70 in the peripheral blood leukocytes at 21 d prepartum compared with the other groups. At calving, the mRNA abundance of HSP70 was greater in HS cows, followed by CLPF, compared with the CL cows. In conclusion, HS during the late gestation period caused metabolism and production differences, which were only partially attributed to reduced feed intake in dairy cows., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

In the dairy calf feeding, supplementation of forage to the starter feed is commonly practiced. However, data are insufficient about how changes in particle size (PS) of forage affect calf performance and behavior in a free-choice forage provision system. This study aimed to assess the effects of supplementing wheat straw varying in PS on performance, skeletal growth characteristics, ruminal pH, nutritional behaviors, and blood metabolites of dairy calves. Forty-eight Holstein calves (43.8 ± 3.2 kg of BW) from d 15 of age were randomly assigned to one of the four treatments (n = 12/treatment; six males and six females): (1) starter without wheat straw supplementation (CON), (2) CON supplemented with wheat straw chopped at 1 mm geometrical mean particle length (GMPL) (fine PS), (3) CON supplemented with wheat straw at 4 mm GMPL (medium PS), and (4) CON supplemented with wheat straw at 7 mm GMPL (long PS). The calves were given ad libitum access to feed and water throughout the study. All calves were weaned on d 56 of age and continued the experiment until d 90. The starter, wheat straw, and total solid feed intakes were not affected by GMPL of wheat straw; however, CON calves had a lower solid feed-, total DM- and NDF-intake than calves offered wheat straw. Further, ADG, weaning, and final BWs as well as feed efficiency were similar between treatments. No difference was observed in growth rate of hip height, hip width, body barrel, wither height, and heart girth among treatments, however, wheat straw supplemented calves tended to have greater body length at weaning. Blood concentration of serum β-hydroxy butyrate was greater in wheat straw supplemented calves compared with CON calves on d 56 and 90. Calves supplemented with wheat straw spent more time eating starter and forage, lying and ruminating and less time for standing and non-nutritional behaviors compared with the CON calves on d 49 and 63 of the study. Moreover, calves offered wheat straw had greater ruminal pH than CON calves at 4 and 8 h after offering starter feed on d 35. In conclusion, supplementing wheat straw as a free-choice increased solid feed intake, rumen pH, and calves' welfare, however, PS of wheat straw had no effect., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

The interrelationships between silage preference of ruminants and compounds that may affect forage choice are not yet fully enlightened. Analysis of the forage metabolome in addition to conventional chemical analysis and preference trials can provide new insights. Six silage treatments each of alfalfa (AL) and red clover (RC), with different dry matter concentrations (222-391 g/kg), silage additives, and intended addition of soil, were produced in quadruplicate to obtain a range of qualities. After 120 d of ensiling, silages were sampled for chemical analysis, vacuum-packed, and refrigerated for subsequent preference trials with goats. Within 21 d, each possible combination of 2 silages and an AL hay that served as control (n = 21) was presented to goats (Saanen-type wethers, n = 8, body weight 105 ± 2.7 kg) for 3 h for ad libitum intake. Comparisons among means for 3-h dry matter intake (DMI) for forages offered in choice situations were made using variance analysis, including terms for treatment and animal and the Waller-Duncan k-ratio t-test to separate means. The most preferred and avoided treatments of AL and RC silage amounted to 863, 858, 226, and 282 g DMI/3 h, respectively. To further explore relations between silage composition and preference, a metabolome analysis of the most preferred and most avoided AL and RC treatments were conducted. Metabolites (all low molecular weight molecules) were analyzed by a nontargeted metabolite profiling in the range of 50-1,700 Da. Metabolites showing the most distinct difference between preferred and avoided silages were identified by partial least squares discriminant analysis. In the 2 selected treatments of each plant species (those that were most different in forage preference), more than 6,400 compounds were detected and 2,010 were identified. Between preferred and avoided treatments, 934 of the detected compounds differed in RC and 1,860 in AL, of which 475 were altered in both plant species (251 were reduced and 186 were increased; only 38 behaved contrarily, meaning that they were increased in one substrate and decreased in the other). The database provides a useful foundation for the approach of explaining silage preference by ruminants., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

A 2 × 2 factorial study was conducted to evaluate the effects of milk allowance and physical forms of starter on growth performance, ruminal fermentation, and blood metabolites of Holstein dairy calves. A total of 48 calves [40.4 ± 1.55 kg of body weight (BW), n = 12 per treatment: 6 males and 6 females] were randomly assigned to 1 of the 4 treatments: (1) calves fed low milk allowance and finely ground (FG) starter feed [low-FG; 1.47 ± 2.12-mm geometric mean particle size (GMLP)], (2) calves fed low milk allowance and textured (TS) starter feed [low-TS, includes steam-flaked grains (corn and barley) with a pelleted supplement, GMLP 4.15 ± 1.77 mm], (3) calves fed high milk allowance and FG starter feed (high-FG); and (4) calves fed high milk allowance and TS starter feed (high-TS). The starter diets were blended with 7% of chopped alfalfa hay as a proportion of diet dry matter (DM). No milk refusal was observed in any treatments, and calves on both treatments were weaned from milk by wk 8 of the study using a gradual weaning procedure. We observed no interaction between milk allowance and physical forms of starter on feed intake, average daily gain, feed efficiency, BW, and structural growth. Calves fed high milk allowance had lower starter feed intake but greater feed efficiency and overall BW compared with those fed low allowance. Total DM intake and average daily gain were not different among treatments. Regardless of the physical form of starter feed, hip height, heart girth, the molar proportion of ruminal acetate, acetate to propionate ratio, plasma cholesterol, and high-density lipoprotein were greater, but ruminal total volatile fatty acids, the molar proportion of propionate, and plasma β-hydroxybutyrate were lower in calves fed high milk allowance compared with those fed low allowance. Regardless of the milk allowance, calves fed the FG starter feeds had greater blood urea nitrogen concentrations compared with calves fed the TS starter diets. In conclusion, both forms of the starter feeds can be used when calves are fed high milk allowance with no negative effect on their performance., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The current study evaluated the effects of step-down weaning implementation time on starter feed intake, growth performance, blood metabolites, and ruminal pH in dairy calves. A total of 48 Holstein dairy calves (24 male and 24 female; 3 ± 1 d old; 41.2 ± 1.8 kg of body weight) were assigned (n = 12 per treatment; 6 male and 6 female) to 4 experimental treatments in a completely randomized block design. All calves were fed 6 L/d of milk from d 3 to 10 of age, and the treatments consisted of the following: calves were offered 8 L/d of milk from d 11 to 28 of age and then 4 L/d from d 29 to 63 (Step-28; total milk offered = 326 L); calves were offered 8 L/d of milk from d 11 to 42 of age and then 4 L/d from d 43 to 63 (Step-42; total milk offered = 382 L); calves were offered 8 L/d of milk from d 11 to 56 of age and 4 L/d from d 57 to 63 (Step-56; total milk intake = 438 L); and calves were fed 8 L/d of milk from d 11 to 63 of age and abruptly weaned (control; total milk offered = 466 L). All calves were housed individually in pens and had ad libitum access to water and solid feed throughout the experiment. All calves were completely milk weaned on d 64, and their performance was measured until d 80 of age. During the experiment, the starter intake (kg/d and % of body weight) was greater in calves in the Step-28 group compared with those in the other groups. However, the total metabolizable energy intake was greater in the Step-56 calves compared with the other calves. Overall, the average daily gain (ADG), feed efficiency (ADG/dry matter intake), and ADG/total metabolizable energy intake were similar across the treatments. Circulating glucose, β-hydroxybutyrate, blood urea nitrogen, albumin, total protein, alanine aminotransferase, and aspartate aminotransferase, as well as ruminal pH, were not affected by the treatments. The implementation of step-down weaning in early life (4-6 wk of age) could stimulate solid feed intake compared with weaning at a later age with no negative effect on performance., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study investigated the effects of substituting beet pulp (BP) for different grains (barley or corn) in the diet of high-producing dairy cows on intake, feeding behavior, nutrient digestibility, ruminal fermentation, milk production, and feed conversion efficiency. Eight second-parity Holstein cows (62 ± 2 d in milk; milk yield = 54 ± 1.2 kg/d; body weight = 624 ± 26; all mean ± SE) were used in a replicated 4 × 4 Latin square design during 4 periods of 21 d. Cows were randomly assigned to 1 of 4 treatments that were a 2 × 2 factorial arrangement of 2 grain sources (corn or barley) and 2 levels of BP inclusion [5 or 15% of dry matter (DM)] in the diet: (1) barley-based diet with BP at 5% of dietary DM; (2) barley-based diet with BP at 15% of dietary DM; (3) corn-based diet with BP at 5% of dietary DM; and (4) corn-based diet with BP at 15% of dietary DM. The increasing amount of BP in the diet was at the expense of decreasing an equal proportion of grain (barley or corn). All diets were high in concentrates (65% of diet DM) and formulated to have similar concentrations of energy and protein. The portion of feedstuffs that is potentially able to be consumed by humans is known as human edible. Accordingly, human-edible protein (HEP) and human-edible energy (HEE) inputs were calculated according to the recommended potential human-edible fraction of each dietary ingredient, and HEP and HEE outputs were determined as the amount of gross energy and true protein in the milk. Feed conversion efficiency (FCE) for HEP and HEE were expressed as output per input of each variable, whereas FCE for the production of fat-corrected milk (FCM) and energy-corrected milk (ECM) were expressed as the amount of each variable per DM intake. Results showed that substituting BP for grain did not affect DM intake, crude protein intake, or nutrient digestibility, whereas starch intake (5.70 vs. 7.43 kg/d for the low-BP vs. high-BP diets, respectively), HEP (2.34 and 1.92 kg/d), and HEE (186 and 147 MJ of gross energy/d) decreased. Treatments did not affect sorting and chewing activities, but increasing BP in the diet increased ruminal pH at 4 h after feeding (6.20 vs. 6.39) and milk fat content (2.92 vs. 3.15%). Similarly, FCE for ECM production (1.44 vs. 1.54) as well as FCE for HEE (0.653 vs. 0.851) and HEP (0.629 vs. 0.702) were greater in high-BP diets compared with low-BP diets. The interaction of BP and grain sources significantly affected FCE for ECM production, where improvements were more evident when BP was substituted for barley than for corn. The improvement in FCE for HEE was greater when BP was substituted for barley (0.236) rather than corn (0.161). In conclusion, the substitution of BP for barley or corn grains in high-concentrate diets of high-producing cows decreased starch intake, increased ruminal pH at 4 h after feeding, and improved FCE for FCM production. Substitution for barley, rather than for corn, promoted greater FCE for ECM production and HEE., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study aimed to investigate the effects of feeding dairy calves starter diets containing corn grain processed by different methods (ground versus steam-flaked; GRC vs. SFC) and either 18% or 21% crude protein (CP) contents (dry matter basis) on growth performance, digestibility, ruminal fermentation, urinary purine derivatives, and blood metabolites of dairy calves. Holstein dairy calves (39.3 ± 1.9 kg of body weight, n = 12 calves per treatment, 6 males and 6 females) were randomly distributed to experimental diets in a 2 × 2 factorial arrangement of treatments. The 4 dietary treatments were (1) starter diet containing GRC and 18% dietary CP (GRC-18CP; geometric mean particle size, GMPS = 0.73 mm); (2) GRC and 21% dietary CP (GRC-21CP; GMPS = 0.71 mm); (3) SFC and 18% dietary CP (SFC-18CP; GMPS = 2.21 mm); and (4) SFC and 21% dietary CP (SFC-21CP; GMPS = 2.16 mm). Calves were weaned on d 63 and remained in the study until d 83 of age. The starter feed intake did not differ among treatments; however, average daily gain and feed efficiency (FE) were improved and final body weight was higher for SFC diets compared with GRC diets. The organic matter and nonfiber carbohydrate digestibilities were greater for calves fed the SFC diets than for those fed the GRC diets. The ruminal total volatile fatty acid concentrations and the molar proportions of propionate and butyrate were greater, and the molar proportion of acetate and NH 3 -N concentrations were lower, for calves fed the SFC diets than for those fed the GRC diets. The excretion of allantoin and total purine derivatives, and subsequently microbial protein synthesis, were greater for calves fed the SFC diets than the GRC diets. The total urinary nitrogen excretion and its proportion of N intake were lower for calves fed the SFC diets than the GRC diets. The blood concentrations of insulin (pre- and postweaning), glucose (postweaning), and β-hydroxybutyrate (preweaning) were greater and blood urea nitrogen (preweaning) was lower for calves fed the SFC diets than the GRC diets. The protein content of the concentrate did not affect feed intake, growth performance, or ruminal fermentation of the calves. The neutral detergent fiber digestibility was greater for calves fed the 21% CP diets than the 18% CP diets. No interaction between main effects was observed regarding the starter intake, average daily gain, body weight, FE, ruminal fermentation, and nutrient digestibility of calves. The interaction between corn grain processing and starter protein content was significant for withers and hip heights with the greatest values found for SFC-21CP treatment. Our results show that steam flaking of corn improved the organic matter and nonfiber carbohydrate digestibilities, weight gain, FE, and ruminal microbial protein synthesis, and reduced urinary nitrogen excretion compared with grinding corn. Regardless of the marginal benefit derived from feeding the diet containing SFC and 21% CP in the height of calves, lower starter protein content (18% CP) may be used efficiently when calves are fed the SFC diets., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

This study evaluated the effects of conditioner retention time during the pelleting process of starter feed on intake, nutrient digestibility, ruminal fermentation, blood metabolites, and growth performance of dairy calves. A total of 30 Holstein female dairy calves [40 ± 1.93 kg of body weight (BW)] were randomly assigned to 1 of 3 treatments: (1) pelleted starter conditioned for 0 min (CON), (2) pelleted starter conditioned for 2 min, and (3) pelleted starter conditioned for 4 min. Three pelleted starter feeds had similar nutritional composition, and the starters were blended with 3% chopped wheat straw and fed to individually housed calves from d 3 to 70 of age. All calves were fed 4 L/d of pasteurized whole milk twice daily at 0800 and 1600 h from d 3 to 50 of calf age, followed by 2 L/d of morning feeding from 51 to 56 d of age. All calves were weaned on d 56 of age and remained in the study until d 70 of age. With the increase of conditioner retention time during pellet processing for 0, 2, and 4 min, the gelatinized starch content of pelleted starter feed linearly increased from 14, 30, and 45%, respectively. Additionally, the pellet durability and hardness also linearly increased with increasing conditioner retention time during pelleting. Feeding pelleted feed prepared using different conditioner retention time did not affect feed dry matter intake, metabolizable energy intake, weaning BW, final BW, or feed efficiency during the study. We observed no differences in the total-tract apparent digestibility of organic matter, neutral detergent fiber, and crude protein around weaning (d 49 to 56 of age) and after weaning (d 63 to 70 of calf cage); however, the digestibility of dry matter and starch after weaning was increased with increasing conditioner retention time during pelleting of starter feed. No difference was found in overall average daily gain (ADG) or growth rates of hip height, withers height, and heart girth. Ruminal volatile fatty acid profile was not affected by pelleting under different conditioner retention times. The ruminal ammonia concentration tended to be lower for calves fed the 4-min diet compared with those fed the CON diet during the postweaning period. The postweaning (d 57-70 of age) ADG was greater for calves fed the 4-min diet compared with those fed the CON diet. In conclusion, the conditioning time during the pelleting process of starter feed increased the gelatinization of starch, durability, and hardness of the pellets but did not influence feed intake, feed efficiency, and skeletal growth during the first 70 d of age. Increasing conditioning time during the pelleting process improved postweaning ADG; however, the final BW of calves was similar among treatments., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to determine the effect of partial replacement of whole milk with colostrum on the growth performance and health status of Holstein dairy calves. Neonatal heifer calves (n = 144; 2 d of age; 39.3 ± 0.82 kg of body weight, BW; mean ± SE) were assigned randomly to 3 groups with partial replacement of pasteurized whole milk with pasteurized colostrum at 0 (C0; 0 kg/d of colostrum + 5 kg/d of whole milk), 350 g (C350; 0.350 kg/d of colostrum + 4.650 kg/d of whole milk), or 700 g (C700; 0.700 kg/d of colostrum + 4.300 kg/d of whole milk) for 14 d; there were no refusals of liquid feed. From d 15 onward, the calves were fed with 5 kg/d of pasteurized whole milk, weaned on d 61, and monitored until d 81 of life. Throughout the study, the calves had free access to fresh clean water and calf starter. Partial replacement of whole milk with colostrum increased liquid feed dry matter intake (DMI) but decreased milk DMI; however, intakes of starter DMI, total DMI, metabolizable energy, crude protein, and ether extract were not affected by treatments. Overall, the C700 calves recorded greater weaning weight, final BW, heart girth change, feed efficiency, and average daily gain (ADG). The calves fed milk had a higher chance of having rectal temperature ≥39.4°C and general appearance score ≥2 compared with those receiving colostrum in their milk. Diarrhea was more prevalent in C0 versus C700 calves. The occurrence of pneumonia tended to be higher in milk-fed calves compared with C350 and C700 animals. Colostrum feeding resulted in fewer days with a rectal temperature ≥39.4°C, general appearance ≥2, diarrhea, and pneumonia. We computed Cliff's delta (effect sizes) of the extended colostrum feeding (C350 vs. C0, C700 vs. C0, and C700 vs. C350) on starter and milk DMI, ADG, BW, and feed efficiency. In C350 calves, the effect sizes (Cliff's delta) for milk DMI, ADG, BW, and feed efficiency were positive and small, but negative in C700 calves. Compared with C350 treatment, C700 treatment resulted in greater final BW with moderate effect size. Positive and moderate effects of feeding colostrum (C700 vs. C0) were observed on postweaning ADG and final BW. The findings showed that the inclusion of 700 g of colostrum in 5 kg of milk may be beneficial to the growth and health of dairy calves., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to evaluate the effects of heat stress in late gestation independent of maternal reduced feed intake on performance, blood hormones and metabolites, and immune responses of dairy calves from birth through weaning. A total of 30 multiparous Holstein cows at 45 d before expected calving were randomly assigned to one of 3 groups: (1) thermal neutral (CL, n = 10) conditions with ad libitum feed intake (10% of refusals on an as-fed basis); (2) pair-fed thermal neutral (CLPF, n = 10) conditions to reduce feed intake to levels similar to the heat stress (HS) group while reared under thermoneutral conditions (80% of the CL group); or (3) heat stress (HS, n = 10) conditions with ad libitum feed intake. Pair-feeding was conducted to quantify the confounding effects of dissimilar feed intake. Calves (10/group) born to cows that were exposed to cooling (IU-CL), pair-feeding (IU-CLPF), or heat stress (IU-HS) were used from birth through weaning. After birth, all the calves were managed under identical conditions. IU-HS calves had lower birth weight, and hip height at birth and 14 d of age. Compared with IU-CL and IU-CLPF calves, IU-HS calves had lower serum concentration of IgG and apparent efficiency of IgG absorption but higher serum insulin concentrations. Cortisol concentration in serum was higher in IU-HS and IU-CLPF calves compared to IU-CL calves. The neutrophil percentage was lower in IU-CL calves than in IU-HS and IU-CLPF calves. Neutrophil-lymphocyte ratio was higher in IU-HS calves compared to IU-CLPF and IU-CL calves. The mRNA expression of TNFα of IU-HS calves was downregulated compared with IU-CL and IU-CLPF calves. In summary, maternal HS during late gestation reduces calf birth weight and dramatically alters blood hormones and metabolites, but its effect on immune system function was not independent of maternal reduced feed intake., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Haptoglobin (Hp), one of the major positive acute phase proteins in cattle, is released in response to proinflammatory cytokines. Colostrum intake might influence the response of the innate immune system, including Hp gene expression. Thus, we hypothesized that plasma concentrations and tissue mRNA expression of Hp in neonatal calves might be influenced by early nutrition in the neonatal calf and would thus be greater if receiving colostrum compared with milk-based formula. Two trials were performed. In trial 1, German Holstein calves were fed either colostrum (COL; n = 7) or milk-based formula (FOR; n = 7) up to 4 d of life. Blood was sampled from d 1 to 4 before morning feeding and before and 2 h after feeding on d 4. Tissue samples from liver, kidney fat, duodenum, and ileum were collected after slaughter on d 4 at 2 h after feeding. In trial 2, calves born preterm (n = 7) or at term (n = 7) received colostrum only at 24 h post natum. Blood was sampled at birth, and before and 2 h after feeding. Tissue samples from liver and kidney fat were collected after slaughter at 26 h after birth. Blood plasma, colostrum, and formula Hp concentrations were determined using a competitive ELISA. Tissue expression of Hp mRNA was quantified by real-time quantitative PCR. The formula contained much less Hp (≤0.5 µg/mL) than colostrum (69.3, 93.9, and 20.4 µg/mL from d 1 to d 3, respectively). In trial 1, before colostrum or formula feeding, plasma concentrations of Hp were comparable in both groups. Plasma Hp increased in FOR after feeding, resulting in greater or a trend for greater plasma Hp concentrations in FOR than in COL calves. The mRNA abundance of Hp in liver and kidney fat was 3- and 2.2-fold greater in FOR than in COL calves, respectively, whereas duodenal and ileal abundance of Hp mRNA did not differ between groups. In trial 2, plasma Hp concentrations decreased slightly over time in term calves, but they did not differ in both groups before and 2 h after feeding on d 2. The abundance of Hp mRNA in liver was 5.3-fold greater in term than in preterm calves, whereas its abundance in kidney fat did not differ between groups. Contrasting our hypothesis, formula, but not colostrum feeding was associated with greater Hp mRNA abundance in liver and adipose tissue, indicating that the response of innate immune system seems to be modulated by formula feeding because of the lack of immunoglobulin intake. The lower hepatic abundance of Hp mRNA in preterm calves than in term calves may indicate lower synthetic capacity of the liver for Hp in preterm calves shortly after birth., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

We aimed to evaluate the effects of feeding sugarcane liquid molasses (LM) with or without a commercial buffer mix (BFM) on ruminal fermentation parameters, milk fatty acid (FA) profile, and milk yield and composition in dairy cows fed high-concentrate diets (35:65 forage-to-concentrate ratio). Eight multiparous Holstein cows (4 ruminally cannulated) averaging 165 ± 12 d in milk at the beginning of the study were randomly assigned to a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Cows were fed the following diets: (1) no LM or BFM supplementation (CTRL), (2) LM without BFM supplementation (MOL), (3) BFM without LM supplementation (BUF), and (4) LM plus BFM supplementation (COMBO). These 4 isonitrogenous and isoenergetic diets were formulated by replacing (dry matter basis) 5% ground corn with LM, whereas BFM replaced wheat bran at 0.8% of the diet. Significant LM × BFM interactions were observed for the duration of ruminal pH below 5.8, molar proportion of propionate, acetate-to-propionate ratio, milk proportions of trans-10 18:1 and total trans FA, and concentration and yield of milk fat. Feeding MOL and BUF alone were effective on reducing the time that ruminal pH remained below 5.8 compared with the CTRL treatment, and the COMBO diet decreased it further. A similar pattern was observed for the ruminal molar proportion of propionate. The milk proportions of trans-10 18:1 and total trans FA dropped significantly with BFM or LM supplementation versus cows fed CTRL, and the COMBO diet decreased these variables further. Note, however, that these changes elicited by the COMBO diet were not in the same magnitude as those caused by MOL or BUF fed alone. The ruminal molar proportion of acetate increased with the BUF diet and that of butyrate increased in cows fed MOL, but mean ruminal pH was not affected by treatments. Diets with LM resulted in increased concentrations of short- and medium-chain FA in milk fat. The yield of 3.5% fat-corrected milk increased significantly in cows fed MOL or BUF due to the improved concentration of milk fat. A trend and a significant increase for energy-corrected milk were observed with feeding MOL or BUF, respectively. Overall, inclusion of LM and BFM appears to reduce milk trans-10 18:1 FA and total trans FA by modulating ruminal pH and volatile FA profile in cows fed high-concentrate diets., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The transition from late gestation to early lactation is associated with extensive changes in metabolic, endocrine, and immune functions in dairy cows. Skeletal muscle plays an important role in maintaining the homeorhetic adaptation to the metabolic needs of lactation. The objective of this study was to characterize the skeletal muscle metabolome in the context of the metabolic changes that occur during the transition period in dairy cows with high (HBCS) versus normal body condition (NBCS). Fifteen weeks antepartum, 38 pregnant multiparous Holstein cows were assigned to 1 of 2 groups, which were fed differently to reach the targeted BCS and back fat thickness (BFT) until dry-off at -49 d before calving (HBCS: >3.75 and >1.4 cm; NBCS: <3.5 and <1.2 cm). During the dry period and the subsequent lactation, both groups were fed identical diets. The differences in both BCS and BFT were maintained throughout the study. The metabolome was characterized in skeletal muscle samples (semitendinosus muscle) collected on d -49, 3, 21, and 84 relative to calving using a targeted metabolomics approach (AbsoluteIDQ p180 kit; Biocrates Life Sciences AG, Innsbruck, Austria), which allowed for the quantification of up to 188 metabolites from 6 different compound classes (acylcarnitines, amino acids, biogenic amines, glycerophospholipids, sphingolipids, and hexoses). On d -49, the concentrations of citrulline and hydroxytetradecadienyl-l-carnitine in muscle were higher in HBCS cows than in NBCS cows, but those of carnosine were lower. Over-conditioning did not affect the muscle concentrations of any of the metabolites on d 3. On d 21, the concentrations of phenylethylamine and linoleylcarnitine in muscle were lower in HBCS cows than in NBCS cows, and the opposite was true for lysophosphatidylcholine acyl C20:4. On d 84, the significantly changed metabolites were mainly long-chain (>C32) acyl-alkyl phosphatidylcholine and di-acyl phosphatidylcholine, along with 3 long-chain (>C16) sphingomyelin that were all lower in HBCS cows than in NBCS cows. These data contribute to a better understanding of the metabolic adaptation in skeletal muscle of dairy cows during the transition period, although the physiological significance and underlying molecular mechanisms responsible for the regulation of citrulline, hydroxytetradecadienyl-l-carnitine, carnosine, and phenylethylamine associated with over-conditioning are still elusive and warrant further investigation. The changes observed in muscle lysophosphatidylcholine and phosphatidylcholine concentrations may point to an alteration in phosphatidylcholine metabolism, probably resulting in an increase in membrane stiffness, which may lead to abnormalities in insulin signaling in the muscle of over-conditioned cows., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The mammalian target of rapamycin (mTOR) is a major regulator of protein synthesis via its main downstream effectors, ribosomal protein S6 kinase (S6K1) and eukaryotic initiation factor 4E binding protein (4EBP1). The ubiquitin-proteasome system (UPS) is the main proteolytic pathway in muscle, and the muscle-specific ligases tripartite motif containing 63 (TRIM63; also called muscle-specific ring-finger protein 1, MuRF-1) and F-box only protein 32 (FBXO32; also called atrogin-1) are important components of the UPS. We investigated 20S proteasome activity and mRNA expression of key components of mTOR signaling and UPS in skeletal muscle of dairy cows during late gestation and early lactation and tested the effects of dietary supplementation (from d 1 in milk) with conjugated linoleic acids (sCLA; 100 g/d; n = 11) compared with control fat-supplemented cows (CTR; n = 10). Blood and muscle tissue (semitendinosus) samples were collected on d -21, 1, 21, and 70 relative to parturition. Dry matter intake increased with time of lactation in both groups. It was lower in sCLA than in CTR on d 21, which resulted in a reduced calculated metabolizable protein balance. Most serum and muscle concentrations of AA followed time-related changes but were unaffected by CLA supplementation. In both groups, serum and muscle 3-methylhistidine (3-MH) concentrations and the ratio of 3-MH:creatinine increased from d -21 to d 1, followed by a decline on d 21. The mRNA abundance of MTOR on d 21 and 70 was greater in sCLA than in CTR. The abundance of 4EBP1 mRNA did not differ between groups but was upregulated in both on d 1. The mRNA abundance of S6K1 on d 70 was greater in CTR than in sCLA, but remained unchanged over time in both groups. The mRNA abundance of FBXO32 (encoding atrogin-1) on d 21 was greater in sCLA than in CTR. The mRNA abundance of TRIM63 (also known as MuRF1) showed a similar pattern as FBXO32 in both groups: an increase from d -21 to d 1, followed by a decline. The mRNA for the α (BCKDHA) and β (BCKDHB) polypeptide of branched-chain α-keto acid dehydrogenase was elevated in sCLA and CTR cows on d 21, respectively, suggesting a role of CLA in determining the metabolic fate of branched-chain AA. For the mTOR protein, no group differences were observed. The abundance of S6K1 protein was greater across all time points in sCLA versus CTR. The antepartum 20S proteasome activity in muscle was elevated in both groups compared with postpartum, probably reflecting the start of protein mobilization before parturition. Plasma insulin concentrations decreased in both groups postpartum but to a greater extent in CTR than in sCLA, resulting in greater insulin concentrations in sCLA than in CTR. Thus, the greater abundance of MTOR mRNA and S6K1 protein in sCLA compared with CTR might be mediated by the greater plasma insulin postpartum. The upregulation of MTOR mRNA in sCLA cows on d 21, despite greater FBXO32 mRNA abundance, may reflect a simultaneous activation of both anabolic and catabolic signaling pathways, likely resulting in greater protein turnover., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of the current study was to investigate the effects of overconditioning around calving on gene expression of key components of the mammalian target of rapamycin (mTOR) pathway and ubiquitin-proteasome system (UPS) in skeletal muscle as well as the AA profiles in both serum and muscle of periparturient cows. Fifteen weeks antepartum, 38 multiparous Holstein cows were allocated to either a high body condition group (HBCS; n = 19) or a normal body condition group (NBCS; n = 19) and were fed different diets until dry-off (d -49 relative to calving) to amplify the difference. The groups were also stratified for comparable milk yields (NBCS: 10,361 ± 302 kg; HBCS: 10,315 ± 437 kg). At dry-off, the NBCS cows (parity: 2.42 ± 1.84; body weight: 665 ± 64 kg) had a body condition score (BCS) <3.5 and backfat thickness (BFT) <1.2 cm, whereas the HBCS cows (parity: 3.37 ± 1.67; body weight: 720 ± 57 kg) had a BCS >3.75 and BFT >1.4 cm. During the dry period and the subsequent lactation, both groups were fed identical diets but maintained the BCS and BFT differences. Blood samples and skeletal muscle biopsies (semitendinosus) were repeatedly (d -49, +3, +21, and +84 relative to calving) collected for assessing the concentrations of free AA and the mRNA abundance of various components of mTOR and UPS. The differences in BCS and BFT were maintained throughout the study. The circulating concentrations of most AA with the exception of Gly, Gln, Met, and Phe increased in early lactation in both groups. The serum concentrations of Ala (d +21 and +84) and Orn (d +84) were lower in HBCS cows than in NBCS cows, but those of Gly, His, Leu, Val, Lys, Met, and Orn on d -49 and Ile on d +21 were greater in HBCS cows than in NBCS cows. The serum concentrations of 3-methylhistidine, creatinine, and 3-methylhistidine:creatinine ratio increased after calving (d +3) but did not differ between the groups. The muscle concentrations of all AA (except for Cys) remained unchanged over time and did not differ between groups. The muscle concentrations of Cys were greater on d -49 but tended to be lower on d +21 in HBCS cows than in NBCS cows. On d +21, mTOR and eukaryotic translation initiation factor 4E binding protein 1 mRNA abundance was greater in HBCS cows than in NBCS cows, whereas ribosomal protein S6 kinase 1 was not different between the groups. The mRNA abundance of ubiquitin-activating enzyme 1 (d +21), ubiquitin-conjugating enzyme 1 (d +21), atrogin-1 (d +21), and ring finger protein-1 (d +3) enzymes was greater in HBCS cows than in NBCS cows, whereas ubiquitin-conjugating enzyme 2 was not different between the groups. The increased mRNA abundance of key components of mTOR signaling and of muscle-specific ligases of HBCS cows may indicate a simultaneous activation of anabolic and catabolic processes and thus increased muscle protein turnover, likely as a part of the adaptive response to prevent excessive loss of skeletal muscle mass during early lactation., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)