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Even with the same energy supply, differences in energy sources may affect calf growth and metabolism. In this study, we evaluated the effects of feeding 3 different milk replacers (MR) with different lactose supplies under isoenergetic conditions on calf digestibility, growth, and metabolism-related markers. Fifteen Holstein bull calves were randomly assigned to one of 3 MR feeding treatments: low lactose (L: 38%), medium lactose (M: 41%), or high lactose (H: 46%). After birth, calves were provided colostrum, and treatment MR were fed from 1 d of age and gradually increased to a maximum feeding rate at 20 d of age (L: 1.16 kg/d, M: 1.21 kg/d, H: 1.26 kg/d DM) without feeding solid feeds during the experimental period. Blood samples were temporarily collected weekly to assess blood concentrations of metabolites and hormones. From 30 to 36 d of age, the calves were euthanized and liver samples were collected to determine growth-related mRNA expression. The L calves showed a greater body length than H calves and the highest growth hormone receptor (GHR) mRNA expression. Plasma concentrations of total cholesterol, urea nitrogen, total protein, albumin, insulin, and IGF-1 were not different, but plasma concentrations of triglycerides were greater in order H, M, and L. These results showed that the difference in lactose content in the MR affected calf metabolism, and the L-MR was suggested to be more likely to enhance growth into the peripheral tissues.

ABSTRACT: The objectives of this study were to determine the farm-level hyperketolactia (HKL) prevalence, as diagnosed from the milk BHB concentration, on dairy farms milking with an automated milking system (AMS) and to describe the farm-level housing, management, and nutritional risk factors associated with increased farm-average milk BHB and the within-herd HKL prevalence in the first 45 DIM. Canadian AMS farms (n = 162; eastern Canada, n = 8; Quebec, n = 24; Ontario, n = 75; western Canada n = 55) were visited once between April and September 2019 to record housing and herd management practices. The first test milk data for each cow under 45 DIM were collected, along with the final test of the previous lactations for all multiparous cows, from April 1, 2019, to September 30, 2020. The first test milk BHB was then used to classify each individual cow as having or not having HKL (milk BHB ≥0.15 mmol/L) at the time of testing. Milk fat and protein content, milk BHB, and HKL prevalence were summarized by farm and lactation group (all, primiparous, and multiparous). During this same time period, formulated diets for dry and lactating cows, including ingredients and nutrient composition, and AMS milking data were collected. Data from the AMS were used to determine milking behaviors and milk production of each herd during the first 45 DIM. Multivariable regression models were used to associate herd-level housing, feeding management practices, and formulated nutrient composition with first test milk BHB concentrations and within-herd HKL levels separately for primiparous and multiparous cows. The within-herd HKL prevalence for all cows was 21.8%, with primiparous cows having a lower mean prevalence (12.2 ± 9.2%) than multiparous cows (26.6 ± 11.3%). Milk BHB concentration (0.095 ± 0.018 mmol/L) and HKL prevalence for primiparous cows were positively associated with formulated prepartum DMI and forage content of the dry cow diet; however, they were negatively associated with formulated postpartum DMI, the major ingredient in the concentrate supplemented through the AMS, and the partially mixed ration to AMS concentrate ratio. Multiparous cows' milk BHB concentration (0.12 ± 0.023 mmol/L) and HKL prevalence were positively associated with the length of the previous lactation, milk BHB at dry-off, prepartum diet nonfiber carbohydrate content, and the major forage fed on farm, while tending to be negatively associated with feed bunk space during lactation. This study is the first to determine the farm-level risk factors associated with herd-level prevalence of HKL in AMS dairy herds. The results may help to optimize management and guide diet formulation and thus promote the reduction of HKL prevalence.

ABSTRACT: The objective of this study was to investigate the effects of feeding surplus dairy calves a milk replacer (MR) or one of 2 different oral rehydration solutions (ORS) during a midtransportation rest period on metabolic and clinical health indicators, growth, and behavioral outcomes after arrival at a calf-raising facility. Surplus dairy calves (n = 128) were transported in 4 cohorts from February to July 2022 for 12 h to a holding facility, rested for 8 h, then transported for an additional 6 h to a calf-raising facility. Upon arrival at the holding facility, calves were randomly assigned to 1 of 3 treatments: MR (n = 43), a high-sodium ORS developed for diarrhea (ORS-D; n = 43), or a high-potassium ORS developed for transportation (ORS-T; n = 42). The exact age of calves at transportation was unknown; however, all calves were less than 14 d of age. Calf BW at enrollment was 43.9 ± 5.9 kg, 43.7 ± 6.5 kg, and 45.0 ± 4.5 kg for calves fed MR, ORS-D, and ORS-T, respectively. Calves were fed 2.0 L of their treatment twice, once upon arrival and once before leaving the holding facility. At unloading and reloading at the holding facility, calves were weighed and blood samples were obtained. Calves were also health scored at unloading at the holding facility. After arrival at the calf-raising facility, calves were weighed, health scored, and blood samples were taken. Blood samples were collected at 24 and 48 h and BW was recorded at 24 h, 48 h, 72 h, 5 d, 7 d, 14 d, and at 8 wk after arrival at the calf-raising facility. Calves were also health scored daily for 14 d; health scoring included fecal consistency scoring and evaluating the presence or absence of respiratory disease. Lying time, lying bouts, and activity index were measured during transportation and from 3 d relative to transportation using accelerometers. At arrival to the calf-raiser, calves fed ORS-D had higher concentrations of nonesterified fatty acids (NEFA) and BHB than calves fed MR. Furthermore, calves fed ORS-T had higher concentrations of BHB at arrival to the calf raiser compared with calves fed MR. In the 14 d after arrival at the calf-raiser, there was evidence that calves fed ORS-T had a higher proportion of days with diarrhea and respiratory disease compared with those fed MR. During transportation, calves fed ORS-T had a lower activity index than calves fed MR, suggesting that ORS-T calves had lower overall activity. In addition, on the day of transportation (d 0), ORS-T and ORS-D calves had a lower activity index than calves fed MR. There were no treatment effects on growth outcomes. The results of this study suggest that feeding MR rather than an ORS during a midtransportation rest period could minimize fat mobilization and can potentially improve diarrhea and respiratory disease but does not affect growth outcomes after arrival at calf-raisers.

ABSTRACT: The objective of this study was to describe the nutritional strategies used on Canadian dairy farms with automated milking systems (AMS), both at the feed bunk and the concentrate offered at the AMS, as well as to determine what dietary components and nutrients, as formulated, were associated with milk production and milking behaviors on those farms. Formulated diets (including ingredients and nutrient content) and AMS data were collected from April 1, 2019, until September 30, 2020, on 160 AMS farms (eastern Canada [East] = 8, Ontario [ON] = 76, Quebec [QC] = 22, and western Canada [West] = 54). Both partial mixed ration (PMR) and AMS concentrate samples were collected from May 1 to September 30, 2019, on 169 farms (East = 12, ON = 63, QC = 42, West = 52). We collected AMS milking data for 154 herds. For each farm (n = 161), milk recording data were collected and summarized by farm to calculate average milk yield and components. Multivariable regression models were used to associate herd-level formulated nutrient composition and feeding management practices with milk production and milking behavior. Milk yield (mean ± SD = 37.0 ± 0.3 kg/d) was positively associated with the PMR ether extract (EE) concentration (+0.97 kg/d per percentage point [p.p.] increase) and with farms that fed barley silage as their major forage source (n = 16; +2.18 kg/d) as compared with haylage (n = 42), whereas farms that fed corn silage (n = 96; +1.23 kg/d) tended to produce more milk than farms that fed haylage. Greater milk fat content (4.09 ± 0.28%) was associated with a greater PMR-to-AMS concentrate ratio (+0.02 p.p. per unit increase) and total diet net energy for lactation (+0.046 p.p. per 0.1 Mcal/kg increase), but a lesser percentage of NFC of the PMR (−0.016 p.p. per p.p. increase of NFC percentage). Milk protein content (3.38 ± 0.14%) was positively associated with the forage percentage of the PMR (+0.003 p.p. per p.p. increase of forage percentage) and the total diet starch percentage (+0.009 p.p. per p.p. increase of starch percentage), but was negatively associated with farms feeding corn silage (−0.1 p.p. compared with haylage) as their major forage. Greater milking frequency (2.77 ± 0.40 milkings/d) was observed on farms with free-flow cow traffic systems (+0.62 milkings/d) and was positively associated with feed push-up frequency (+0.013 milkings/d per additional feed push-up), but negatively associated with PMR NFC content and forage percentage of the total ration (−0.017 milkings/d per p.p. increase of forage percentage). Lastly, greater milking refusal frequency (1.49 ± 0.82 refusals/d) was observed on farms with free-flow cow traffic systems (+0.84 refusals/d) and farms feeding barley silage (+0.58 refusals/d) than with guided flow and farms feeding either corn silage or haylage, respectively. These data give insight into the ingredients, nutrient formulations and type of diets fed on AMS dairy farms across Canada and the association of those factors with milk production and milking behaviors.

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

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

ABSTRACT: The preweaning period for a dairy calf is characterized by high morbidity and mortality rates, leading to financial losses for producers. Identifying strategies to improve the health and welfare of calves while reducing antimicrobial use continues to be crucial to the success of the dairy industry. The objective of this study was to determine the effects of feeding colostrum replacer (CR) to dairy heifer calves beyond d 1 of life on growth, serum IgG, the incidence of diarrhea and bovine respiratory disease (BRD), and the risk of mortality in the preweaning period. At birth, Holstein heifer calves (n = 200; 50/treatment) weighing 40.7 ± 0.35 kg (mean ± SE) were fed 3.2 L of CR (205 g IgG/feeding) at 0 h and 12 h of life. Calves were then randomly assigned to 1 of 4 treatments: 450 g of milk replacer (MR) from d 2 to 14 (control, CON), 380 g of CR + 225 g of MR from d 2 to 3, then 450 g of MR from d 4 to 14 (transition, TRAN), 45 g of CR + 450 g of MR from d 2 to 14 (extended, EXT); or 380 g of CR + 225 g of MR from d 2 to 3, then 45 g of CR + 450 g of MR from d 4 to 14 (transition + extended, TRAN+EXT). Each treatment was reconstituted to 3 L and fed twice daily. All CR treatments were fed using bovine-derived CR containing 27% IgG. From d 15 to 41, all calves were fed 600 g of MR reconstituted to 4 L twice daily. Body weight was recorded at birth and every 7 d until study completion on d 49. Blood samples were taken daily until d 7 to evaluate serum IgG and then every 7 d until d 49. A health assessment was performed daily to evaluate calves for BRD and diarrhea. Data were analyzed using mixed linear regression, mixed logistic regression, and survival analysis models in SAS 9.4. Serum IgG concentrations were not affected by treatment for the study period. The EXT and TRAN+EXT groups had greater average daily gain (ADG) from d 7 to 14 (0.14 kg/d) and the TRAN group had greater ADG from d 14 to 21 (0.11 kg/d), compared with CON. There was no association of treatment with the odds or the duration of a diarrhea bout. However, provision of CR to the TRAN and EXT calves was associated with a reduced hazard of diarrhea compared with CON calves. Furthermore, TRAN and EXT calves have a lower hazard of mortality compared with CON calves, with TRAN and EXT calves had a 2.8- and 3.8-times lower hazard of mortality, respectively. Our findings suggest that the supplementation of CR to dairy calves positively affects ADG, and reduces the hazard of diarrhea and mortality during the preweaning period. Future research should look to further refine the supplementation strategy of CR to calves and explore the mechanism of action.

ABSTRACT: This graduate student literature review provides an examination of the ontological adaptations of the calf's immune system and how the intestinal microbiota influences calf immune function in health and disease. Within dairy rearing systems, various nutritional and management factors have emerged as critical determinants of development influencing multiple physiological axes in the calf. Furthermore, we discuss how multiple pre- and postnatal maternal factors influence the trajectory of immune development in favor of establishing regulatory networks to successfully cope with the new environment, while providing early immune protection via immune passive transfer from colostrum. Additionally, our review provides insights into the current understanding of how the intestinal microbiota contributes to the development of the intestinal and systemic immune system in calves. Lastly, we address potential concerns related to the use of prophylactic antimicrobials and waste milk, specifically examining their adverse effects on intestinal health and metabolic function. By examining these factors, we aim to better understand the intricate relationship between current management practices and their long-term effect on animal health.

ABSTRACT: The objective of this study was to quantify the effects of supplementing a low level of dry glycerol product pre- and postpartum on the feeding behavior, lying behavior, and reticulorumen pH of dairy cows. Multiparous Holstein dairy cows (n = 60) were enrolled in a 2 × 2 factorial design study. Twenty-one days before expected parturition, cows individually received a dry cow diet with (1) 250 g/d glycerol supplementation (GLY; 66% pure glycerol, United States Pharmacopeia grade), or (2) no supplementation (CON). Following parturition, cows were individually assigned to either (1) 250 g/d glycerol product (GLY; 66% pure glycerol), or (2) no supplementation (CON) to their partial mixed ration (PMR) for the first 21 d in milk (DIM). All cows were milked by an automated milking system and offered a target of 5.4 kg/d pellet (23% of target total dry matter intake [DMI]). For both treatment periods, cows were individually assigned to automated feed bins to measure PMR feeding behavior. Rumination time and lying behavior were monitored with electronic sensors for the whole study (−21 to 21 DIM). Reticulorumen pH boluses were administered to a subset of cows (n = 40) where pH was recorded every 10 min from 21 d prepartum to 21 d postpartum. Prepartum, cows fed GLY had fewer, larger meals and spent 20.2% more time feeding than CON while consuming feed at a similar rate. Cows on the CON diet prepartum spent more time lying down in more frequent bouts in the 21 d before calving. Following parturition, cows that received GLY prepartum continued to devote more time to eating, while tending to spend less time ruminating per kilogram of DMI. Cows receiving CON postpartum had larger meals with longer intervals between meals. In the first 21 DIM, cows receiving CON prepartum tended to have shorter, but significantly more frequent, lying bouts than cows fed GLY prepartum. Glycerol supplementation pre- and postpartum resulted in less time spent lying down following parturition. Minimal differences between treatments were observed for pre- and postpartum sorting behavior or reticulorumen pH. Overall, supplementation of glycerol pre- and postpartum altered cow time budgets, with cows spending more time eating pre- and postpartum, less time lying pre- and postpartum, and having fewer, larger meals prepartum when receiving glycerol prepartum, and with cows having slower feeding rates and smaller meals following parturition with postpartum glycerol supplementation.

ABSTRACT: This study aimed to characterize the development of systemic and colon tissue resident B and γδ T cells in newborn calves from birth until weaning. At birth, calves have limited capacity to initiate immune responses, and the immune system gradually matures over time. Gamma delta (γδ) T cells are an important lymphocyte subset in neonatal calves that confer protection and promote immune tolerance. A total of 36 newborn calves were enrolled in a longitudinal study to characterize how systemic and colon tissue resident B and γδ T cells develop from birth until weaning. Blood and colon biopsy samples were collected on d 2, 28, and 42 to determine the proportions of various B and γδ T cell subsets by flow cytometry. We classified γδ T cells into different functional subsets according to the level of expression intensity of the coreceptors WC1.1 (effector function) and WC1.2 (regulatory function). Furthermore, naive B cells were classified based on the expression IgM receptor, and activation state was determined based on expression of CD21 and CD32, 2 receptors with opposing signals involved in B cell activation in early life. Additional colon biopsy samples were used for 16S sequencing, and microbial diversity data are reported. At birth, γδ T cells were the most abundant lymphocyte population in blood, accounting for 58.5% of the lymphocyte pool, after which the proportions of these cells declined to 38.2% after weaning. The proportion of γδ T cells expressing WC1.1 decreased by 50% from d 2 to d 28, whereas no change was observed in the expression of WC1.2. In the colon, there was a 50% increase of γδ T cells after weaning and the proportion of WC1.2+ γδ T cells doubled from d 28 to 42. The proportion of IgM+ B lymphocytes in blood increased from 23.6% at birth to 30% after weaning, were the proportion of B cells expressing CD21 increased by 25%, while the proportion of B cells expressing CD32 decreased by 30%. While no changes were observed for the overall proportion of IgM+ B lymphocytes in the colon, there was a 6-fold increase in the proportion of CD21+ B cells from pre- (d 28) to postweaning (d 42). Microbial diversity increased from d 2 of life to 28 and declined abruptly after weaning. The reduction in microbial diversity during weaning was negatively correlated with the increase in all γδ T cell subsets and CD21+ B cells. These data suggest that developmental adaptations after birth coordinate expansion of γδ T cells to provide early systemic protection, as well as to steer immune tolerance, while B cells mature over time. Additionally, the increase of colonic γδ T cells on d 42 suggests a protective role of these cells during weaning.

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

ABSTRACT: The objective of this study was to quantify the effects of supplementing transition dairy cows with a low inclusion dry glycerol product in the pre- and postpartum periods on feed intake, metabolic markers, and milk yield and components. Multiparous Holstein dairy cows (n = 60) were enrolled in a 2-by-2 factorial design study. Starting 21 d before expected parturition, cows individually received a dry cow diet with (1) 250 g/d glycerol product supplementation [66% pure glycerol (United States Pharmacopeia grade); GLY], or (2) no supplementation (CON) mixed to their total mixed ration. After parturition, cows, again, were individually assigned to either GLY, or (2) no supplementation (CON) to their partial mixed ration for the first 21 d in milk (DIM). Cows were milked by an automated milking system and offered a target of 5.4 kg DM/d pellet (23% of target total dry matter intake, DMI) in the automated milking system and followed for 42 d into lactation. Blood samples were collected 6.3 ± 3.47 d before calving for all blood measures and 3, 7, 10, and 14 DIM for analysis of glucose and β-hydroxybutyrate, as well as 3 and 7 DIM for nonesterified fatty acids (NEFA) and haptoglobin. Initial dry cow body weight (BW), calf birth weight, previous 305-d milk, and month of parturition were used as covariates in the statistical model. Cows supplemented with GLY prepartum lost less BW and consumed more DMI pre- and postpartum, as well as had lower postpartum blood β-hydroxybutyrate and NEFA concentrations compared with those fed the CON treatment prepartum. Cows supplemented with GLY postpartum had lesser DMI in the first 42 DIM than cows fed CON postpartum, but also had reduced blood NEFA concentrations, odds of a high haptoglobin test, odds of a low blood glucose test, and lesser preformed fatty acid concentrations and yields in their milk. Cows supplemented glycerol both pre- and postpartum lost the least total BW from −21 to 21 DIM. No treatment effects were detected for milk yield; however, cows receiving GLY postpartum had lower milk fat. Overall, glycerol supplementation during the transition period, particularly during the 21 d before calving, was associated with markers of improved metabolic status.

Investigating the immune responses of the intestine in response to different insults is predominantly limited to indirect methods such as circulating markers of intestinal health or gene expression from dissections. We describe here a validated protocol for the isolation and subsequent flow cytometry analysis of intestinal intraepithelial lymphocytes (IEL) from colonic biopsy samples. Colon biopsy samples were collected with endoscopy forceps from Holstein dairy bull calves at d 2, 28, and 42 of life. The biopsies were put into an isolation solution of Hanks' balanced salt solution, and fetal bovine serum followed by digestion solution. The solution was filtered and the flow-through, containing IEL, was stained with fluorescent antibodies for flow cytometry analysis. Density gradient separation of the isolate yielded higher viability and cleaner samples for flow cytometry analysis. Anti-bovine γ chain of the T cell receptor was used to identify populations of gamma delta (γδ) T cells via flow cytometry. In addition, γδ T cell subsets were identified using an anti-bovine antibody against the coreceptor workshop cluster 1. This method allowed for the precise identification of lymphocyte populations and evaluation of the proportion of different subsets of γδ T cells from intestinal IEL over time. The technique described here will allow the research community to characterize intestinal immune function over time and improve our understanding of how different management and nutritional strategies affect intestinal health.

Since insulin has been demonstrated to suppress IgG absorption in other neonatal species, we had the objective to delineate how colostral insulin concentrations affect IgG absorption in neonatal bovines. We enrolled Holstein bull calves (n = 48; body weight = 46.3 ± 0.84 kg) at birth and randomized them by birth order to receive (1) colostrum that contained basal insulin concentrations (12.9 μg/L; n = 16), or colostrum that had been supplemented with an exogenous insulin to increase the insulin concentration to either (2) 5 times (70.0 μg/L; n = 16) or (3) 10 times (149.7 μg/L; n = 16) that of the basal colostrum. Gross colostrum composition (crude fat: 4.1 ± 0.06%; crude protein: 11.7 ± 0.05%; lactose: 1.9 ± 0.01%; IgG: 63.9 ± 1.19 g/L) was similar between treatments and calves were fed (7% body weight, 3.1 ± 0.06 L) their treatments at 2, 14, and 26 h postnatal. Serum was collected at 0, 30, 60, 90, 120, 180, 240, 360, 480, and 600 min postprandial respective to the first and second colostrum feeding and analyzed for IgG concentration. The incremental area under the curve (I-AUC) and apparent efficiency of absorption (AEA) were calculated for the 10-h periods following the first and second colostrum meal. Serum IgG concentrations over time, I-AUC, and AEA were statistically analyzed as a complete randomized design. Colostrum insulin concentration did not affect serum IgG concentrations or the I-AUC or AEA after calves were fed colostrum at 2 and 14 h postnatal. High colostral insulin content is not detrimental or promotive to IgG absorption in neonatal Holstein bulls.

ABSTRACT: Our objectives were to evaluate the effects of complete replacement of inorganic salts of trace minerals (STM) with organic trace minerals (OTM) in both pre- and postpartum diets on ovarian dynamics, estrous behavior measured by sensors, preimplantation conceptus development, and reproductive performance in dairy cows. Pregnant cows and heifers (n = 273) were blocked by parity and body condition score and randomly assigned to either STM or OTM diets at 45 ± 3 d before their expected calving. Pre- and postpartum diets were formulated to meet 100% of recommended levels of each trace mineral in both treatments, taking into consideration both basal and supplemental levels. The final target concentrations of Co, Cu, Mn, Se, and Zn were, respectively, 0.25, 13.7, 40.0, 0.3, and 40.0 mg/kg in the prepartum diet, and 0.25, 15.7, 40.0, 0.3, and 63.0 mg/kg in the postpartum diet. The STM group was supplemented with Co, Cu, Mn, and Zn sulfates and sodium selenite, while the OTM group was supplemented with Co, Cu, Mn, and Zn proteinates and selenized yeast. Treatments continued until 156 d in milk (DIM) and were assigned to individual cows using automatic feeding gates. Starting at 21 DIM, ultrasonography examinations of the ovaries were performed weekly to determine the presence of a corpus luteum and postpartum resumption of ovarian cyclicity. Cows were presynchronized with 2 injections of PGF2α at 42 and 56 DIM. Estrous behavior was monitored using electronic activity tags that indirectly measured walking activity. Cows detected in estrus after the second PGF2α were inseminated, and those not detected in estrus by 67 DIM were enrolled in a synchronization program. Cows that returned to estrus after artificial insemination (AI) were reinseminated. Pregnancy diagnosis was performed 33 d after AI, and nonpregnant cows were resynchronized. Transcript expression of interferon-stimulated genes in peripheral blood leukocytes was performed in a subgroup of cows (STM, n = 67; OTM, n = 73) on d 19 after AI. A different subgroup of cows (28 STM, 29 OTM) received uterine flushing 15 d after AI for recovery of conceptuses and uterine fluid for analyses of transcriptomics and metabolomics, respectively. In addition, dominant follicle diameter, luteal size and blood flow, and concentration of progesterone in plasma were measured on d 0, 7, and 15 relative to AI. After flushing, PGF2α was given and the dominant follicle was aspirated 2 d later to measure the concentration of trace minerals by mass spectrometry. Estrous behavior, size of the dominant follicle and corpus luteum, concentration of progesterone, time to pregnancy, and proportion of cows pregnant by 100 d of the breeding period did not differ between treatments. A greater proportion of cows supplemented with OTM had a corpus luteum detected before presynchronization (64.3 vs. 75.2%), and primiparous cows supplemented with OTM tended to resume cyclicity earlier than their STM counterparts. Cows supplemented with OTM had a greater concentration of Cu in follicular fluid than cows supplemented with STM (0.89 vs. 0.77 µg/mL, respectively). In pregnant multiparous cows, expression of receptor transporter protein 4 in peripheral blood leukocytes was 42% greater in the OTM group. Conceptuses of the 2 treatments had 589 differentially expressed transcripts, with many indicating advanced conceptus elongation and greater transcript expression of selenoproteins in the OTM group. In pregnant cows, 24 metabolites were more abundant in the uterine fluid of OTM, including spermidine, sucrose, and cholesterol. In conclusion, replacing STM with OTM caused modest improvements to resumption of ovarian cyclicity and important changes in preimplantation conceptus development, but it did not alter conception risk and pregnancy rate.

ABSTRACT: The objectives of this study were to evaluate how varying colostral insulin concentrations influenced small intestinal development and peripheral metabolism in neonatal Holstein bulls. Insulin was supplemented to approximately 5× (70.0 μg/L; n = 16) or 10× (149.7 μg/L; n = 16) the basal colostrum insulin (12.9 μg/L; BI, n = 16) concentration to maintain equivalent macronutrient intake (crude fat: 4.1 ± 0.06%; crude protein: 11.7 ± 0.05%; and lactose: 1.9 ± 0.01%) among treatments. Colostrum was fed at 2, 14, and 26 h postnatal and blood metabolites and insulin concentration were measured at 0, 30, 60, 90, 120, 180, 240, 360, 480, and 600 min postprandial respective to the first and second colostrum meal. At 30 h postnatal, a subset of calves (n = 8/treatment) were killed to excise the gastrointestinal and visceral tissues. Gastrointestinal and visceral gross morphology and dry matter and small intestinal histomorphology, gene expression, and carbohydrase activity were assessed. Insulin supplementation tended to linearly reduce the glucose clearance rate following the first meal, whereas after the second meal, supplementation linearly increased the rate of glucose absorption and nonesterified fatty acid clearance rate, decreased the time to maximum glucose concentrations, and decreased the time to reach minimum nonesterified fatty acid concentrations. Additionally, insulin clearance rate was linearly increased by insulin supplementation following the second colostrum feeding. However, there were no overall differences between treatments in the concentrations of glucose, nonesterified fatty acids, or insulin in plasma or serum. With respect to macroscopic intestinal development, dry rumen tissue mass linearly decreased when insulin was supplemented in colostrum, and supplementation linearly increased duodenal dry tissue density (g dry matter/cm) while tending to increase duodenal dry tissue weight. Increasing the colostrum insulin concentration improved small intestinal histomorphological development in the distal small intestine, as ileal villi height and mucosal-serosal surface area index were increased by supplementing insulin. Lactase enzymatic activity linearly increased in the proximal jejunum while ileal isomaltase activity linearly decreased with insulin supplementation. These data indicate that changes in colostrum insulin concentrations rapidly affect gastrointestinal growth prioritization and carbohydrase activity. The changes in gastrointestinal ontology result in minor changes in postprandial metabolite availability and clearance.

ABSTRACT: Echinacea purpurea (EP) is an herb that has demonstrated immunostimulatory and anti-inflammatory effects with the potential to improve immunity, health, and performance in animals. The objective of this study was to investigate how supplementing calves with EP affects their blood immunity marker profile, health, intake, and growth. Male Holstein calves (n = 240), sourced from local dairy farms or auction, arrived at a rearing facility between 5 and 14 d of age and were kept in individual pens in 1 of 3 rooms (80/room) for 56 d, and then put into groups for the remaining 21 d of the trial. Calves received milk replacer (MR) 2× per day for 56 d (total = 36 kg of MR) and had ab libitum water and starter access. Within room, calves were randomly assigned to 1 of 3 treatments: (1) control (n = 80), (2) 3g of dried (powder) EP extract per day split over 2 milk feedings from experiment d 14–28 (n = 80), and (3) 3 g of dried (powder) EP extract per day split over 2 milk feedings from experiment d 1–56 (E56; n = 80). The powdered EP treatments were mixed into the liquid MR. On d 1, 14, 28, and 57 rectal temperatures and blood were collected from a subset of calves (n = 117; 39 calves/treatment), and blood serum was assessed for serum total protein (d 1), haptoglobin, white blood cells, and cytokines. Failed transfer of passive immunity was defined as serum total protein

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

ABSTRACT: Our objectives were to evaluate the effect of the form of supplementary trace minerals—inorganic salts (STM: Co, Cu, Mn, and Zn sulfates and Na selenite) or organic (OTM: Co, Cu, Mn, Zn proteinates, and selenized yeast)—fed at 100% of recommended levels in both pre- and postpartum diets on in vitro phagocytic activity of neutrophils, and in vivo IgG responses to an ovalbumin challenge during the transition period. In addition, we investigated the associations of these immunological responses with incidence of postpartum clinical diseases and the dynamic changes of metabolic markers during the transition period. Pregnant heifers and cows (n = 273) were enrolled at 45 ± 3 d before expected calving, blocked by parity and body condition score, and allocated randomly to STM or OTM supplementation. Cows in both treatments were fed the same diet, except for the form of supplementary trace minerals. Automatic feeding gates were used to assign treatments to individual cows. Blood was collected on d −7 ± 3 and 7 ± 3 relative to calving in a subgroup of cows (n = 131 and 133, respectively) to measure phagocytic activity of neutrophils in vitro using flow cytometry. Subcutaneous immunization with 0.5 mg of chicken egg ovalbumin was performed in a subgroup of cows (n = 181) on d −45, −21, and 3 relative to calving. Concentration of anti-ovalbumin IgG in serum was measured by ELISA on d −45, −21, 3, 7, and 21. Trace mineral concentrations in blood were measured by inductively coupled plasma mass spectrometry on d −45, −21, −7, 0, 7, and 21 relative to calving. Selected metabolites were measured on d −21, −10, −3, 0, 3, 7, 10, 14, and 21 relative to calving. Treatment did not affect the percentage of neutrophils performing phagocytosis on d −7 or 7 but the median fluorescence intensity of phagocytosis on d 7 was greater for OTM than STM. We found no differences between treatments in the level of anti-ovalbumin IgG in serum on any of the sampling days. Changes in neutrophil function from prepartum to postpartum were associated with incidence of postpartum clinical disease, postpartum feed intake and milk production, concentrations of Ca, K, Se, Mn, Co, and total protein in serum. Immunoglobulin G responses to ovalbumin injections were not associated with incidence of postpartum clinical disease but were associated with body weight, feed intake, energy balance, and concentrations of nonesterified fatty acids, aspartate aminotransferase, gamma-glutamyl transpeptidase, albumin, Na, P, and Cu in serum. In conclusion, replacement of STM by OTM improved one measure of phagocytic capacity of neutrophils in vitro, which was also greater in cows that did not develop postpartum clinical disease. The associations of innate and acquired immune responses with feed intake, energy balance, and circulating concentrations of key macro and micronutrients reinforce the importance of nutritional management for the health of dairy cows during the transition period.

ABSTRACT: Diarrhea is the primary cause of morbidity and mortality in dairy calves. Many cases of diarrhea in calves are treated with antimicrobials, increasing the risk of antimicrobial resistance, therefore, creating a need for alternative therapies. The objective of this study was to evaluate the effects of feeding spray-dried maternal derived bovine colostrum replacer at the onset of diarrhea on calf growth and duration and severity of the disease in preweaning dairy calves. At a calf-raising facility in southern Ontario, calves were scored for fecal consistency twice daily on a scale of 0 to 3 and enrolled into the trial when they had 2 consecutive fecal scores of 2 (runny or spreads readily) or one fecal score of 3 (liquid consistency, splatters). Calves were then randomly allocated to receive one of the following 3 treatments: (1) control (CON; n = 35): 8 feedings over 4 d of 2.5 L of milk replacer at a concentration of 130 g/L (26% crude protein and 17% fat); (2) short-term colostrum supplementation (STC; n = 35): 4 feedings over the first 2 d of 2.5 L of a mixture of milk replacer at 65 g/L and bovine colostrum replacer at 65 g/L (26% IgG and 14.5% fat) followed by 4 feedings over 2 d of 2.5 L of milk replacer at a concentration of 130 g/L; or (3) long-term colostrum supplementation (LTC; n = 38): 8 feedings over 4 d of 2.5 L of a mixture of milk replacer at 65 g/L and bovine colostrum replacer at 65 g/L. Serum IgG was determined at arrival to the facility and body weight, days to enrollment since facility arrival, and severity of diarrhea were recorded at enrollment. Daily health exams evaluating fecal consistency were performed for 28 consecutive days after enrollment and body weight was measured at d 0, 1, 2, 3, 4, 7, 14, 21, 28, 42 and 56 after enrollment. The median days to resolution of a case of diarrhea was 3.5 d (range: 0.5–11.5 d), 2.75 d (range: 0.5–11.0 d), and 2.75 d (range: 0.5–7.0 d) in CON, STC, and LTC, respectively. Using a Cox proportional hazards model, it was found that calves in LTC group had faster resolution of diarrhea compared with calves in the CON group. In addition, there was an association between both days to enrollment since facility arrival and body weight and resolution of diarrhea, where calves who were at the facility longer before enrollment and heavier at the onset of diarrhea, resolved diarrhea quicker. In addition, calves with a fecal score of 3 at enrollment took longer to resolve their case of diarrhea. With respect to body weight, a linear regression model was built and found that over the 56 d following enrollment calves in the LTC treatment grew 98 g/d more than calves in the CON group. These results suggest that bovine colostrum may be an effective therapy for diarrhea in preweaning calves.

ABSTRACT: Subacute ruminal acidosis (SARA) is assumed to be a common disease in high-yielding dairy cows. Despite this, the epidemiological evidence is limited by the lack of survey data. The prevalence of SARA has mainly been determined by measuring the pH of ruminal fluid collected using rumenocentesis. This may not be sufficiently accurate, because the symptoms of SARA are not solely due to ruminal pH depression, and ruminal pH varies among sites in the rumen, throughout a 24-h period, and among days. The impact of SARA has mainly been studied by conducting SARA challenges in cows, sheep, and goats based on a combination of feed restriction and high-grain feeding. The methodologies of these challenges vary considerably among studies. Variations include differences in the duration and amount of grain feeding, type of grain, amount and duration of feed restriction, number of experimental cows, and sensitivity of cows to SARA challenges. Grain-based SARA challenges affect gut health. These effects include depressing the pH in, and increasing the toxin content of, digesta. They also include altering the taxonomic composition of microbiota, reducing the functionality of the epithelia throughout the gastrointestinal tract (GIT), and a moderate inflammatory response. The effects on the epithelia include a reduction in its barrier function. Effects on microbiota include reductions in their richness and diversity, which may reduce their functionality and reflect dysbiosis. Changes in the taxonomic composition of gut microbiota throughout the GIT are evident at the phylum level, but less evident and more variable at the genus level. Effects at the phylum level include an increase in the Firmicutes to Bacteroidetes ratio. More studies on the effects of a SARA challenge on the functionality of gut microbiota are needed. The inflammatory response resulting from grain-based SARA challenges is innate and moderate and mainly consists of an acute phase response. This response is likely a combination of systemic inflammation and inflammation of the epithelia of the GIT. The systemic inflammation is assumed to be caused by translocation of immunogenic compounds, including bacterial endotoxins and bioamines, through the epithelia into the interior circulation. This translocation is increased by the increase in concentrations of toxins in digesta and a reduction of the barrier function of epithelia. Severe SARA can cause rumenitis, but moderate SARA may activate an immune response in the epithelia of the GIT. Cows grazing highly fermentable pastures with high sugar contents can also have a low ruminal pH indicative of SARA. This is not accompanied by an inflammatory response but may affect milk production and gut microbiota. Grain-based SARA affects several aspects of gut health, but SARA resulting from grazing high-digestible pastures and insufficient coarse fiber less so. We need to determine which method for inducing SARA is the most representative of on-farm conditions.

ABSTRACT: This study was designed to develop a protocol for repeated intramuscular indomethacin injections to replicate leaky-gut-like symptoms in male Holstein calves to model and study the detrimental effects of leaky gut on gut tissue function and inflammatory response. A generalized randomized block design was used to evaluate how repeated indomethacin intramuscular injections affected the development of leaky gut in 18 male Holstein calves. Animals were enrolled at 3 ± 1 d of life, and after 21 d of adaptation, they were randomly assigned to 1 of 3 treatments consisting of intramuscular saline or indomethacin injections every 12 h for 48 h: (1) control (CTL), saline injection, (2) low intramuscular indomethacin (INDO-L) dosed at 1.2 mg/kg of body weight (BW), and (3) high intramuscular indomethacin (INDO-H) dosed at 2.4 mg/kg of BW. During the challenge, milk intake, starter intake, fecal scores, and rectal temperature were measured daily, and BW was measured at the beginning and at the end of the challenge. Plasma samples were used to measure the recovery of markers of intestinal permeability before and after the challenge by dosing lactulose, d-mannitol, and chromium-EDTA. In addition, several cytokines were measured in plasma during the challenge. Calves were dissected at the end of the challenge to obtain tissue and digesta samples from the gastrointestinal tract and liver. No treatment differences were observed for starter and milk intakes, fecal scores, BW, and rectal temperature. The difference in marker concentrations between pre and post challenges was higher for INDO calves compared with CTL calves in the case of lactulose and chromium-EDTA. In addition, chemokine ligand 2 and 4 and IL-6 were higher for INDO-H calves compared with CTL. Both doses of indomethacin resulted in reductions in villus length and surface area in the distal jejunum and ileum and reductions in crypt depth and width in the colon. We showed that repeated indomethacin injections over a 48-h period induced leaky-gut-like symptoms in a region-specific manner, affecting mainly the distal section of the intestine. This outcome was characterized by histomorphological changes in the distal jejunum, ileum, and colon and by increased gut permeability. Interestingly, changes in liver morphology and immune function also occurred, possibly due to the increased translocation of foreign antigens breaching the epithelial cell wall. The leaky gut challenge model described here could be used to improve understanding of the pathogenesis of intestinal disorders in cattle and provide a reliable alternative for testing feed additives with intestinal health benefits.

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

ABSTRACT: Our objectives were to evaluate the effects of complete replacement of supplementary inorganic salts of trace minerals (STM) by organic trace minerals (OTM) in both pre- and postpartum diets on feeding behavior, ruminal fermentation, rumination activity, energy metabolism, and lactation performance in dairy cows. Pregnant cows and heifers (n = 273) were blocked by parity and body condition score and randomly assigned to either STM or OTM diets at 45 ± 3 d before their expected calving date. Both groups received the same diet, except for the source of trace minerals (TM). The STM group was supplemented with Co, Cu, Mn, and Zn sulfates and Na selenite, whereas the OTM group was supplemented with Co, Cu, Mn, and Zn proteinates and selenized yeast. Treatments continued until 156 days in milk and pre- and postpartum diets were formulated to meet 100% of recommended levels of each TM in both treatments, taking into consideration both basal and supplemental levels. Automatic feed bins were used to assign treatments to individual cows and to measure feed intake and feeding behavior. Rumination activity was monitored by sensors attached to a collar from wk −3 to 3 relative to calving. Blood metabolites were evaluated on d −21, −10, −3, 0, 3, 7, 10, 14, 23, and 65 relative to calving. Ruminal fluid samples were collected using an ororuminal sampling device on d −21, 23, and 65 relative to calving, for measurement of ruminal pH and concentration of volatile fatty acids. Cows were milked twice a day and milk components were measured monthly. Cows supplemented with OTM tended to have longer daily feeding time (188 vs. 197 min/d), and greater dry matter intake (DMI; 12.9 vs. 13.3 kg), and had a more positive energy balance (3.6 vs. 4.2 Mcal/d) and shorter rumination time per kg of dry matter (DM; 40.1 vs. 37.5 min/kg of DM) than cows supplemented with STM during the prepartum period. In the postpartum period, OTM increased DMI in multiparous cows (24.1 vs. 24.7 kg/d) but not in primiparous cows (19.1 vs. 18.7 kg/d). The difference in DMI of multiparous cows was more evident in the first 5 wk of lactation, when it averaged 1 kg/d. Milk yield was not affected by treatment in multiparous cows (44.1 vs. 44.2 kg/d); however, primiparous cows supplemented with OTM had lesser yields than primiparous cows supplemented with STM (31.9 vs. 29.8 kg/d). Cows supplemented with OTM had a greater percentage of protein in milk (3.11 vs. 3.17%), reduced concentration of nonesterified fatty acids in serum (0.45 vs. 0.40 mmol/L), and rumination activity (30.1 vs. 27.8 min/kg of DM) than cows supplemented with STM. At the end of the transition period, cows supplemented with OTM had reduced molar proportion of acetate, reduced pH, and tended to have a greater concentration of total volatile fatty acids in ruminal fluid. In conclusion, complete replacement of STM by OTM caused modest changes in rumen fermentation, feeding behavior, energy metabolism, and performance of dairy cows, improving postpartum DMI in multiparous cows and reducing circulating levels of nonesterified fatty acids. The pre-absorptive effects of TM source and the parity specific responses on performance warrant further research.

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

The objective of this cross-sectional, diagnostic accuracy study was to validate a human blood glucose meter (Contour Next One Meter, Ascensia Diabetes Care) for accuracy and precision when measuring blood glucose, and for diagnostic accuracy for hypoglycemic status in dairy calves using whole blood and blood plasma. A total of 49 male dairy calves [body weight (BW): 46.3 ± 0.8 kg] had jugular catheters placed within 75 min after birth. Thereafter, blood was withdrawn from the catheter at specific time points (−10, 10, 20, 30, 45, 60, 90, 120, 180, 240, 360, 480, and 600 min) relative to the first and second colostrum feedings (2 h 15 min and 14 h 5 min postnatal; feeding rate: 7% of BW wt/wt). The reference standard method for plasma glucose concentration was determined colorimetrically and in duplicate using the glucose oxidase-peroxidase reaction. Data were assessed for agreement between the glucose meter and the reference standard using Lin's concordance correlation coefficients (CCC), coefficients of determination (precision), and Bland-Altman plots. In addition, a mixed linear regression model was built using the reference method as the outcome, with the glucose meter and repeated measures of time as the explanatory variables and calf as a random effect. The sensitivity (Se), specificity (Sp), and area under the curve (AUC) for the glucose meter using calf whole-blood and plasma were calculated at a threshold of

ABSTRACT: Calves might experience an upper limit of IgG absorption from colostrum ingestion at birth, but it is not clear whether the total IgG mass fed in the first meal or feeding frequencies can saturate the IgG transport mechanism and therefore limit IgG absorption. The objective of this study was to determine whether different colostrum replacer (CR) feeding frequencies affect serum IgG levels or apparent efficiency of absorption (AEA) in neonatal calves. Male Holstein calves (n = 40) were separated from their dams immediately after parturition and randomly assigned to receive CR [12% of birth body weight (BW)], following either (1) a low-frequency (LF; n = 20) or (2) a high-frequency (HF; n = 20) feeding protocol. Low-frequency calves received 2 CR meals (8% and 4% birth BW within 1 h after birth and 12 h after first CR feeding, respectively), whereas HF calves received 3 CR meals (4% of BW for each meal; within 1 h after birth, 6, and 12 h after first CR feeding). The CR powder fed had a dry matter IgG concentration of 30% and an IgG concentration of 70.5 g/L when reconstituted. All CR was fed via esophageal tube within 1 h after birth. Calves were bottle-fed pasteurized milk (5% birth BW) at 24, 36, and 48 h after the first CR feeding. Blood was collected before first CR feeding and at the following intervals post-CR feeding: every 2 h until 18 h; every 3 h from 18 to 30 h; and every 6 h from 30 to 48 h after the first CR feeding. Serum IgG values at 24 h did not differ between LF and HF (25.79 ± 0.93 and 25.66 ± 0.88 g/L, respectively). In the first meal, calves fed LF ingested a higher total IgG mass than HF (257.98 ± 4.16 g and 126.72 ± 4.05 g, respectively); however, AEA at 24 h did not differ for calves fed HF or LF (27.68 ± 1.16% and 27.63 ± 1.26%, respectively). The IgG area under the curve (AUC) at 24 h was greater for calves fed LF than HF (443.13 ± 15.17 and 379.59 ± 13.99 g of IgG/L × h, respectively). Additionally, AUC at 6 h, 12 h, and 48 h were greater for calves fed LF than HF. These results indicate that, although LF calves had a greater AUC, HF calves were still able to absorb IgG in the second and third meal, allowing HF calves to achieve serum IgG levels similar to those of LF calves at 24 h. In addition, the provision of 3 meals at 70.5 g/L of IgG within the first 12 h of life did not result in added benefits to serum IgG or AEA levels.

ABSTRACT: The specific fatty acid (FA) profile of colostrum may indicate a biological requirement for neonatal calves. The objective of this study was to characterize the FA profile and yields in colostrum, transition milk, and mature milk in primiparous (PP) and multiparous (MP) cows. Colostrum was milked from 10 PP and 10 MP Holstein cows fed the same pre- and postpartum rations. Milkings (M) 2 to 5 and 12 were respectively termed transition and mature milk. Overall, short-chain FA (C4:0 and C6:0) were 61 and 50% lower in colostrum than mature milk, respectively. A parity by milking interaction was also present, with higher C4:0 for PP cows at M2 and for MP cows at M12. Additionally, higher concentrations of C6:0 were present for PP cows at M2 through M4 and for MP cows at M12. Palmitic (C16:0) and myristic (C14:0) acids were 38% and 19% higher in colostrum than mature milk, respectively. However, total saturated FA remained relatively stable. Branched-chain FA were 13% lower in colostrum than mature milk and higher in PP than MP cows throughout the milking period. The proportion of trans-monounsaturated FA (MUFA) was 72% higher in PP cows throughout the milking period, as well as 13% lower in colostrum than mature milk. In contrast, cis-MUFA and total MUFA were not affected by milking nor parity. Linoleic acid (LA) was 25% higher in colostrum than transition and mature milks, but α-linolenic acid (ALA) did not differ. Consequently, the ratio of LA to ALA was 29% higher in colostrum than mature milk and 33% higher in MP cows. Linoleic acid was also 15% higher in MP cows, whereas ALA was 15% higher in PP cows. Conjugated linoleic acid (CLA, cis-9,trans-11) was 2.7-fold higher in PP cows, and no differences between colostrum and mature milk were detected. Overall, polyunsaturated FA (PUFA) from the n-6 and n-3 series were over 40% higher in colostrum compared with transition and mature milk. Milking by parity interactions were present for arachidonic acid (ARA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), docosahexaenoic acid (DHA), and total n-3 PUFA, translating to higher proportions in PP cows in M1 to M3, whereas proportions remained relatively stable throughout the milking period in MP cows. Despite increasing milk yields throughout the subsequent milkings, higher yields of EPA, ARA, DPA, and DHA were present in colostrum than in mature milk. Greater proportions and yields of n-3 and n-6 FA in colostrum may translate to specific requirements for newborn calves. Differences were also observed between PP and MP cows and may reflect different nutrient requirements and partitioning.

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

ABSTRACT: The objective of this study was to investigate the effects of a milk byproduct–based starter pellet, high in lactose, on feed intakes, growth, and rumen development when calves were fed either a high or low level of milk. This was done by using a 2 × 2 factorial design, with 120 Holstein heifer calves randomly assigned to 1 of 2 milk treatments and 1 of 2 feed treatments upon enrollment at birth. Calves were either assigned to a low (LM) or high (HM) level of milk replacer by an automated milk-feeding system for individually-housed calves. The LM treatment calves were offered 6 L/d (150 g/L; 1.2 L, 5×/d), whereas HM treatment calves were offered 15 L/d (150 g/L; 3 L, 5×/d) of milk replacer. Calves were also assigned to a milk byproduct- (whey) based starter feed treatment (WF), or a grain-based starter ration (95% concentrate starter pellet, 5% chopped wheat straw) feed treatment (GF). All calves were offered their treatment feed starting on d 5, with water offered ad libitum from d 0. The WF treatment calves were offered 150 g/d of the milk byproduct–based pellet until they consumed the entirety for 2 out of 3 consecutive days. At this point, WF treatment calves received 150 g of the milk byproduct–based pellet top dressed onto the grain-based ration to allow for ad libitum feed consumption until weaned, when they only received the grain-based ration postweaning (d 64–77). Calves on the GF feed treatment received only the grain-based ration. All calves were gradually weaned from d 43 to 63. By design, calves on the HM treatment consumed greater levels of milk than LM treatment calves for the majority of the days in the preweaning phase (8.9 ± 0.11 vs. 5.5 ± 0.11 L/d; SE). Calves on the GF and WF treatments consumed similar amounts of milk in the preweaning phase (7.2 ± 0.11 vs. 7.2 ± 0.11 L/d); however, there was a milk and feed treatment interaction detected for milk consumption in the weaning phase, with HM-GF calves drinking more milk than the other treatments and HM-WF calves drinking more milk than both LM-GF and LM-WF calves. The WF treatment calves had greater solid feed intake during the preweaning phase than GF treatment calves, with no feed intake differences in the weaning or postweaning phases. The WF calves drank more water in the preweaning phase than GF calves. Average daily gain was similar throughout the study between the GF and WF treatment calves. Rumen physiological measures were all similar by the end of the trial (d 77) regardless of feed or milk treatment, suggesting similar rumen development postweaning. Overall, regardless of the level of milk fed, offering the milk byproduct–based starter pellet may increase preweaning feed intake, along with water consumption. However, both feed and milk treatments had similar performance and rumen development measures by the end of the trial.

Optimizing feeding regimens in early life to maximize lifelong growth and production are essential in the dairy industry. This study investigated the effects of milk replacer (MR) feeding frequency and calf age on behavior, and glucose and insulin kinetics of pre- and post-weaned calves fed an elevated plane of MR. Ten male Holstein calves (42.2±1.8 kg BW) were blocked by BW and randomly assigned to two treatments offering 8 l MR/day (150 g/l) in two (2×; meal size 4 l) or four (4×; meal size 2 l) feedings via an automated calf feeder. Milk replacer was gradually stepped down by 1 l/day during week 8, with calves being weaned by week 9. Water and pelleted calf starter were offered ad libitum. Individual intake of MR and starter were recorded daily, and BW was recorded weekly. The number of visits to the MR feeder (rewarded and unrewarded), and behaviors such as lying, cross-sucking, non-nutritive sucking and occupancy time in the feeder were recorded for individual calves from weeks 4 to 10. Jugular catheters were placed on weeks 4, 7 and 10 to facilitate postprandial blood sampling and glucose tolerance tests. Statistical analysis was conducted using the PROC GLIMMIX procedure (SAS) for behavioral observations, and the MIXED procedure (SAS) with repeated measures for BW, intake, plasma glucose and plasma insulin data. Final BW, starter and MR intake did not differ between treatments. There were no differences in observed calf behaviors; with the exception that 2× calves visited the MR feeder more often (P

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

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

Characterizing ruminal parameters in the context of sampling routine and feed efficiency is fundamental to understand the efficiency of feed utilization in the bovine. Therefore, we evaluated microbial and volatile fatty acid (VFA) profiles, rumen papillae epithelial and stratum corneum thickness and rumen pH (RpH) and temperature (RT) in feedlot cattle. In all, 48 cattle (32 steers plus 16 bulls), fed a high moisture corn and haylage-based ration, underwent a productive performance test to determine residual feed intake (RFI) using feed intake, growth, BW and composition traits. Rumen fluid was collected, then RpH and RT logger were inserted 5.5±1 days before slaughter. At slaughter, the logger was recovered and rumen fluid and rumen tissue were sampled. The relative daily time spent in specific RpH and RT ranges were determined. Polynomial regression analysis was used to characterize RpH and RT circadian patterns. Animals were divided into efficient and inefficient groups based on RFI to compare productive performance and ruminal parameters. Efficient animals consumed 1.8 kg/day less dry matter than inefficient cattle (P⩽0.05) while achieving the same productive performance (P⩾0.10). Ruminal bacteria population was higher (P⩽0.05) (7.6×1011 v. 4.3×1011 copy number of 16S rRNA gene/ml rumen fluid) and methanogen population was lower (P⩽0.05) (2.3×109 v. 4.9×109 copy number of 16S rRNA gene/ml rumen fluid) in efficient compared with inefficient cattle at slaughter with no differences (P⩾0.10) between samples collected on-farm. No differences (P⩾0.10) in rumen fluid VFA were also observed between feed efficiency groups either on-farm or at slaughter. However, increased (P⩽0.05) acetate, and decreased (P⩽0.05) propionate, butyrate, valerate and caproate concentrations were observed at slaughter compared with on-farm. Efficient had increased (P⩽0.05) rumen epithelium thickness (136 v. 126 µm) compared with inefficient cattle. Efficient animals also spent 318% and 93.2% more time (P⩽0.05) in acidotic (4.14% v. 1.30%) (pH⩽5.6) and optimal (5.6