The effect of supplementary selenium source on apparent and true absorption, retention, performance, and selenium status in lactating Holstein cows.

Selenium is an essential trace mineral for dairy cattle and can be provided in the diet in various forms that may differ in bioavailability. The objective of this study was to determine how source of Se affects animal performance, Se status, retention, and apparent and true absorption. Multiparous Holstein cows (n = 24; 597 ± 49 kg body weight) were blocked by days in milk (DIM; 161 ± 18) and randomly assigned to receive 0.3 mg Se/kg of dry matter (100% of NASEM requirements) of either organic Se (ORG; selenized yeast) or inorganic Se (INO; sodium selenite). The Se premix was top-dressed on a common total mixed ration fed daily and mixed into the top 15 cm directly before feeding. Following an 11-wk adaptation period, cows received simultaneous infusions of an intraruminal isotope dose of 77Se in the same chemical form as the premix, and an intravenous dose of 82Se in an inorganic form. Infusions were followed by a 4-d period of blood and rumen fluid sampling, and total collection of feces, urine, and milk. Daily dry matter intake (23 ± 0.6 kg), milk yield (35 ± 1.2 kg), and serum Se (0.11 ± 0.003 µg/g) were not different between treatments during the adaptation period, but milk Se concentrations were greater for ORG compared with INO. Serum 77Se maximum concentration (Cmax) and area under the curve (AUC) were not different between treatments for 72 h following infusion, but rumen fluid 77Se AUC was higher for ORG than INO. Apparent absorption (64 ± 1.4%), and retention (44 ± 1.5%) of the 77Se dose did not differ between treatments. True absorption was calculated using 82Se enrichment in serum and feces and was determined to be 69 ± 1.3% and did not differ between treatments. Fecal excretion of the 77Se dose was not different between treatments (36 ± 1.4%), but ORG had lower urinary excretion and higher milk excretion compared with INO. These results indicate that organic Se resulted in greater Se concentration of milk and lower urinary Se excretion into the environment, but absorption, Se status, and performance of the cow were not affected by Se source at this supplementation level.

Farm-level nutritional factors associated with milk production and milking behavior on Canadian farms with automated milking systems.

The objective of this study was to describe nutritional strategies utilized 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). AMS milking data were collected for 154 herds. For each farm (n = 160), 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 (37.0 ± 0.3 kg/d) was positively associated with the PMR ether extract (EE) concentration (PMR % EE; +0.97 kg/d per percentage point (p.p.) increase) and with farms that fed barley silage as their major forage source on farm (n = 16; +2.18 kg/d) compared with haylage (n = 42), while 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 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 lesser % non-fiber carbohydrates (NFC) of the PMR (-0.016 p.p. per p.p. increase of % NFC). Milk protein content (3.38 ± 0.14%) was positively associated with forage % of the PMR (+0.003 p.p. per p.p. increase of % forage) and total diet % starch (+0.009 p.p. per p.p. increase of % starch), but 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 positively associated with feed push-up frequency (+0.013 milkings/d per additional feed push-up), while being negatively associated with PMR NFC content and % forage of the total ration (-0.017 milkings/d per p.p. increase of % forage). 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 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.

Investigating nutritional strategies during a rest period to improve health, growth, and behavioral outcomes of transported surplus dairy calves.

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 mid-transportation rest period on metabolic and clinical health indicators, growth, and behavioral outcomes after arrival to 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 to 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 under 14 d of age. Calf body weight 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 sampled. 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 sampled. Blood samples were collected at 24 and 48 h and body weight (BW) was recorded at 24 h, 48 h, 72 h, 5 d, 7 d, 14 d, and at 8 wks after arrival to the calf-raising facility. Calves were also health scored daily for 14 d, which 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 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 to 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. Additionally, 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 mid-transportation rest period could minimize fat mobilization and can potentially improve diarrhea and respiratory disease but does not affect growth outcomes after arrival to calf-raisers.

Effects of supplementing colostrum beyond the first day of life on growth and health parameters of preweaning Holstein heifers.

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.

Graduate Student Literature Review: Developmental adaptations of immune function in calves and the influence of the intestinal microbiota in health and disease.

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.

Invited review: "Probiotic" approaches to improving dairy production: Reassessing "magic foo-foo dust".

The gastrointestinal microbial consortium in dairy cattle is critical to determining the energetic status of the dairy cow from birth through her final lactation. The ruminant's microbial community can degrade a wide variety of feedstuffs, which can affect growth, as well as production rate and efficiency on the farm, but can also affect food safety, animal health, and environmental impacts of dairy production. Gut microbial diversity and density are powerful tools that can be harnessed to benefit both producers and consumers. The incentives in the United States to develop Alternatives to Antibiotics for use in food-animal production have been largely driven by the Veterinary Feed Directive and have led to an increased use of probiotic approaches to alter the gastrointestinal microbial community composition, resulting in improved heifer growth, milk production and efficiency, and animal health. However, the efficacy of direct-fed microbials or probiotics in dairy cattle has been highly variable due to specific microbial ecological factors within the host gut and its native microflora. Interactions (both synergistic and antagonistic) between the microbial ecosystem and the host animal physiology (including epithelial cells, immune system, hormones, enzyme activities, and epigenetics) are critical to understanding why some probiotics work but others do not. Increasing availability of next-generation sequencing approaches provides novel insights into how probiotic approaches change the microbial community composition in the gut that can potentially affect animal health (e.g., diarrhea or scours, gut integrity, foodborne pathogens), as well as animal performance (e.g., growth, reproduction, productivity) and fermentation parameters (e.g., pH, short-chain fatty acids, methane production, and microbial profiles) of cattle. However, it remains clear that all direct-fed microbials are not created equal and their efficacy remains highly variable and dependent on stage of production and farm environment. Collectively, data have demonstrated that probiotic effects are not limited to the simple mechanisms that have been traditionally hypothesized, but instead are part of a complex cascade of microbial ecological and host animal physiological effects that ultimately impact dairy production and profitability.

Fat composition of milk replacer influences growth performance, feeding behavior, and plasma fatty acid profile in ad libitum-fed calves.

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.

Developmental adaptations of γδ T cells and B cells in blood and intestinal mucosa from birth until weaning in Holstein bull calves.

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.

Effect of pre- and postpartum supplementation of a pure glycerol product to dairy cows on feeding behavior, lying behavior, and reticulorumen pH.

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.

Postprandial metabolism and gut permeability in calves fed milk replacer with different macronutrient profiles or a whole milk powder.

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.

Fat composition of milk replacer influences postprandial and oxidative metabolisms in dairy calves fed twice daily.

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.

A novel method for isolation and flow cytometry analysis of intraepithelial lymphocytes from colon biopsies.

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.

Colostrum insulin supplementation does not influence immunoglobulin G absorption in neonatal Holstein bulls.

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.

Effect of pre- and postpartum supplementation of a pure glycerol product to dairy cows on feed intake, metabolic markers, and milk yield and components.

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 ß-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 ß-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.

Colostrum insulin supplementation to neonatal Holstein bulls affects small intestinal histomorphology, mRNA expression, and enzymatic activity with minor influences on peripheral metabolism.

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.

Effects of Echinacea purpurea supplementation on markers of immunity, health, intake, and growth of dairy calves.

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 <5.2 g/dL. Calves were health scored 2× per day, receiving fecal and respiratory scores until d 28 and 77, respectively. Calves were weighed on arrival and then weekly until d 77. Milk replacer and feed refusals were recorded. Supplementation of EP was associated with lower haptoglobin levels, segmented neutrophil counts, segmented neutrophil per lymphocyte ratio, respiratory scores in auction derived calves, and higher lymphocyte counts and d 28 rectal temperature. Of calves with heavier arrival body weight, E56 calves had greater postweaning weekly body weight. There was no detected effect of EP supplementation on total white blood cells, band neutrophil, monocyte, and basophil counts, IL-10, IL-6, and TNF-α levels, fecal scores, risk of receiving diarrhea and respiratory treatment, risk of bovine respiratory disease (calves were deemed at risk for bovine respiratory disease if they had at least 1 respiratory score ≥5), risk of mortality, MR and feed intake, average daily gain, and feed conversion ratio. Overall, EP supplementation to dairy calves was associated with immunomodulation and reduced inflammation, evidenced through blood markers, although only few minor health and growth improvements were observed. Benefits were observed particularly when fed across the whole milk feeding period.

Effects of transportation duration on lying behavior in young surplus dairy calves.

Surplus dairy calves are commonly transported long distances from dairy farms to calf-raising facilities and livestock auctions. Current calf transportation research mainly describes physiological changes resulting from transportation. However, few studies have described the effects of transportation on calf behavior. The main objective of this study was to determine the effects of different durations of transportation (6, 12, and 16 h) on lying time and bouts in surplus dairy calves. A secondary objective of this study was to investigate whether calf age affected lying behavior around transportation. Surplus dairy calves (n = 175) were transported in 7 cohorts from 5 commercial dairy farms in Ontario to a single veal facility. On the day of transportation (d 0), calves were randomly assigned to 1 of 3 treatment groups: (1) 6 h (n = 60), (2) 12 h (n = 58), or (3) 16 h (n = 57) of continuous transportation by road. Calf lying and standing behaviors were recorded using HOBO data loggers (Hobo Pendant G Acceleration Data Logger, Onset Computer Corporation). Daily lying time (h/d) and bouts (no./d) were assessed from -1 to 3 d relative to transportation. The total time spent lying during transportation was assessed as the percentage of time lying (min lying/total min on the trailer × 100) from the time each calf was loaded onto the trailer until the time each calf was unloaded at the veal facility (n = 167). On the day of transportation (d 0), calves transported for 12 and 16 h spent less time lying (6 h: 17.1 h/d; 12 h: 15.9 h/d; 16 h: 15.0 h/d) and had more lying bouts (6 h: 21.9 bouts/d; 12 h: 25.8 bouts/d; 16 h: 29.8 bouts/d) compared with those transported for 6 h. On the day after transportation (d 1), calves transported for 16 h spent more time lying down than calves transported for 6 h (19.9 h/d vs. 18.8 h/d, respectively). In addition, during transportation, calves transported for 12 h and 16 h spent 5.8% and 7.6% more time lying down, respectively, than calves transported for 6 h. On each day relative to transportation (d -1 to 3), younger calves (2 to 5 d of age) spent a greater amount of time lying down than older calves (6 to 19 d of age) and, overall, had a greater number of lying bouts. The results of this study suggest that longer durations of transportation influence the lying behavior of surplus dairy calves, resulting in more fatigue during and after the journey and, therefore, potentially have negative implications for calf welfare. Additionally, longer durations of transportation may have greater influence on younger calves than older calves.

Effects of source of supplementary trace minerals in pre- and postpartum diets on reproductive biology and performance in dairy cows.

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.

Effects of replacing inorganic salts of trace minerals with organic trace minerals in the diet of prepartum cows on quality of colostrum and immunity of newborn calves.

Our objectives were to evaluate the impact of supplementary trace mineral (TM) form-inorganic salts (STM; Co, Cu, Mn, Zn sulfates, and Na selenite) or organic (OTM; Co, Cu, Mn, Zn proteinates, and selenized yeast)-in the prepartum diet on quantity and quality of colostrum, passive immunity, antioxidant biomarkers, cytokine responses to lipopolysaccharide (LPS), health, and growth of newborn calves. Pregnant heifers (n = 100) and cows (n = 173) were enrolled at 45 d before calving, blocked by parity and body condition score, and allocated randomly to STM (50 heifers; 86 cows) or OTM (50 heifers; 87 cows) supplementation. Cows in both treatments were fed the same diet, except for the source of supplementary TM. Within 2 h of calving, dams and calves were separated, colostrum was harvested, the yield was measured, and a sample was saved for posterior analyses of colostrum quality. A subgroup of calves (n = 68) had a blood sample collected before colostrum feeding. After colostrum feeding, all samples and data collection were limited to 163 calves (STM = 82; OTM = 81) fed 3 L of good quality (Brix% >22) maternal colostrum via nipple bottle minutes after harvesting. Concentration of IgG in colostrum and serum was determined 24 h after colostrum feeding using radial immunodiffusion. Concentration of TM in colostrum and serum were performed by inductively coupled plasma mass spectrometry. Activity of glutathione peroxidase, ferric reducing ability of plasma, and concentration of superoxide dismutase were evaluated in plasma by colorimetric assays. Ex vivo whole blood stimulation with LPS was performed on d 7 of life to evaluate cytokine responses in a subgroup of 66 calves. Health events were recorded from birth to weaning, and body weight was recorded at birth (all calves) and on d 30 and 60 (heifers only). Continuous variables were analyzed by ANOVA and binary responses were analyzed by logistic regression. Complete replacement of STM by OTM in prepartum diet resulted in greater concentration of Se (461 vs. 543 ± 7 µg/g; ± SEM) but did not alter the concentration or total mass of other TM and IgG in colostrum. Female calves of the OTM group had greater concentration of Se in serum at birth (0.23 vs. 0.37 ± 0.05 µg/mL), were lighter in weight at birth (40.9 vs. 38.8 ± 0.6 kg) and weaning (93.2 vs. 89.7 ± 1.6 kg) than those of the STM group. Maternal treatments did not affect passive immunity or antioxidant biomarkers. On d 7, basal concentrations (log10 of concentration in pg/mL) of IFNγ (0.70 vs. 0.95 ± 0.083) and LPS-stimulated concentrations of CC chemokine ligand 2 (CCL2; 2.45 vs. 2.54 ± 0.026), CC chemokine ligand 3 (CCL3; 2.63 vs. 2.76 ± 0.038), IL-1α (2.32 vs. 2.49 ± 0.054), and IL-1ß (3.62 vs. 3.86 ± 0.067) were greater in OTM than in STM. Supplementation with OTM in pregnant heifers, but not in pregnant cows, reduced the incidence of preweaning health problems in their calves (36.4 vs. 11.5%). Complete replacement of STM by OTM in the prepartum diet did not cause major changes in colostrum quality, passive immunity, and antioxidant capacity, but increased cytokine and chemokine responses to LPS on d 7 of life and benefited preweaning health of calves born to primiparous cows.

Effects of enriching IgG concentration in low- and medium-quality colostrum with colostrum replacer on IgG absorption in newborn Holstein calves.

Ingestion and absorption of greater quantities of IgG are required to increase serum IgG levels in newborn calves. This could be achieved by adding colostrum replacer (CR) to maternal colostrum (MC). The objective of this study was to investigate whether low and high-quality MC can be enriched with bovine dried CR to achieve adequate serum IgG levels. Male Holstein calves (n = 80; 16/treatment) with birth body weights (BW) of 40 to 52 kg were randomly enrolled to be fed 3.8 L of the following combinations: 30 g/L IgG MC (C1), 60 g/L IgG MC (C2), 90 g/L IgG MC (C3), C1 enriched with 551 g of CR (60 g/L; 30-60CR), or C2 enriched with 620 g of CR (90 g/L: 60-90CR). A subset of 40 calves (8/treatment) had a jugular catheter placed and were fed colostrum containing acetaminophen at a dose of 150 mg/kg of metabolic body weight, to estimate abomasal emptying rate per hour (kABh). Baseline blood samples were taken (0 h), followed by sequential samples at 1, 2, 3, 4, 5, 6, 8, 10, 12, 24, 36, and 48 h relative to initial colostrum feeding. Results for all measurements are presented in the following order, unless otherwise stated: C1, C2, C3, 30-60CR, and 60-90CR. Serum IgG levels at 24 h were different among calves fed C1, C2, C3, 30-60CR, and 60-90CR: 11.8, 24.3, 35.7, 19.9, and 26.9 mg/mL ± 1.02 (mean ± SEM), respectively. Serum IgG at 24 h increased when enriching C1 to 30-60CR, but not from C2 to 60-90CR. Similarly, apparent efficiency of absorption (AEA) values for calves fed C1, C2, C3, 30-60CR, and 60-90CR were different: 42.4, 45.1, 43.2, 36.3, and 33.4% ± 1.93, respectively. Enriching C2 to 60-90CR reduced AEA, and enriching C1 to 30-60CR tended to decrease AEA. The kABh values for C1, C2, C3, 30-60CR, and 60-90CR were also different: 0.16, 0.13, 0.11, 0.09, and 0.09 ± 0.005, respectively. Enriching C1 to 30-60CR or C2 to 60-90CR reduced kABh. However, 30-60CR and 60-90CR have similar kABh compared with a reference colostrum meal (90 g/L IgG, C3). Even though kABh was reduced for 30-60CR, results indicate that C1 has the potential to be enriched and achieve acceptable serum IgG levels at 24 h without affecting AEA.