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The majority of Salmonella serovars cause no clinical disease in cattle, while some are associated with severe disease. The objective of the current study was to determine the innate immune responses of bovine peripheral blood leukocytes exposed to Salmonella enterica serovar Dublin (bovine-specific), Salmonella typhimurium (murine adapted, but zoonotic), and Salmonella enteritidis (poultry host-adapted) in 3-week-old calves. All Salmonella exposures increased cell surface CD14 and CD18 regardless of serovar. The greatest CD14 marker mean fluorescence was in monocytes and the greatest mean fluorescent of the marker mean was in neutrophils. Phagocytosis increased with all serovars, but was not different among them. Neutrophils had the greatest marker mean fluorescence for phagocytosis, with all serovars being equal. Oxidative burst increased in all serovars compared to control cells, but were not different among the serovars. Neutrophils and monocytes were similar in the oxidative burst, with limited oxidative burst detected in the primarily lymphocyte population. mRNA expression of TNF-α, IL-8, and IL-12, increased above the control cells whereas none of these serovars affected mRNA expression of TLR4. TNF-α was greatest in S. enterica and S. typhimurium, compared to Salmonella dublin. In contrast, IL-8 was expressed more in S. dublin than S. typhiurium, with S. Enteriditus intermediary. These results show while cell surface markers, phagocytosis, and oxidative burst were largely unaffected by serovar, cytokine and chemokine expression differed among the Salmonella serovars. It appears that internal responses of the cells differ, rather than cell recognition, creating pathogenicity differences among of the serovars, even in the neonate with developing immunity., (Published by Elsevier B.V.)

Chronic stressors are a major health and well-being issue in animals. Immune status of animals under chronic stress is compromised, thus reducing disease resistance and compromising well-being of the animal. The objective of this study was to determine the influence of group size of veal calves on immune status and leukocyte mRNA expression of acute phase cytokines, toll-like receptor 4 (TLR4) and tachykinin 1 (TAC1) over a five-month finishing period. Holstein bull calves (n=168), 44±3 days of age were assigned to one of three treatments; 2, 4, or 8 calves/pen (pen space allowance of 1.82m(2)/calf). Jugular blood samples were collected at the day of grouping and then monthly for 4 months. The differential leukocyte counts were determined and mRNA was extracted from the leukocytes. Reverse transcription-qPCR was used to measure the gene expression of interleukin-1 (IL-1β), IL-1 receptor antagonist (IL-1Ra), tumor necrosis factor (TNF-α), TLR4, and TAC1 in leukocytes. Health was evaluated before grouping and monthly for 4 months. On the 1st month after grouping, veal calves that were housed in groups of 8 have greater expression of IL-1β mRNA than calves housed in groups of 4 or 2 (treatment×month, P=0.04). Also at 1 month, groups of 8 had greater TAC1 expression (P<0.05) than calves housed in groups of 4 or 2. However, the expression of IL-1Ra, TNF-α, and TLR4 were not influenced by group size. In the first month of the trial, calves in groups of 8 coughed more (P<0.05) than calves in groups of 2 and coughed more than calves in groups of 4 and 2 during the 2nd month (treatment×month, P=0.03). Calves housed in groups of 8 tended to have greater neutrophil percentage (P=0.09), neutrophil to lymphocyte ratio (P=0.06), and had lower lymphocyte percentage (P=0.06) than those housed in groups of 4 or 2. In conclusion, the number of veal calves in a group, given the same space during the finishing period did not alter IL-1Ra, TNF-α, and TLR4 mRNA expression. However, housing of calves in groups of 8 was associated with greater expression of IL-1β and TAC1 mRNA in peripheral blood leukocytes, and coughing during the first 2 months after grouping. Therefore, housing of veal calves in larger groups may lead to greater susceptibility to respiratory disease and stress., (Published by Elsevier B.V.)

Feed sorting is a natural behavior of dairy cows that can result in inconsistencies in the nutritive value of a total mixed ration (TMR). The objective of this study was to determine the effects of replacing high-moisture corn with glycerol on feed sorting and the feed intake pattern of transition dairy cows. Multiparous Holstein cows (n=26) were paired by expected calving date, housed in individual tie stalls, and fed diets containing either glycerol or high-moisture corn once daily from d -28 to +56 relative to calving. Glycerol was included at 11.5 and 10.8% of the ration dry matter for the pre- and postpartum diets, respectively. The feed consumption pattern was determined by measuring TMR disappearance during the intervals from 0 to 4 h, 4 to 8 h, 8 to 12 h, and 12 to 24 h relative to feed delivery. Feed sorting was determined on d -16, -9, 9, 16, and 51 relative to calving at 4, 8, 12 and 24 h after feeding. The TMR particle size profile was determined at feed delivery and at 4, 8, 12, and 24 after feed delivery by using the Penn State Particle Separator (Nasco, Fort Atkinson, WI) to yield long (>19 mm), medium (<19 mm, >8 mm), short (<8 mm, >1.18 mm), and fine (<1.18 mm) particles. Overall feed intake did not differ between diets and was 14.7±0.4 and 20.2±0.5 kg/d for the pre- and postpartum intervals, respectively. During the prepartum period, glycerol decreased the amount of feed consumed during the first 4h after feed delivery (7.22 vs. 5.59±0.35 kg; control vs. glycerol, respectively) but increased feed consumed from 12 through 24 h after feed delivery (2.22 vs. 3.82±0.35 kg; control vs. glycerol, respectively). Similar effects on the feed consumption pattern were observed after calving. During the prepartum period, cows fed the control diet sorted against long particles, whereas cows fed glycerol did not sort against long particles (77.2 vs. 101.5±3.50% of expected intake for control vs. glycerol; significant treatment effect). The data indicate that addition of glycerol to the TMR alters the feed consumption pattern to increase feed consumption late in the day at the expense of feed consumed immediately after feeding, and it reduces sorting behavior against long particles. Together, these may reduce diurnal variations in the rumen environment to promote greater rumen health in transition cows., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to determine the effects of maternally supplemented natural- or synthetic-source vitamin E on suckling calf performance and immune response. In a 2-yr study, one hundred fifty-two 2- and 3-yr-old, spring-calving, Angus-cross beef cows were blocked by age, BW, and BCS into 1 of 3 isocaloric, corn-based dietary supplements containing 1) no additional vitamin E (CON), 2) 1,000 IU/d of synthetic-source vitamin E (SYN), or 3) 1,000 IU/d of natural-source vitamin E (NAT). Maternal supplementation began approximately 6 wk prepartum and continued until the breeding season. Colostrum from cows and blood from calves was collected 24 h postpartum for analysis of IgG concentration as an indicator of passive transfer and circulating alpha-tocopherol concentration. At 19 d of age, blood was collected from calves to determine the expression of CD14 and CD18 molecules on leukocytes. At 21 and 35 d of age, humoral immune response was measured by a subcutaneous injection, in the neck, with ovalbumin (20 mg; OVA) and blood samples collected weekly until d 63 of age to determine antibodies produced against OVA. At d 63 of age, calves were administered an intradermal injection of OVA (1 mg) in the neck to assess cell-mediated immunity, which was determined on d 65 of age by measuring nodule size with calipers. Circulating alpha-tocopherol concentrations were increased at both 24 h (P = 0.001) and at the day of initial OVA challenge (P < 0.001) in SYN and NAT compared with CON calves. No differences were detected (P > 0.05) for calf birth BW, ADG, or weaning BW. There were no differences (P > 0.05) in calf serum total IgG or cow colostrum total IgG at 24 h or presence of CD14 and CD18 receptors at d 19 of age. The NAT calves had a greater antigen response to OVA at d 63 than SYN calves (P = 0.01; treatment x day interaction). As an indicator of cell-mediated immunity to OVA, nodule size at 65 d of age was not affected (P = 0.92) by maternal dietary supplementation. In conclusion, calves suckling cows supplemented with natural- and synthetic-source vitamin E had increased circulating concentrations of alpha-tocopherol at 24 h, which appeared to continue throughout maternal supplementation; however, calf immune function and performance were not affected.

The objectives were to ascertain whether a yeast cell-wall derivative that was 1.8% beta-glucan in combination with ascorbyl-2-polyphosphate could improve innate immunity and mediate transportation stress in neonatal calves, and to compare the 1.8% beta-glucan yeast cell-wall derivative with a more purified yeast cell-wall derivative (70% beta-glucan). Treatments were 1) an unsupplemented control (CNT); 2) 113 g of a 1.8% (approximately 2%) beta-glucan derivative of yeast cell walls plus 250 mg of l-ascorbic acid phosphate (BG2); or 3) 150 mg of a purified beta-glucan fraction from yeast cell walls (approximately 70% beta-glucan) plus 250 mg/feeding of l-ascorbic acid phosphate (BG70). Calves (n = 39) were transported for 4 h, placed in outdoor hutches, and randomly assigned to treatments. Treatments (mixed with a milk replacer) were individually fed twice daily for 28 d. Calves were offered calf starter, free choice, throughout the study. Weekly starter intake and BW were measured, and fecal samples were collected for Salmonella Typhimurium and Escherichia coli O157:H7 PCR analysis. Blood was collected immediately before transport (d 0) and on d 3, 7, 10, 14, 21, and 28 after transport. Starter intake and DMI were less (P < 0.05) at d 28 for the BG2 and BG70 treatments compared with the CNT treatment. Hematocrit percentages increased (P = 0.002) throughout the experiment. White blood cell counts (treatment x time interaction, P = 0.066) were less for the calves supplemented with BG70 than for those supplemented with BG2 (P = 0.01) or for CNT calves (P = 0.04) on d 28. Granulocyte counts changed (P = 0.04) throughout the experiment. A trend (P = 0.077) for a treatment x time interaction was detected for peripheral blood mononuclear cell counts (PBMC). Counts of PBMC were greater (P = 0.006) for the BG2 treatment compared with the CNT treatment on d 3. Calves given the BG70 supplement had fewer PBMC than those given the BG2 supplement on d 21 (P = 0.03) and 28 (P = 0.05). Fibrinogen concentrations were affected only by time (P = 0.002). Time effects were detected for phagocytosis (P = 0.005), oxidative burst (P < 0.001), expression of cluster of differentiation 18 (P = 0.001), and increased cluster of differentiation 18 (P = 0.006). Phagocytosis was less (P = 0.05) for calves in the BG70 group than for those in the CNT group. Percentage of calves positive for E. coli O157:H7 was greatest (P

Time spent lying by lactating Holstein-Friesian cows of varying body condition scores (BCS) and milk yield was measured using an animal activity monitor. A 3-week average BCS was calculated for each cow; and in total, 84 cows were selected with 28 cows each among three BCS categories (Thin: BCS<2.75; Moderate: 2.75 > or = BCS<3.25; Heavy: BCS> or = 3.25) and two stage of lactation categories (<150 days in milk or >150 days in milk). Cows were kept in two management systems: parlour/freestall (n=60) or automated milking system/freestall (n=24). Behaviour was recorded for 5.3+/-0.1 d for each cow. Production levels were considered using a 28-d rolling average of daily milk production. Cows that exhibited clinical lameness before or during the observation period were excluded from analyses. For cows exhibiting oestrus, the day prior to, day of, and day following breeding were removed. The final analysis included 77 cows (408 d of observation). A mixed model was fitted to describe average daily hours spent lying. Results demonstrated that lying time increased as days in milk (DIM) increased (P=0.05). Variables that were tested but not significant (P>0.05) were BCS category, parity category (1 or 2) and 28-d rolling average daily milk production. Although a numerical trend for increasing hours spent lying with increasing BCS was observed, after accounting for other factors in the mixed model, BCS did not significantly impact lying time. Continued investigation of these management factors that impact lying time and bouts, using new technologies, more cows, and more herds will help dairy owners better manage facilities and cow movements to optimize this essential behaviour.

This study evaluated the effect of 2 dairy cow housing systems on cow locomotion, immune status, and expression of genes associated with lameness during the dry and periparturient periods. Cows were assigned to freestall housing with either rubber (RUB; n = 13) or concrete (CON; n = 14) at the feed-bunk and alley immediately after their first calving, and managed on this system during all subsequent lactations. At dry off, cows were moved to a straw bedded-pack dry cow pen, and remained there until about 2 d before subsequent calving. To investigate whether greater exposure to RUB or CON increased the differences between cows on each treatment, cows at the end of either their first (n = 16) or second (n = 11) lactations were included in the experiment. Locomotion scores and blood samples were obtained at -60 (beginning of the dry period), -30, 0 (after calving), +10 and +18 d relative to calving. Leukocyte counts were obtained by using an automated cell counter. Phagocytic activity, and cells positive for CD14 and CD18 expression were measured by flow cytometry using labeled microbeads and antibodies. Expression of tachikinin 1(TAC1), histamine receptor 1 (H1), and metalloproteinase (MMP)13 in blood leukocytes was estimated using quantitative real-time PCR. Treatment effects were determined using a repeated measures model. Provision of rubber flooring did not improve dairy cow locomotion during the subsequent study period. However, time relative to calving had an effect on locomotion score and speed, which were worst on d 0, probably because of the discomfort associated with calving. An interaction occurred between treatment and time for neutrophil and lymphocyte counts. The RUB cows had greater neutrophil and lesser lymphocyte numbers postpartum than CON. These cows also had more cells positive for CD14 postpartum compared with prepartum. Moreover, RUB cows showed upregulation of MMP13 and TAC1 compared with CON. These genes are associated with lameness and pain detection respectively. Greater neutrophil to lymphocyte ratios and CD14 expression are associated with physiological stress or with activated immunity. Rubber flooring is associated with an increase in activity and standing. This may have resulted in indications of physiological stress and upregulation of genes associated with lameness and pain for RUB cows. However, this study did not take into account the long-term effects of concrete or rubber flooring; for instance, occurrence of lameness or survivability within the herd.

Transition of primiparous heifers to the milking herd is a period with multiple stressors. The objective of these studies was to determine effects of parlour experience and prepartum milking (pre-milking) on behavioural and physiological indicators of stress after calving. Two experiments were conducted, one was in a free-stall housing confinement system and the second was in a modified grazing system. Forty-eight first-calf heifers were assigned to three treatments: control; experienced heifers taken through the parlour without milking; or pre-milk heifers milked for 3 weeks prior to estimated parturition. Blood was collected within 24 h of parturition and on days 3, 5, 7, 10 and 14 following parturition for cortisol and acute phase protein determination. In the grazing system, 20 heifers were assigned to a prepartum milked or control group as in the confinement system and behaviour observations included days -21, -14, -7, -5, -3 and -1 relative to calving and days 1, 3, 7, 9, 14, and 16 post-calving. Milk production was greatest for prepartum milked heifers in both housing systems. However, somatic cell score was reduced by prepartum milking only in the confinement system. Balking occurred least in parlour-experienced heifers. In confinement housing, shifting while in the parlour was the only behaviour that was greater at first milking in control heifers. Kicking was most frequent for parlour experienced heifers on day 2. Grazing system pre-milked heifers shifted more at their first milking (day -21) than did the controls at their first milking (day 1). Shifting within cow was greatest on day -21 compared with day -5 (P<0.05). Pre-milked heifers shifted more on day 1 post-calving than did the control heifers (P<0.05). These results showed that shifting was the most indicative behaviour of restlessness, was transient, and decreased by day 5 prior to calving. Cortisol and alpha1-acid glycoprotein concentrations were not different; however, haptoglobin increased for all treatments up to and including day 3 and haptoglobin concentrations of pre-milked heifers began to decrease by day 5 post-calving. Pre-milked heifers had lower haptoglobin concentrations than the control heifers and tended to have lower concentrations than experienced heifers on day 10 post partum. By day 14 post partum, all haptoglobin concentrations were <200 microg/ml, but the haptoglobin concentration of control heifers was greater than that of pre-milked and experienced heifers. These results showed that prepartum milking and parlour experience shorten some acute phase protein responses, but minimally affect early parlour behaviours.

Docking the tails of dairy cattle causes mild to moderate behavior changes and physiological indicators of acute pain, but no studies have investigated the possibility that tail docking may lead to chronic pain. In human amputees, an incidence of increased limb surface temperature is associated with phantom limb pain, a central nervous system representation that survives peripheral loss. The objectives of this study were to assess indicators of sensitivity or chronic pain in heifers by using behavioral indicators and thermography. We tested 14 Holstein heifers, 7 docked and 7 intact, from a previous neonatal tail-docking experiment. All 14 animals were videotaped during a test sequence of alternating cold (-9 degrees C), hot (54 degrees C), and neutral packs applied to the underside of the tail. Packs were placed approximately 30.5 cm from the tail head on all animals. A thermal image of the tail was taken using infrared imagery prior to and after temperature sensitivity testing. Docked heifers tended to have greater changes in surface temperatures following the test sequence than did nondocked heifers. In docked heifers, temperatures on the underside of the tail were higher than those at the tip of the tail, both prior to and following the test sequence. Docked heifers also showed substantially higher stomping activity following application of the cold pack. Shifting increased in intact heifers after application of the hot pack, but shifting of the docked heifers did not change. Greater changes were observed in the tail surface temperatures of the docked heifers following temperature manipulation, similar to human amputees who are experiencing phantom limb pain, indicating that similar mechanisms are present in the stump of the docked tail. The behaviors of docked heifers indicated changes in their sensitivity to heat and cold.

Cattle are exposed to growth hormone stimulants and to stressors that cause cortisol release. Both of these hormones affect immune responses which may reduce disease resistance. Toll-like receptors are the pattern recognition molecules of pathogens that are on immune cells. They then orchestrate the induction of the appropriate acute phase cytokines of the early innate response. The objective of this study was to determine changes in toll-like receptors and acute phase cytokines following treatment with a synthetic glucocorticoid (dexamethasone) and growth hormone (GH). Twenty-eight calves were given the control (Cnt), dexamethasone (DEX), GH, or dexamethasone and GH (Both) treatments from 3 until 56 days of age. Blood was collected by jugular venipuncture on days 14, 28, 42, and 56. On day 56, a lung lavage was performed and spleen and thymus tissues collected. Total RNA was extracted from blood leukocytes, lung lavage cells, spleen and thymus cells. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was used to quantify interleukin-1 (IL-1), IL-1 receptor antagonist (IL-1Ra), tumor necrosis factor (TNF)-alpha, toll-like receptor 2 (TLR2), and toll-like receptor 4 (TLR4). Blood leukocytes had a time effect for IL-1Ra (P < 0.01), with a trend for a treatment effect (P = 0.07) and had a treatment by time interaction (P < 0.05). IL-1, TNF, and TLR2 and TLR4 were greatest (P < 0.05) for Cnt only at day 14. IL-1 expression of lung lavage cells was greatest (P < 0.05) for calves on the Both treatment compared to the other three treatments. However, IL-1Ra was not different among the treatments. Toll-like receptor 2 expression was enhanced with Both compared to either DEX (P < 0.05) or GH (P < 0.05) and tended to be greater than Cnt expression (P = 0.07). Expression of TLR4 tended to be reduced by Both compared to Cnt (P = 0.06). Tumor necrosis factor-alpha was greatly enhanced by Both compared to the other three treatments (P < 0.05). Spleen cell tended to have different IL-1 expression between GH and Both (P < 0.10). Interleukin-1 receptor antagonist and TLR2 and TLR4 were not different among treatments. However, TNF-alpha expression was enhanced by the DEX treatment alone compared to the GH treatment (P < 0.05), and tended (P < 0.10) to be greater than Cnt expression. None of the gene expressions were different among treatments for thymus cells. Lung lavage cell expression appears to be most susceptible to these hormones while blood leukocyte expression was only slightly affected, and thymus cells were not affected at all. These data demonstrate that TLR2 and TLR4 and acute phase cytokine expression can be altered by stress and growth hormones, which may decrease resistance of those animals to disease.

Previous work showed that the banding process of docking minimally affected mature cows' behavior and physiology, but cutting off the necrotic tail increased haptoglobin. Additionally the docked cows had more flies on the rear legs and exhibited more fly avoidance behaviors. Because many producers dock young calves while they are in hutches where fly problems are more pronounced, we investigated changes in behavior and physiology of young calves following docking by banding. Twenty calves (3 to 5 wk of age) were assigned to a docked or control group, at each of two locations (Indiana and Wisconsin). After applying a band to dock the tail, calves were tested every 15 min for sensitivity to heat below the band at the Indiana location. Calf behavior was recorded for 2 h postbanding and analyzed continuously for that period. After 3 wk, tails were removed and then 1 wk later, fly counts and fly avoidance behaviors were observed at both locations. Tails were sensitive to heat below the banding site, for 60 to 120 min postbanding (mean 87 min). Banded calves were more active than control calves during the 2 h following banding. Percentage of time spent lying was greater for control calves, and the percentage of time spent walking was greater for docked than control calves. More importantly, movements of the head to touch the tail were increased for banded calves (eight-fold more movements). Fly avoidance behaviors directed toward the rear of the calf were evident at noon or in the afternoon. Ear twitches were more frequent for the docked calves and less frequent in the morning for all calves. Licking was more frequent for the docked calves at 1200 and 1600 h. Tail swings were most frequent at 1200 and more frequent for control calves. Two acute phase proteins, haptoglobin and alpha1 acid-glycoprotein, were not different at any time. In this study, calves that were banded at 3-wk-of-age showed behaviors indicative of discomfort for 2 h, were attacked by more flies, and showed increased fly avoidance behaviors when docked.

Tail docking is an animal well-being issue not only regarding the docking procedures but also because of concerns during fly season. To address the latter question, we selected eight cows that had been tail-docked in a previous experiment and eight nondocked cows matched by stage of lactation. Physiological, immunological, and behavioral measures were used to evaluate the well being of those cows housed in a tie-stall barn during fly season for 5 consecutive days. Behavior was observed for 5-min interval instantaneous scan samples for 1 h each at 0800, 1200, and 1600 h. Flies were counted before behavior observations. Blood samples were taken daily for plasma and leukocyte separation. Cows were scored on d 5 for cleanliness on a five-point scale. Docked cows were cleaner, but fly counts of docked cows were greater for total fly counts and rear leg counts. However, counts were not different on front legs. Time of day was significant, so each time of day was analyzed separately. Docked cows were observed to exhibit fewer tail swings at 0800 h, but docked cows tended to ruminate more at that time. Docked cows tended to stand less at the 1200 h observation. Total fly-avoidance behaviors were greater for all cows at the 1600-h observation. Only tail swings tended to be more frequent with docked cows, but foot stomps occurred only in the docked cows. Lymphocyte phenotypes, acute-phase proteins, and immunoglobulin concentrations did not differ. In conclusion, although docked cows were cleaner, as the fly numbers increase throughout the day, fly-avoidance behaviors also increased and foot stomping appeared as an alternative method for fly avoidance by docked cows.

Behavioral and physiological changes were measured following tail-docking in primiparous heifers. One month before projected first parturition, 21 heifers were assigned to control (nondocked), docked, or docked with lidocaine groups. Heifers were banded to initiate tail-docking and the necrotic tail was removed after 144 h. Physiological, immunological, and behavioral measures were taken for 240 h following banding. Cortisol was not different for control and treated heifers. Haptoglobin increased for docked heifers by 168 h postbanding (24 h postdocking). Alpha1-acid glycoprotein decreased as haptoglobin increased, and alpha1-acid glycoprotein increased until 240 h postbanding. Tumor necrosis factor-alpha increased only with lidocaine and did not show an effect of docking by 240 h postbanding. Lymphocyte phenotyping demonstrated increased CD4+ and CD8+ peripheral blood mononuclear cells for docked plus lidocaine heifers and gammadelta+ cells of those heifers tended to be reduced compared with docked heifers. Eating was the only maintenance behavior affected by banding in both docked groups (increased with banding and decreased with docking). The initial banding procedure did not alter heifer physiology and altered only eating behavior, but the cutting of the tail (docking) increased haptoglobin in response to the tissue damage and returned eating behavior to baseline. The use of lidocaine to anesthetize the tail before banding affected lymphocyte phenotypes and TNF-alpha (banding alone did not alter these parameters).

Two experiments were conducted to examine the effects of the form of alpha-tocopherol or interactions of alpha-tocopherol with vitamin A on its bioavailability. In Experiment 1, Holstein steers were fed a diet that was low in vitamins A and E for 1 mo; then, steers were blocked by body weight (X = 97.5 kg) and assigned randomly to one of four oral treatments: 1) no added vitamins, 2) 442 mg of retinyl acetate, 3) 1342 mg of D-alpha-tocopherol, or 4) 442 mg of retinyl acetate and 1342 mg of D-alpha-tocopherol. Each treatment was given as a pulse dose. Blood was sampled over a 36-h period. Concentrations of plasma retinyl palmitate peaked at 2 to 6 h postsupplementation for all calves and then peaked again at 22 to 28 h for calves receiving vitamin supplements. Concentrations of plasma alpha-tocopherol peaked earliest with D-alpha-tocopherol supplementation alone at 12 to 20 h after supplementation, but simultaneous supplementation with retinyl acetate resulted in lower plasma alpha-tocopherol concentrations. Plasma retinyl palmitate decreased during peak alpha-tocopherol concentrations. In Experiment 2, blood and tissue were analyzed after a single gastric tube administration of a powder (DL-alpha-tocopheryl acetate) or a liquid (D-alpha-tocopherol) form of vitamin E to Holstein calves. Plasma and kidney concentrations of alpha-tocopherol were higher when calves were fed D-alpha-tocopherol than when calves were fed the DL-alpha-tocopherol acetate form. Concentrations in the liver, spleen, adipose tissue, heart, muscle, cellular blood fraction, and gut did not differ between the two forms.

Holstein calves (n = 44) were fed milk replacers from d 3 to 45 with low or high concentrations of vitamin A (7000 or 87,000 IU/kg) and vitamin E (11.2 or 57 IU/kg) to examine the influence of vitamin concentrations on vitamin bioavailability and leukocyte functions. Concentrations of alpha-tocopherol in plasma reflected increased vitamin E but were unaffected by increased vitamin A. Plasma retinol concentrations were greatest with high supplementation of vitamins A and E at wk 3; however, at wk 6, plasma of calves fed high vitamin A and low vitamin E contained the most retinol. Increased supplementation of vitamin A improved fecal consistency compared with that for calves fed the low vitamin A diet at wk 3 and 4. At wk 3, calves that received increased supplementation of vitamins E and A had enhanced neutrophil bactericidal activity compared with that of calves with increased individual vitamins. Lymphocyte DNA synthesis and chemotactic index were unaffected by dietary treatments. Results indicate that increased vitamin A in milk replacer did not affect plasma vitamin E concentrations, benefited fecal scores regardless of vitamin E concentrations, but was only beneficial to bactericidal activity of neutrophils in conjunction with increased vitamin E.

Blood neutrophils and pulmonary alveolar macrophages, isolated from calves at 3 and 6 wk of age, were cultured in medium without added vitamins or supplemented with 100 micrograms/dl of vitamin A, 1000 micrograms/dl of vitamin E, 100 micrograms/dl of vitamin A plus 1000 micrograms/dl of vitamin E, or .25 micrograms/dl of beta-carotene plus 1000 micrograms/dl of vitamin E. Macrophage bactericidal activity improved with supplementation of vitamins A plus E compared with supplementation of beta-carotene plus E or vitamin E at wk 3. Neutrophil bactericidal activity decreased with all vitamin E treatments at wk 3 and with vitamins E or A plus E at wk 6. Neutrophil phagocytosis improved at wk 3 with supplementations of vitamins A, E, and A plus E. The chemotactic index improved with beta-carotene and vitamin E compared with vitamin E alone at wk 3 and at wk 6 with vitamin E compared with vitamin A and control treatments. Retinol content of neutrophils varied at wk 3, but, by wk 6, cells supplemented with vitamins A, E, or A plus E had greater retinol concentrations than control cells. Neutrophil alpha-tocopherol concentrations at wk 3 increased from those of controls with supplementation of vitamin E or beta-carotene and vitamin E, but, at wk 6, vitamin E-supplemented cells were different only from vitamin A-supplemented cells. These data suggest that optimal plasma concentrations of vitamins A and E exist for leukocyte function.

To determine the effect of age at grouping on behavior, health, and production of dairy bull calves, 90 Holstein-Friesian bull calves were housed in individual pens until moved to 1 of 3 treatments. Calves were housed in groups of 3 calves at 3 d old (GH3), 7 d old (GH7), or 14 d old (GH14) until 7 wk of age. Ten groups of 3 calves for each treatment were used, with 5 pens/treatment in each of 2 replications (10 pens/treatment, 3 treatments, 3 calves/treatment; 90 calves total). Direct behavioral observations using instantaneous scan sampling every 10 min were conducted twice per week for 7 wk. At the same times, video data were recorded for continuous observations at feeding time to observe the overall activity of group-housed calves. Hip height, heart girth, and health scores were recorded weekly and body weight was recorded at the start and end of the study. Calves in GH3 spent more time playing and but more time cross-sucking and displacing other calves from milk bottles. Calves engaged in social interaction as early as 3 d of age, and social interactions between 3 to 6 wk of age increased markedly. Calves housed in GH14 vocalized more than did calves in GH7 and GH3. No difference was found between treatments in growth performance. Calf fecal, cough, and nasal and ocular discharge scores, differential leukocyte counts, and plasma cortisol concentrations were not affected by age at grouping. However, during the first week of grouping, when calves were moved from individual pens to group pens, some calves were unable to find their milk bottles and required guidance. In conclusion, these data show no adverse effects on health or performance and some benefits on social behavior for early (d 3) grouping of calves.