Effects of mixed tocopherols added to milk replacer and calf starter on intake, growth, and indices of stress.

Vitamin E comprises 8 fat-soluble isoforms: α-, ß-, γ-, and δ-tocopherol and α-, ß-, γ-, and δ-tocotrienol. Yet the body preferentially uses α-tocopherol, and only α-tocopherol supplementation can reverse vitamin E deficiency symptoms. However, other isoforms influence many biological functions in the body, including inflammation and stress. Therefore, the study objective was to determine metabolic and performance responses in young calves fed diets containing a constant amount of α-tocopherol and increasing amounts of soybean oil-derived mixed γ- and δ-tocopherols. Holstein calves [n = 48; 2-3 d of age; 40.2 kg of initial body weight (BW), standard error = 0.54] were assigned to receive approximately 0, 5, 10, or 15 mg/kg of BW daily (treatments T0, T1, T2, and T3, respectively) of mixed tocopherols (TMIX) provided in milk replacer (MR) and calf starter. The TMIX liquid contained 86% γδ-tocopherols and 9% α-tocopherol. Milk replacers were formulated to contain approximately 0, 400, 800, or 1,200 mg of TMIX/kg for treatments T0, T1, T2, and T3, respectively. Calf starters were formulated to contain approximately 0, 250, 500, or 750 mg of TMIX/kg for treatments T0, T1, T2, and T3, respectively. Mean consumption of γδ-tocopherols was 0.0, 6.5, 14.3, and 20.5 mg/kg of BW, respectively. Milk replacer contained 24% crude protein (CP) and 20% fat on a dry matter (DM) basis. Calf starters were pelleted and offered for ad libitum consumption from 0 to 56 d. Starters contained 18 to 20% CP and 9 to 12% starch in the DM. On d 28, 4 calves per treatment were randomly selected for slaughter, and necropsy was performed. Samples of liver, duodenum, ileum, and trapezius muscle were collected and stored before analysis for α-, ß-, γ-, and δ-tocopherols and δ-tocotrienol. Data were analyzed using a completely randomized design using mixed model ANOVA with orthogonal polynomials to determine linear and quadratic effects of TMIX. Repeated-measures analyses were performed for data collected over time. Increasing dietary TMIX increased or tended to increase change in hip width at 28 and 56 d, respectively, and improved average daily BW gain and gain-to-feed ratio at 56 d. Increasing TMIX reduced plasma xanthine oxidase at 0 h and tended to reduce concentrations at 24 h following vaccination with 2 commercial vaccines on d 28; however, we detected no effect of TMIX following vaccination on d 56. Concentration of α-tocopherol in skeletal muscle declined quadratically with increasing TMIX, whereas ileal and liver γ-tocopherol increased linearly with increasing TMIX. The number of mucin-2 cells in the ileum increased more than 2-fold in calves fed T3. Addition of mixed tocopherols to diets of young dairy calves improved animal growth and altered indices of antioxidant metabolism.

The effects of feeding mixed tocopherol oil on whole-blood respiratory burst and neutrophil immunometabolic-related gene expression in lactating dairy cows.

The 4 major tocopherol isoforms differ in their biochemical reactivity and cellular effects due to basic chemical structural differences. Alpha-tocopherol has been well studied regarding effects on bovine polymorphonuclear leukocyte (PMN) function and its involvement in respiratory burst. However, no studies to date have identified the effects of supplementing a mixed tocopherol oil (Tmix) particularly enriched in non-α tocopherol isoforms (i.e., γ- and δ-isoforms) on fundamental immunometabolic changes in dairy cows. Therefore, the objectives of this study were to determine whether short-term feeding of vegetable oil-derived Tmix alters specific biomarkers of metabolism, whole-blood leukocyte populations, respiratory burst, immunometabolic-related gene expression of PMN, or gene expression of isolated PMN when challenged with lipopolysaccharides (LPS). Clinically healthy multiparous lactating Holstein cows (n = 12; 179 ± 17 d in milk, 40.65 ± 3.68 kg of milk yield) were fed Tmix (620 g/d) for 7 consecutive days. Jugular blood (EDTA anticoagulant) was collected from all cows on d 0 before treatment initiation and again on d 7 after Tmix feeding. Total stimulated respiratory burst activity (RBA) and leukocyte populations were assessed in whole blood, and tocopherol isoform concentrations, metabolites, and hormones were measured in plasma. For gene expression analysis, isolated PMN from cows before and after Tmix feeding were incubated with LPS at a final concentration of either 0.0 or 1.5 µg/mL. Feeding of Tmix for 7 d increased the concentrations of α- and γ-tocopherol. The Tmix did not alter plasma insulin but decreased cholesterol. The Tmix did not alter whole-blood RBA or the leukocyte populations. The LPS challenge increased the expression of proinflammatory genes TNFA and IL6. However, Tmix treatment did not alter the patterns of LPS-affected expression of genes (e.g., TNFA, ITGB2, PPARA, and RXRA) associated with the immune or metabolic response. In conclusion, short-term feeding of Tmix may have no negative effect on animal health as Tmix increased α- and γ-tocopherol concentrations in blood and did not impair whole-blood RBA or alter leukocyte populations. The data provide further support that the α- and γ-tocopherol isoforms do not interfere with normal immune or metabolic function.

Short communication: Amino acid supplementation and stage of lactation alter apparent utilization of nutrients by blood neutrophils from lactating dairy cows in vitro.

Glutamine is the preferred AA used by polymorphonuclear leukocytes (PMN) during the inflammatory response. However, the effect of other AA on bovine PMN response during inflammation and how this is altered by stage of lactation has not been fully elucidated. The objective of this study was to determine the effect of additional AA supplementation (pool of AA excluding Gln) on AA profiles, gene expression, and inflammatory function of PMN from dairy cows in early and mid lactation in vitro. We used 18 Holstein cows for this study. Polymorphonuclear leukocytes were isolated. Working solutions of AA (0 or 4 mM) and LPS (0 or 50µg/mL) were added to cell populations suspended in RPMI and incubated for 2h at 37°C. We used a subset of samples for gene and protein expression. Concentrations of AA in medium were determined using gas chromatography-mass spectrometry with norleucine as an internal standard. Apparent AA and glucose utilization were calculated by subtracting the concentration after from that of before incubation. Data were analyzed as a randomized block design. Challenge with LPS increased the expression of proinflammatory genes and AA supplementation decreased both the expression of some proinflammatory genes and the media concentrations of tumor necrosis factor-α. Neither stage of lactation, LPS challenge, nor AA supplementation altered the chemotactic or phagocytic abilities of PMN in vitro. Polymorphonuclear leukocytes supplemented with AA had greater concentrations and apparent utilization of most of the supplemented AA, whereas the unsupplemented group had greater apparent utilization of glucose. Alanine was not provided in the media but was present in spent media, and Ile, Gly, and Pro were greater in spent media than in media before incubation indicating synthesis of these AA. Regarding expression of genes involved in nutrient metabolism, the expression of G6PD, coding for the enzyme glucose 6-phosphate dehydrogenase, was increased and that of PDHA1, coding for the enzyme pyruvate dehydrogenase α 1, tended to increase with AA supplementation. Due to the lower concentration of tumor necrosis factor-α in media coupled with a downregulation of several proinflammatory genes, we concluded that AA, rather than Gln, alter the inflammatory response of bovine blood PMN. Independent from Gln, blood PMN from cows in early lactation may use certain AA as their primary carbon source for energy than cows in later lactation. Evaluating cows during the early postpartum period will provide additional information on the effect of stage of lactation and nutrient supplementation on PMN function.

Glucose supplementation has minimal effects on blood neutrophil function and gene expression in vitro.

During early lactation, glucose availability is low and the effect of glucose supply on bovine polymorphonuclear leukocyte (PMNL) function is poorly understood. The objective of this study was to determine the effect of glucose supplementation on the function and transcriptomic inflammatory response of PMNL from cows in early and mid-lactation in vitro. Twenty Holstein cows in early (n=10; days in milk=17±3.1) and mid-lactation (n=10; days in milk=168±14.8) were used for this study. Jugular blood was analyzed for serum concentrations of nonesterified fatty acids, ß-hydroxybutyrate, and glucose. Polymorphonuclear leukocytes were isolated and diluted using RPMI (basal glucose concentration was 7.2 mM) to different concentrations of PMNL/mL for phagocytosis, chemotaxis, gene expression, and medium analyses. Working solutions of glucose (0 or 4 mM of d-glucose) and lipopolysaccharide (0 or 50µg/mL) were added and tubes were incubated for 120 min at 37°C. Media were analyzed for concentrations of glucose and tumor necrosis factor-α (TNF-α). Data were analyzed in a randomized block (stage of lactation) design. Challenge with lipopolysaccharide increased the expression of the genes encoding for nuclear factor kappa B (NFKB1), IL-10 (IL10), IL1B, IL6, IL8, TNF-α (TNFA), glucose transporter 3 (SLC2A3), and the concentration of TNF-α in medium (147.3 vs. 72.5 pg/mL for lipopolysaccharide and control, respectively). Main effect of stage of lactation was minimal where the expression of IL10 increased for cows in early compared with cows in mid-lactation. After lipopolysaccharide challenge, cows in early lactation experienced more marked increases in the expression of IL6, TNFA, and IL8 when compared with cows in mid-lactation. Glucose supplementation had minimal effects on gene expression where glucose supplementation increased the expression of lysozyme (LYZ). Glucose supplementation increased PMNL phagocytosis but did not alter chemotaxis, morphology, or concentration of TNF-α in the medium. Under the conditions of the experiment, stage of lactation had minimal effects on PMNL response to glucose supply where only the expression of NFKB1 and the production of TNF-α were greater for cows in mid-lactation when compared with early lactation. Metabolic profiles for cows in early lactation did not parallel those for cows during the early postpartum period and may partly explain results for this study. Future studies investigating the effect of glucose supply on bovine PMNL function in vivo and how this may be altered by stage of lactation are warranted.

The effect of citrus-derived oil on bovine blood neutrophil function and gene expression in vitro.

Research on the use of natural products to treat or prevent microbial invasion as alternatives to antibiotic use is growing. Polymorphonuclear leukocytes (PMNL) play a vital role with regard to the innate immune response that affects severity or duration of mastitis. To our knowledge, effect of cold-pressed terpeneless Valencia orange oil (TCO) on bovine PMNL function has not been elucidated. Therefore, the objective of this study was to investigate the effect of TCO on bovine blood PMNL chemotaxis and phagocytosis capabilities and the expression of genes involved in inflammatory response in vitro. Polymorphonuclear leukocytes were isolated from jugular blood of 12 Holstein cows in mid-lactation and were incubated with 0.0 or 0.01% TCO for 120min at 37°C and 5% CO2, and phagocytosis (2×10(6) PMNL) and chemotaxis (6×10(6) PMNL) assays were then performed in vitro. For gene expression, RNA was extracted from incubated PMNL (6×10(6) PMNL), and gene expression was analyzed using quantitative PCR. The supernatant was stored at -80°C for analysis of tumor necrosis factor-α. Data were analyzed using a general linear mixed model with cow and treatment (i.e., control or TCO) in the model statement. In vitro supplementation of 0.01% of TCO increased the chemotactic ability to IL-8 by 47%; however, migration of PMNL to complement 5a was not altered. Treatment did not affect the production of tumor necrosis factor-α by PMNL. Expression of proinflammatory genes (i.e., SELL, TLR4, IRAK1, TRAF6, and LYZ) coding for proteins was not altered by incubation of PMNL with TCO. However, downregulation of TLR2 [fold change (FC=treatment/control)=-2.14], NFKBIA (FC=1.82), IL1B (FC=-2.16), TNFA (FC=-9.43), and SOD2 (FC=-1.57) was observed for PMNL incubated with TCO when compared with controls. Interestingly, expression of IL10, a well-known antiinflammatory cytokine, was also downregulated (FC=-3.78), whereas expression of IL8 (FC=1.93), a gene coding for the cytokine IL-8 known for its chemotactic function, tended to be upregulated in PMNL incubated with TCO. Incubation of PMNL with TCO enhanced PMNL chemotaxis in vitro. The expression of genes involved in the inflammatory response was primarily downregulated. Results showed that 0.01% TCO did not impair the function of PMNL in vitro. Future studies investigating the use of TCO as an alternative therapy for treatment of mastitis, including dose and duration, for cows during lactation are warranted.

Acute regulation of IGF-I by alterations in post-exercise macronutrients.

This investigation sought to examine the contributions of exercise and nutrient replenishment on in vivo regulation of the insulin-like growth factor-I (IGF-I) axis components. Eight college-aged males completed three high-intensity interval training (HIIT) protocols followed by three post-exercise nutritional protocols: (1) placebo (EX); (2) carbohydrate only (CHO); and (3) essential amino acid/carbohydrate (EAA/CHO). Samples were analyzed for growth hormone (GH), free IGF-I, IGFBP-1, IGFBP-2, insulin, hematocrit, hemoglobin, serum leucine, matrix metalloproteinase-9 (MMP-9) proteolytic activity, and presence of IGFBP-3 protease activity. No evidence for IGFBP-3 proteolysis was observed. Significant increases in [free IGF-I] and [leucine] were observed in the EAA/CHO group only. Significant differences were noted in [IGFBP-1] and [IGFBP-2] across conditions. Significant increases in [GH] and MMP-9 activity were observed in all groups. These results indicate that post-exercise macronutrient ratio is a determinant of [free IGF-I], [IGFBP-1 and -2] and may play a role in modulating the IGF-I axis in vivo.

Interleukin-1beta and tumor necrosis factor-alpha mediation of endotoxin action on growth hormone.

In humans and sheep, endotoxin (LPS) administration results in increased growth hormone (GH) concentrations. To determine the role of cytokines in the effect of LPS on GH, sheep were challenged with IL-1beta or TNF-alpha. GH data were compared with results with LH, where the major effects of LPS are known to act via the hypothalamus. Intracerebroventricular (icv) administration of IL-1beta or TNF-alpha did not alter plasma concentrations of GH. Endotoxin was then administered intravenously (iv) in combination with icv injection of IL-1 receptor antagonist (IL-1RA), TNF antagonist (sTNF-R1), or saline. Administration of LPS increased GH (P < 0.0001), although coadministration of IL-1ra or sTNF-R1 icv did not alter GH response to LPS. In contrast, plasma concentrations of LH were profoundly inhibited by icv administration of either cytokine (P < 0.03), but the LH response to LPS was not altered by cytokine antagonists. Intravenous administration of either IL-1beta or TNF-alpha increased plasma concentrations of GH (P < 0.0001). Administration of IL-1RA and sTNF-R1 iv prevented LPS-induced increases in GH. Although LH was suppressed by high iv doses of IL-1beta (P = 0.0063), the antagonists did not alter the LH response to LPS. To determine whether LPS might directly activate GH release, confocal microscopy revealed colocalization of CD14, the LPS receptor, with GH and, to a lesser extent, LH and some prolactin (PRL)-containing cells, but not ACTH or TSH. These data are consistent with the effects of LPS on GH secretion originating through peripheral cytokine presentation to the pituitary, as well as a potential to act directly on selective populations of pituitary cells via CD14.

Effect of endotoxin challenge on hepatic 5'-deiodinase activity in cattle.

Thyroid status is compromised in a variety of acute and chronic infections and toxin-mediated disease states. Conversion of thyroxine (T4) into the metabolically active hormone, triiodothyronine (T3), is catalyzed by 5'-deiodinase (5'D). Our objective was to determine the effect of endotoxin (LPS) challenge with and without L-arginine (Arg) infusion on hepatic activity of 5'D and plasma concentrations of T4 and T3. In a 2 x 2 factorial, beef heifers (275-310 kg b.wt.) were fed low (8% CP; 6.5 kg/d) or high (14% CP; 7.2 kg/d) isocaloric protein diets (1.96 Mcal/kg DM) for 10 d before LPS challenge. L-Arginine in saline (0.5 g/kg b.wt.) or saline alone was infused i.v. throughout an 8 hr period starting 2 hr before bolus LPS injection (Escherichia coli, 055: B5; 0.2 microg/kg; i.v.). Blood samples were collected at -2, 0, 3, 6, 12, and 24 hr relative to LPS injection. Liver samples were obtained 20 hr before, and then 6 and 24 hr after LPS challenge using a biopsy needle. Plasma T4 and T3 concentrations were not affected by dietary CP or Arg. Compared with levels at 0 hr, LPS challenge decreased plasma T4 (P < 0.01) and T3 (P < 0.001), respectively, 8.4% and 28.9% at 6 hr and 19.7% and 31.3% at 24 hr. Consistent with these changes, the T3:T4 ratio was lower than that at 0 hr (P < 0.001) 22.0% at 6 hr and 13.5% at 24 hr. Hepatic 5'D activities 20 hr before LPS injection were 2.80 +/- 0.11 nmol I- x hr(-1) x mg protein(-1) and decreased 24 hr after LPS, respectively, 45.4% (P < 0.01) and 17.6% (P < 0.05) in saline- and Arg-infused heifers. The results indicate that mild LPS challenge in cattle inhibits hepatic generation of T3 and decreases plasma concentrations of thyroid hormones. The data also suggest that the impact of LPS on 5'D activity in liver can be altered by Arg supplementation.

Performance and digestibilities of beef cattle fed diets supplemented with either soybean meal or roasted soybeans and implanted with Synovex.

Two 160-d feedlot experiments, each consisting of 20 Angus-Hereford steers (216 +/- 5 kg BW, Exp. 1; 258 +/- 5 kg BW, Exp. 2) and 20 Angus-Hereford heifers (208 +/- 5 kg BW, Exp. 1; 236 +/- 5 kg BW, Exp. 2), were used to investigate the effects of supplementing diets with either roasted soybeans (RSB, roasted at 127 degrees C for 10 min) or soybean meal (SBM) and implanting or not implanting with an estrogenic growth promoter (SYN; Synovex-S, 20 mg of estradiol benzoate plus 200 mg of progesterone or Synovex-H, 20 mg of estradiol benzoate plus 200 mg of testosterone) on performance. The cattle were fed a basal diet of 15% orchardgrass silage, 15% corn silage, and 70% corn-based concentrate. Treatments were 1) no SYN and fed a SBM-supplemented diet, 2) no SYN and fed a RSB-supplemented diet, 3) SYN and SBM, and 4) SYN and RSB. Cattle in the SYN groups were reimplanted at 80 d. Four additional Angus-Hereford steers were used in a digestion and nitrogen balance experiment conducted during the first half of Exp. 1. For the total 160-d feedlot experiments, DMI for RSB compared with SBM was lower (P < .01; 8.5 vs 9.2 kg/d, SEM = .07) and ADG/DMI tended to be higher (P < .10; 165 vs 157 g/kg, SEM = 1.3). Final BW of steers fed RSB was similar (P > .10) to that of steers fed SBM (473 vs 478 kg, SEM = 5.6), as was ADG (1.39 vs 1.43 kg/d, SEM = .02). Dry matter intake for SYN-implanted steers was higher (P < .01) than for steers not implanted (9.2 vs 8.5 kg/d). Likewise, final BW (491 vs 460 kg) and ADG (1.49 vs 1.33 kg/d) were higher (P < .01), and ADG/DMI (166 vs 157 g/kg) tended to be higher (P < .10), for SYN-implanted steers than for steers not implanted. During the more rapid muscle growth period (0 to 80 d), DMI for RSB compared with SBM was lower (P < .01; 7.8 vs 8.6 kg/d, SEM = .07) and ADG/DMI was similar (P > .10; 181 vs 172 g/kg, SEM = 1.8). Dry matter intake for SYN-implanted steers was higher (P < .05) than for steers not implanted (8.4 vs 8.0 kg/d), as was ADG/DMI (P < .01, 182 vs 171 g/kg). During this more rapid growth period, the supplement x implant interaction for ADG was significant (P < .05; 1.35, 1.36, 1.59, and 1.44 kg/d for Treatments 1, 2, 3, and 4, respectively, SEM = .04). There were no differences in digestibilities or N balance. The results suggest that there is no improvement in performance under feedlot conditions when RSB replaces SBM in the diet of beef cattle, and, in young cattle, RSB may reduce the response expected by an estrogenic growth promoter.

Nutritional regulation of plasma tumor necrosis factor-alpha and plasma and urinary nitrite/nitrate responses to endotoxin in cattle.

Effects of dietary protein level with and without L-arginine (Arg) infusion on plasma tumor necrosis factor-alpha (TNF-alpha) response to endotoxin (lipopolysaccharide [LPS]) as well as plasma concentration and urine output of nitrite and nitrate (NOx), the stable end products of nitric oxide radical (NO), were studied in beef heifers (275-310 kg body wt). The animals were fed low- (LP; 7.96%) or high- (HP; 13.94%) protein diets for 10 days before LPS administration (Escherichia coli; 0.2 microgram/kg, iv). L-Arginine in saline (0.5 g/kg body wt) or saline was infused for 8 hr with one-third of total Arg infused before LPS administration. Plasma TNF-alpha concentrations increased in all heifers after LPS injection (peak at 1 hr and return to baseline at 4 hr); however, concentrations were lower in HP- than in LP-fed heifers at 1, 2, and 3 hr. Infusion of Arg did not affect plasma TNF-alpha response to LPS. Plasma NOx concentrations increased in all heifers after LPS challenge; compared with saline, Arg infusion increased the total response (integrated area under concentration curve) in LP- but not in HP-fed heifers. Relative to pretreatment period, the rate of NOx output in urine collected 2-6 hr after LPS administration increased in all heifers regardless of dietary protein level and was further amplified by Arg infusion. The rate of NOx output in urine collected 6-24 hr after LPS challenge was even higher in LP-fed heifers infused with Arg but returned to the basal values in other groups. Activity of hepatic inducible NO synthase was not affected by LPS, Arg, or dietary protein level at the time points studied. The data suggest that dietary protein levels can modulate both TNF-alpha and NO responses to LPS in cattle; high dietary protein intake decreases TNF-alpha response and attenuates the conversion of supplemental Arg to NO.

Supplemental dietary chromium does not influence ACTH, cortisol, or immune responses in young calves inoculated with bovine herpesvirus-1.

Twelve Holstein bull calves (6 to 8 wk of age) were used to determine the influence of supplemental dietary Cr on ACTH, cortisol, and immune responses of calves experimentally inoculated with bovine herpesvirus-1 (BHV-1). Calves supplemented with Cr received 3 mg Cr/d (Chromium, n = 6) of a high-Cr-yeast product. Following 53 d of treatment, all calves were fitted with jugular catheters, and blood samples were collected every 4 h into tubes containing ETDA. Twenty-four hours later, all calves were inoculated intranasally with BHV-1 (1 x 10(7) plaque-forming units in each naris). Serial blood collection continued at 4-h intervals for 6 d. Plasma was harvested, immediately frozen in liquid nitrogen, and stored at -20 degrees C. Individual rectal temperatures and urine samples were collected at the same time each day. Rectal temperatures were elevated (P < .05) on d 2, 3, 4, and 5 but were not affected by Cr treatment. Treatment with Cr did not affect secretion of ACTH, cortisol, or plasma tumor necrosis factor-alpha, although clear circadian variation in ACTH and cortisol occurred. No differences were detected in the concentrations of trace minerals excreted daily in the urine, lymphocyte proliferative response to mitogen stimulation, and neutrophil bactericidal function. The acute phase proteins, ceruloplasmin and fibrinogen, also were not affected by treatment or viral challenge. These data suggest the Cr supplementation using high-Cr yeast (3 mg/d) did not alter stress responses of calves experimentally inoculated with BHV-1.

Dietary fatty acids modulate hormone responses in lactating cows: mechanistic role for 5'-deiodinase activity in tissue.

Supplemental dietary fat provides excess fatty acids (FA), which can alter circulating concentrations of several hormones. To test the effects of fatty acid isomer type and possible sites of regulation, we abomasally infused fat mixtures high in cis-C18:1 FA (iCIS), high in trans-C18:1 FA (iTRS) or no infusion (NI) and performed intravenous arginine (ARG) and intramuscular thyrotropin-releasing hormone (TRH) challenges. The experimental design was a replicated 3 x 3 Latin square. Challenges were conducted on Days 10 (ARG) and 12 (TRH) after initiation of fat infusion on each of three 4-wk experimental periods. Plasma concentrations of IGF-I were lower (P < 0.01) when cows received iCIS or iTRS compared with NI. Plasma insulin concentrations increased with ARG but responses were not affected by FA. Plasma growth hormone (GH) was unchanged after ARG. Peak plasma GH and thyroid-stimulating hormone (TSH) responses to TRH were blunted (P < 0.05 and P < 0.1, respectively), whereas thyroxine (T4) and triiodothyronine (T3) responses were augmented post-TRH (P < 0.01) when cows received either FA isomer. Prolactin responses to TRH were not different between infusion treatments, although basal plasma concentrations before TRH were higher in cows infused with iTRS (P < 0.05). To focus on fat regulation of the thyroid axis, we tested directly in vitro the ability of fatty acids dissolved with sodium taurocholate to affect Type-I 5'-deiodinase (5'D) activity in bovine liver homogenates. Homogenate 5'D was not affected by C2:0-C10:0 fatty acids, but decreased linearly (P < 0.01) with increasing concentrations of C12:0-C16:0 and C18:1 isomers. Cis C18:1 decreased 5'D more than the trans-isomer (P < 0.01), but the difference was only apparent at concentrations greater than 0.25 mM. The data suggest that various aspects of pituitary hormone regulation are differentially affected by FA composition. Fatty acid infusion may accentuate end organ responses in the thyroid axis and decrease IGF-I in the somatotropic axis. The data also suggest that FA isomer may alter patterns of extrathyroidal generation of thyroid hormones via direct influences on 5'D.

Effects of supplemental chromium on production of cytokines by mitogen-stimulated bovine peripheral blood mononuclear cells.

This study determined whether supplementing the diets of dairy cows during the peripartum period with organic trivalent Cr influenced the capacity of their peripheral blood mononuclear cells to produce activation cytokines in response to stimulation with mitogens in vitro. Nine cows were fed 0.5 ppm of Cr/d per cow from 6 wk prepartum to 16 wk postpartum; 10 other periparturient cows served as unsupplemented controls. Mononuclear leukocytes, enriched from peripheral blood during wk 0, 2, 4, and 6 of lactation, were cultured with or without the T-lymphocyte mitogen, concanavalin A. Culture supernatants, harvested at 24, 48, or 72 h, were assayed for interleukin-2, interferon-gamma, and tumor necrosis factor-alpha. The cytokines were barely detectable in the supernatants from the unstimulated cultures, but supernatants from mitogen-stimulated cultures contained higher concentrations of each cytokine. For cows fed Cr, concentrations of all three cytokines in the culture supernatants of the mitogen-stimulated mononuclear cells decreased significantly relative to values for unsupplemented cows, particularly around peak lactation for the 24- and 48-h cultures. Theses results extended our previous observations and supported the hypothesis that organic Cr is immunomodulatory in high producing cows.

Immunomodulatory activity of blood serum from chromium-supplemented periparturient dairy cows.

Our previous research showed enhanced immune responses, including mitogen-induced blastogenesis of peripheral blood mononuclear cells from feedlot calves and periparturient dairy cows supplemented with dietary chromium (Cr). The objective of the present study were to test whether blood sera from Cr-supplemented periparturient cows contained immunomodulatory activity for mitogen-stimulated peripheral blood mononuclear cells and, if so, to determine if this activity was explicable by differences in blood profiles of some glucose-regulating hormones (insulin, cortisol, growth hormone, insulin-like growth factor I, and tumor necrosis factor-alpha) between Cr-supplemented and unsupplemented (control) animals. Blood sera from ten unsupplemented cows and nine Cr-supplemented cows (0.5 ppm day-1) were collected weekly from 2 weeks before to 6 weeks after parturition, and were used to supplement (1, 10, and 20% vol/vol) culture medium supporting concanavalin A (Con A) stimulated mononuclear cells enriched from blood of four nulliparous donor cows. Hormone concentrations were determined using radioimmunoassays. Con A-induced blastogenesis was enhanced when 1, 10, and 20% sera from Cr-supplemented cows was added to the mononuclear cell cultures, and this was particularly evident around parturition. Conversely, peripartum sera from unsupplemented cows depressed Con A-induced blastogenesis. Except for a marginal rise in blood cortisol 2-4 weeks after parturition, no significant effects of Cr supplementation on other hormones (insulin, growth hormone, insulin-like growth factor I, tumor necrosis factor-alpha) were observed. These observations suggest that factors in peripheral blood serum from Cr-supplemented cows, other than absolute concentrations of the glucose-regulating hormones studied, modulate Con A-induced blastogenesis of mononuclear leukocytes.

Nitrogen metabolism of lactating ewes fed rumen-protected methionine and lysine.

Twenty multiparous, crossbred, black-faced ewes and their newborn twin lambs were assigned to one of four dietary treatments in a 2 x 2 factorial experiment to examine the effects of increased supply of CP or a mixture of encapsulated methionine and lysine or both on the performance of ewes and their nursing twin lambs. Ewes were fed ad libitum amounts of either a 10.2% low CP diet or a 16.2% moderate CP diet with or without additional encapsulated amino acids. Nitrogen metabolism trials were conducted simultaneously on both ewes and lambs at wk 2, 4, and 8 of lactation. Analyses were conducted for blood urea N, plasma 3-hydroxybutyrate, lactate, NEFA, insulin, and amino acids (plasma, feed, and milk). Ewe DMI, BW, BW gain, and milk yield were not changed by dietary treatments. Balance of N and N digested were increased by moderate CP treatment. The portion of retained N used for milk synthesis was increased by low CP treatment. Methionine and total branched-chain amino acids were increased by encapsulated amino acids and by protein treatment. Gains in BW and N balance were increased in lambs nursing ewes fed protected amino acids. Increased growth of nursing lambs would be an important beneficial effect of supplementing diets of ewes with encapsulated methionine and lysine.