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The objective of this study was to determine the effect of feeding different combinations of dietary vitamin A supplementation (0 or 110 IU/kg body weight), protein (10.3% or 12.2%), and an ionophore (monensin at 0 or 400 mg/day) on retinoid metabolism and immune function of dairy cows. Eighty multiparous Holstein dairy cows were studied from d −35 to +21 relative to expected parturition in a complete randomized block design with a 2 × 2 × 2 factorial arrangement of treatments. The significance of treatments was declared at p ≤ 0.05. Dairy cows receiving high crude protein (CP) diets with monensin had a greater retinol-binding protein serum concentration than cows receiving high CP diets without monensin (p = 0.04). Animals supplemented with vitamin A showed lower SCC (p = 0.04) and a higher thiobarbituric acid reactive substances concentration (p = 0.06) than cows non-supplemented. Moreover, cows receiving low crude protein diets had a greater haptoglobin concentration (p = 0.01). In addition, cows fed a high crude protein diet had a greater TNF-α expression in peripheral blood mononuclear cells (p = 0.04). Animals fed diets without monensin had a greater serum haptoglobin on day 3 postpartum than those fed monensin (p = 0.01). Moreover, dietary vitamin A increased serum 13-cis retinoic acid postpartum. We conclude that vitamin A, crude protein levels, and monensin fed during the close-up period affect milk somatic cell count, some vitamin statuses, and inflammatory markers during early lactation.

During the periparturient period, elevated circulating nonesterified fatty acids (NEFA) from excessive lipid mobilization affect not only the circulating fatty acid (FA) composition, but also that of the peripheral blood mononuclear cells (PBMC) and polymorphonuclear leukocytes (PMNL). However, the changes to specific lipid fractions remain unknown. We hypothesized that elevated lipid mobilization will alter FA profiles and gene expression of selected proinflammatory mediators in PBMC and PMNL. Starting -28 d relative to expected calving (d 0), treatment cows (n = 18) received a dry cow ration plus an additional 10 kg of corn/head per day, while the control cows (n = 16) received the dry cow ration (no additional corn) supplemented with 400 mg of monensin/head per day to minimize lipid mobilization. Postpartum, treatment cows were feed deprived for 8 h on d +3. For FA analysis, serum was collected on d -28 and -7 relative to expected parturition and d +1, +3, +6, +15, and +21 postpartum, in addition to milk samples. Immune cells, PBMC and PMNL, were isolated on d -28, +3, +12, and +21 for FA analysis and gene expression analysis by reverse-transcription PCR. Serum, PBMC, and PMNL lipids were fractionated into NEFA and phospholipids (PL). The FA composition of milk, serum, PBMC, and PMNL was analyzed by gas chromatography. Data were analyzed as repeated measures ANOVA using mixed model procedures in SAS (9.3) with significance declared at P ≤ 0.05. Several FA varied by treatment and across time and parity. Within the serum PL fraction, FA associated with altered immune function, C18:3n-6, C20:4, C20:5, total n-3, and the ratio of n-6 to n-3 varied significantly by a treatment × parity × time interaction. Overall, FA composition of NEFA and PL fractions from PBMC and PMNL did not significantly reflect FA of serum. Gene expression for IL-1β in PBMC was greater for control, whereas ICAM, IL-1β, IL-6, and TNF-α were greater in primiparous than multiparous cows, without a detectable treatment effect. Whereas gene expression of CASP, IL-8R, and SELL in PMNL changed over time, no treatment effect was detected. In summary, high-energy prepartal diets altered FA profile in serum, milk, PBMC, and PMNL lipids; however, gene expression of selected proinflammatory mediators was not significantly affected.

Our objective was to study the effect of dietary vitamin A and protein on retinoid metabolism and retinoid binding protein (RBP) in vivo. Before the initiation of treatments (confirmed pregnancy), 4 rats were euthanized with blood and tissue samples obtained to establish baseline measures. Female rats, ~21 d before parturition, were fed rations altered in protein (13 or 22%) and vitamin A contents (3 or 5 IU/g). Samples were collected at d −14 (before parturition), d +3, and +10 postpartum via cardiac puncture under anesthesia (n=3–4/ group). Serum RBP showed no significant differences among treatments. Serum and liver 13‐cis retinoic acid (13‐cis), all‐trans retinoic acid (all‐trans) and retinol contents showed no differences, whereas serum all‐trans and retinol concentrations differed at −14 and +3 relative to baseline. The liver 13‐cis and all‐trans were significantly altered on d +3 and +10, and on d −14 relative to baseline, respectively. Treatments and time affected small intestine content of 13‐cis on d −14 and +3, and on d −14 and +10, respectively. The liver mRNA expression of RBP tended to differ for High Protein‐ High Vitamin group than other groups on d −14, whereas the liver TNF‐α gene expression tended to be lower for High Protein‐Low Vitamin group than other groups on d +3. Physiological state and dietary treatments affected the status of retinoids and RBP differently around the time of parturition.