Prepartum nutrition alters fatty acid composition in plasma, adipose tissue, and liver lipids of periparturient dairy cows.

The fatty acyl profile of phospholipids (PL) determines the fluidity of cell membranes and affects cell function. The degree to which long-chain fatty acid (LCFA) composition of PL and triacylglycerols (TG) in liver and total lipids in adipose tissue can be altered by prepartum nutrition in peripartal dairy cows is unclear. Multiparous Holsteins (n = 25) were assigned to 1 of 4 prepartal diets: 1) CA, the control diet fed to meet 120% of energy requirements; 2) CR, a control diet fed to meet 80% of requirements; 3) S, a diet supplemented with mostly saturated free fatty acids (47% 16:0, 36% 18:0, 14% cis-18:1) and fed to meet 120% of requirements; or 4) U, a diet similar to S except that cows were abomasally infused with soybean oil so that the diet plus infused fat would meet 120% of requirements. Diets were fed for 40 d prepartum; all cows received a lactation diet postpartum. Groups CR and U had lower prepartum intakes of dry matter and net energy, but glucose concentrations in plasma were similar among treatments. Cows fed S, U, or CR had greater nonesterified fatty acids in plasma prepartum, but cows fed U had decreased beta-hydroxybutyrate postpartum. Postpartal concentrations of total lipids and glycogen in liver tissue were similar among treatments. Cows in group U had a greater percentage of 18:2 but less 16:0, 18:0, and 20:4 in plasma total lipids than cows fed S. Treatment U increased 18:2 and 18:3 and decreased 18:1 in subcutaneous adipose tissue at 1 d postpartum. Across diets, percentages of 16:0 and trans-18:1 were increased, and 18:0, 20:3, and 20:5 were decreased, in hepatic PL at d 1 postpartum. Significant treatment x time interactions indicated that treatment U increased 18:2 in hepatic PL at the expense of 18:1, 20:3, 20:4, 22:6, and 24:0 on d 1 postpartum, but changes were normalized by d 65 postpartum. The unsaturation index of hepatic PL was lower at d 1 than at d -45 or 65, which implies that hepatic membrane fluidity decreased around parturition. The unsaturation index at d 1 was greater for cows fed S than those fed CA or U. Percentages of 16:0, 18:1, and 22:0 were increased, and 18:0, 20:3, 20:4, 20:5, 24:0, and 26:0 were decreased, in hepatic TG at d 1. Prepartal feed restriction modestly affected tissue LCFA profiles. The LCFA profile of adipose tissue, liver PL, and liver TG can be altered by dietary LCFA supply prepartum; changes in liver are normalized by 65 d postpartum.

Prepartal plane of nutrition, regardless of dietary energy source, affects periparturient metabolism and dry matter intake in Holstein cows.

Previous research in our laboratory showed that dietary fat supplementation during the dry period was associated with decreased peripartum hepatic lipid accumulation. However, fat supplementation decreased dry matter (DM) intake and thereby confounded results. Consequently, 47 Holstein cows with body condition scores (BCS) < or = 3.5 at dry-off were used to determine whether source or amount of energy fed to dry cows was responsible for the decreased hepatic lipid content. Moderate grain- or fat-supplemented diets [1.50 Mcal of net energy for lactation (NE(L))/kg] were fed from dry-off (60 d before expected parturition) to calving at either ad libitum (160% of NE(L) requirement) or restricted (80% of NE(L) requirement) intakes. Postpartum, cows were fed a single lactation diet for ad libitum intake and performance was measured for 105 d. Prepartum intakes of DM and NE(L) were significantly lower for feed-restricted cows as designed. During the first 21 d postpartum, previously restricted cows had higher intakes of DM and NE(L). Body weights and BCS were lower prepartum for restricted cows but groups converged to similar nadirs postpartum. Restricted-fed cows had lower concentrations of glucose and insulin and increased concentrations of NEFA in plasma during the dry period. Peripartum NEFA rose markedly for all treatments but were higher postpartum for cows previously fed ad libitum. Plasma concentrations of NEFA and BHBA remained lower in cows restricted-during the dry period. Postpartum concentrations of total lipid and triglyceride in liver were lower in cows previously feed-restricted. Across dietary treatments, activity of carnitine palmitoyltransferase (CPT) in hepatic mitochondria was lowest at - 21 d, highest at 1 d, and decreased at 21 and 65 d relative to parturition. The activity of CPT at d 1 tended to be higher for previously feed-restricted cows; thereafter, CPT activity declined more rapidly than in cows fed ad libitum. Nutrient intake during the dry period had more pronounced effects on peripartal lipid metabolism and DMI than did composition of the prepartum diet.

Peripartal metabolism and production of holstein cows fed diets supplemented with fat during the dry period.

Previous research from our laboratory demonstrated that cows fed supplemental fat throughout the dry period in an attempt to increase body condition score (BCS) had little hepatic lipid accumulation at d 1 postpartum compared with cows fed an isocaloric high-grain diet or a lower energy control diet. However, results were confounded by lower dry matter intake and loss of BCS by cows fed the fat-supplemented diet. Here, cows were fed a control diet (C) moderately high in nonfiber carbohydrates (NFC) or an isocaloric fat-supplemented, low NFC (F) diet to reassess the effects of supplemental fat throughout the dry period on peripartal lipid accumulation in liver. A more energy-dense, high-NFC diet supplemented with fat (CF) was also fed to test the efficacy of supplemental fat in a diet with similar carbohydrate composition but higher energy density. Intakes of dry matter and net energy for lactation were similar among treatments throughout the experiment, although diet x day interactions during the last 21 d before parturition indicated that cows fed CF decreased intakes more slowly. Cows gained similar amounts of BCS and body weight among diets prepartum, but cows fed C tended to lose more BCS and body weight around parturition. Milk production and milk components did not differ among treatments. Prepartum concentrations of glucose, insulin, total protein, nonesterified fatty acids, and mu-hydroxybutyrate in plasma were similar among treatments. Supplemental fat increased prepartum concentrations of urea and cholesterol in plasma. Postpartum concentrations of metabolites and insulin in plasma were similar among treatments. Concentrations of total lipid and triglyceride in liver increased at parturition, whereas hepatic glycogen concentration decreased, but concentrations were not different among treatments. Supplemental fat fed prepartum did not affect peripartal lipid accumulation in liver tissue and did not benefit postpartum milk production.

Technical note: a technique for multiple liver biopsies in neonatal calves.

Our objective was to develop a rapid and safe liver biopsy technique that could be repeated on multiple occasions in individual neonatal calves. A pilot study was performed to verify the efficacy of sedation and restraint procedures and to evaluate different biopsy instruments. Following the pilot experiment, a biopsy trocar was fabricated and an experiment was conducted using this procedure. Liver biopsies were performed in neonatal calves on d 4, 9, 15, 21, and 28 of life to evaluate the effect of vitamin A intake on liver vitamin A concentrations. On these days, a single injection of ceftiofur sodium was administered i.m. 1 to 2 h prior to the procedure. Calves were lightly sedated with xylazine and placed on a surgical table in left-lateral recumbency. The right caudo-thoracic area was clipped and scrubbed with an iodophor agent. Following administration of a local anesthetic (lidocaine), a small incision was made in the skin between the 12th and 13th ribs approximately 15 cm from the dorsal midline. The biopsy trocar was inserted through the body wall and peritoneum and introduced into the liver parenchyma, and a liver sample was collected. Following the biopsy, the cutaneous incision was sutured and an antiseptic agent was applied to prevent infection. An i.m. injection of an analgesic was administered 1 h following the procedure to alleviate postsurgical discomfort. Most calves were able to stand within 2 h after the biopsy. The entire procedure, which could be performed by a single individual, usually required about 20 min from initial sedation until skin closure. Although liver samples of up to 500 mg were obtained, most samples weighed 75 to 150 mg (wet weight). A total of 156 liver biopsies were performed on 33 calves. Complications due to the biopsy procedure were observed in only two calves. Therefore, this procedure can be useful for studies designed to monitor changes in liver composition or enzyme activities over time.

Partial characterization of a T cell-derived factor that suppresses the initiation of the humoral immune response in vitro.

Specific anamnestic stimulation of spleen cells from mice immunized 7 days earlier with horse erythrocytes (HRBC) generated the release of a soluble factor that was capable of suppressing the initiation of the in vitro primary gammaM immune response to sheep red blood cells (SRBC), as well as to the immunogen that elicited its formation. Moreover, the suppressive macromolecule (mol. wt yields to 34,000), derived from antigen-activated, HRBC-primed T lymphocytes (but not B cells), inhibited the secondary gammaM and gammaG anti-SRBC plaque-forming cell responses of SRBC-primed spleen cells. The active material was resistant to treatment with DNase and RNase, but was inactivated by protease (10 microgram/ml, 30 min) or exposure to mild heat (56 degrees, 30 min). The antibody initiation suppressor factor (AISF) was concentrated and partially purified by gel filtration, followed by poly-acrylamide gel electrophoresis.

Non-specific suppression of the initiation of the immune response to a heterologous immunogen by supernatants from specifically stimulated, primed lymphoid cells.

In cultures of spleen cells from mice immunized with horse erythrocytes (HRBC) 7 days earlier, the simultaneous addition of sheep erythrocytes (SRBC) and the priming antigen on day 0 resulted in the suppression of the anti-SRBC plaque-forming cell (PFC) response by day 5 compared to the response of similar cultures that received only SRBC. The cell-free culture fluid from specifically-stimulated, HRBC-primed cells, but not normal cells, contained a factor that nonspecifically inhibited the anti-SRBC PFC response of cultures of normal spleen cells to which SRBC and diluted supernatant aliquots were added at the beginning of culture. Suppressor activity was not manifest unless active supernatants were added on day 0 of a 5-day culture period. Inhibition of the reference plaque response was not due to cytotoxicity of the active material, decreased immunogenicity of the SRBC, or switchover from IgM to IgG plaque formation. The soluble mediator was released slowly into the culture fluid, with linear kinetics, from specifically activated, primed cells, with maximum suppression obtained with the 120-h supernatant. When active supernatants were fractionated by gel filtration over Sephadex G-150, the inhibitory factor eluted with molecules of about 34,000 mol. wt.