Longitudinal liver proteome profiling in dairy cows during the transition from gestation to lactation: Investigating metabolic adaptations and their interactions with fatty acids supplementation via repeated measurements ANOVA-simultaneous component analysis.

Repeated measurements analysis of variance - simultaneous component analysis (ASCA) has been developed to handle complex longitudinal omics datasets and combine novel information with existing data. Herein, we aimed at applying ASCA to 64 liver proteomes collected at 4-time points (day -21, +1, +28, and + 63 relative to parturition) from 16 Holstein cows treated from 9 wk. antepartum to 9 wk. postpartum (PP) with coconut oil (CTRL) or a mixture of essential fatty acids (EFA) and conjugated linoleic acid (CLA) (EFA + CLA). The ASCA modeled 116, 43, and 97 differentially abundant proteins (DAP) during the transition to lactation, between CTRL and EFA + CLA, and their interaction, respectively. Time-dependent DAP were annotated to pathways related to the metabolism of carbohydrates, FA, and amino acid in the PP period. The DAP between FA and the interaction effect were annotated to the metabolism of xenobiotics by cytochrome P450, drug metabolism - cytochrome P450, retinol metabolism, and steroid hormone biosynthesis. Collectively, ASCA provided novel information on molecular markers of metabolic adaptations and their interactions with EFA + CLA supplementation. Bioinformatics analysis suggested that supplemental EFA + CLA amplified hepatic FA oxidation; cytochrome P450 was enriched to maintain metabolic homeostasis by oxidation/detoxification of endogenous compounds and xenobiotics. SIGNIFICANCE: This report is among the first ones applying repeated measurement analysis of variance-simultaneous component analysis (ASCA) to deal with longitudinal proteomics results. ASCA separately identified differentially abundant proteins (DAP) in 'transition time', 'between fatty acid treatments', and 'their interaction'. We first identified the molecular signature of hepatic metabolic adaptations during postpartum negative energy balance; the enriched pathways were well-known pathways related to mobilizing fatty acids (FA) and amino acids to support continuous energy production through fatty acid oxidation, TCA cycle, and gluconeogenesis. Some of the DAP were not previously reported in transition dairy cows. Secondly, we provide novel information on the mechanisms by which supplemented essential FA and conjugated linoleic acids interact with hepatic metabolism. In this regard, FA amplified hepatic detoxifying and oxidation capacity through ligand activation of nuclear receptors. Finally, we briefly compared the strengths and weaknesses of the ASCA model with PLS-DA and outlined why these methods are complementary.

Maternal Supply of Ruminally-Protected Lysine and Methionine During Close-Up Period Enhances Immunity and Growth Rate of Neonatal Calves.

The objective of this study was to evaluate the effect of supplying ruminally-protected lysine (RPL), methionine (RPM), or the two in combination (RPML) to transition dairy cows on the immunity and performance of their offspring. Eighty heifer calves (n = 20 calves per group) were assigned to four treatments based on their dam diet; basal diet (CON), a basal diet with lysine [RPL, 0.33% of dry matter (DM)], a basal diet with methionine (RPM, 0.16% DM), or with the combination (RPML). Calves were fed colostrum from their dams within 2 h of birth. Calves were then fed milk only (d 2-22), a combination of milk and milk replacer (d 23-25), and milk replacer (d 25-60). Starter feed was fed to the calves twice daily after liquid feeding. Calves blood samples were collected after calving on 0, 12, 24, and 48 h and 5 and 7 d after birth. Data were analyzed by SAS software v9.4. Providing ruminally-protected amino acids (RPAA) to transition cows improved colostrum quality compared to the CON (Brix; P < 0.01). Serum total protein concentrations were higher in calves from supplemented cows than in calves from unsupplemented cows (P < 0.01). Calves born to dams in the RPM, RPL, and RPML groups had higher plasma immunoglobulin G (IgG) concentrations 0, 12, 24, and 48 h and 7 d after birth than those born to dams in the CON group (P < 0.05). The percentage of calves with adequate passive immunity transfer was increased with RPM and RPL or the two in combination (P < 0.01). However, there was no difference in the percentage of calves with adequate passive immunity transfer between the RPM and RPL groups (P = 0.21). Calves from cows that receive supplemental RPAA have a greater average daily gain (ADG) than those born to cows in the CON group (P < 0.01). These results indicate that maternal supplementation with RPM or RPL or the two in combination during the periparturient period could be an alternative strategy to improve the performance of calves, especially in accelerated growth programs in calves.

Hepatic effects of rumen-protected branched-chain amino acids with or without propylene glycol supplementation in dairy cows during early lactation.

Essential amino acids (EAA) are critical for multiple physiological processes. Branched-chain amino acid (BCAA) supplementation provides energy substrates, promotes protein synthesis, and stimulates insulin secretion in rodents and humans. Most dairy cows face a protein and energy deficit during the first weeks postpartum and utilize body reserves to counteract this shortage. The objective was to evaluate the effect of rumen-protected BCAA (RP-BCAA; 375 g of 27% l-leucine, 85 g of 48% l-isoleucine, and 91 g of 67% l-valine) with or without oral propylene glycol (PG) administration on markers of liver health status, concentrations of nonesterified fatty acids (NEFA) and ß-hydroxybutyrate (BHB) in plasma, and liver triglycerides (TG) during the early postpartum period in dairy cows. Multiparous Holstein cows were enrolled in blocks of 3 and randomly assigned to either the control group or 1 of the 2 treatments from calving until 35 d postpartum. The control group (n = 16) received 200 g of dry molasses per cow/d; the RP-BCAA group (n = 14) received RP-BCAA mixed with 200 g of dry molasses per cow/d; the RP-BCAA plus PG (RP-BCAAPG) group (n = 16) received RP-BCAA mixed with 200 g of dry molasses per cow/d, plus 300 mL of PG, once daily from calving until 7 d in milk (DIM). The RP-BCAA and RP-BCAAGP groups, on average (± standard deviation), were predicted to receive a greater supply of metabolizable protein in the form of l-Leu 27.4 ± 3.5 g/d, l-Ile 15.2 ± 1.8 g/d, and l-Val 24.2 ± 2.4 g/d compared with the control cows. Liver biopsies were collected at d 9 ± 4 prepartum and at 5 ± 1 and 21 ± 1 DIM. Blood was sampled 3 times per week from calving until 21 DIM. Milk yield, dry matter intake, NEFA, BHB, EAA blood concentration, serum chemistry, insulin, glucagon, and liver TG and protein abundance of total and phosphorylated branched-chain ketoacid dehydrogenase E1α (p-BCKDH-E1α) were analyzed using repeated measures ANOVA. Cows in the RP-BCAA and RP-BCAAPG groups had lower liver TG and lower activities of aspartate aminotransferase and glutamate dehydrogenase during the first 21 DIM, compared with control. All cows, regardless of treatment, showed an upregulation of p-BCKDH-E1α at d 5 postpartum, compared with levels at 21 d postpartum. Insulin, Met, and Glu blood concentration were greater in RP-BCAA and RP-BCAAPG compared with control during the first 35 DIM. Therefore, the use of RP-BCAA in combination with PG might be a feasible option to reduce hepatic lipidosis in dairy cows during early lactation.

Supplementation with beta-1,3-glucan improves productivity, immunity and antioxidative status in transition Holstein cows.

Dairy cows undergo dramatic physiological changes during the transition from late pregnancy to early lactation, which make them vulnerable to metabolic stress and immune dysfunction. The objective of this study was to evaluate the effects of a commercial beta-1,3-glucan product (Aleta™, containing 50% beta-1,3-glucan) on productivity, immunity and antioxidative status in transition cows. Fifty-four multiparous Holstein cows received a control diet or a diet supplemented with 5 or 10 g of beta-1,3-glucan per cow per day from 21 days before expected calving to 21 days after parturition. Blood samples were collected at day -21, 1, and 21 relative to calving. Colostrum and milk were collected at day 1 and 21 after calving, respectively. Data showed that supplementation with beta-1,3-glucan had no effect on milk composition, but increased milk production. Beta-1,3-glucan treatment also improved the milk quality, as shown by reduced milk somatic cell count and increased immunoglobulin levels in colostrum. Notably, beta-1,3-glucan markedly reduced serum levels of pro-inflammatory cytokines and C-reactive protein, while elevated serum immunoglobulin levels, indicating its immunity enhancement in transition cows. Moreover, beta-1,3-glucan addition reduced the serum malondialdehyde level and enhanced the activities of serum superoxide dismutase and catalase, which enhanced the antioxidative capacity in transition cows. In summary, supplementation with beta-1,3-glucan improves productivity, immunity and antioxidative status in transition dairy cows.

Supplementation effects of pomegranate by-products on oxidative status, metabolic profile, and performance in transition dairy cows.

Recent studies demonstrated a high antioxidant capacity for pomegranate components due to their rich bioactive compounds, such as conjugated fatty acids and phenolics. The objective of the present study was, therefore, to assess whether pomegranate seed or pomegranate seed pulp (peel + seed) supplementation could be effective to improve antioxidant status, and hence metabolic profile and performance in periparturient dairy cows. After a 1-wk pretreatment period, Holstein cows (primiparous n = 12, multiparous n = 18) were assigned to 3 dietary treatments from 25 d before expected calving through 25 d postcalving. The dietary treatments included (1) control (CON); (2) diet supplemented with pomegranate seeds (PS; 400 g/cow per day); and (3) diet supplemented with pomegranate seed pulp (PSP; 400 g of seeds/cow per day + 1200 g of peels/cow per day). Compared with CON, supplementation with either PS or PSP had no effects on dry matter intake, rumen fermentation, and plasma concentrations of cholesterol, total protein, globulin, and aspartate amino transferase, but enhanced plasma total antioxidant activity, and lowered triacylglycerol, free fatty acids (FFA), and ß-hydroxybutyrate at both pre- and postpartum periods. Plasma concentration of glucose, albumin, malondialdehyde (MDA) and blood superoxide dismutase (SOD) activity were not affected by dietary treatments at prepartum, whereas SOD activity increased and glucose, albumin, MDA, and FFA-to-albumin ratio decreased by feeding both by-products at postpartum period. In contrast to PS, supplementing PSP resulted in a greater decrease in plasma glucose and triacylglycerol concentration and higher increase in SOD activity. Energy- and fat-corrected milk yields were higher in cows fed PSP diet compared with those fed CON or PS diets, but content of milk fat, protein, and lactose were similar across the dietary treatments. These findings indicated that dietary pomegranate by-products supplementation, in particular PSP, could improve antioxidant status, which was associated with a decline in lipid oxidation (FFA and ß-hydroxybutyrate) and peroxidation (MDA) and an enhancement in glucose utilization as well as fat-corrected milk yield.

Physiologic, health, and production responses of dairy cows supplemented with an immunomodulatory feed ingredient during the transition period.

This study compared physiological, health, and productive parameters in dairy cows supplemented or not with Omnigen-AF (OMN; Phibro Animal Health, Teaneck, NJ) during the transition period. Thirty-eight nonlactating, multiparous, pregnant Holstein × Gir cows were ranked by body weight (BW) and body condition score (BCS), and assigned to receive (n=19) or not (CON; n=19) OMN at 56 g/cow daily (as-fed basis) beginning 35 d before expected date of calving. Before calving, cows were maintained in single drylot pen with ad libitum access to corn silage, and received (as-fed basis) 3kg/cow daily of a concentrate. After calving, cows were moved to an adjacent drylot pen, milked twice daily, offered (as-fed basis) 35kg/cow daily of corn silage, and individually received a concentrate formulated to meet their nutritional requirements after both milkings. Cows received OMN individually as top-dressing in the morning concentrate feeding. Before calving, cow BW and BCS were recorded weekly and blood samples were collected every 5 d beginning on d -35 relative to expected calving date. After calving and until 46 d in milk, BW and BCS were recorded weekly, individual milk production was recorded, and milk samples were collected daily for total solids and somatic cell count analyses. Blood was sampled daily from 0 to 7 d in milk, every other day from 9 to 21 d in milk, and every 5 d from 26 to 46 d in milk. On 30 and 46 d in milk, cows were evaluated for endometritis via cytobrush technique, based on % of polymorphonuclear (PMN) cells in 100 total cell count (PMN + endometrial cells). On 48.7±1.6 d in milk, 9 cows/treatment received a lipopolysaccharide (LPS) injection (0.25µg/kg of BW), and blood was sampled hourly from -2 to 8 h, at 12-h intervals from 12 to 72 h, and at 24-h intervals form 96 to 120 h relative to LPS administration. No treatment differences were detected on BW, BCS, serum concentrations of cortisol, fatty acids, insulin, glucose, haptoglobin, cortisol, and insulin-like growth factor-I. Cows receiving OMN had greater milk yield (30.3 vs. 27.1kg/d) and percentage of PMN cells in endometrial cell population (12.2 vs. 3.9%) compared with CON cows. After LPS administration, cows receiving OMN had greater mean serum haptoglobin (212 vs. 94 µg/mL), as well as greater serum concentration of tumor necrosis factor α at 1, 2, and 3 h relative to LPS injection compared with CON cows. In conclusion, OMN supplementation during the transition period enhanced innate immunity parameters and increased milk production in dairy cows.