Palmitic Acid Supplementation Research Articles

Abstract 3130: Effect Of Fatty Acids And Berberine On Macrophages Lipids Metabolism And Inflammation

Dietary fatty acids play a significant role in cardiometabolic health. Recent research indicates that genetic alterations of unsaturated fatty acid metabolism, linked to modulated macrophage function contribute to the pathogenesis of atherosclerosis. Numerous studies have explored the dietary effects of fatty acids supplementation on macrophage functions. Berberine (BBR) is an isoquinoline alkaloid found in various medicinal herbs. Aims and objectives: The current study is designed to determine the effect of Palmitic acid (PA) and Linoleic acid (LA) supplementation on the macrophage’s lipids metabolism and inflammation. Additionally, this study determines the effect of BBR on the fatty acid’s supplementation on macrophage modulation. Method: Mouse macrophages J774 were cultured using media made of DMEM, 10%FBS and 1% PNS. PA, LA and berberine were dissolved in ethanol separately. Three different concentrations 5ug/ul, 25ug/ul and 50ug/ul of PA and LA were used. Macrophages were first treated with three different concentrations of PA LA each in 6 well plates and then 5uM of berberine was introduced in another set of PA and LA treated cells. Cells and the media were collected after 12hrs and 24hrs for GCMS analysis to determine the uptake of fatty acids and the effect of BBR on the PA and LA metabolism in macrophages. The media and cell samples of only fatty acid treated samples and cells treated with both fatty acids and BBR were processed in a Shimadzu GCMS-QP2010 SE. RNA was isolated from lysed cells and Real Time - qPCR analysis was processed in Bio-Rad RT-PCR to determine the effect of saturated (PA) and polyunsaturated fatty acids on lipids packaging and signaling as well as the resultant inflammation associated with these fatty acids’ uptake and metabolism. Results and Conclusion: BBR differentially inhibited fatty acids uptake by macrophages. The GCMS analysis has shown that BBR aggressively inhibited PA uptake compared to LA uptake by macrophages indicating a selective and preferential mode of action for BBR towards saturated fatty acids linked to atherosclerosis compared to unsaturated fatty acids.

Effects of oleogels (alpha-linolenic acid plus beeswax) extracted supplementation for approaching the therapeutic food ingredient: in vitro model

The dietary fatty acid intake for vascular disease has been highlighted for a decade. This study aimed to determine the effect of palmitic acid and fatty acid-incorporated beeswax (oleogels: OG) supplementation on anti-atherosclerotic effects through vascular smooth muscle cell (VSMC) activity. The A7r5 cells were cultured, cells were treated with three treatments, including 1) control, 2) palmitic acid (PA), and 3) alpha-linolenic acid (ALA) + beeswax, ratio 1:1 (OG) at different concentrations of 0, 25, 50 and 100 μM for 48 hrs. Cell proliferation, cell apoptosis, wound repair, and relative quantity of mRNA level of inflammatory cytokines and angiogenetic transcription factors were determined. The results showed there was a significantly reduced VSMCs proliferation with concentrationdependent (p < 0.05). In the presence of OG at 100 μM, the wound area had healed after 24 hrs (59.2%) compared to PA (28.0%) group treatments. OG and PA supplementation significantly up-regulated eNOS and VEGF but down-regulated the TNF-α, CRP, CD36, iNOS, and NF-κB mRNA expression levels, while PA was a contrasting pattern (p < 0.05). Therefore, supplementation of OG reduced the pro-inflammatory effects of inflammatory cytokines in macrophages and induced VEGF expression in VSMCs, contributing to anti-atherosclerotic effect.

Functional ability of mitochondria and mitochondrial genome polymorphism as factors affecting arrhythmogenesis in chronic coronary artery disease

Aim. To investigate functional state of mitochondria and mitochondrial DNA (mtDNA) polymorphism in coronary artery disease (CAD) patients with life-threatening cardiac rhythm disorders (CRD).Methods. We investigated venous blood samples of 45 patients with uncomplicated CAD and 120 CAD patients with CRD. Oxygen consumption rate of mitochondrias of leukocytes in V3 and V4 states were determined in pyruvate-malate and succinate buffers, as well as in the presence of palmitic acid (PA). In patients with complicated CAD, mtDNA haplogroup and substitutions in gene encoding proteins of the respiratory chain complexes and mitochondrial rRNA were determined. Statistical analysis was performed using Mann-Whitney, Wilcoxon tests and Chi-square test with Yates’ correction.Results. In CAD and CAD with CRD, oxygen consumption rate of intact mitochondria did not different in either pyruvate-malate or succinate buffers. In uncomplicated CAD, PA supplementation increases oxygen consumption rate by mitochondria in both succinate and pyruvate-malate buffers. The majority of patients (41%) with CAD and CRD were carriers of the haplogroup «H» and, in this indicator, the sample did not differ from patients with uncomplicated CAD. However, mtDNA of patients with complicated CAD was characterized by a more frequent combined carriage of two and more missense substitutions in genes of respiratory chain and rRNA.Conclusion. Mitochondria of patients with coronary artery disease and life-threatening cardiac rhythm disorders have reduced functional reserve. The distribution of frequencies of main mtDNA haplogroups of patients with coronary artery disease with life threatening cardiac rhythm disorders corresponds to the population. The mtDNA of such patients is characterized by a high frequency of carriage of combined polymorphisms in gene of electron transport chain proteins and rRNA.

73 Effects of Fatty Acids Sources and Lysophospholipids on Nutrient Digestibility in Lactating Cows

Abstract Fatty acids (FA) are a common supplement for lactating cows to increase energy supply, potentially improving feed efficiency and milk and milk fat yields. However, different FA can cause varying responses probably because of different FA digestibility and interactions with other nutrients (e.g., fibers). Lysophospholipids (LPL) are natural emulsifiers and LPL as a feed additive have shown increases in FA digestibility and nutrient absorption in non-ruminant animals. This study aimed to investigate the effects of FA sources with LPL supplementation on nutrient digestibility in lactating cows. Mid-lactation cows (n = 48) were allocated into 12 blocks, and cows in each block were randomly assigned to the following dietary treatments:1) a diet with a FA supplement [1.8% of dietary dry matter (DM)] mainly containing palmitic acid (PA); 2) the PA diet with LPL (0.003% of dietary DM); 3) a diet with a FA supplement (1.8% of dietary DM) mainly containing stearic acid (SA); and 4) the SA diet with LPL (0.003% of DM). The experiment was conducted for 6 weeks, and spot samples of feces and urine were collected over 3 days (every 3 hours within a 24-hour cycle) during the last week of the experiment to determine nutrient digestibility and utilization. Data were analyzed using the mixed model with FA source, LPL, and their interaction as fixed effects and block as a random effect. The digestibility of DM, organic matter, and crude protein were not affected by the FA source and no interaction between FA and LPL was observed. However, PA had greater neutral detergent fiber (NDF) digestibility (41.4 vs. 38.8%; P = 0.04) than SA with no interaction between the FA source and LPL. In addition, as expected, PA decreased (P < 0.01) the digestibility of 16-carbon FA compared with SA due to the greater C16:0 supply, and SA decreased (P < 0.01) that of 18-carbon FA compared with PA due to the greater C18:0 supply. Supplementation of LPL did not affect digestibility of DM, OM, CP, and NDF, but increased total FA digestibility (64.7 vs. 60.5%; P = 0.02) compared with no LPL. In addition, LPL increased 18-carbon FA digestibility (68.2 vs. 74.1%; P < 0.01) compared with no LPL. An interaction between LPL and FA was observed for 16-carbon digestibility because LPL increased 16-carbon digestibility with SA but not with PA. In conclusion, PA supplementation improved fiber digestibility compared with SA, which could be a factor for increased productivity (e.g., milk fat). Supplementation of LPL was effective in increasing FA digestibility and may have potential to increase dietary energy supply or provide a similar energy supply with less dietary fat content in practice. However, more studies are needed to understand different interactions of LPL with FA sources.

027 In vitro embryo development and extracellular vesicle signaling during bovine oocyte maturation in response to palmitic acid and tumour necrosis factor-α supplementation

027 In vitro embryo development and extracellular vesicle signaling during bovine oocyte maturation in response to palmitic acid and tumour necrosis factor-α supplementation

Nutrient intake, digestibility, and serum metabolites in dairy cows fed diets differing in starch concentration with palmitic acid or stearic acid supplementation postpartum.

The present study determined the effect of supplementing palmitic acid (PA) and stearic acid (SA) on the nutrient intake, digestibility, and serum metabolites of dairy cows fed two different starch levels during the postpartum period. Forty-four multiparous Holstein cows were used in a completed randomized block based on their parity and previous milk yield. Dietary treatments were arranged in a 2 × 2 arrangement with two dietary starch levels (HS: 260g/kg of diet dry matter (DM) vs LS: 210g/kg of diet DM) and two fat supplements rich in PA or SA at 15g/kg of diet DM. Increasing the starch concentration of the postpartum diet improved organic matter (OM), ether extract (EE), crude protein (CP), and starch intake. Moreover, HS diets resulted in higher apparent digestibility of OM and CP but lower starch digestibility than LS diets. Feeding HS diets increased fecal starch output compared with LS diets. There was starch levels and FA supplements interaction for serum albumin and total antioxidant capacity (TAC), with higher concentrations in HSSA and LSPA compared to HSPA and LSSA. Significant correlations between performance and blood metabolites were observed in weeks 3 and 4. In week 3, a negative correlation was observed between serum TAC with milk protein (r = - 0.51) and lactose percentage (r = - 0.49) in the HS diet. However, non-esterified FA was correlated with the fat to protein ratio in the LS diet (r = 0.54). Moreover, in week 4, serum TAC was negatively related to the body condition score of the cows fed LS diet(r= 0.50), while there was no relationship for cows fed HS diets. In conclusion, feeding HS diets to postpartum cows increased nutrient intake and the digestibility of OM and CP compared with LS diets. The addition of SA to the HS diet may be morebeneficial than PAin improving the oxidative status of dairy cows in the postpartum period.

Lysosome Inhibition Reduces Basal and Nutrient-Induced Fat Accumulation in Caenorhabditis elegans.

A long-term energy nutritional imbalance fundamentally causes the development of obesity and associated fat accumulation. Lysosomes, as nutrient-sensing and lipophagy centers, critically control cellular lipid catabolism in response to nutrient deprivation. However, whether lysosome activity is directly involved in nutrient-induced fat accumulation remains unclear. In this study, worm fat accumulation was induced by 1 mM glucose or 0.02 mM palmitic acid supplementation. Along with the elevation of fat accumulation, lysosomal number and acidification were also increased, suggesting that lysosome activity might be correlated with nutrient-induced fat deposition in Caenorhabditis elegans. Furthermore, treatments with the lysosomal inhibitors chloroquine and leupeptin significantly reduced basal and nutrient-induced fat accumulation in C. elegans. The knockdown of hlh-30, which is a critical gene in lysosomal biogenesis, also resulted in worm fat loss. Finally, the mutation of aak-2, daf-15, and rsks-1 showed that mTORC1 (mechanistic target of rapamycin complex-1) signaling mediated the effects of lysosomes on basal and nutrient-induced fat accumulation in C. elegans. Overall, this study reveals the previously undescribed role of lysosomes in overnutrition sensing, suggesting a new strategy for controlling body fat accumulation.

O144 Effect of dietary palmitic acid supplementation and increased milking frequency on dairy cow performance and butter properties

O144 Effect of dietary palmitic acid supplementation and increased milking frequency on dairy cow performance and butter properties

Carotenoid Biosynthesis: Genome-Wide Profiling, Pathway Identification in Rhodotorula glutinis X-20, and High-Level Production.

Rhodotorula glutinis, as a member of the family Sporidiobolaceae, is of great value in the field of biotechnology. However, the evolutionary relationship of R. glutinis X-20 with Rhodosporidiobolus, Sporobolomyces, and Rhodotorula are not well understood, and its metabolic pathways such as carotenoid biosynthesis are not well resolved. Here, genome sequencing and comparative genome techniques were employed to improve the understanding of R. glutinis X-20. Phytoene desaturase (crtI) and 15-cis-phytoene synthase/lycopene beta-cyclase (crtYB), key enzymes in carotenoid pathway from R. glutinis X-20 were more efficiently expressed in S. cerevisiae INVSc1 than in S. cerevisiae CEN.PK2-1C. High yielding engineered strains were obtained by using synthetic biology technology constructing carotenoid pathway in S. cerevisiae and optimizing the precursor supply after fed-batch fermentation with palmitic acid supplementation. Genome sequencing analysis and metabolite identification has enhanced the understanding of evolutionary relationships and metabolic pathways in R. glutinis X-20, while heterologous construction of carotenoid pathway has facilitated its industrial application.

Effect of dietary supplementation with fatty acids on growth, survival, and fatty acid patterns in Procambarus clarkii and Procambarus virginalis: the first comparison of two invasive crayfish species

The red swamp crayfish, Procambarus clarkii, and the marbled crayfish, Procambarus virginalis, are invasive species expanding their ranges on a global scale. Their spread is favored by the fact that these crayfish are also aquaculture species of great economic importance (P. clarkii) or promising prospects in this regard (P. virginalis). This study represents the first attempt to assess the impact of diet supplementation with fatty acids on growth, survival, and fatty acid content in these crayfish species, as well as the first record of fatty acid patterns in the obligate parthenogenetic marbled crayfish. Our results showed that supplementation with saturated palmitic acid increased P. clarkii survival, which has not previously been reported for crustaceans. This positive effect on the part of palmitic acid supplementation was not observed for P. virginalis individuals. Marbled crayfish had significantly higher survival rates than red swamp crayfish regardless of the diet used. The growth of both species was not affected by fatty acid supplementation. Our study indicated that young marbled crayfish had a poorer fatty acid profile than juvenile conspecifics and red swamp crayfish. Fatty acid supplementation and type of diet significantly affected the fatty acid composition in the crayfish tissues. There was an increase in unsaturated α-linolenic acid for P. clarkii and palmitic acid for P. virginalis. Palmitic acid quantitatively dominated in both species regardless of the experimental treatment, with the exception of juvenile P. virginalis, which was fed an oligochaete diet rich in EPA.

Feeding diets varying in starch concentration supplemented with palmitic acid or stearic acid: Effects on performance, milk fatty acid profile, and metabolic parameters of postpartum dairy cows

The objective of this study was to investigate the effects of dietary fat supplement rich in palmitic acid or stearic acid on milk yield and composition, fatty acid profile, energy partitioning, and metabolic parameters of dairy cows fed low or high starch diets from d 1 to 28 of the postpartum period. Forty-four multiparous Holstein cows were used in a complete randomized block (based on parity, and previous 305-d milk yield); 2 × 2 arrangement with two dietary starch levels [high starch (HS); 260 g/kg of diet dry matter (DM) versus low starch (LS); 210 g/kg of diet DM] with either palmitic acid (PA) or stearic acid (SA) supplementation at the level of 15 g/kg of diet DM. A basal close-up diet (6.19 MJ/kg of diet DM, NEL; 200 g/kg starch) was fed to animals prior to calving. Regardless of the fatty acid (FA) type, cows fed HS diets had a higher dry matter and NEL intake, milk protein and lactose content, and a more positive energy balance than cows fed LS diets during the postpartum period. Compared with HS diets, LS diets reduced energy toward body reserves and maintenance as a fraction of the NEL intake. Although there was no main effect of PA or SA supplement on intake and energy balance, the effect of fat supplementation depended on dietary starch levels for milk yields and milk energy as a fraction of the NEL intake. Stearic acid increased milk yield in HS diets while PA tended to increase yield in LS diets. Supplementation of PA partitioned less energy toward milk (and more energy for maintenance) in HS diets than LS diets. Moreover, the C16:0 FA concentration in milk fat was higher, and the polyunsaturated FA (PUFA) and C18:1 cis-9 FA were lower in PA supplemented cows fed HS diets but not cows fed LS diets. High starch rations increased de novo FA concentrations in milk and decreased C18:0 and preformed FA concentrations compared with LS diets. Also, HS diets tended to have higher levels of insulin and lower levels of beta-hydroxybutyrate (BHBA) than LS diets. In conclusion, the HS diet was more beneficial to improving the energy balance (EB) of dairy cows than LS diets during the early postpartum period. Also, interactions between dietary starch levels and FA supplement for the variables measured in the current study (e.g. milk yield or FA profile) indicate that the response to fat supplements rich in specific FA differed on the basis of dietary starch concentrations.

Supplementation of Prill Fat Derived from Palm Oil on Nutrient Digestibility and Dairy Cow Performance

Prill fat is a hydrogenated vegetable oil which contains more than 85% palmitic acid with a high melting point. Due to this reason, it does not melt at rumen temperature and bypass rumen degradation. This study was aimed to determine the optimum level of prill fat (86% palmitic acid) supplementation in dairy ration on fermentation characteristics, feed digestibility, milk production, milk components and milk fatty acid profiles. In vitro analysis used Randomized Block Design with 4 treatments and 4 replications and the in vivo analysis using T-Test. The in vitro result showed prill fat supplementation was significantly affected the total Volatile Fatty Acids (VFA) concentration (p<0.05). The addition of prill fat significantly decreased dry matter and organic matter digestibility (p<0.01), Neutral Detergent Fiber (NDF) and Acid Detergent Fiber (ADF) digestibility (p<0.05), total protozoa biomass (p<0.01) and population of cellulolytic bacteria (p<0.05). Furthermore, supplementation of prill fat in the level of 2% had no effect on pH, ammonia (NH3)N concentration and total rumen bacteria activity. The prill fat treatment at the level of 2% had no effect on dry matter intake, milk component and milk yield on in vivo analysis. In addition, milk yield, milk component and milk fatty acids were elevated after the prill fat treatment. This research concluded, the optimum level of prill fat addition in dairy ration was at the level of 2%. Supplementation of prill fat had no effect on rumen fermentation and fiber digestibility. Prill fat supplementation can be used to augment the milk yield, milk component, fatty acid composition in milk and had no effect on Atherogenicity Index (AI) and Hypocholesterolemic/Hypercholesterolemic (HH).

Abstract B07: NUAK2 inhibition for prostate cancer

Abstract Obesity is a US and global epidemic that exacerbates various cancers including prostate cancer (PC). A recent meta-analysis of men with PC found a 20% increased risk of PC-specific mortality for each 5 kg/m2 increase in BMI. To identify ways to exploit this, we performed an in vivo screen to identify kinases that were causally involved in tumor/growth in obese mice. This resulted in the identification of the AMPK-related kinase SNARK a.k.a NUAK2, a YAP1 target gene and positive feedback regulator by inhibition LATS1 kinase. To identify obesity essential kinases LAPC-4 cells were inoculated with an shRNA library of ~5,000 lentivirus targeting 513 kinases. 5 × 106 cells were grafted to chronically obese mice. Tumors were established to ~200 mm3 and a portion collected for reference. Remaining mice were randomized to continue on ad lib WD or 25% CR diet. Genome-integrated shRNA inserts were amplified using nested barcoded primers and sequenced using Illumina Hi-Seq 2000 and quantified. NAUK2 shRNAs (3 of 10) were significantly depleted in LAPC4 xenograft tumors from mice on a Western diet (WD), but not tumors from mice on an isocaloric 25% calorie-restricted (CR) diet. Analysis of NUAK2 gene expression in human PCs shows NUAK2 expression is higher in PC tissue vs. normal benign. In addition, Cox regression analysis showed that NUAK2 is associated with a 3-fold increased chance of developing metastasis disease in high NUAK2 group. Proteomic data from metastatic PC rapid autopsy biopsies shows NUAK2 phospho-target LATS1 Ser464 is higher in metastatic PCs vs. benign localized prostatic tissues. Western blot analysis of PC cell lines shows NUAK2 is elevated in comparison to primary prostate epithelial cells. To test if NUAK2 is an actionable target in PC cells, we utilized two narrow-spectrum kinase inhibitors from the literature, WZ4003 and HTH-02-006. Pharmacologic inhibition of NUAK2 slows 2D PC cell growth and 3D PC spheroid growth of various PC cell lines with an IC50 range between 1-10 uM, with HTH-02-006 being the more potent. In addition, NUAK2 kinase inhibition with WZ4003 and HTH-02-006 significantly slowed migration and Matrigel invasion of PC3 and DU145 cells. To test if NUAK2 is associated with obesity-enhanced PC progression, we tested NUAK2 inhibition in combination with palmitic acid supplementation. Our preliminary studies show that pharmacologic inhibition of NUAK2 sensitized PC cells to fatty acid-induced cell death. In conclusion, our in vivo shRNA screen identified NUAK2 as an essential kinase for tumors grown in obese mice. NUAK2 expression correlates with worse outcomes in PC and NUAK2 kinase activity is elevated in metastatic vs. localized PCs. Our preliminary data show that pharmacologic inhibition of NUAK2 slows PC cell growth and migration/invasion. Lastly, our preliminary studies show that NUAK2 may lower the threshold for fatty acid-induced toxicity. Together, these preliminary data suggest NUAK2 is a relevant actionable target in PC and may synergize with obese host physiology to slow PC tumor growth. Citation Format: Weiwei Fu, Megan Zhao, Amelia Schirmer, Erick J. Maravilla, Junhee Yoon, Sungyong You, Stephen J. Freedland, Everardo Macias. NUAK2 inhibition for prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr B07.

Palmitic acid supplementation does not improve performance of pre-ruminant calves

Performance and feeding behavior of pre-ruminant calves were examined in an experiment during winter in response to the incremental levels of a high-palmitic acid fat supplement (>86% C16:0) in the starter diet. Forty-eight 3-d-old male Holstein calves (40.3 ± 3.85 kg of body weight) were assigned randomly to 1 of 4 starter diets with the incremental levels of palm fat powder (PF) as 0 (PF0), 3% (PF3), 5% (PF5), or 7% (PF7). The calculated metabolizable energy of starter diet increased from 2.53 to 2.69 Mcal/kg as PF level increased. Calf starter intake and growth rate were not different as PF increased in the starter diet. Treatment had no effect on the pre-weaning calf behavior; however, the post-weaned calves spent more time on ruminating and laying while less time on standing as dietary PF increased. Overall, it appears that pre-ruminant calves would not benefit from the inclusion of palmitic acid in their starter diet.

Pre-fermentative supplementation of fatty acids alters the metabolic activity of wine yeasts

Fatty acids play important roles in the maintenance of cell membrane, viability and overall metabolism of wine yeasts (particularly Saccharomyces cerevisiae) during adverse winemaking conditions. We previously showed that linoleic acid supplementation markedly affect aroma compound production of S. cerevisiae wine strains. However, very little is known about how other commonly found fatty acids in grape juice modulate the growth and metabolism of S. cerevisiae. We aimed to determine the individual effect of five fatty acids on fermentation patterns and metabolism of two wine yeast strains (S. cerevisiae EC1118 and X5). Microvinification was performed at 15 °C by supplementing a grape juice (individually) with three different concentrations of saturated (palmitic acid), unsaturated (oleic, linoleic and γ-linolenic acids) and short-chain (hexanoic acid) fatty acids. Metabolite profiles of the resulting wines were determined using Gas-chromatography coupled to Mass-spectrometry (GC–MS). Our data show that the addition of γ-linolenic acid to the juice caused the production of higher amounts of amino and organic acids (except isoleucine and 2-oxoglutaric acid) in wines when fermented by EC1118, while palmitic acid supplementation showed similar trends when fermented by X5. The effect of linoleic acid was independent of yeast strains and we observed a global reduction of amino and organic acids (except pyruvic acid) while increased production of most of the fatty acids other than the supplemented ones. Our data clearly suggest that pre-fermentative supplementation of different fatty acids indeed influenced the growth and metabolism of wine yeasts in a different way. Thus, attention needs to be paid not only to the wine yeast strain used during the winemaking but also to the overall grape juice composition, including fatty acids, to obtain the desired wine characteristics.

Effects of timing of palmitic acid supplementation during early lactation on nutrient digestibility, energy balance, and metabolism of dairy cows

The objective of our study was to evaluate the effects of timing of palmitic acid (C16:0) supplementation during early lactation on nutrient digestibility, energy intake and balance, and metabolic responses of dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh (FR) period (1-24 d in milk) cows were assigned to either a control diet containing no supplemental fat (CON) or a C16:0-supplemented diet [PA; 1.5% of diet dry matter (DM)]. During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA diet (1.5% of diet DM) in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, compared with CON, PA increased DM digestibility by 3.0 percentage units and neutral detergent fiber (NDF) digestibility by 4.4 percentage units, and the increase in these variables was consistent over time. Although PA did not affect 18-carbon fatty acid (FA) digestibility, it decreased 16-carbon FA digestibility by 10.8 percentage units and total FA digestibility by 4.7 percentage units compared with CON. We observed a tendency for an interaction between treatment and time for total FA digestibility and 16-carbon FA digestibility due to the difference in FA digestibility between PA and CON reducing over time. Compared with CON, PA increased digestible energy intake by 3.9 Mcal/d, metabolizable energy intake by 3.5 Mcal/d, and net energy for lactation intake by 2.5 Mcal/d. The PA diet also increased milk energy output, negative energy balance, and plasma nonesterified fatty acid concentration and reduced plasma insulin concentration. We also observed a tendency for an interaction between treatment and time for energy balance due to cows receiving the PA treatment being in a greater negative energy balance over time. During the PK period, PA increased DM digestibility by 2.9 percentage units and NDF digestibility by 3.5 percentage units compared with CON. Although PA decreased 16-carbon FA digestibility by 7.0 percentage units, PA did not affect 18-carbon FA digestibility or total FA digestibility. Feeding PA during the PK period increased energy intake and milk energy output and did not affect energy balance. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased DM and NDF digestibilities and energy intake compared with a control diet containing no supplemental fat. Feeding C16:0 markedly increased milk energy output in both the FR and PK periods but increased negative energy balance only in the FR period.

Effects of timing of palmitic acid supplementation on production responses of early-lactation dairy cows

The objective of our study was to evaluate the effects of timing of palmitic acid (C16:0) supplementation on production responses of early-lactation dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh period (FR; 1-24 d in milk) cows were assigned to either a control diet containing no supplemental fat (CON) or a diet supplemented with C16:0 (palmitic acid, PA; 1.5% of diet dry matter). During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA (1.5% of diet dry matter) diet in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, we did not observe treatment differences for dry matter intake or milk yield. Compared with CON, PA increased the yield of 3.5% fat-corrected milk by 5.30 kg/d, yield of energy-corrected milk (ECM) by 4.70 kg/d, milk fat content by 0.41% units, milk fat yield by 280 g/d, and protein yield by 100 g/d. The increase in milk fat associated with the PA treatment during the FR period occurred due to an increase in yield of 16-carbon milk fatty acids (FA) by 147 g/d (derived from both de novo synthesis and extraction from plasma) and preformed milk FA by 96 g/d. Compared with CON, PA decreased body weight (BW) by 21 kg and body condition score (BCS) by 0.09 units and tended to increase BW loss by 0.76 kg/d. Although PA consistently increased milk fat yield and ECM over time, a treatment × time interaction was observed for BW and BCS due to PA inducing a greater decrease in BW and BCS after the second week of treatments. Feeding PA during the PK period increased milk yield by 3.45 kg/d, yield of 3.5% fat-corrected milk by 4.50 kg/d, yield of ECM by 4.60 kg/d, milk fat content by 0.22% units, milk fat yield by 210 g/d, protein yield by 140 g/d, and lactose yield by 100 g/d but tended to reduce BW by 10 kg compared with CON. Also, during the PK period we observed an interaction between diet fed in the FR and PK periods for milk fat yield due to feeding PA during the PK period increasing milk fat yield to a greater extent in cows that received the CON diet (+240 g/d) rather than the PA diet (+180 g/d) during the FR period. This difference was associated with the yield of preformed FA because feeding PA during the PK period increased the yield of preformed milk FA only in cows that received the CON diet during the FR period. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased the yield of ECM in both the FR and PK periods compared with a control diet. For some variables, the effect of feeding C16:0 was affected by timing of supplementation because milk yield increased only during the PK period and BW decreased to a greater extent in the FR period. Regardless of diet fed in the FR period, feeding a C16:0 supplement during the PK period increased yields of milk and milk components.

Long-term palmitic acid supplementation interacts with parity in lactating dairy cows: Production responses, nutrient digestibility, and energy partitioning

The objective of our study was to evaluate the effects of long-term palmitic acid (C16:0) supplementation and parity on production, nutrient digestibility, and energy partitioning of mid-lactation dairy cows. Forty mid-lactation Holstein cows (18 primiparous and 22 multiparous) were used in a block design. Cows were assigned to receive either a control diet containing no supplemental fat (CON) or a C16:0-enriched supplemented diet (PA; 1.5% diet dry matter) fed for 10 wk. Compared with CON, PA increased dry matter intake, milk yield, cumulative milk yield, milk fat content, milk fat yield, 16-carbon milk fatty acid (FA) yield, 3.5% fat-corrected milk yield, and energy-corrected milk yield. Additionally, PA increased body weight change, but did not affect body condition score change compared with CON. A tendency for a treatment by parity interaction was observed for milk yield due to PA increasing milk yield in multiparous but not in primiparous cows. In addition, we observed interactions between treatment and parity for fat-corrected milk, energy-corrected milk, and milk fat yield due to PA increasing these variables to a greater extent in multiparous compared with primiparous cows. Interestingly, we observed an interaction between treatment and parity for body weight change, due to PA increasing body weight change in primiparous but not in multiparous cows. The PA treatment increased dry matter and neutral detergent fiber digestibilities compared with CON. Although PA did not affect 18-carbon FA digestibility, compared with CON, PA decreased 16-carbon and total FA digestibilities and increased total FA intake by 470 g/d and absorbed total FA by 316 g/d. We also observed an interaction between treatment and parity for total absorbed FA due to PA increasing it to a greater extent in multiparous than in primiparous cows. Compared with CON, PA increased apparent energy intake and milk energy output. We observed an interaction between treatment and parity for milk energy output due to PA increasing milk energy output to a greater extent in multiparous than primiparous cows. Additionally, an interaction between treatment and parity was observed for energy output in body reserves due to PA increasing energy output in body reserves in primiparous but not in multiparous cows. In conclusion, production responses of dairy cows to PA were consistent throughout the 10-wk treatment period. In addition, PA supplementation interacted with parity, with production responses increased to a greater extent in multiparous than primiparous cows and energy partitioned to body reserves only increased in primiparous cows.

Hot topic: Ceramide inhibits insulin sensitivity in primary bovine adipocytes

In nonruminants, the sphingolipid ceramide inhibits insulin sensitivity by inactivating protein kinase B (AKT) within the insulin-signaling pathway. We have established that ceramide accrual develops with impaired systemic insulin action in ruminants during the transition from gestation to lactation, dietary palmitic acid supplementation, or controlled nutrient restriction. We hypothesized that ceramide promotes AKT inactivation and antagonizes insulin sensitivity in primary bovine adipocytes. Stromal-vascular cells were grown from bovine adipose tissue explants and cultured in differentiation media. To modify ceramide supply, we treated differentiated adipocytes with (1) myriocin, an inhibitor of de novo ceramide synthesis, or (2) cell-permeable C2:0-ceramide. Insulin-stimulated AKT activation (i.e., phosphorylation) and 2-deoxy-D-[3H]-glucose (2DOG) uptake were measured. Treatment of adipocytes with myriocin consistently decreased concentrations of ceramide, monohexosylceramide, and lactosylceramide. The insulin-stimulated ratio of phosphorylated AKT to total AKT was increased with myriocin but decreased with C2:0-ceramide. Moreover, adipocyte insulin-stimulated 2DOG uptake was decreased with C2:0-ceramide and increased with myriocin. We conclude that ceramide inhibits insulin-stimulated glucose uptake by downregulating AKT activation in primary bovine adipocytes.

Nutrient digestibility and milk production responses to increasing levels of palmitic acid supplementation vary in cows receiving diets with or without whole cottonseed

Nutrient digestibility and milk production responses to increasing levels of palmitic acid supplementation vary in cows receiving diets with or without whole cottonseed