Non-esterified Fatty Acid Concentrations Research Articles

β-sitosterol Alleviates High Fatty Acid-Induced Lipid Accumulation in Calf Hepatocytes by Regulating Cholesterol Metabolism

A significant reduction in plasma concentration of cholesterol during early lactation is a common occurrence in high-yielding dairy cows. An insufficient synthesis of cholesterol in the liver has been linked to lipid accumulation caused by high concentrations of fatty acids during negative energy balance (NEB). As ruminant diets do not provide quantitative amounts of cholesterol for absorption, phytosterols such as β-sitosterol may serve to mitigate the shortfall in cholesterol within the liver during NEB. To gain mechanistic insights, primary hepatocytes were isolated from healthy female 1-day old calves for in vitro studies with or without 1.2mM fatty acids (FA) to induce metabolic stress. Furthermore, hepatocytes were treated with 50μM β-sitosterol with or without FA. Data were analyzed by one-way ANOVA with subsequent Bonferroni correction. Results revealed that calf hepatocytes treated with FA had greater content of non-esterified fatty acids (NEFA) and triacylglycerol (TAG), and greater mRNA and protein abundance of the lipid synthesis-related SREBF1 and FASN. In contrast, mRNA and protein of CPT1A (fatty acid oxidation) and the cholesterol metabolism-related targets SREBF2, HMGCR, ACAT2, APOA1, ABCA1 and ABCG5 was lower. Content of the antioxidant-related glutathione (GSH) and activities of superoxide dismutase (SOD) also was lower. Compared with FA challenge alone, 50μM β-sitosterol led to greater mRNA and protein abundance of SREBF2, HMGCR, ACAT2 and ABCG5, and greater content of GSH and activity of SOD. In contrast, compared with the FA group, the mRNA and protein abundance of SREBF1 and ACC1 and the content of TAG and NEFA in the β-sitosterol + FA group were lower. Overall, β-sitosterol can promote cholesterol metabolism and reduce oxidative stress while reducing lipid accumulation in hepatocytes challenged with high concentrations of fatty acids.

L-histidine attenuates NEFA-induced inflammatory responses by suppressing Gab2 expression

Non-esterified fatty acids (NEFAs), key to energy metabolism, may become pathogenic at elevated levels, potentially eliciting immune reactions. Our laboratory's findings of reduced L-histidine in ketotic states, induced by heightened NEFA concentrations, suggest an interrelation with NEFA metabolism. This observation necessitates further investigation into the mitigating role of L-histidine on the deleterious effects of NEFAs. Our study unveiled that elevated NEFA concentrations hinder the proliferation of Bovine Mammary Epithelial Cells (BMECs) and provoke inflammation in a dose-responsive manner. Delving into L-histidine's influence on BMECs, RNA sequencing revealed 2124 genes differentially expressed between control and L-histidine-treated cells, with notable enrichment in pathways linked to proliferation and immunity, such as cell cycle and TNF signaling pathways. Further analysis showed that L-histidine treatment positively correlated with an increase in EdU-555-positive cell rate and significantly suppressed IL-6 and IL-8 levels (p < 0.05) compared to controls. Crucially, concurrent treatment with high NEFA and L-histidine normalized the number of EdU-555-positive cells and cytokine expression to control levels. Investigating the underlying mechanisms, Gab2 (Grb2-associated binder 2) emerged as a central player; L-histidine notably reduced Gab2 expression, while NEFA had the opposite effect (p < 0.05). Gab2 overexpression escalated nitric oxide (NO) production and IL6 and IL8 expression. However, L-histidine addition to Gab2-overexpressing cells resulted in NO concentrations indistinguishable from controls. Our findings collectively indicate that L-histidine can counteract NEFA-induced inflammation in BMECs by inhibiting Gab2 expression, highlighting its therapeutic potential against NEFA-related metabolic disturbances.

Lactation performance, feed efficiency, and blood metabolites of dairy cows treated with recombinant bovine somatotropin: A systematic review and meta-analysis

A systematic review and meta-analysis were conducted to assess the impact of recombinant bovine somatotropin (rbST) on lactation performance, feed efficiency, and blood metabolites in dairy cows. In the systematic review, articles were selected based on the following criteria: (1) Data focusing on the influence of bovine somatotropin doses on milk production; (2) Submission of original data; (3) Articles published in journals; and (4) Articles in English or Portuguese. The analysis of variance was used with a completely randomized design and mixed models methodology. Polynomial regression was applied to significant fixed effects (rbST dose). The use of rbST resulted in increased milk yield and 4% fat-corrected milk yield, while fat, protein, and lactose contents remained unaffected. Dry matter and metabolizable energy intakes, as well as milk/feed efficiency, exhibited a linear increase, but body condition score (BCS) was negatively impacted. The administration of rbST led to higher blood concentrations of triglycerides and insulin. Cows treated with rbST showed a 23% increase in non-esterified fatty acid (NEFA) concentrations compared to non-treated cows. Additionally, growth factors IGF-1 and IGF-2 displayed a linear increase with rbST treatment. In summary, rbST administration increased milk yield and fat-corrected milk yield without affecting milk components. However, despite increasing intake, it resulted in BCS losses and alterations in blood parameters such as NEFA, IGF-1, and IGF-2.

Effects of Summer Heat on Adipose Tissue Activity in Periparturient Simmental Cows.

Hot climate is one of the major factors affecting the dairy industry. Heat stress could be responsible for decreased feed intake and consequently leads to alterations in energy metabolism, particularly during late pregnancy and early lactation. This study aimed to assess the effects of summer heat on adipose tissue activities during the periparturient period in Simmental cows. Two groups of cows were involved: heat-stressed cows (n = 12) that calved from June to August and thermoneutral cows (n = 12) that calved from October to December. Blood samples were taken from each cow during the periparturient period: 21 and 7 days before calving and 8, 16, 24, 32, and 40 days after calving. Glucose, beta-hydroxy butyrate (BHB), non-esterified fatty acids (NEFA), leptin (LP), and adiponectin (ADP) were measured in serum samples by commercial kits. Thermoneutral cows expressed higher degrees of lipomobilization syndrome than heat-stressed cows, indicated by significantly higher serum NEFA and BHB concentrations in the early lactation. Leptin levels were significantly decreased, while adiponectin was increased in heat-stressed cows compared to thermoneutral ones. The results indicated that heat-stressed cows during the periparturient period mobilized less fat from adipose tissue to reduce the heat generation by fatty acid oxidation.

Capsaicin as a Dietary Additive for Dairy Cows: A Meta-Analysis on Performance, Milk Composition, Digestibility, Rumen Fermentation, and Serum Metabolites.

This study aimed to evaluate the effects of dietary supplementation with capsaicin (CAP) on productive performance, milk composition, nutrient digestibility, ruminal fermentation, and serum metabolites of dairy cows using a meta-analytical approach. The database included 13 studies, from which the response variables of interest were obtained. Data were analyzed using a random effects model, and results were expressed as weighted mean differences between treatments supplemented with and without CAP. Dietary supplementation with CAP increased (p < 0.05) dry matter intake, milk yield, feed efficiency, milk fat yield, and milk fat content. However, CAP supplementation did not affect (p > 0.05) milk protein and lactose yield, milk urea nitrogen, or milk somatic cell count. Greater (p < 0.05) apparent digestibility of dry matter and crude protein was observed in response to the dietary inclusion of CAP. Likewise, supplementation with CAP increased (p < 0.05) the rumen concentration of total volatile fatty acids. In contrast, CAP supplementation did not affect (p > 0.05) ruminal pH or the ruminal concentration of ammonia nitrogen, acetate, propionate, and butyrate. In blood serum, CAP supplementation increased (p < 0.05) the glucose concentration and decreased (p < 0.05) the concentration of non-esterified fatty acids. However, CAP supplementation did not affect (p > 0.05) the serum concentration of urea and beta-hydroxybutyrate. In conclusion, capsaicin can be used as a dietary additive to improve the productive performance, milk composition, and nutrient digestibility in dairy cows and, at the same time, improve the ruminal concentration of total volatile fatty acids and serum levels of glucose and non-esterified fatty acids.

Feed restriction of lactating cows triggers acute downregulation of mammary mTOR signaling and chronic reduction of mammary epithelial mass

While there is generally no consensus about how nutrients determine milk synthesis in the mammary gland, it is likely that the mechanistic target of rapamycin complex 1 (mTORC1) plays a role as a key integrator of nutritional and mitogenic signals that can influence a multitude of catabolic and anabolic pathways. The objectives of this study were to evaluate acute changes (<24 h) in translational signaling, in addition to chronic changes (14 d) in mammary gland structure and composition, in response to a severe feed restriction. Fourteen lactating Holstein dairy cows were assigned to either ad libitum feeding (n = 7), or a restricted feeding program (n = 7). Feed-restricted cows had feed removed after the evening milking on d 0. Mammary biopsies and blood samples were collected 16 h after feed removal, after which cows in the restricted group were fed 60% of their previously observed ad lib intake for the remainder of the study. On d 14, animals were sacrificed and mammary glands dissected. In response to feed removal, an acute increase in plasma nonesterified fatty acid concentration was observed, concurrent to a decrease in milk yield. In mammary tissue, we observed downregulation of the mTORC1-S6K1 signaling cascade, in addition to reductions in mRNA expression of markers of protein synthesis, endoplasmic reticulum biogenesis, and cell turnover (i.e., transcripts associated with apoptosis or cell proliferation). During the 14 d of restricted feeding, animals underwent homeorhetic adaptation to 40% lower nutrient intake, achieving a new setpoint of 14% reduced milk yield with 18% and 29% smaller mammary secretory tissue dry matter and crude protein masses, respectively. On d 14, no treatment differences were observed in markers of protein synthesis or mammary cell turnover evaluated using gene transcripts and immunohistochemical staining. These findings implicate mTORC1-S6K1 in the early phase of the adaptation of the mammary gland's capacity for milk synthesis in response to changes in nutrient supply. Additionally, changes in rates of mammary cell turnover may be transient in nature, returning to basal levels following brief alterations that have sustained effects.

Rumen-Protected Choline Improves Metabolism and Lactation Performance in Dairy Cows.

Choline is required for the synthesis of phosphatidylcholine, an important constituent of lipoproteins. Early lactation cows presumably synthesize insufficient phosphatidylcholine, and choline supplementation in a rumen-protected form might benefit metabolism and lactation. The objectives of this study were to determine the effects of feeding rumen-protected choline (RPC) on lactation and metabolism in dairy cows. In experiment 1, 369 nulliparous and parous Holstein cows housed in four pens per treatment were fed 12.9 g/day of choline as RPC from 25 days prepartum until 80 days postpartum. In experiment 2, 578 nulliparous cows housed in five pens/treatment were fed 12.9 g/day of choline as RPC in the last 22 days of gestation only. In both experiments, blood was sampled and analyzed for concentrations of nonesterified fatty acids (NEFAs) and glucose at 1, 14, and 21 days postpartum and of choline at 1 and 14 days postpartum. Blood from all cows was sampled and analyzed for concentrations of β-OH butyrate (BHB) at 1 and 14 days postpartum. Cows with BHB > 1.2 mmol/L were classified as having hyperketonemia. Hepatic tissue was collected from 46 cows from the eight pens in experiment 1 at 9 days postpartum and analyzed for concentrations of glycogen and triacylglycerol. Milk yield and components were measured for 80 days postpartum in experiment 1, whereas only milk yield was measured in experiment 2. The pen was the experimental unit of analysis. Supplementing RPC tended to increase dry matter intake (DMI) prepartum in experiments 1 and 2 and postpartum in experiment 1. Feeding cows with RPC increased yields of 3.5% fat-corrected milk (42.8 vs. 44.8 kg/day), energy-corrected milk (38.5 vs. 40.3 kg/day), milk fat (1.52 vs. 1.61 kg/day), and true protein (1.16 vs. 1.21 kg/day) in experiment 1. Milk yield tended to be greater with RPC (26.4 vs. 27.4 kg/day) in experiment 2. Supplementing RPC increased plasma choline concentrations on day 14 postpartum in experiment 1 (3.32 ± 0.27 vs. 4.34 ± 0.28 µM) and on day 1 in experiment 2 (3.35 ± 0.16 and 13.73 ± 0.15 µM). Treatment did not affect the concentrations of glucose, NEFAs, or BHB in plasma, but the incidence of hyperketonemia was less in multiparous cows fed RPC than those fed the control in experiment 1. Feeding cows with RPC reduced hepatic triacylglycerol content and tended to reduce the ratio of triacylglycerol to glycogen and the risk of hepatic lipidosis in cows in experiment 1. The concentrations of hepatic triacylglycerol on day 9 postpartum were inversely related to those of choline in plasma on day 1 postpartum. Feeding cows with RPC improved lactation and metabolism, but more benefits were noted when it was fed before and after calving.

Prepartum behaviors as early indicators for postpartum energy associated biomarkers status in Holstein dairy cows.

Our main aim was to investigate the predictive value of prepartum behaviors such as total daily rumination (TDR), total daily activity (TDA) and dry matter intake (DMI) as early indicators to detect cows at risk for hyperketonemia (HYK), hypoglycemia (HYG) or high non-esterified fatty acid (NEFA) status in the first (wk1) and second week (wk2) postpartum. In a case control study, 64 Holstein cows were enrolled 3 weeks before the expected time of calving and monitored until 15 days in milk (DIM). Postpartum blood samples were taken at D3 and D6 for wk1 and at D12 and D15 for wk2 to measure beta-hydroxybutyrate, NEFA and glucose concentration. Ear-mounted accelerometers were used to measure TDR and TDA. DMI and milk yield were obtained from farm records. Relationships between the average daily rate of change in prepartum TDR (ΔTDR), TDA (ΔTDA), and DMI (ΔDMI) with postpartum HYK, HYG and NEFA status in wk1 and wk2 post-partum were evaluated using linear regression models. Models were adjusted for potential confounding variables, and covariates retained in the final models were determined by backward selection. No evidence was found to support the premise that prepartum ΔTDR, ΔTDA or ΔDMI predicted postpartum HYK, HYG or NEFA status in wk1 or in wk2. Overall, prepartum ΔTDR, ΔTDA and ΔDMI were not effective predictors of HYK, HYG or NEFA status in the first 2 weeks postpartum.

Fat supplement for dairy cows during early lactation – potentials, challenges, and risks – a meta-analysis

ABSTRACT The most critical challenge in early lactation is to meet the increasing energy demand for milk production, which results in a negative energy balance (EB). The objective of this study was to evaluate the productive performance and plasma metabolite responses of early lactating dairy cows fed diets supplemented with increasing levels of different fat sources. Diets were categorized as diets supplemented with saturated fatty acid rich fat sources (SFAR), with Ca-soaps of fatty acids (CaFA), and with polyunsaturated fatty acid rich fat sources (PUFAR). There was an interaction between crude fat level and source for dry matter intake (DMI); each percentage-unit increase reduced DMI by 0.38 and 0.77 kg/d in CaFA and PUFAR, respectively, and increased DMI by 0.55 kg/d in SFAR. Corrected milk yield increased by 0.44 kg/d for each percentage-unit increase in crude fat level. Plasma non-esterified fatty acid (NEFA) concentration increased with 0.04 mmol/L per percentage-unit increase. In conclusion, the increased crude fat level increased corrected milk yield. Fat supplementation in early lactation does not seem to cause negative effects on productive performance or metabolism, and despite a small increase, both plasma NEFA and β-hydroxybutyrate (BHB) concentrations stayed below the threshold for incidence of subclinical ketosis.

Hydrogen-rich water supplementation improves metabolic profile during peripartum period in Gurcu goats and enhances the health and survival of kids

In this study, the effect of intaking hydrogen-rich water (HRW) on the metabolic profile of Gurcu goats during the peripartum period and the survival/growth performance of kids were evaluated. Twenty-three pregnant goats were divided into two groups 21–23 days before the due date. Group 1 (G1, n = 10) was given HRW from day 21 before delivery until day 21 after delivery. Group 2 (G2, n = 13) served as the control. Blood samples were weekly taken from 21 days before delivery until 21 days after delivery. Hydrogen-rich water increased serum glucose concentration on the delivery day more than in G2 (P = 0.016). Hydrogen-rich water decreased serum total cholesterol (P = 0.02) and creatinine (P = 0.05) concentration at delivery. Group effect and time effect were significant in triglyceride (P < 0.001, P = 0.001, respectively) and albumin (P < 0.001, P = 0.002, respectively) concentration. Aspartate transaminase decreased towards the delivery day in G1 (P < 0.05). Serum non-esterified fatty acids concentration was lower in G1 than in G2, but there was no significant differences (P > 0.05). Beta-hydroxybutyric acid concentration an increased in both groups during the prepartum period, although there was no significance (P > 0.05). Hydrogen-rich water did not affect the birth weight and growth performance of the kids (P > 0.05), but it increased their survival rates and overall health, although there was no significance (P > 0.05). In conclusion, HRW may have an impact on the metabolic profiles during the peripartum period and have a positive effect on lipid profiles. Additionally, intaking HRW to goats during the peripartum period may improve the health and survival of kids and reduce their mortality.

85 Evaluation of a Saccharomyces cerevisiae fermentation product on performance, plasma metabolites, rumen parameters, and fecal pathogen shedding in feedlot steers

Abstract The objective of the experiment was to evaluate a commercial Saccharomyces cerevisiae fermentation product (SCFP; NaturSafe, Dimond V, Cedar Rapids, IA) on performance, plasma metabolites, rumen parameters, and fecal pathogen shedding in feedlot steers. Crossbred steers (n = 61; 271 ± 48.1 kg) were sourced from 3 different research units at Oklahoma State University and assigned to 1 of 3 experimental treatments in a randomized complete block design. Steers were group housed in pens equipped with an automated individual feed intake system. Treatments consisted of a negative control (CON), CON with tylosin phosphate (TYL; 9.9 mg · kg-1 of diet DM), or CON with SCFP (1.63 g · kg-1 of diet DM). Body weight and feed efficiency did not differ among treatments (P ≥ 0.12). However, steers fed SCFP tended to have greater ADG from d 35 to 56 (P = 0.10). In addition, steers fed SCFP exhibited increased DMI from d 56 to 112 (P = 0.03), a tendency for increased DMI from d 112 to final (P = 0.07), and increased DMI overall (P = 0.05). There was no treatment × time interaction for pH, pH change, or lactate concentrations in rumen fluid (P ≥ 0.11), but all were impacted by time (P &amp;lt; 0.01). A treatment × time interaction (P &amp;lt; 0.01) was detected for fecal pH and fecal pH change, with the pH being greater in TYL fed steers on d 56 and in CON fed steers on the final day. Steers that consumed CON had a reduced fecal pH (P &amp;lt; 0.01) and steers consuming SCFP tended to have a reduced fecal pH (P = 0.08) from d 35 to 56 compared with TYL fed steers, while both CON and SCFP fed steers experienced an increase in fecal pH from d 56 to 112 (P &amp;lt; 0.01). A time effect was detected (P ≤ 0.04) for plasma glucose, lactate, urea nitrogen, and non-esterified fatty acid concentrations among treatment groups. A treatment × time interaction was observed (P &amp;lt; 0.02) for stx2 genes of Shiga toxin producing Escherichia coli (STEC) in individual fecal samples where SCFP and TYL fed steers tended to exhibit an increased concentration compared with CON fed steers on d 14, while on d 190 CON and SCFP fed steers exhibited an increased concentration compared with TYL fed steers. Overall, STEC, E. coli O157:H7, and Salmonella concentrations were all impacted by time (P &amp;lt; 0.001). This experiment suggests that SCFP could be utilized as a potential alternative to TYL in feedlot cattle without adversely affecting cattle performance. However, SCFP did not impact pathogen shedding. In addition, CON fed steers performed similarly to both SCFP and TYL fed steers.

Dry period length affects rumen adaptation in dairy cattle precalving and during the first weeks after calving

Omitting or shortening the dry period may result in a fairly constant ration throughout the transition period of dairy cows, reducing the need for adaptation of cow metabolism and rumen function to a new lactation. The objective of this study was to determine the effect of dry period length on rumen adaptation and cow metabolic state during the transition period. Twelve pregnant, rumen-cannulated Holstein Friesian dairy cows at the end of their first lactation were assigned to one of 3 treatments: a conventional (60 d), short (30 d) or no dry period (0 d). At dry-off, cows received a dry cow ration until calving. Lactating cows received a lactation ration. Cows were monitored from 8 wk before calving until 8 wk after calving for milk yield and dry matter intake (DMI). Rumen biopsies were taken from 3 locations in the rumen at 60, 40 and 10 d before calving and 3, 7, 14, 28 and 56 d after calving to assess papillae dimensions. Blood was sampled weekly from 3 wk before until 8 wk after calving, and liver biopsies were taken at wk -2, wk 2 and wk 4 relative to calving. Prepartum, DMI and milk yield were greater for cows with a short or no dry period, compared with cows with a conventional dry period. Postpartum, DMI was greater for cows with a short dry period compared with cows with a conventional dry period. Plasma glucose concentration was greater for cows without a dry period, compared with the other dry period lengths postpartum. Plasma concentrations of nonesterified fatty acids and β-hydroxybutyrate, and liver triglyceride content, did not differ among dry period. Rumen papillae differed in size based on biopsy location, but there was no interaction between biopsy location and the effect of dry period length. Rumen papillae surface area for cows managed for a 30 d or 60 d dry period decreased toward calving. At 40 d prepartum, papillae surface area was greater for short and no dry period treatment compared with a conventional dry period. At 10 d prepartum, papillae surface area was greater for the no dry period treatment compared with both other treatments, and this difference was still present 3 d postpartum. Cows managed for a short dry period showed faster increase in papillae dimensions after calving compared with cows managed for a conventional dry period. From d 28 onwards, no differences in papillae surface area were observed. The faster rumen adaptation postpartum may be related to the increased DMI during the first weeks postpartum for cows managed for a short dry period. However, this did not result in improved metabolic status or milk yield. The results from the present study demonstrate that the dietary changes related to a conventional dry period length affected rumen papillae development, not only prepartum but also early postpartum. Further optimization of dry period length as well as dietary composition throughout the transition period may support cows in their adaptation to a new lactation.

Ionophore antibiotics and hop cones as regulators of digestion and metabolism in ruminants

The general characteristics of ionophore antibiotics and the mechanisms of their antimicrobial action were analyzed. Two types of ionophore antibiotics are known: those that transport ions across the membrane, and those that form a channel in the cell membrane through which ions pass. Ionophore antibiotics used in animal husbandry belong to the former group (monensin, lasalocid, salinomycin, narasin). They are synthesized by bacteria of the Streptomyces genus. Bacterial cells and rumen fluid differ in ionic composition, which is regulated by active ion transport. As a result, the cytoplasm of bacteria contains more potassium ions, while the rumen fluid, on the contrary, has more sodium ions. Ionophores transport potassium inside the cell and remove sodium outside. The bacteria try to correct this imbalance and require ATP energy to carry out this process. Eventually, energy deficit develops in the bacterial cell and it dies. Ionophore antibiotics affect only the Gram-positive bacteria in the rumen of ruminants, because they cannot penetrate through the cell wall of Gram-negative bacteria. Thus, there is a selective destruction of some types of bacteria, the most sensitive among which are the so-called hyper-ammonia-producing bacteria (HAB). Ionophore antibiotics, which change the breakdown of protein and carbohydrates, change the ratio of volatile fatty acids by increasing the proportion of propionic acid and thus inhibit methanogenesis in the rumen. Ionophore antibiotics are an important antiketotic agent because they reduce the concentration of β-hydroxybutyrate and non-esterified fatty acids in the blood of ruminants. During the transition period, cows have metabolic disorders so they are more prone to infectious diseases due to a decrease in resistance. The use of ionophore antibiotics affects the immune function and resistance to inflammatory processes, which is caused by an indirect influence associated with a lower pathological effect of ketosis and steatosis. Ionophore antibiotics reduce the negative energy balance and its pathological impact on metabolism. The influence of ionophore antibiotics on cow’s milk productivity is mostly absent or insignificant. The study presents characteristics of biologically active substances of hop cones. Hop cones contain biologically active components similar to ionophore antibiotics by action. These are prenylated flavonoids: humulone (α-acid), lupulone (β-acid) and their derivatives. These components of hop cones can be regarded as a potential substitute for ionophoric antibiotics. In particular, lupulone and some other components of hop cones inhibit the activity of Gram-positive bacteria, causing reactions similar to ionophore antibiotics. In addition, hop cones have many other biologically active compounds: phenols, essential oils, and resins, which have antimicrobial, antioxidant, sedative, phytoestrogen, insulin stimulating, immunomodulatory, and antitumor effects.

Effect of high NEFA concentration on lipid metabolism disorders in hepatocytes based on lipidomics.

Introduction: High concentrations of nonesterified fatty acids (NEFA) is the key of characteristic of fatty liver in dairy cows. Therefore, the aim of this study was to investigate the effect of high concentration of NEFA on lipid metabolism in hepatocytes through the lipidomic approach and molecular biology techniques. Methods: Stimulate AML-12 cells with different concentrations of NEFA, observe the cellular lipid accumulation, and select 0.6mM NEFA stimulation concentration for subsequent experiments. Collect cells for lipidomics analysis. Results: High concentration of NEFA (0.6-2.4mM) significantly reduced the cell viability in a concentration-dependent manner, indicating that high concentrations of NEFA have lipotoxicity on hepatocytes. In addition, NEFA promoted triglycerides (TAG) accumulation, increased the mRNA expression of the lipogenic molecules SREBP1c and FASN, and decreased the mRNA expression of lipolytic molecules CPT1A and HSL in hepatocytes. Mechanistically, high concentration of NEFA induced lipid metabolism disorders in hepatocytes by regulating metabolic pathways such as glycerol phospholipid metabolism, glycosyl phosphatidylinositol anchored biosynthesis, triglyceride metabolism, sphingolipid metabolism, and inositol phosphate metabolism. Discussion: High concentration of NEFA is lipotoxic to cells, promoting lipid accumulation. LPE (18:2), LPE (18:3), LPE (18:1) via glycerophospholipid metabolism, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, glycerolipid metabolism, sphingolipid metabolism, and inositol phosphate metabolism, indicating their potential regulation role in the pathogenesis of fatty liver.

Effect of two recombinant bovine somatotropin formulations on feeding behavior and metabolism of Holstein cows

This study evaluated the effect of two commercial forms of recombinant bovine somatotropin (rbST; Later® and Fast®; 500 mg) on feed variability, feeding behavior, zootechnical parameters, and milk production of Holstein cows after peak production. Eighteen cows were randomly divided into two groups and treated with injectable of rbST every 14 days for 70 days. Blood samples were collected on day 1, 4, and 7 of each cycle, to assess circulating levels of non-esterified fatty acids (NEFA). Weight and body condition score (BCS) assessments were performed. Milk yields were measured daily using the DelPro™ software (DeLaval®). Feed consumption and feeding behavior was obtained daily using automatic individual feeders (Intergado®). No difference was observed in dry matter intake (DMI) between groups (P=0.07), and no impact was noted on dietary variability (P=0.64). In addition, animals treated with rbST-Fast exhibited more frequent visits to feeders, longer time in consumption, and higher relative DMI (per percentage of live weight) (P&lt;0.01). Animals in the rbST-Fast group produced more milk than those in the rbST-Later group (P=0.03). Furthermore, the weight of the cows was significantly different (P=0.05) between groups. The weight of the rbST-Fast animals was smaller and varied more than thot of the rbST-Later animals (P &lt;0.01). Nonetheless, no difference was noted in BCS (P=0.30) between the groups. The animals in the rbST-Fast group had higher concentrations of NEFA than those in the rbST-Later group (P&lt;0.01). In conclusion, behavioral and metabolic shifts that sustained greater milk production in the cows receiving rbST-Fast. Nonetheless, these shifts are likely linked to lipomobilization and not dietary variability or consumption, as evidenced by the increase in NEFA and greater weight loss in that group.

Effect of Myrtus communis L. Plant Extract as a Milk Supplement on the Performance, Selected Blood Parameters and Immune Response of Holstein Calves.

This research aimed to understand the effects of adding myrtle plant extract obtained from its leaves (MPEL) and roots (MPER) to the milk fed to suckling female Holstein calves, focusing on performance, reproduction, selected blood parameters and immune response. The 50 Holstein female calves, one week of age, were divided into five groups: one group received no plant extract (Control), while the others were supplemented with myrtle plant extracts at doses of 25 mL/day leaf extract (MPEL-25), 25 mL/day root extract (MPER-25), 50 mL/day leaf extract (MPEL-50) and 50 mL/day root extract (MPER-50) for each calf in each treatment group. The extracts were given along with the milk to the experimental groups for 60 days, and for an additional 12 days post-weaning. The results reveal that the feed consumption and live weights increased significantly. Significantly higher leukocyte counts were observed in the 50 mL/head × day myrtle groups, and a higher IgG concentration was also noted in the MPER-50 group compared to the other groups. The serum non-esterified fatty acid (NEFA) concentration significantly decreased in the MPEL-50 and MPER-50 groups, whereas the betahyrdoxy butyric acid (BHBA) concentration increased and the serum glucose concentration significantly decreased with myrtle supplementation. In conclusion, it was determined that the performance, immune system and negative energy balance compensation of female Holstein calves were positively affected by administering extracts obtained from the leaves and roots of the Myrtus communis L. plant at dose levels of 25 and 50 mL/head × day for 72 days, without causing any side effects.

The serine protease 2 gene regulates lipid metabolism through the LEP/ampkα1/SREBP1 pathway in bovine mammary epithelial cells

Molecular breeding has brought about significant transformations in the milk market and production system during the twenty-first century. The primary economic characteristic of dairy production pertains to milk fat content. Our previous transcriptome analyses revealed that serine protease 2 (PRSS2) is a candidate gene that could impact milk fat synthesis in bovine mammary epithelial cells (BMECs) of Chinese Holstein dairy cows. To elucidate the function of the PRSS2 gene in milk fat synthesis, we constructed vectors for PRSS2 overexpression and interference and assessed intracellular triglycerides (TGs), cholesterol (CHOL), and nonesterified fatty acid (NEFA) contents in BMECs. Fatty acid varieties and components were also quantified using gas chromatography‒mass spectrometry (GC‒MS) technology. The regulatory pathway mediated by PRSS2 was validated through qPCR, ELISA, and WB techniques. Based on our research findings, PRSS2 emerges as a pivotal gene that regulates the expression of associated genes, thereby making a substantial contribution to lipid metabolism via the leptin (LEP)/Adenylate-activated protein kinase, alpha 1 catalytic subunit (AMPKα1)/sterol regulatory element binding protein 1(SREBP1) pathway by inhibiting TGs and CHOL accumulation while potentially promoting NEFA synthesis in BMECs. Furthermore, the PRSS2 gene enhances intracellular medium- and long-chain fatty acid metabolism by modulating genes related to the LEP/AMPKα1/SREBP1 pathway, leading to increased contents of unsaturated fatty acids C17:1N7 and C22:4N6. This study provides a robust theoretical framework for further investigation into the underlying molecular mechanisms through which PRSS2 influences lipid metabolism in dairy cows.

The impact of forearm immobilization and acipimox administration on muscle amino acid metabolism and insulin sensitivity in healthy, young volunteers.

Although the mechanisms underpinning short-term muscle disuse atrophy and associated insulin resistance remain to be elucidated, perturbed lipid metabolism might be involved. Our aim was to determine the impact of acipimox administration [i.e., pharmacologically lowering circulating nonesterified fatty acid (NEFA) availability] on muscle amino acid metabolism and insulin sensitivity during short-term disuse. Eighteen healthy individuals (age: 22 ± 1 years; body mass index: 24.0 ± 0.6 kg·m-2) underwent 2 days forearm immobilization with placebo (PLA; n = 9) or acipimox (ACI; 250 mg Olbetam; n = 9) ingestion four times daily. Before and after immobilization, whole body glucose disposal rate (GDR), forearm glucose uptake (FGU; i.e., muscle insulin sensitivity), and amino acid kinetics were measured under fasting and hyperinsulinemic-hyperaminoacidemic-euglycemic clamp conditions using forearm balance and l-[ring-2H5]-phenylalanine infusions. Immobilization did not affect GDR but decreased insulin-stimulated FGU in both groups, more so in ACI (from 53 ± 8 to 12 ± 5 µmol·min-1) than PLA (from 52 ± 8 to 38 ± 13 µmol·min-1; P < 0.05). In ACI only, and in contrast to our hypothesis, fasting arterialized NEFA concentrations were elevated to 1.3 ± 0.1 mmol·L-1 postimmobilization (P < 0.05), and fasting forearm NEFA balance increased approximately fourfold (P = 0.10). Forearm phenylalanine net balance decreased following immobilization (P < 0.10), driven by an increased rate of appearance [from 32 ± 5 (fasting) and 21 ± 4 (clamp) preimmobilization to 53 ± 8 and 31 ± 4 postimmobilization; P < 0.05] while the rate of disappearance was unaffected by disuse or acipimox. Disuse-induced insulin resistance is accompanied by early signs of negative net muscle amino acid balance, which is driven by accelerated muscle amino acid efflux. Acutely elevated NEFA availability worsened muscle insulin resistance without affecting amino acid kinetics, suggesting increased muscle NEFA uptake may contribute to inactivity-induced insulin resistance but does not cause anabolic resistance.NEW & NOTEWORTHY We demonstrate that 2 days of forearm cast immobilization in healthy young volunteers leads to the rapid development of insulin resistance, which is accompanied by accelerated muscle amino acid efflux in the absence of impaired muscle amino acid uptake. Acutely elevated fasting nonesterified fatty acid (NEFA) availability as a result of acipimox supplementation worsened muscle insulin resistance without affecting amino acid kinetics, suggesting increased muscle NEFA uptake may contribute to inactivity-induced insulin resistance but does not cause anabolic resistance.

Herd-level associations between the proportion of elevated prepartum nonesterified fatty acid concentrations and postpartum diseases, reproduction, or culling on dairy farms

The objectives of this herd-level prospective observational cohort study were to describe the proportion of cows with elevated prepartum nonesterified fatty acid concentrations (PropElevNEFA) in dairy herds and to assess the herd-level associations between PropElevNEFA and postpartum diseases, reproductive performance, and culling. From November 2018 to December 2020, a convenience sample of 49 herds was enrolled in this study. Blood sampling (16 to 29 cows per herd) was performed during the week before and during the 2 wk following calving to quantify the concentration of nonesterified fatty acids (NEFA) and β-hydroxybutyrate acids (BHBA), respectively. Elevated NEFA was defined as ≥280 µmol/L and hyperketonemia as BHBA ≥1.4 mmol/L. Retained placenta, metritis, purulent vaginal discharge, endometritis, and mastitis were diagnosed on-farm following standardized definitions, and success at first artificial insemination (AI) and culling events were recorded. The associations between PropElevNEFA and each individual disease, success at first AI, and culling were evaluated using Bayesian aggregated binomial regression models with weakly informative priors, from the which odds ratio (OR) and the 95% credible intervals (BCI) were obtained. A total of 981 cows were included in the statistical analyses representing 16 to 29 (median = 19) cows per herd. Cows were enrolled in the prepartum period of their first to tenth (median = third) lactation, and 41% of them had an elevated prepartum NEFA concentration. At the herd level, PropElevNEFA varied between 11% and 78% (median = 39%). The odds of metritis (OR = 1.37, 95% BCI = 1.13–1.67) increased for every 10-point increase in PropElevNEFA, whereas the odds of success at first AI decreased (OR = 0.69, 95% BCI = 0.59–0.80). The PropElevNEFA was not associated with the other tested diseases or culling. Our results suggest that the herd-level proportion of cows having elevated prepartum NEFA concentrations is associated with metritis and poor success at first AI in dairy herds.

Investigation of the Alternations in Lipid Oxidation and Lipase Activity in Air-Dried Hairtail (Trichiurus lepturus) during Chilled Storage.

The effects of water content and water activity on the lipid stability of air-dried hairtail (Trichiurus haumela) were investigated during chilled storage. Air-dried hairtail samples with high and low water contents were comparatively analyzed over 8 days of storage at 4 °C. The results indicated that the decreases in water activity and increases in the NaCl content significantly inhibited lipid oxidation in the air-dried hairtail samples. The peroxidation value (PV), conjugated diene value (CD), thiobarbituric acid reactive substance (TBARS) value, and p-anisidine value (p-AnV) of the air-dried hairtail significantly increased with the extension of storage time. The low water content significantly inhibited the activity of neutral and alkaline lipase, in addition to lipoxygenase, and retarded the rapid increases in the non-esterified fatty acid (NEFA) content in the hairtail samples. The correlation analysis results showed that the TBARS, p-AnV, and lipase activity were positively correlated in the air-dried hairtail samples, and the lower water content significantly inhibited the progress of lipid oxidation. This study offers a theoretical framework for the industrial processing and storage of air-dried hairtail products.