NEFA Levels Research Articles

Effect of supplementation of phytoadditive mixtures on live weight, blood indices, hormonal profile and reproductive performance in Black Bengal goats

The study was conducted to evaluate the effect of phytoadditive mixtures containing equal proportions of Curcuma longa & Allium sativum (PAM-1) and Curcuma longa & Ocimum sanctum (PAM-2) on live weight, blood parameters, hormonal profile and reproductive performance in Black Bengal goats. A total of 21 post weaned female kids of 3–4 months age and similar body weights maintained at Goat Research Unit of Eastern Regional Station (ERS), National Dairy Research Institute (NDRI), Kalyani, India during the year 2021–2022 were randomly distributed in three groups namely, Control, PAM-1 and PAM-2 for 10 months. Blood parameters i.e. glucose, urea, total protein, albumin, globulin, ALT, AST, ALP, total lipids, cholesterol, and Triglycerides, WBC, RBC, Hb and PCV %; NEFA levels and hormones i.e. prolactin, cortisol, T3 and T4 were estimated in all three experimental groups. Higher (P < 0.001) live weights were investigated in supplemented goats. Blood parameters including glucose, urea, total protein, albumin, globulin, ALT, AST, ALP, WBC, RBC, Hb and PCV% were similar (P > 0.05) in all groups. Lower (P < 0.001) blood lipids were found in supplemented groups. Higher Prolactin, T3 and T4 levels and lower Cortisol levels were reported in PAM-1 and PAM-2 groups (P < 0.001) than control group. Age at first estrus, first conception, first kidding and post kidding first estrus days were lower (P < 0.001) in supplemented groups. Therefore, it was concluded that supplementation of PAM-1 and PAM-2 improved reproductive performance, hormonal profile and lipid profile through improved live weights in Black Bengal goats.

The role of the hypothalamus-gut microbiota in the pathogenesis of periparturient fatty liver disease in dairy cows.

During the periparturient period, dairy cows experience negative energy balance due to reduced feed intake, leading to adipose tissue breakdown, liver damage, and fat accumulation. This study examined the gut-liver-brain axis to explore the link between fatty liver disease, changes in hypothalamic appetite-related neurons, and microbiome shifts in dairy cows. Thirty cows were monitored, with daily DMI recordings and blood sampling. Postpartum brain, liver, and ileal contents were collected from 10 selected cows, divided into two groups: H-DMI (slight DMI decrease) and L-DMI (severe DMI decrease). The L-DMI group of cows exhibited higher plasma NEFA, BHBA, ALT, and AST levels, along with severe hepatic steatosis and lipid accumulation. Transcriptome sequencing of the hypothalamic arcuate nucleus (ARC) revealed decreased expression of Hypocretin Neuropeptide Precursor (HCRT), orexin-A (OX-A), Orexin Receptor Type 1 (OX1R), and Cannabinoid Receptor 1 (CB1) in the L-DMI group, while Pro-opiomelanocortin (POMC) and Melanocortin 4 Receptor (MC4R) expression increased. Metagenomic analysis of ileal contents showed reduced abundance of Ruminococcus spp. in the L-DMI group, which may be associated with fatty liver disease (FL). Integrated omics analysis showed that increased MC4R expression was correlated with the elevated abundance of bacteria such as Akkermansia glycaniphila, and reduced abundance of species such as Methanobrevubacter thaueri and Ruminococcus spp. Decreased HCRT expression was also linked to Akkermansia glycaniphila. In conclusion, these changes may affect DMI through the OX-A/POMC pathway, with neurological and gut microbiome alterations potentially leading to appetite suppression, negative energy balance, and the development of fatty liver disease.

Therapeutic Potential of Asparagus Racemosus Root in Managing Hyperglycemia, Dyslipidemia, and Platelet Aggregation in Neonatal Streptozotocin-induced Type 2 Diabetic Rats

Lipid abnormalities frequently accompany hyperglycemia and thus a primary goal in diabetes therapy is the management of dyslipidemia. Asparagus racemosus root has previously been shown to reduce postprandial blood glucose in diabetic rats by delaying carbohydrate absorption and enhancing insulin secretion. In the present study, the chronic effects of A racemosus root on serum glucose, fructosamine, lipids, and platelet aggregation were assessed in rats with type 2 diabetes induced by neonatal streptozotocin injection. The type 2 diabetes model was created by injecting 48-hour-old pups with a single intraperitoneal dose of streptozotocin (STZ). Platelet aggregation was measured by optical aggregometry. Daily oral administration of ethanol extract of A racemosus to diabetic rats (n = 10) lowered serum glucose by 21% (p&lt;0.01) and fructosamine by 11% (p&lt;0.05) after 28 days. Total cholesterol (p&lt;0.05), triglyceride (p&lt;0.05), and NEFA (p&lt;0.01) levels were also lowered by 9%, 16% and 38% respectively. No difference in HDL cholesterol or body weights was observed compared to control rats but platelet aggregation was significantly reduced by 18% (p&lt;0.05). Food and water intake, stool formation, water content of stools, and urine formation were unchanged in extract-treated rats in a 24-hour acute observational study in Nalgene Metabolic Cages. In conclusion, this study reveals that ethanol extract of A racemosus root lowers circulating glucose, and atherogenic blood lipids and decreases platelet aggregation. Thus, A racemosus is the source of glucose-lowering bioactive agents and a useful dietary adjunct in the management of diabetes, dyslipidemia, and related complications.

Interpretation of 25-OH-D3, NEFA, and calcium correlations among cow and calf pairs

In the present study, the aim was to determine alteration of NEFA, calcium and vitamin D3 levels in cow and calf pairs at parturition as well as correlation between each parameter levels. For this purpose, a cow-side device employing the enzymatic colorimetric method was used for measurement of NEFA and calcium levels. On the other hand, serum 25(OH)D3 analysis was performed using the fluorescence immunochromatographic method at the laboratory in the Faculty. Blood samples were taken from Vena jugularis of 15 Simmental cow and calf pairs immediately after parturition and placed in serum and heparinized tubes. In cow and calf pairs, concentration of vitamin D3 (15.6-120 and 31.8-120 ng/mL, respectively), NEFA (0.12-1.2 and 0.09-0.8 mmol/L, respectively) and calcium (1.8 ± 0.9 and 2.2 ± 0.6 mmol/L, respectively) were determined. There was no significant correlation between NEFA, Ca, and vitamin D3 in cows and calf pairs. Taking into account several co-factors that influenced test results, which could not easily be excluded, further studies may be warranted with larger cow-calf pair populations. In conclusion, vitamin D3 concentration in calves is not affected by the negative energy balance of dams in the parturition period.

M1 polarization of hepatic macrophages in cows with subclinical ketosis is an important cause of liver injury.

Subclinical ketosis (SCK) is highly prevalent and easily overlooked, with insidious and slow progression of hepatic injury, often characterized by an imbalance in immune homeostasis. In nonruminants, macrophage polarization plays an important regulatory role in hepatic lipid accumulation, fibrosis and inflammatory processes. Thus, we aimed to investigate the status of hepatic macrophage polarization in SCK cows and to corroborate its association with liver injury and inflammation. Twelve Holstein dairy cows (parity, 2-4) were selected, and liver biopsy and blood were collected on the second week postpartum (10-14 d days in milk). On the basis of serum BHBA concentrations., selected cows were categorized into healthy (n = 6; BHBA <1.0 mM) and SCK (n = 6; 1.2 mM ≤ BHBA < 3.0 mM) groups. Serum biochemical parameters were measured using an automatic biochemical analyzer, which indicated higher serum levels of BHBA and NEFA and an upregulation of liver injury indicators (AST, ALT, TP, GLB) in SCK cows compared with healthy cows. ELISA assays revealed that SCK cows displayed systemic low-grade inflammation, as demonstrated by increased serum levels of HP, SAA, TGF-β, IFN-γ, and IL-1β. Liver biopsies revealed pathological histological alterations, hepatic inflammation, and macrophage polarization status. Oil red staining indicated steatosis, while Sirius red staining demonstrated mild extracellular matrix deposition in the liver of SCK cows. The expression of inflammatory response-related proteins (TLR4, p-NFκB, p-I-κB, NLRP3, and Caspase-1) was elevated in the liver of SCK cows, with the increased mean fluorescence intensity of NFκB further confirming the activation of the inflammatory pathway. Furthermore, the levels of pro-inflammatory factors, TNF-α and IFN-γ, were elevated in the tissue homogenate. Macrophage phenotypic changes in SCK cows were further explored based on the results of liver injury and inflammation. Compared with healthy cows, the protein and mRNA abundance of the macrophage marker CD68 in the liver of SCK cows was higher, along with an increased mean fluorescence intensity of CD68. SCK cows also exhibited reduced mRNA expression of the Kupffer cell marker CLEC4F and elevated chemokine levels (CXCL1 and CCL2). As evidenced by greater protein and mRNA abundance of macrophage M1 polarization markers (iNOS, IL-1β, CD86, IL-6, IL-12b, and CCL3), higher fluorescence intensity of iNOS and CD86, and an increased number of CD68+/CD86+-positive cells observed via immunofluorescence, the macrophage polarization phenotype in the liver of SCK cows was predominantly M1. In contrast, the protein and mRNA abundances of M2 polarization markers (CD206, IL-10, and Arg1) were lower in SCK cows, accompanied by a reduced fluorescence intensity of CD206 and a lower number of CD68+/CD206+-positive cells. Overall, the present study revealed that the number of macrophages in liver is enhanced during subclinical ketosis and is dominated by pro-inflammatory macrophages (M1 macrophages). This could partly explain the increased risk of steatosis, fibrosis, and inflammatory response processes in these cows.

Largemouth bronze gudgeon (Coreius guichienoti) responds positively to liver damage induced by high feeding rates via enhancing self-repair capability

Overfeeding intermittently induces metabolic liver diseases in aquaculture practice. Currently, less information regarding the impact of feeding rates on hepatic lipid metabolism was reported. Therefore, this study investigated the effects of feeding rates (2 %, 3 %, 4 %, and 5 % of body weight per day) on growth performance, hepatic lipid metabolism, immunity, inflammation, and apoptosis in juvenile largemouth bronze gudgeon (Coreius guichenoti) over 8 weeks. The results demonstrated that increasing feeding rates significantly enhanced weight gain rate and specific growth rate. Furthermore, the lipid content of the whole body increased progressively, with the highest levels observed in the 5 % group. High feeding rates also resulted in significant increases in hepatic TG, TC, and NEFA levels. This was accompanied by upregulated mRNA levels and enzyme activities of lipid synthesis-related genes (ACC1α and FASN). However, no significant differences were observed in mRNA levels and enzyme activities of lipolysis-related genes (HSL and CPT1). Histological observations identified two distinct hepatic phenotypes: no obvious abnormality liver and fatty liver. Overall, the proportion of fatty liver phenotype increased with the increasing feeding rate. Additionally, feeding rates significantly increased IgG, IgM, and lysozyme levels, and upregulated mRNA levels of inflammatory cytokines (TNFα and IL1β), apoptosis-related genes (CASP3, CASP8, and CASP9), and hepatic repair factors (YAP1, FGF10, FGF13, FGF18, FGF19, and PCNA). Nevertheless, hepatic function indices, including ALT, AST, and AKP levels, remained unchanged. In conclusion, these findings indicated that a feeding rate exceeding 3 % led to hepatic lipid metabolism disorders, characterized by the accumulation of TG and TC, thus promoting the development of fatty liver, inflammation response, and cellular apoptosis in largemouth bronze gudgeon. Moreover, the largemouth bronze gudgeon may exhibit self-repair capability to mitigate liver damage induced by lipid metabolism disorders. Consequently, we infer that largemouth bronze gudgeon possesses a robust capability for lipid accumulation and utilization.

Gender and age-related variations in rumen fermentation and microbiota of Qinchuan cattle.

Our study aimed to investigate the gender and age-related variations in rumen fermentation, serum metabolites, and microbiota in Qinchuan cattle. A total of 38 Qinchuan beef cattle were selected and maintained on a uniform diet for three months. Rumen fluid and blood samples were collected to determine rumen fermentation, serum metabolites, and microbial 16S rRNA sequencing. The results revealed that the concentration of rumen butyrate in female Qinchuan cattle was significantly higher than in males (P<0.05). Isobutyrate, butyrate, and isovalerate exhibited significant age-related differences. Females exhibited lower serum GLU and higher TG, NEFA levels compared to males (P<0.05). Serum ALB and UREA levels increased with age (P<0.05). Furthermore, the alpha diversity of rumen bacteria improved with age (P<0.05), with no gender differences observed. Males had higher relative abundances of Bacteroidota, Verrucomicrobiota, and Cyanobacteria, while females had higher Firmicutes and Desulfobacterota (P<0.05). The cellulose-degrading genus Ruminococcus and propionate-producing genus Succiniclasticum were more abundant in females, whereas the anti-inflammatory genus Lachnospiraceae_NK4A136_group and the hemicellulose-degrading genus Prevotella were more abundant in males (P<0.05). Age-related differences in bacteria were found in Pseudobutyrivibrio and several members of the Lachnospiraceae. Functional prediction indicated that "Amino acid metabolism" and "Lipid metabolism" were mainly enriched in females, whereas "Carbohydrate metabolism" and "Glycan biosynthesis and metabolism" were enriched in males (P<0.05). RDA analysis highlighted butyrate as a key factor influencing the rumen bacterial community. NK4A214_group and Ruminococcus were positively correlated with butyrate, while Prevotella and Pseudobutyrivibrio were negatively correlated with butyrate (P<0.05). We observed a significant improvement in the diversity and stability of rumen microbiota as age increased. Ruminococcus, NK4A214_group, and Prevotella were likely contributors to variations in energy utilization and fat deposition between male and female Qinchuan cattle.

Mulberry galacto-oligosaccharides ameliorates non-alcoholic fatty liver disease in mice induced by a high-fat diet

A type of oligosaccharide named mulberry galacto-oligosaccharides (MGO), was isolated from mulberry with a galactose-only, weighs 987 Da and displays diverse biological properties. The objective of this work was to investigate the protective effects of MGO in non-alcoholic fatty liver disease (NAFLD) mice. The results showed that MGO can reduce the body weight and liver weight of NAFLD mice, and reduce the levels of NEFA, TG, TC and LDL-C in serum and liver as well as the activities of AST and ALT. Increase HDL-C content, SOD, and GSH activity in serum and improve pathological damage. MGO regulates the AMPK lipid metabolism pathway, upregulates the expression of ATGL, downregulates the expression of SREBP-1, inhibits lipogenesis, and promotes lipolysis. MGO alleviates liver function damage in NAFLD mice, improves blood lipids, liver metabolism, and increases the body’s antioxidant capacity, suggesting MGO potential as a dietary supplement and functional food to combat HFD-induced NAFLD.

Effect of Gestational Direct-Fed Microbials Supplementation on the Metabolic Profile in Periparturient Dairy Cows.

This study was conducted at a commercial dairy farm in Queensland, Australia to evaluate the effects of feeding a lactic acid bacteria-based direct-fed microbial (DFM) during gestation on the metabolic profile of periparturient dairy cows and its effects on milk production and body weight. A total of 150 multiparous Holstein cows were randomly selected based on parity (2.3) and days in milk (130 DIM) and divided into two groups of 75 cows each (control and DFM). The control cows were assigned to a basal diet consisting of a silage-based partial mixed ration (PMR), concentrate fed in the dairy twice a day, and ad libitum pasture. The DFM group received the same basal diet supplemented with three strains of Lactobacillus top-dressed in the feed. The DFM supplementation continued during both the dry period and the subsequent lactation. A subset of 82 cows (40 control and 42 DFM) were monitored during the calving season (March to July 2022) to assess the metabolic profile and postpartum performance. Blood samples were collected during the periparturient period (-4 to -2 w prepartum, around calving, and at weeks 1, 3, and 6 postpartum) to measure the levels of metabolites, enzymes, and minerals. Overall, the serum glucose, NEFA, and chloride levels were higher, while protein and urea were lower in cows supplemented with the DFM (p < 0.005). The pre-calving levels of glucose were higher and the total bilirubin, urea, and BHB were lower in cows supplemented with DFM than in the control (p < 0.05). The post-calving levels of glucose and Mg were also higher in the DFM cows than in the control cows (p < 0.05). Average milk production at 110 DIM was significantly higher in the DFM cows compared to control cows (p = 0.03). Although the total milk production over 305 days was numerically greater in the DFM cows, the difference was not statistically significant (p = 0.3), whereas the milk protein percentage was higher in the control cows (p = 0.03). The body weight of the DFM cows was greater during the periparturient period (p = 0.001) than that of the control cows. In the DFM cows, glucose levels had a positive correlation (r = 0.16) with milk yield, at 110 DIM, while serum total protein had a positive correlation with body weight (r = 0.32) (p < 0.05). In conclusion, feeding Lactobacillus-based DFM during gestation can positively influence the metabolic profile of periparturient cows, which, in turn, may affect the milk production and body weight of postpartum dairy cows.

Associations of the rs12504538 and rs6824447 Polymorphisms of the Elovl6 Gene with Estimated Elongase and Desaturase Activity and Fatty Acid Concentrations in Mexican Women with Gestational Diabetes Mellitus

Background: This study aimed to investigate the possible associations of the rs12504538 and rs6824447 polymorphisms of the Elovl6 gene with estimated elongase and desaturase activity and saturated fatty acid concentrations in Mexican women with Gestational Diabetes Mellitus (GDM). Methods: We recruited 172 women in the second and third trimesters of pregnancy who had undergone an oral glucose tolerance test, including 66 who had diagnosed with GDM according to the 2016 ADA criteria and 106 who had normal glucose tolerance test results, from the General Hospital and Health Centers of the Health Ministry of Guanajuato, México. Participants were matched by gestational week. Data on age, gestational week and anthropometric characteristics were collected. Blood samples were drawn after an overnight fast for the measurement of serum glucose, lipid, NEFA, serum fatty acid and insulin levels; SNP genotyping and quantification of fatty acids was performed and elongase and desaturase activity was estimated. Findings: With the exception of HDL-cholesterol, all variables, including NEFA levels, were significantly greater in the GDM group than in the non-GDM group. The rs6824447 polymorphism of the Elovl6 gene, an age &gt;25 years and HOMA-IR levels were associated with the development of GDM (OR=5.1, 95% CI 1.56-17.1, p=0.006; OR=4.89, 95% CI 1.65-14.4, p=0.003; OR=34.1 95% CI 8.7-133, p&lt;0.000001, respectively). Palmitic acid (OR=1.08; 95% CI: 1.06-1.11; p&lt;0.001) and oleic acid (OR=1.17; 95% CI: 1.14-1.2; p&lt;0.001) concentrations were also associated with the development of GDM. No differences in the estimated elongase and desaturase activity among the non-GDM and GDM groups were found and only the activity of Δ9D SCD18 desaturase was marginally high in GDM patients (p=0.050). Conclusion: The rs6824447Elovl6 polymorphism is associated with the development of GDM, as are high serum palmitic acid, oleic acid and stearic acid concentrations and estimated Δ9D SCD18 desaturase activity.

The Effect of Subclinical Ketosis on the Peripheral Blood Mononuclear Cell Inflammatory Response and Its Crosstalk with Depot-Specific Preadipocyte Function in Dairy Cows.

During the periparturient period, cows undergo heightened energy demands at lactation onset, paired with reduced dry matter intake, leading to negative energy balance (NEB). Excessive lipolysis-driven adipose tissue remodeling, triggered by NEB, significantly contributes to ketosis in periparturient dairy cows. However, the role of peripheral blood mononuclear cells (PBMCs) in the pathogenesis of ketosis and in modulating adipose tissue function remains poorly understood. Here, we investigated how ketosis affects the transcriptional profile and secretome of PBMCs and its influence on preadipocyte function in visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT). Twenty-one postpartum Holstein dairy cows were categorized as either subclinical ketosis (SCK; BHB ≥ 1.0 mM) or control (CON; BHB < 0.8 mM) based on blood beta-hydroxybutyrate (BHB) concentration screening. Blood samples were collected intravenously for the isolation of PBMCs and serum metabolic profiling. Ketosis elevated circulating NEFA and BHB levels but reduced total WBC and neutrophil counts. Isolated PBMCs were evaluated for gene expression and used to produce conditioned media (PBMC-CM), during which PBMCs were stimulated with 10 ng/mL LPS. The overall phenotype of PBMCs was largely consistent between SCK and CON cows, with minimal differences detected in immunomodulatory cytokine expression and PBMC-CM composition following stimulation. Preadipocytes isolated from non-ketotic cows were treated with PBMC-CM to assess the effect of PBMC secretomes on adipose cell function. Preadipocytes treated with SCK PBMC-CM showed reduced lipid accumulation compared to those treated with CON PBMC-CM regardless of the depot. SAT preadipocytes had heightened expression of lipid metabolism-related genes, including DGAT1, LIPE, and FASN, compared to VAT when treated with SCK PBMC-CM. Preadipocytes treated with CM from PBMC stimulated by LPS exhibited upregulation in IL1B and IL6 regardless of the depot or source of PBMCs. Together, these results indicate that although PBMC profiles showed minimal differences, preadipocytes treated with PBMC-CM may be influenced by additional factors, leading to altered preadipocyte function and gene expression that may contribute to adipose cellular dysfunction.

DHA-enriched phosphatidylserine alleviates bisphenol A-induced liver injury through regulating glycerophospholipid metabolism and the SIRT1-AMPK pathway

To investigate the alleviating effect and mechanism of the docosahexaenoic acid-enriched phosphatidylserine (DHA-PS) on bisphenol A (BPA)-induced liver injury in mice, the murine liver injury model was established by gavage of BPA (5 mg/kg) or co-administration of BPA and DHA-PS (50 mg/kg or 100 mg/kg) for 6 weeks. The results showed that after administration of 100 mg/kg DHA-PS, the liver index, serum levels of AST, ALT, TC, TG, NEFA, and LDL-C in mice were significantly decreased, while HDL-C was significantly increased. The LPS, IL-6, IL-1β, TNF-α, and MDA levels in liver tissues were effectively down-regulated, and IL-10, SOD, GSH-Px, and CAT levels were effectively up-regulated. The H&E and Oil Red O staining results showed that liver damage was notably repaired and lipid deposition was notably reduced after DHA-PS administration. Furthermore, metabolomics and immunohistochemical studies revealed that DHA-PS mainly regulates glycerophospholipid metabolism and the SIRT1-AMPK pathway to improve metabolic disorders of the liver caused by BPA. Therefore, DHA-PS could potentially alleviate BPA-induced murine liver injury through suppressing inflammation and oxidative stress, and modulating lipid metabolism disorders.

UFL1 regulates cellular homeostasis by targeting endoplasmic reticulum and mitochondria in NEFA-stimulated bovine mammary epithelial cells via the IRE1α/XBP1 pathway

Elevated levels of NEFA caused by negative energy balance in transition cows induce cellular dyshomeostasis. Ubiquitin-like modifier 1 ligating enzyme 1 (UFL1) can maintain cellular homeostasis and act as a critical regulator of stress responses besides functioning in the ubiquitin-like system. The objective of this study was to elucidate the UFL1 working mechanism on promoting cellular adaptations in bovine mammary epithelial cells (BMECs) in response to NEFA challenge, with an emphasis on the ER and mitochondrial function. The results showed that exogenous NEFA and UFL1 depletion resulted in the disorder of ER and mitochondrial homeostasis and the damage of BMEC integrity, overexpression of UFL1 effectively alleviated the NEFA-induced cellular dyshomeostasis. Mechanistically, our study found that UFL1 had a strong interaction with IRE1α and could modulate the IRE1α/XBP1 pathway of unfolded protein response in NEFA-stimulated BMECs, thereby contributing to the modulation of cellular homeostasis. These findings imply that targeting UFL1 may be a therapeutic alternative to relieve NEB-induced metabolic changes in perinatal dairy cows.

Assessing Serum Vaspin Dynamics in Dairy Cows during Late Pregnancy and Early Lactation in Relation to Negative Energy Balance

The periparturient period, which spans late pregnancy to early lactation in dairy cows, is a crucial phase characterized by complex metabolic and endocrine adjustments necessary for sustained milk production. This research focused on the relationship between serum vaspin, inflammatory cytokines (IL-1, TNF), and markers of negative energy balance (NEB) in 100 primiparous and multiparous Holstein dairy cows. The results demonstrated that one month post-calving, both groups had a significant decrease in serum vaspin levels but increased NEFA levels, indicating possible consequences for lipid metabolism and energy balance. Multiparous cows showed significant elevations in cholesterol, IL-1, and TNF concentrations after calving, indicating increased inflammatory responses. Primiparous cows, on the other hand, responded differently, indicating the role of parity in metabolic adjustments. The study acknowledges limitations such as sample size and its observational nature. Future research should investigate the long-term effects of these metabolic changes on herd health and lactational performance, using advanced technologies to gain a molecular understanding. Despite limitations, this study provides valuable insights into how adipokines, inflammatory markers, and energy balance interact during the periparturient period, offering the potential for improved dairy cow management and productivity while ensuring animal welfare.

Effects of antioxidant vitamins (A, D, E) and trace elements (Cu, Mn, Se, Zn) administration on gene expression, metabolic, antioxidants and immunological profiles during transition period in dromedary camels

BackgroundNutrition has a primary role for optimum expression of genetic potential, and most of the farmers have limited resources of green fodder. Hence, a fat-soluble vitamin, especially vitamin A and E and trace elements remained most critical in the animal’s ration and affects their productive and reproductive performance adversely. Animals cannot be able to produce these vitamins in their bodies; hence, an exogenous regular supply is needed to fulfil the physiological needs and to maintain high production performance. This study elucidated effects of antioxidant vitamins (A, D, E) and trace elements (Cu, Mn, Se, Zn) administration on gene expression, metabolic, antioxidants and immunological parameters in dromedary camels during transition period.ResultsAt 0 day, there were no appreciable differences in the expression patterns of the metabolic (IGF-I, ACACA, SCD, FASN, LPL, and BTN1A1) genes between the control and treatment groups, despite lower levels. A substantial variation in the mRNA levels of SOD1, SOD3, PRDX2, PRDX3, PRDX4, PRDX6, and AhpC/TSA was observed between the control and treatment groups, according to the antioxidant markers. In comparison to the control group, the treatment group displayed a significant up-regulation at 0 and 21 days. The treatment and control groups exhibited substantial differences in the mRNA values of IL-1α, IL-1β, IL-6, and TNFα, as indicated by immunological markers. In comparison to the control group, there was a noticeable down-regulation in the treatment group at 0 and + 21 days. But IL10 produced the opposite pattern. No significant difference was observed in glucose, cholesterol, triglyceride, HDL, total protein, NEFA, BHBA, cortisol and IGF-1 levels between control and treatment group. The activity of serum GPx, SOD and TAC was significantly affected by time and treatment x time in supplemented groups as compared with control group. IL-1, IL-1, IL-6, and TNF were noticeably greater in the control group and lower in the treatment group. Additionally, in all groups, the concentration of all pro-inflammatory cytokines peaked on the day of delivery and its lowest levels showed on day 21 following calving. The IL-10 level was at its peak 21 days prior to calving and was lowest on calving day.ConclusionThe results demonstrated a beneficial effect of antioxidant vitamins and trace elements on the metabolic, antioxidant and immunological markers in dromedary camels throughout their transition period.

Effect of a Dietary Essential Oil Blend in Dairy Cows during the Dry and Transition Period on Blood and Metabolic Parameters of Dams and Their Calves.

Cows face severe challenges of immunosuppression and negative energy balance during transition periods. The current study was designed to investigate the effects of essential oil blend supplementation in dairy cow's ration during dry periods on the health of the cow and calf. In the current study, 45 dry Holstein dairy cows were divided into three groups, each consisting of 15 animals. The control group was without any supplementation; the positive control group was only injected with 0.2 mg/kg levamisole (intramuscular) at 2 months before parturition and 1 month before parturition; and the treatment group was supplemented with 3 g/day for each cow essential oil blend mixed in total mixed ration (TMR). A mixed vaccine against E. coli, Rotavirus, and Coronavirus was administered to all cows before 42 days and after 21 days of calving. The day of the calving of the animal was accepted as day 0. Blood samples were collected from the coccygeal vein of all cows at -60, -45, -30, -15, -7, 0, 1 and 2 days, and the colostrum samples were taken on days 0, 1, and 2. Blood samples were also collected from the jugular vein (V. Jugularis) of the newborn calves on days 0, 1, and 2. The results of the hematological parameters revealed no difference in the total red blood cell count, hemoglobin amount, hematocrit, MCV, MCH, MCHC, RDW, PLT, MPV, and PCT values in both prenatal and postnatal blood of the cows (p > 0.05). In terms of immunological parameters, the total IgG level was significantly higher in the experimental group on the 7th day before birth compared to the other two groups, while the total leukocyte count, lymphocyte count, monocyte count and granulocyte counts were found to be lower after birth (p < 0005). Blood NEFA and BHBA levels were also lower in the experimental group compared to the other two groups (p < 0.005), and the blood glucose level was not different (p > 0.05). In calves, no difference was found between the groups in any of the parameters examined (p > 0.05). It is concluded that the dietary addition of an essential oil blend during the transition period enhanced the immune status and energy metabolism of cows without any effect on the health status of newborn calves.

Analysis of the relationship between energy balance and properties of rumen fermentation of primiparous dairy cows during the perinatal period.

Short-chain fatty acids (SCFAs) produced in the rumen are key factors affecting dairy cows' energy balance (EB). This study aimed to quantitatively evaluate the effects of SCFAs production on EB in dairy cows. Primiparous dairy cows were divided into high non-esterified fatty acid (NEFA; group H) and low NEFA (group L) groups based on their blood NEFA levels at week 3 postpartum, which served as an indicator of EB. The amounts of SCFAs produced in the rumen, including acetate, propionate, and butyrate (SCFAsP), were calculated using the predicted rumen volume. Because there were no differences between the groups in SCFAsP/dry matter intake, whereas 4% fat-corrected milk (FCM)/SCFAsP was significantly higher in group H, it was suggested that more body fat was mobilized for milk production in group H. However, group L, which showed better EB, had propionate dominant and lower FCM/SCFAsP and milk energy/SCFAs energy at 3 and 7weeks postpartum, indicating that group L had a better energy supply for milk production. These results suggest that SCFAsP produced by rumen fermentation and the composition of SCFAs in the rumen affect milk production and EB.

The determinants of fasting and post-load non-esterified fatty acids in older adults: The cardiovascular health study

AimNon-esterified fatty acids (NEFA) are potential targets for prevention of key cardiometabolic diseases of aging, but their population-level correlates remain uncertain. We sought to identify modifiable factors associated with fasting and post-load NEFA levels in older adults. MethodsWe used linear regression to determine the cross-sectional associations of demographic, anthropometric, and lifestyle characteristics and medication use with serum fasting and post-load NEFA concentrations amongst community-dwelling older adults enrolled in the Cardiovascular Health Study (n = 1924). ResultsFasting NEFA levels generally demonstrated a broader set of determinants, while post-load NEFA were more consistently associated with metabolic factors. Waist circumference and weight were associated with higher fasting and post-load NEFA. Cigarette smoking and caffeine intake were associated with lower levels of both species, and moderate alcohol intake was associated with higher fasting levels whereas greater consumption was associated with lower post-load levels. Unique factors associated with higher fasting NEFA included female sex, higher age, loop and thiazide diuretic use and calcium intake, while factors associated with lower fasting levels included higher educational attainment, beta-blocker use, and protein intake. Hours spent sleeping during the daytime were associated with higher post-load NEFA, while DASH score was associated with lower levels. ConclusionFasting and post-load NEFA have both common and unique modifiable risk factors, including sociodemographics, anthropometric, medications, and diet. Post-load NEFA were particularly sensitive to metabolic factors, while a broader range of determinants were associated with fasting levels. These factors warrant study as targets for lowering levels of NEFA in older adults.

In-Line Registered Milk Fat-to-Protein Ratio for the Assessment of Metabolic Status in Dairy Cows.

This study endeavors to ascertain alterations in the in-line registered milk fat-to-protein ratio as a potential indicator for evaluating the metabolic status of dairy cows. Over the study period, farm visits occurred biweekly on consistent days, during which milk composition (specifically fat and protein) was measured using a BROLIS HerdLine in-line milk analyzer (Brolis Sensor Technology, Vilnius, Lithuania). Clinical examinations were performed at the same time as the farm visits. Blood was drawn into anticoagulant-free evacuated tubes to measure the activities of GGT and AST and albumin concentrations. NEFA levels were assessed using a wet chemistry analyzer. Using the MediSense and FreeStyle Optium H systems, blood samples from the ear were used to measure the levels of BHBA and glucose in plasma. Daily blood samples were collected for BHBA concentration assessment. All samples were procured during the clinical evaluations. The cows were categorized into distinct groups: subclinical ketosis (SCK; n = 62), exhibiting elevated milk F/P ratios without concurrent clinical signs of other post-calving diseases; subclinical acidosis (SCA; n = 14), characterized by low F/P ratios (<1.2), severe diarrhea, and nondigestive food remnants in feces, while being free of other post-calving ailments; and a healthy group (H; n = 20), comprising cows with no clinical indications of illness and an average milk F/P ratio of 1.2. The milk fat-to-protein ratios were notably higher in SCK cows, averaging 1.66 (±0.29; p < 0.01), compared to SCA cows (0.93 ± 0.1; p < 0.01) and healthy cows (1.22). A 36% increase in milk fat-to-protein ratio was observed in SCK cows, while SCA cows displayed a 23.77% decrease. Significant differences emerged in AST activity, with SCA cows presenting a 26.66% elevation (p < 0.05) compared to healthy cows. Moreover, SCK cows exhibited a 40.38% higher NEFA concentration (p < 0.001). A positive correlation was identified between blood BHBA and NEFA levels (r = 0.321, p < 0.01), as well as a negative association between BHBA and glucose concentrations (r = -0.330, p < 0.01). Notably, AST displayed a robust positive correlation with GGT (r = 0.623, p < 0.01). In light of these findings, this study posits that milk fat-to-protein ratio comparisons could serve as a non-invasive indicator of metabolic health in cows. The connections between milk characteristics and blood biochemical markers of lipolysis and ketogenesis suggest that these markers can be used to check the metabolic status of dairy cows on a regular basis.

THU047 Growth Hormone (GH) Signaling In Hepatocytes Controls Hepatic Insulin Sensitivity And Nutrient Disposition

Abstract Disclosure: M. del Rio-Moreno: None. M. Vazquez-Borrego: None. J. Cordoba-Chacon: None. O.P. McGuinness: None. R.D. Kineman: None. It is commonly accepted that GH reduces the actions of insulin. However, we reported plasma GH levels within a physiological range are negatively associated with insulin-mediated glucose uptake, but positively associated with insulin-mediated suppression of hepatic glucose production (HGP) [Cordoba-Chacon J. et al. 2014. AJP Endocrinol Metab]. These findings suggest that GH may promote insulin’s actions on the liver. To determine the direct contribution of GH signaling on hepatic insulin sensitivity, hyperinsulinemic-euglycemic clamps were performed with stable isotope tracers in chow-fed adult-onset, hepatocyte-specific GHR knockdown (aHepGHRkd) mice. aHepGHRkd mice show reduced IGF1 and increased GH levels [Cordoba-Chacon J. et al 2015, Diabetes] which indirectly impacts systemic insulin sensitivity and substrate flux to the liver. To normalize plasma IGF1 and GH levels, aHepGHRkd mice were co-treated with adeno-associated virus (AAV) that express IGF1 or a constitutively active form of STAT5b (STAT5bCA). These AAVs have a thyroxin binding globulin promoter, leading to hepatocyte-specific expression. Under both basal and clamped conditions, aHepGHRkd mice exhibited hyperinsulinemia, that was normalized by AAV-mediated IGF1 and STAT5bCA expression. Interestingly, glucose disappearance (Rd), tissue-specific glucose uptake, or HGP (gluconeogenesis and glycogenolysis) did not differ across all groups. However, higher insulin tone in aHepGHRkd mice was required to achieve the same level of Rd and suppression of HGP in the steady-state, which was indicative of systemic and hepatic insulin resistance. These observations are consistent with our previous findings that GH reduces tissue-specific glucose uptake but promotes insulin-mediated control of HGP. In contrast, aHepGHRkd mice exhibited an increased rate of hepatic de novo lipogenesis, and an increase in plasma levels of newly formed triglyceride and triglyceride-bound palmitate, that was normalized by IGF1 and STAT5bCA expression. Taken together, these results indicate loss of hepatocyte GH signaling leads to pathway selective hepatic insulin resistance. This pathway selective hepatic insulin resistance has also been reported in mouse models due to altered substrate delivery to the liver as consequence of excess dietary intake and/or increased adipose tissue lipolysis. However, chow-fed aHepGHRkd mice show hyperinsulinemia that was sufficient to normalize systemic glucose uptake and suppress WAT lipolysis, as noted by normal plasma NEFA levels under both basal and clamped conditions. Therefore, changes in substrate delivery to the liver cannot readily explain the changes observed, suggesting hepatocyte GHR-signaling plays a direct role in partitioning of hepatic nutrients in response to insulin. Presentation: Thursday, June 15, 2023