Higher Milk Fat Research Articles

Association of STAT1 gene with milk fat and protein yield in Holstein Friesian crossbred cattle maintained in the sub-tropical climate of India.

Signal transducers and activators of transcription (STAT) genes are involved in signal mediation of various hormones and cytokines. STAT1 located on chromosome number 2 is involved in mammary gland development and is associated with milk composition traits in bovines. This study aimed to find any relationship and impact of STAT1/BspHI gene with milk fat and protein yields in a herd of Holstein Friesian (HF) crossbred cattle of sub-tropical climate of Northern India. Milk composition data of 535 adult HF crossbred cows for a period of 12 years was collected from the records maintained at Livestock Farm, Guru Angad Dev Veterinary and Animal Sciences University. First lactation data of 222 animals was chosen for further analysis. After data correction for non-genetic factors (season of calving, period of calving, interaction effect of season and period of calving and age at first calving) these animals were categorised into two groups based on corrected high and low milk fat and protein yields. Forty animals were then selected for blood collection and further laboratory analysis. Amplified using PCR-RFLP technique, the 314 bp STAT1 gene was digested using BspHI restriction enzyme. C-T polymorphism at nucleotide position 201 and 260 of the STAT1 amplicon was observed. At 201, for genotype AA and Aa, the genotypic frequencies were 0.80 and 0.20%. At 260, for genotype BB and Bb, the genotypic frequencies were 0.25 and 0.75%. Least square analysis showed a significant association of all genotypes with milk fat and protein yields. Hence, STAT1 can be used as a potential candidate gene to aid in better animal selection in breeding programmes.

Bta-miR-224 regulates milk fat metabolism by targeting FABP4 in bovine mammary epithelial cells

Milk fat is produced and secreted by the mammary gland, which is mainly regulated by diet and gene-molecule network. Therefore, understanding the molecular mechanism of milk fat synthesis is of practical significance for improving milk quality. Fatty acid-binding protein 4 (FABP4) is a candidate messenger RNA (mRNA) closely linked to milk fat metabolism obtained from transcriptomic analysis of mammary epithelial cells of cows in the pre-existing high- and low-milk-fat groups, and its expression pattern and function are still unclear. The qRT-PCR results depicted that FABP4 was highly expressed in bovine mammary epithelial cells (BMECs) in breast tissues and the high milk fat group. Subsequently, the regulatory effects of FABP4 on BMECs were analyzed by CCK8, EdU, and flow cytometry, and the results demonstrated that FABP4 inhibited the proliferation and viability of BMECs and promoted their apoptosis. Next, the effect of FABP4 on milk lipid metabolism was explored. pEGFP-N1-FABP4 was transfected into BMECs, and FABP4 upregulated the expression levels of the milk lipid marker genes XDH, PPARG, and ACSS2, and promoted the formation of triglycerides (TGs), cholesterol, lipid droplets, and β-casein. Strong interactions between FABP4 and PPARG were identified using STRING prediction. Western blotting revealed that FABP4 interacted with PPARG to promote PPARG expression, while the opposite result was observed after interfering with FABP4. The gene regulation of microRNA (miRNA) is essential for fatty acid metabolism and synthesis. Predicted by website and combined with pre-miRNA transcriptome sequencing results, we hypothesized that FABP4 might be the target gene of bta-miR-224. The results of the dual-luciferase reporter gene and qRT-PCR revealed that bta-miR-224 negatively regulated FABP4 expression by targeting the 3′-UTR of FABP4. By exploring the function of bta-miR-224, we observed that bta-miR-224 mimics downregulated the expression of the milk fat marker genes AGPAT6, ACSS2, and XDH and inhibited TG synthesis and lipid droplet secretion. However, the bta-miR-224 inhibitor depicted the opposite results. In conclusion, FABP4 plays a crucial role in regulating BMEC proliferation and differentiation. Bta-miR-224 targeting FABP4 may promote biological processes such as TG synthesis and lipid droplet formation through PPARG, which lays a solid foundation for further analysis of the functional mechanism of milk lipid metabolism in dairy cows from a miRNA-mRNA perspective.

Dairy consumption and risk of prediabetes and type 2 diabetes in the Fenland study

Background & AimsLimited observational evidence suggests that a higher intake of high-fat dairy may be associated with lower prediabetes risk, while opposite associations have been observed for low-fat milk intake. This study aimed to examine associations between baseline and changes in dairy consumption, risk of prediabetes, and glycaemic status. Methods7,521 participants from the prospective UK Fenland study were included (mean age 48.7±2.0 years, 51.9% female). Dairy intake was measured using self-reported food frequency questionnaires. Associations with prediabetes risk and glycaemic status were analysed using Poisson regression models adjusted for social demographics, health behaviours, family history of diabetes and food group intake. ResultsAt a mean follow-up of 6.7±2.0 years, 290 participants developed prediabetes (4.3%). Most dairy products were not significantly associated with prediabetes risk. A higher baseline intake of high-fat dairy (RRservings/day 1.20, 95%CI 1.03-1.39) and high-fat milk (RRservings/day 1.22, 1.01-1.47) were associated with higher prediabetes risk. Conversely, low-fat milk was associated with lower prediabetes risk (RRservings/day 0.86, 0.75-0.98). In the analyses evaluating dietary changes over time, increases in high-fat milk were inversely associated with risk of progressing from normoglycaemia to prediabetes or type 2 diabetes. ConclusionsThis population-based study showed that most dairy products not associated with prediabetes risk or progression in glycaemic status. Positive associations of high-fat dairy, high-fat milk, and the inverse association of low-fat milk with prediabetes risk found were inconsistent with prior literature and suggestive of the need for future research on environmental, behavioural, and biological factors that explain the available evidence.

How dairy food affects prostate cancer - a literature overview

Introduction: Prostate cancer is one of the most common type of tumors. Potential factors influencing this neoplasm are subject of many scientific studies as a scarce number of risk variables have been well-established so far. Purpose of the work: The objective of this review is to collect and interpret recent findings regarding dairy food and its influence on prostate cancer. Materials and method: A systematic research was conducted using the PubMed database. Articles in English published from 2015 to 2024 were selected using keywords: dairy products, dairy, milk, prostate cancer. State of knowledge: There is an overall positive association between total dairy products, milk and prostate cancer risk. Although whole milk is not considered as a risk factor, it has been reported that high-fat milk has a positive association with fatality rate and recurrence. Several studies presented a new potential pathomechanisms of dairy’s influence on prostate tumors. Casein, IGF-1 and exosomes are labeled as mediators between high dairy food intake and prostate cancer occurrence. Summary: As of now, there is not strong enough evidence to limit dairy food intake in the whole population. Further research is needed to explore the relationship between dairy consumption and prostate cancer. Men with diagnosed prostate tumors are suggested to eat food according to the Mediterranean diet as it has been reported that it decreases cancer-specific and overall mortality.

Production Responses of Multiparous Dairy Cattle with Differing Prepartum Muscle Reserves and Supplementation of Branched-Chain Volatile Fatty Acids

Periparturient dairy cattle undergo physiological adaptations to support fetal growth and colostrum synthesis in late gestation and milk production in early lactation. To support energy and protein demands dairy cattle mobilize body tissue reserves. The objective of this study was to determine the effects of prepartum skeletal muscle reserves and supplementation of branched-chain volatile fatty acids (BCVFA) on body composition measurements, metabolic markers related to health, protein, and energy status, and subsequent milk yield in multiparous dairy cows. Skeletal muscle reserves were assessed by 3 ultrasounds of the longissimus dorsi muscle depth (LDD) measured 42 d before expected calving (BEC), and cows (n = 48) were assigned to either high muscle (HM; > 4.6 cm) or low muscle (LM; ≤ 4.6 cm) groups. Cows were then randomly assigned to either control (CON) of soyhull pellets (80 g/d) or BCVFA treatment which contained isobutyrate (40 g/d), isovalerate (20 g/d), and 2-methylbutyrate (20 g/d) calcium salt products. Treatments were top dressed from 42 BEC until parturition, resulting in 4 combinations of muscle groups and treatments: HM-CON (n = 13), HM-BCVFA (n = 13), LM-CON (n = 11), and LM-BCVFA (n = 11). Measurements of the LDD, BW, and BCS were taken on the following days relative to calving -42, -35, -21, -7, 0, 7, 14, 21, 28. Weekly blood samples were taken to measure glucose, BHB, and insulin concentrations, and 5 of the blood sample time points were utilized to determine 3-methylhistidine and creatinine blood concentrations. Milk yield was recorded daily for the first 28 d of lactation, and samples were taken from both milkings once a wk for the first 4 wk to determine components. The statistical model included the fixed effects of treatment, group, time, their interactions, and the random effect of cow nested within group and treatment. Prepartum muscle mobilization varied between muscle groups, as LM cows accreted muscle prepartum, and HM cows mobilized muscle. The HM cows had higher milk fat, protein, lactose, and energy corrected milk yields. The BCVFA supplementation tended to increase blood glucose concentrations both prepartum and postpartum and decreased milk urea nitrogen concentrations. Higher prepartum skeletal muscle reserves improve productivity of early lactation cows likely due to differences in muscle mobilization, and BCVFA supplementation improves glucose dynamics during the transition period, which may improve the metabolic health of the periparturient dairy cow.

Evaluation of Rumen Microbes For Milk Fat Yield in Cattle Based on 16S rRNA Amplicon Metagenomic Sequencing

Background: Ruminal microbes support growth, lactation and maintenance of animals by providing energy and protein yielding nutrients. Metagenomics is the study of microbial communities directly in their natural environments such as soil, water, gut samples etc. The 16S rRNA gene sequencing is one of the basic genetic markers for the characterisation of prokaryote microorganism community. Current study exploited complex gut microbial communities present in cattle rumen based on variation in milk fat yield. Methods: The study was conducted in cattle reared at University Livestock Farm, Kerala Veterinary and Animal Sciences University. Twelve healthy cattle in the initial stages of lactation (less than three months) were selected and divided into two groups of six each based on high and low milk fat composition. Rumen samples were collected and DNA was isolated. Amplicons of 16S rRNA has been generated from metagenomic DNA targeting V3 and V4 hyper variable regions. Qiime2 was used for analysis for 16S rRNA amplicon sequencing. Result: Results revealed that a total of 38,28,241 sequences were obtained after demultiplexing. Majority of sequence length ranged around 295 to 301 nucleotides for forward reads and 216 to 300 nucleotides for reverse reads. After denoising, a total of 6,62,498 sequences were obtained. Clustering (de-replication) produced 711 unique sequence features. In kingdom level of taxonomy, for bacterial kingdom, there was no significant difference in abundance of organisms for high milk fat yield and low milk fat yield cows. For archaea, significant difference observed between abundance of high milk fat and low milk fat groups at five per cent level in the current study.

SGLT2 inhibition leads to a restoration of hepatic and circulating metabolites involved in the folate cycle and pyrimidine biosynthesis.

Inhibition of sodium-glucose cotransporter 2 (SGLT2) by empagliflozin (EMPA) and other "flozins" can improve glycemic control under conditions of diabetes and kidney disease. Though they act on the kidney, they also offer cardiovascular and liver protection. Previously, we found that EMPA decreased circulating triglycerides and hepatic lipid and cholesterol esters in male TallyHo mice fed a high-milk-fat diet (HMFD). The goal of this study was to determine whether the liver protection is associated with a change in metabolic function by characterizing the hepatic and circulating metabolic and lipidomic profiles using targeted LC-MS. In both male and female mice, HMFD feeding significantly altered the circulating and hepatic metabolome compared with low-fat diet (LFD). Addition of EMPA resulted in the restoration of circulating orotate (intermediate in pyrimidine biosynthesis) and hepatic dihydrofolate (intermediate in the folate and methionine cycles) levels in males and acylcarnitines in females. These changes were partially explained by altered expression of rate-limiting enzymes in these pathways. This metabolic signature was not detected when EMPA was incorporated into an LFD, suggesting that the restoration requires the metabolic shift that accompanies the HMFD. Notably, the HMFD increased expression of 18 of 20 circulating amino acids in males and 11 of 20 in females, and this pattern was reversed by EMPA. Finally, we confirmed that SGLT2 inhibition upregulates ketone bodies including β-hydroxybutyrate. Collectively, this study highlights the metabolic changes that occur with EMPA treatment, and sheds light on the possible mechanisms by which this drug offers liver and systemic protection.NEW & NOTEWORTHY Sodium-glucose cotransporter 2 (SGLT2) inhibitors, including empagliflozin, have emerged as a new treatment option for individuals with type 2 diabetes that have positive impacts on kidney and cardiovascular disease. However, less is known about their impact on other tissues, including the liver. Here, we report that empagliflozin reduces hepatic steatosis that is associated with restoring metabolic intermediates in the folate and pyrimidine biosynthesis pathways. These changes may lead to new approaches to treat nonalcoholic fatty liver disease.

Продуктивность коров при разной комбинации линий

The purpose of the study was to study the milk productivity of cows with different combinations of lines. The object of the study were first-calf cows of the black-and-white breed of the line Vis Back Aidiala 101341, Reflection Soveringa 198998, Montvik Chiefteina 95679. Cows obtained by intraline selection of the Aidial line have a high level of milk productivity, with milk yield of 5571 kg of milk, milk fat yield of 226 kg and protein yield of 171 kg, while significantly (P<0.05-0.01) superior to first-calf heifers, belonging to the Sovering line for 102 kg, 4 kg of fat and 4 kg of protein, and to the Chieftain line - for 259 kg of milk, 16 kg of fat and 11 kg of protein, respectively. When crossing the Ideal-Sovering lines, the highest milk yield is observed (5667 kg), with good milk fat content (4.01 %), high yield of milk fat (227 kg) and milk protein (172 kg). With the reverse cross Sovering-Idial, slightly lower milk productivity rates were obtained. When using bulls of the Aidial line on cows of the Governer line, high milk fat content (4.07 %) and milk fat yield (228 kg) were obtained, with average values of milk yield (5596 kg), protein mass fraction (3.01 %) and its yield (168 kg). Unsuccessful combinations of lines in both forward and reverse directions are crosses of the Idial and Chieftain lines. The descendants of such combinations have the lowest milk productivity. Thus, the use of line manufacturers R. Sovering and V.B. Idiala, both with intraline selection and interline combination, gives the best milk productivity indicators. Consequently, it is necessary to evaluate bulls according to the compatibility of lines; this has a great influence on the productivity of the offspring, since sires of the same line, when inseminating cows of the same lines, produce offspring of different productivity.

Non-targeted metabolomics identifies biomarkers in milk with high and low milk fat percentage

Milk is widely recognized as an important food source with health benefits. Different consumer groups have different requirements for the content and proportion of milk fat; therefore, it is necessary to investigate the differential metabolites and their regulatory mechanisms in milk with high and low milk fat percentages (MFP). In this study, untargeted metabolomics was performed on milk samples from 13 cows with high milk fat percentage (HF) and 13 cows with low milk fat percentage (LF) using ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS). Forty-eight potential differentially labeled compounds were screened using the orthogonal partial least squares-discriminant analysis (OPLS-DA) combined with the weighted gene co-expression network analysis (WGCNA) method. Amino acid metabolism was the key metabolic pathway with significant enrichment of L-histidine, 5-oxoproline, L-aspartic acid, and L-glutamic acid. The negative correlation with MFP differentiated the HF and LF groups. To further determine the potential regulatory role of these amino acids on milk fat metabolism, the expression levels of marker genes in the milk fat synthesis pathway were explored. It was noticed that L-histidine reduced milk fat concentration primarily by inhibiting the triglycerides (TAG) synthesis pathway. L-aspartic acid and L-glutamic acid inhibited milk fat synthesis through the fatty acid de novo and TAG synthesis pathways. This study provides new insights into the mechanism underlying milk fat synthesis and milk quality improvement.

Associations of diet quality and food consumption with serum biomarkers for lipid and amino acid metabolism in Finnish children: the PANIC study.

To investigate the associations of overall diet quality and dietary factors with serum biomarkers for lipid and amino acid metabolism in a general population of children. We studied 194 girls and 209 boys aged 6-8years participating in the Physical Activity and Nutrition in Children study. Food consumption was assessed by 4-day food records and diet quality was quantified by the Finnish Children Healthy Eating Index (FCHEI). Fasting serum fatty acids, amino acids, apolipoproteins, as well as lipoprotein particle sizes were analyzed with high-throughput nuclear magnetic resonance spectroscopy. Data were analyzed using linear regression adjusted for age, sex, and body fat percentage. FCHEI was directly associated with the ratio of polyunsaturated (PUFA) to saturated fatty acids (SFA) (PUFA/SFA), the ratio of PUFA to monounsaturated fatty acids (MUFA) (PUFA/MUFA), the ratio of PUFA to total fatty acids (FA) (PUFA%), the ratio of omega-3-fatty acids to total FA (omega-3 FA%), and inversely associated with the ratio of MUFA to total FA (MUFA%), alanine, glycine, histidine and very-low density lipoprotein (VLDL) particle size. Consumption of vegetable oils and vegetable-oil-based margarine (≥ 60% fat) was directly associated with PUFA/SFA, PUFA/MUFA, PUFA%, the ratio of omega-6 FA to total FA (omega-6 FA%), and inversely associated with SFA, MUFA, SFA to total FA (SFA%), MUFA%, alanine and VLDL particle size. Consumption of high-fiber grain products directly associated with PUFA/SFA, PUFA/MUFA, omega-3 FA%, omega-6 FA%, PUFA% and inversely associated with SFA and SFA%. Fish consumption directly related to omega-3 FA and omega-3 FA%. Consumption of sugary products was directly associated with histidine and VLDL particle size. Vegetable, fruit, and berry consumption had direct associations with VLDL particle size and the ratio of apolipoprotein B to apolipoprotein A1. Consumption of low fat (< 1%) milk was directly associated with phenylalanine. A higher consumption of high-fat (≥ 1%) milk was associated with lower serum MUFA/SFA and higher SFA%. Sausage consumption was directly related to SFA% and histidine. Red meat consumption was inversely associated with glycine. Better diet quality, higher in intake of dietary sources of unsaturated fat and fiber, and lower in sugary product intake were associated with more favorable levels of serum biomarkers for lipid and amino acid metabolism independent of adiposity. ClinicalTrials.gov: NCT01803776, registered March 3, 2013.

Choline regulation of triglycerides synthesis through ubiquintination pathway in MAC-T cells.

This study aims to investigate the regulatory mechanism of choline (CH) on triglyceride (TG) synthesis in cows, with a specific focus on its potential association with high milk fat percentage in the gut of the Zhongdian yak. By employing combined metagenomics and metabolomics analysis, we establish a correlation between CH and milk fat production in yaks. Bovine mammary epithelial cells (MAC-T) were exposed to varying CH concentrations, and after 24 h, we analyzed the expression levels of key proteins (membrane glycoprotein CD36 (CD36); adipose differentiation-related protein (ADFP); and ubiquintin (UB)), cellular TG content, lipid droplets, and cell vitality. Additionally, we evaluated the genes potentially related to the CH-mediated regulation of TG synthesis using real-time qPCR. CH at 200 μM significantly up-regulated CD36, ADFP, UB, and TG content. Pathway analysis reveals the involvement of the ubiquitination pathway in CH-mediated regulation of TG synthesis. These findings shed light on the role of CH in controlling TG synthesis in MAC-T cells and suggest its potential as a feed additive for cattle, offering possibilities to enhance milk fat production efficiency and economic outcomes in the dairy industry.

Effects of Hemp Seed on the Production, Fatty Acid Profile, and Antioxidant Capacity of Milk from Goats Fed Hay or a Mixed Shrubs-Grass Rangeland.

The research objective was to evaluate the effect of dietary incorporation of hemp seeds in goats fed with hay or mixed shrubs-grass rangeland on the production, FA profile and health-related lipid indices, antioxidant content and total antioxidant capacity of milk, with the purpose to increase the content of beneficial ingredients in milk and to improve its functional activity. Forty indigenous Carpathian goats were allocated into two groups according to the type of basic forage in their diet: hay (H) or mixed shrubs-grass rangeland (SG); each of them was further divided into two subgroups according to the presence of Hs in the concentrate mixture (250 g/kg) or not. Milk production was determined, and milk samples were collected and analyzed for fat, protein, lactose, and cholesterol content, as well as FA profile, lipophilic antioxidant content (α-tocopherol and retinol), and milk TAC. SG goats gave less milk (p < 0.01) but with higher milk fat (p < 0.001) and lower cholesterol content (p < 0.01) than H goats, while milk protein and lactose contents were not affected. Supplementing the diet with Hs caused a significant increase in milk production (p < 0.05) and milk fat content (p < 0.001) and a decrease in cholesterol content (p < 0.05). Grazing compared to indoor feeding but also supplementing the diet with Hs had the effect of decreasing the proportion of SFAs and increasing the concentrations of polyunsaturated FA (PUFA) in goat milk fat (p < 0.01). Fats in the milk of SG goats compared to H, but also in those supplemented with Hs, showed significantly higher proportions of vaccenic acid (VA), rumenic acid (CLA c-9,t-11) and omega-3 FA (α-linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosapentaenoic acid (DPA)) which are considered healthy for consumers. The feeding system based on SG and the diet supplementation with Hs ensured the best nutritional and functional quality of milk, confirmed by the FA profile, antioxidant content, and by the value of health-related lipid indices (n-6/n-3 FA ratio and hypo-/hypercholesterolemia, atherogenic index (AI), thrombogenic index (TI), and health promotion index (HPI)). The results of our work will be useful for the development of optimal nutritional strategies that improve the FA profile and the antioxidants content in goat milk, with beneficial effects on human health.

Relationship between rumen bacterial community and milk fat in dairy cows.

Milk fat is the most variable nutrient in milk, and recent studies have shown that rumen bacteria are closely related to milk fat. However, there is limited research on the relationship between rumen bacteria and milk fatty. Fatty acids (FAs) are an important component of milk fat and are associated with various potential benefits and risks to human health. In this experiment, forty-five healthy Holstein dairy cows with alike physiological and productive conditions were selected from medium-sized dairy farms and raised under the same feeding and management conditions. The experimental period was two weeks. During the experiment, raw milk and rumen fluid were collected, and milk components were determined. In this study, 8 high milk fat percentage (HF) dairy cows and 8 low milk fat percentage (LF) dairy cows were selected for analysis. Results showed that the milk fat percentage in HF group was significantly greater than that of the dairy cows in the LF group. 16S rRNA gene sequencing showed that the rumen bacterial abundance of HF dairy cows was significantly higher than that in LF dairy cows; at the genus level, the bacterial abundances of Prevotellaceae_UCG-001, Candidatus_Saccharimonas, Prevotellaceae_UCG-003, Ruminococcus_1, Lachnospiraceae_XPB1014_group, Lachnospiraceae_AC2044_group, probable_genus_10 and U29-B03 in HF group were significantly higher than those in the LF group. Spearman rank correlation analysis indicated that milk fat percentage was positively related to Prevotellaceae_UCG-001, Candidatus_Saccharimonas, Prevotellaceae_UCG-003, Ruminococcus_1, Lachnospiraceae_XPB1014_group, Lachnospiraceae_AC2044_group, probable_genus_10 and U29-B03. Furthermore, Prevotellaceae_UCG-001 was positively related to C14:0 iso, C15:0 iso, C18:0, Ruminococcus_1 with C18:1 t9, Lachnospiraceae_AC2044_group with C18:1 t9 and C18:1 t11, U29-B03 with C15:0 iso. To sum up, rumen bacteria in dairy cows are related to the variation of milk fat, and some rumen bacteria have potential effects on the deposition of certain fatty acids in raw milk.

MicroRNA-19a regulates milk fat metabolism by targeting SYT1 in bovine mammary epithelial cells

MicroRNAs (miRNAs) are important post-transcriptional factors involved in the regulation of gene expression and play crucial roles in biological processes related to milk fat metabolism. Our previous study revealed that miR-19a expression was significantly higher in the mammary epithelial cells of high-milk fat cows than in those of low-milk fat cows. However, the precise molecular mechanisms underlying these differences remain unclear. In this study, we found a high expression of miR-19a in the mammary tissues of dairy cows. The regulatory effects of miR-19a on bovine mammary epithelial cells (BMECs) were analyzed using cell counting kit-8 and 5-ethynyl-2′-deoxyuridine assays, which demonstrated that miR-19a significantly inhibited BMEC proliferation. Transfection of the miR-19a mimic into BMECs significantly upregulated the expression of milk fat marker genes LPL, SCAP, and SREBP1, promoting triglyceride (TG) synthesis and lipid droplet formation, whereas the miR-19a inhibitor exhibited the opposite function. TargetScan and miRWalk predictions revealed that synaptotagmin 1 (SYT1) is a target gene of miR-19a. A dual luciferase reporter gene assay, RT-qPCR, and western blot analyses revealed that miR-19a directly targets the 3′-untranslated region (UTR) of SYT1 and negatively regulates SYT1 expression. Functional validation revealed that overexpression of SYT1 in BMECs significantly downregulated the expression of LPL, SCAP, and SREBP1, and inhibited TG synthesis and lipid droplet formation. Conversely, the knockdown of SYT1 had the opposite effect. Altogether, miR-19a plays a crucial role in regulating the proliferation and differentiation of BMECs and regulates biological processes related to TG synthesis and lipid droplet formation by suppressing SYT1 expression. These findings provide a strong foundation for further research on the functional mechanisms underlying milk fat metabolism in dairy cows.

Association between hyperketolactia and production in early-lactating dairy cows

Study aims were to investigate associations of hyperketolactia (HYKL) status of Holstein dairy cows between 6 and 60 d in milk (DIM), defined by milk acetone (mACE) and β-hydroxybutyrate (mBHB) content, with daily milk yield and composition. Milk samples (∼5.0 million) were collected over a 5-yr period (2014-2019) within the milk recording system in Poland. Concentrations of mACE and mBHB determined by Fourier-transform infrared spectroscopy were used to categorize samples into 4 ketolactia groups. Based on threshold values of ≥0.15 mmol/L mACE and ≥0.10 mmol/L mBHB, ketolactia groups were normoketolactia (NKL; mACE <0.15 mmol/L and mBHB <0.10 mmol/L), BHB hyperketolactia (HYKLBHB; mACE <0.15 mmol/L and mBHB ≥0.10 mmol/L), ACE hyperketolactia (HYKLACE; mACE ≥0.15 mmol/L and mBHB <0.10 mmol/L), and ACE and BHB hyperketolactia (HYKLACEBHB; mACE ≥0.15 mmol/L and mBHB ≥0.10 mmol/L). To investigate ketolactia association with production outcomes, a linear model was developed, including ketolactia group, DIM, parity, their interactions, year-season as fixed effects, and random effects of herd and cow. Among all milk samples, 31.2% were classified as HYKL, and of these, 52.6%, 39.6%, and 7.8% were HYKLACEBHB, HYKLBHB, and HYKLACE, respectively. Ketolactia groups differed for all traits studied in all parities and DIM. Among HYKL groups, lowest milk yield was found in HYKLACEBHB cows, except for 6 to 30 DIM in first- and second-lactation cows. Milk yield of HYKLBHB cows was higher than that of NKL cows until 20 to 30 DIM, and then it was lower than NKL cows. Milk yield of HYKLACE cows was mostly lower than NKL cows. Energy-corrected milk (ECM) yield of HYKLACEBHB cows was higher than that of NKL cows until 30 to 35 DIM for second lactation and third lactation or greater, and in the whole study period for first lactation. The yield of ECM for HYKLBHB cows was mostly higher than that of NKL cows, whereas HYKLACE cows had higher ECM than NKL cows until 15 to 25 DIM and then was lower for the HYKLACE group. Milk composition differed among HYKL groups. Highest milk fat (MF) and lowest milk lactose (ML) contents were observed in HYKLACEBHB cows. Cows in HYKLACEBHB and HYKLBHB groups had higher MF and lower milk protein (MP; except in 6-8 DIM in first lactation) and ML content than NKL cows. Milk fat content was higher in HYKLACE than NKL cows in first lactation and during the first 30 to 40 DIM in older cows. Lactose content was lower in HYKLACE than in NKL cows within 30 to 40 DIM; afterward it was higher in NKL cows. Lower MP content was found in HYKLACE than in NKL cows, except during 6 to 9 DIM for cows in first lactation and third lactation or greater. In conclusion, HYKL is associated with altered milk production in all parities, but a range of these negative relations depends on ketone status addressing both ACE and BHB contents. Further research is needed to ascertain underpinning biochemical defects of HYKL from elevated ACE, alone or in combination with BHB, during early lactation.

Feed intake and milk yield and composition of lactating dairy goats in response to partial substitution of soybean meal for formaldehyde-treated sesame meal in the diet

ABSTRACT This study was conducted to evaluate the effect of substitution of soybean meal (SBM) for formaldehyde-treated sesame meal (FTSM) on nutrient intake and digestibility, ruminal and blood parameters and milk production and composition in lactating Murciano-Granadina goats. Forty lactating goats were randomly assigned to one of the following four treatments: (1) diet with 16.5% CP, containing SBM (CON); (2) diet with 16.5% CP, containing untreated SM (USM); (3) diet with 16.5% CP, containing FTSM (FT); and (4) diet with 14.5% CP containing FTSM (LPFT). The results showed that nutrient intake was highest in the FT group (p < 0.001), while it was similar between the CON and LPFT groups, except for the intake of CP, which was higher in the CON group. The FT and LPFT had lower ruminal pH compared to CON and USM groups (p < 0.001), with goats in group FT having the highest volatile fatty acids (VFA) production (p < 0.001). The highest propionate concentration was observed in the LPFT treatment (p < 0.001), followed by the FT, CON, and USM treatments. Goats offered USM and LPFT treatments presented the highest and lowest acetate: propionate values, respectively, among the experimental groups (p < 0.001). The results also showed that LPFT goats had the lowest blood urea nitrogen (BUN) level (p = 0.004), while FT goats presented a lower non-esterified FA (NEFA) level compared with CON and LPFT goats (p = 0.01). Goats offered the FT diet had the highest milk yield (p = 0.002) and energy-corrected milk yield (p < 0.001) among all dietary groups. The highest milk fat (p < 0.001), protein (p = 0.001), lactose (p = 0.007), total solids (p = 0.003), and solids-not-fat (SNF) (p = 0.003) contents were observed in FT goats, which didn’t differ from USM goats. The inclusion of formaldehyde-treated SM increased the percentage of C18:3 (p < 0.001) and C20:1 (p = 0.04) FAs compared with USM and CON treatments. Milk from USM, FT, and LPFT goats had lower levels of saturated (p < 0.001) and medium-chain FAs (p = 0.014) compared with CON goats, whereas milk from CON goats had lower levels of unsaturated, monounsaturated, and long-chain FAs compared to other groups (p < 0.001). The lowest and the highest concentrations of polyunsaturated FAs were observed in CON and LPFT goats, respectively (p = 0.001). It can be concluded that SBM can be advantageously replaced by formaldehyde-treated SM in the diet as a feasible alternative to improve feed intake and production performance of dairy goats.

МОЛОЧНАЯ ПРОДУКТИВНОСТЬ КОРОВ, ПРОИСХОДЯЩИХ ИЗ ПЕРСПЕКТИВНЫХ ВЕТВЕЙ ГОЛШТИНСКОЙ ПОРОДЫ

The aim of the study was to study the milk productivity of cows originating from different promising branches and lines of the Holstein breed. The population studied was Wis Back Idial 101341, Reflection Sovering 198998, Montvik Chieftain 95679, Siling Traijun Rokita 252803 and Roseife City 267150. The best indicators were established in cows of the line V.B. Ideal for the 1st and 3rd lactation (7086 and 7431 kg of milk with the amount of milk fat 271 and 285 kg and the amount of milk protein 225 and 238 kg), and the difference is statistically significant (P&lt;0.05-0.001) in relation to other lines. In second place are the animals of the Reflection Sovering line. Cows of the line S.T. Rokita had a high milk fat content (3.85 and 3.94%) at P&lt;0.05. In V. Aidial's line, the best milk yield indicators are shown by first-calf heifers of the SWEET-HAVEN TRADITION branch (7143 kg). In the R. Sovering line, cows of the TO-MAR BLACKSTAR-ET branch showed the greatest increase in milk yield by 89-1226 kg, milk fat yield by 4-42 kg, milk protein yield by 2-37 kg (P&lt;0.001), and high milk fat content (3.86%) of the cow of the S-W-D VALIANT line (P&lt;0.01-0.001). According to the 3rd lactation in the line of R. Sovering, cows of the S-W-D VALIANT branch show an advantage in milk yield by 353-831 kg of milk, mass fraction of fat - by 0.05-0.08%, milk fat yield - by 20-36 kg and milk protein yield - by 12-26 kg (P&lt;0.05-0.01). As a result of the study, it was found that the use of animal branches of SWEET-HAVEN TRADITION, ROCKALLI SON OF BOVA, TO-MAR BLACKSTAR-ET and WALKWAY CHIEF MARK makes it possible to obtain high milk productivity from them.

SGLT2 Inhibition Alters the Hepatic and Circulating Metabolome of Mice Under a High-Milk-Fat Diet

Recent studies have shown that the inhibition of Sodium-glucose cotransporter 2 (SGLT2), which reabsorbs glucose in the renal proximal tubule, can aid in glycemic control under conditions of type 2 diabetes (T2D). Though it acts directly on the kidney, these inhibitors also lead to great improvement in liver function. Previously, we challenged TallyHo mice, an inbred polygenetic mice model that recapitulates T2D, with a high milk fat diet (HMFD) for 12 weeks in the continued absence or presence of Empagliflozin (EMPA), an FDA-approved SGLT2 inhibitor. Male mice showed a decrease in circulating triglycerides, hyperglycemia, and hepatic lipid and cholesterol esters (Kurtz et al 2022). However, the mechanisms that underlie this improved liver phenotype is unknown. Thus, we sought to characterize the hepatic and circulating metabolic and lipidomic profiles that are altered and/or restored in the presence of EMPA using targeted and untargeted LC-MS. Control, HMFD, and HMFD + EMPA liver tissues and plasma samples were dissolved in a methanol/water 50/50 extraction buffer that contained debrisoquine (DBQ) for the positive mode standard and 4-nitrobenzoic acid for the negative mode standard. For the untargeted liver and plasma samples, a UPLC-QTOF analysis was performed. MetaboAnalyst was used to quantify metabolite changes with inclusion criteria set to a fold change of 2 and a p-value &lt;0.05. Of note, previous studies have shown that EMPA promotes the upregulation of ketone bodies including β-Hydroxybutyrate (BHB). We noted a similar upregulation of BHB in our metabolomic analysis and confirmed that EMPA led to a ~1.5-fold increase (p &lt; 0.022) in this circulating ketone body. In addition, of those metabolites that met the inclusion criteria, we identified 12 that were downregulated and 9 that were upregulated +EMPA in the male samples. Out of these, orotate, sarcosine, tartrate, and dihydrofolate levels were restored to those of control levels. In the female samples, 6 were downregulated and 3 were upregulated +EMPA. Indoxyl sulfate, butylcarnitine, and valerylcarnetine were all restored to control levels. Of particular interest is the finding that EMPA treatment restores levels of key metabolites that are involved in the folate and pyrimidine biosynthesis pathways. Previous studies have determined that folate deficiency can promote insulin resistance and the urea cycle, a pathway that is integrated with pyrimidine biosynthesis, is upregulated under conditions of T2D and non-alcoholic fatty liver disease. Future studies are aimed at identifying the role of EMPA's hepatic protection in these metabolic pathways. NIDDK This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Characterizing the impact of SGLT2 inhibition on hepatic inflammation and cellular response in a type 2 diabetic mouse model

Type 2 diabetes (T2D) is associated with several comorbidities including nonalcoholic fatty liver disease (NAFLD). NAFLD is characterized by the presence of steatosis with or without hepatic fibrosis and is associated with insulin resistance, inflammation, and hyperlipidemia. Recently, sodium-glucose co-transport 2 inhibitors (SGLT2i) such as Empagliflozin (Empa), have emerged as a treatment option for patients with T2D by blocking renal glucose reabsorption and improving glycemic control. Whether these inhibitors may attenuate T2D-associated NAFLD is still under investigation. To explore this possibility, we challenged TallyHo (TH) mice with a 60% high-milk fat diet (HMFD) with or without Empa (10mg/kg BW) for 12 weeks. Like humans, TH mice possess several quantitative trace loci associated with diabetic/obese phenotypes. In this study, we found that Empa treatment decreased hepatic steatosis in both male and female TH mice. However, the 12-week dietary challenge was not sufficient to induce fibrosis and inflammation (Kurtz et al. 2022). Thus, we have extended the trial to 24-weeks with the hypothesis that Empa promotes anti-inflammatory responses through macrophage infiltration and cellular signaling. Protein expression of the cGAS-STING pathway (STING, pTBK1, TBK1, pIRF3, IRF3), inflammasome pathway (AIM2, ASC/TMS1) and apoptosis signaling pathway (Caspase-3) showed no significant changes between diet or treatment. Additionally, activity of p50 and p65 of NF-κB (measured through nuclear expression) was not changed between treatment groups.While Empa does not appear to alter cellular-mediated inflammatory pathways, we did note changes in cytokine expression. Using a large-scale screen to probe for more than 60 inflammatory markers, we observed elevated levels of three pro-inflammatory cytokines (IL-3, CCL2, M-CSF; p&lt;0.05) in livers from Empa-treated mice while there was downregulation of p40/p70 (p&lt;0.01). The screen of plasma samples identified elevated levels of MIP-1γ (p&lt;0.01) and identified decreased levels of three cytokines (IL-9, TIMP1, TPO; p&lt;0.05) in Empa-treated mice. To further understand the role of macrophage infiltration as a response to increased inflammation, immunofluorescence on liver section with F4/80, a macrophage marker, was performed. There were no significant differences in the density or localization of Kupffer cells between treatments and diets for both sexes. Thus, it is hypothesized that Empa promotes the polarization of macrophages from M1 (pro-inflammatory) to M2 (anti-inflammatory) to promote tissue regeneration and repair. Staining of M1 and M2 macrophage markers are currently underway. Additionally, it has been determined that the upregulation of TIMP1 leads to matrix deposition. Efforts are currently underway to quantify portal inflammation using QuPath. Collectively, the data shows that Empa treatment for 24-weeks does not alter cellular signaling, and potentially alters macrophage signaling to combat NAFLD. NIDDK This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Dairy Product Consumption and Incident Prediabetes in the Australian Diabetes, Obesity, and Lifestyle Study With 12 Years of Follow-Up

BackgroundInvestigating modifiable risk factors for the early stages of the development of type 2 diabetes is essential for effective prevention. Some studies show protective associations between dairy and prediabetes; however, associations are heterogeneous by the type and fat content of dairy foods. ObjectiveTo examine the relationship between the consumption of dairy, including different types of dairy products and risk of prediabetes. MethodsThe study included 4891 participants with normal glucose tolerance (aged 49.0 ± 12.3 y, 57% female) of the Australian Diabetes, Obesity, and Lifestyle (AusDiab) study, a longitudinal population-based study. Dairy intake was measured at baseline using a food frequency questionnaire. Prediabetes at the 5-y and 12-y follow-ups was defined according to the WHO criteria as fasting plasma glucose levels of 110–125 mg/dL or 2-h plasma glucose levels of 140–199 mg/dL. Associations were analyzed using Poisson regression, adjusted for social demographics, lifestyle behaviors, a family history of diabetes, and food group intake. ResultsIn total, 765 (15.6%) incident cases of prediabetes were observed. The mean intake of dairy foods was 2.4 ± 1.2 servings/d, mostly consisting of low-fat milk (0.70 ± 0.78 servings/d) and high-fat milk (0.47 ± 0.72 servings/d). A higher intake of high-fat dairy (RRservings/d: 0.92; 95% CI: 0.85, 1.00), high-fat milk (0.89; 95% CI: 0.80, 0.99), and total cheese (0.74; 95% CI: 0.56, 0.96) was associated with a lower risk of prediabetes. Low-fat milk intake was associated nonlinearly with prediabetes risk. Low-fat dairy foods, total milk, yogurt, low-fat cheese, and ice cream were not associated with prediabetes risk. ConclusionIn this large Australian cohort, protective associations were found for high-fat dairy types, whereas neutral associations were seen for low-fat dairy types. Studies with more detail on sugar content of types of dairy foods and products eaten with dairy foods (e.g., cereals or jam), and studies into potential causal mechanisms of the health effects of dairy intake are required.