Characterization of coconut oil and CLA induced lipolysis

Obesity is a main health concern and leads to many other health complications. Conjugated linoleic acid (CLA) has been shown to cause a reduction in obesity in several species. CLAinduced body fat loss is enhanced when mice are fed coconut oil (CO). The objectives were to determine if the CLA-induced lipolysis in different oil source-fed mice was time-dependent and to determine the effect of cell signaling inhibitors on CO+CLA-induced lipolysis. Study 1: Male mice (ICR; n=80; 3wk) were fed 7% soybean oil (SO) or CO diets for 6wk and then supplemented with 0 or 0.5% CLA for 3, 7, 10 or 14d. Body fat index (BFI) was calculated as [(epididymal fat pad + retroperitoneal fat pad)/ body weight] and lipolysis was determined by non-esterified fatty acid (NEFA) and glycerol release in 3hr ex vivo cultures. BFI was reduced by CO on d7 (P\u3c0.01) and CLA tended (P=0.09) to decrease BFI in CO-fed mice on d10. BFI was reduced in both CO and SO-fed mice (P\u3c0.05) in response to CLA on d14. NEFA release was increased by CLA in CO-fed mice (P\u3c0.01) but not in SO-fed mice on d7 and 10 but on d14 CLA increased NEFA release in both CO and SO-fed mice (P\u3c0.0001). Glycerol release was also increased by CLA in CO-fed mice but not in SO-fed mice on d3 and d7 (P\u3c0.05). We then determined expression and activation level of proteins involved in lipolysis and lipogenesis. CLA tended to decrease (P=0.06) p-perilipin protein expression in CO-fed mice. There was also a trend for CLA (P=0.06) to decrease adipose triglyceride lipase (ATGL) protein. CO-fed mice had greater fatty acid synthase, stearoyl CoA desaturase 1 mRNA expression and less acetyl CoA carboxylase mRNA expression (P\u3c0.01). Sterol regulatory binding protein 1c and malic enzyme expression was least in CO+CLA-fed mice. Study 2: 3T3-L1 cells were differentiated, exposed to CO or SO for 10d, serum starved overnight, and pre-loaded with 3[H]-oleic acid for 12 hrs. Cells were treated with 50 muM CLA or linoleic acid (LA) with/without cell signaling inhibitors for 12-24 hrs. Lipolysis was measured as the 3-hr release of 3[H]-oleic acid. Without inhibitors, CO+CLA treatment caused more lipolysis (P\u3c0.01) than all other treatments, which did not differ. None of the inhibitors tested reduced lipolysis in CO+CLA treated cells. Cyclooxegenase-2 inhibitor increased lipolysis of SO+CLA treated cells (P=0.05) to the level of CO+CLA. Phospholipase C inhibitor increased lipolysis in all treatments (P\u3c0.0001) except that of CO+CLA. Peroxisome proliferator-activated receptor alpha inhibitor also increased lipolysis of CO+LA (P\u3c0.05) to the level of CO+CLA treated cells and in SO+CLA treated cells. There was no effect of the p42 mitogen-activated protein kinase or protein kinase A inhibitor, compared to absence of inhibitor. Therefore CLA-induced lipolysis occurs more rapidly in CO vs SO-fed mice and the CLA enhanced lipolysis in CO group could involve the PLC pathway

The role of microbes in rumen lipolysis and biohydrogenation and their manipulation

Despite the fact that the ruminant diet is rich in polyunsaturated fatty acids (PUFA), ruminant products - meat, milk and dairy - contain mainly saturated fatty acids (SFA) because of bacterial lipolysis and subsequent biohydrogenation of ingested PUFA in the rumen. The link between SFA consumption by man and coronary heart disease is well established. In contrast, ruminant products also contain fatty acids that are known to be beneficial to human health, namely conjugated linoleic acids (CLAs). The aims of research in this field have been to understand the microbial ecology of lipolysis and biohydrogenation and to find ways of manipulating ruminal microbes to increase the flow of PUFA and CLA from the rumen into meat and milk. This review describes our present understanding of the microbial ecology of ruminal lipid metabolism, including some apparently anomalous and paradoxical observations, and the status of how the metabolism may be manipulated and the possible consequential effects on other aspects of ruminal digestion. Intuitively, it may appear that inhibiting the ruminal lipase would cause more dietary PUFA to reach the mammary gland. However, lipolysis releases the non-esterified fatty acids that form the substrates for biohydrogenation, but which can, if they accumulate, inhibit the whole process. Thus, increasing lipase activity could be beneficial if the increased release of non-esterified PUFA inhibited the metabolism of CLA. Rumen ciliate protozoa do not carry out biohydrogenation, yet protozoal lipids are much more highly enriched in CLA than bacterial lipids. How could this happen if protozoa do not metabolise PUFA? The answer seems to lie in the ingestion of plant organelles, particularly chloroplasts, and the partial metabolism of the fatty acids by contaminating bacteria. Bacteria related to Butyrivibrio fibrisolvens are by far the most active and numerous biohydrogenating bacteria isolated from the rumen. But do we misunderstand the role of different bacterial species in biohydrogenation because there are uncultivated species that we need to understand and include in the analysis? Manipulation methods include dietary vegetable and fish oils and plant-derived chemicals. Their usefulness, efficacy and possible effects on fatty acid metabolism and on ruminal microorganisms and other areas of their metabolism are described, and areas of opportunity identified

Efectos de los conjugados del ácido linoleico sobre la incorporación tisular de ácidos grasos y metabolismo lipídico en ratas deficientes en ácidos grasos esenciales

El objetivo del presente estudio fue investigar si los conjugados del ácido linoleico (CLA) podrían atenuar o potenciar algunas alteraciones observadas en ratas con deficiencia marginal de ácidos grasos esenciales (AGE). Para ello, ratas Wistar macho fueron alimentadas 60 días con dietas suficientes o deficientes en AGE, suplementadas o no con CLA. Los CLA presentaron distinto grado de incorporación en los tejidos, mostrando una alta retención en tejido adiposo y una marcada resistencia en cerebro; asimismo en los animales con dietas suficientes o deficientes en AGE, los CLA modificaron en forma diferencial los marcadores tisulares de deficiencia de AGE. En animales con dietas deficientes en AGE, los CLA redujeron los triglicéridos circulantes y el tamaño del tejido adiposo; no obstante, estas modificaciones y el incremento del hígado permitirían postular una similitud con ciertas alteraciones observadas en los inicios de la lipodistrofia presente en otros modelos experimentales.Effects of conjugated linoleic acid on tissular fatty acid incorporation and lipid metabolism in essential fatty acid– deficient rats The aim of this work was to investigate whether conjugated linoleic acid (CLA) may attenuate or enhance some alterations observed in rats with marginal deficiency of essential fatty acids (EFA). For this purpose, male Wistar rats were fed sufficient or deficient in EFA diets, supplemented or not with CLA, for 60 days. CLA showed different degree of incorporation into the tissues, showing a high retention in adipose tissue and noticeable resistance in brain. Moreover, independently of the EFA deficiency, CLA differentially modified the tissue biomarkers of EFA deficiency. In animals with EFA deficient diets, CLA reduced circulating triacylglycerides and adipose tissue weight; however, these changes and the increased liver size would allow postulating that a similarity with some alterations observed in the beginning of the lipodystrophy syndrome in other experimental models might be present in these rats.Fil: Fariña, Ana Clara. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Departamento de Ciencias Biologicas. Catedra de Bromatologia y Nutrición; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Gonzalez, Marcela Aída. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Departamento de Ciencias Biologicas. Catedra de Bromatologia y Nutrición; ArgentinaFil: Latorre, Maria Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Bernal, Claudio Adrian. Universidad Nacional del Litoral. Facultad de Bioquimica y Ciencias Biologicas. Departamento de Ciencias Biologicas. Catedra de Bromatologia y Nutrición; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

PPARγ Modulates Long Chain Fatty Acid Processing in the Intestinal Epithelium.

Nuclear receptor PPARγ affects lipid metabolism in several tissues, but its role in intestinal lipid metabolism has not been explored. As alterations have been observed in the plasma lipid profile of ad libitum fed intestinal epithelium-specific PPARγ knockout mice (iePPARγKO), we submitted these mice to lipid gavage challenges. Within hours after gavage with long chain unsaturated fatty acid (FA)-rich canola oil, the iePPARγKO mice had higher plasma free FA levels and lower gastric inhibitory polypeptide levels than their wild-type (WT) littermates, and altered expression of incretin genes and lipid metabolism-associated genes in the intestinal epithelium. Gavage with the medium chain saturated FA-rich coconut oil did not result in differences between the two genotypes. Furthermore, the iePPARγKO mice did not exhibit defective lipid uptake and stomach emptying; however, their intestinal transit was more rapid than in WT mice. When fed a canola oil-rich diet for 4.5 months, iePPARγKO mice had higher body lean mass than the WT mice. We conclude that intestinal epithelium PPARγ is activated preferentially by long chain unsaturated FAs compared to medium chain saturated FAs. Furthermore, we hypothesize that the iePPARγKO phenotype originates from altered lipid metabolism and release in epithelial cells, as well as changes in intestinal motility

Evaluating the benefits and risks of organic raw milk cheese. Challenges in the production of organic cheeses mad from raw milk

Raw milk offered for sale within and into the European Union has to be produced according to the requirements of Commission Directive 89/362/EEC and to meet quality standards described in Council Directive 92/46/EEC. Together the regulations say many things about the quality of milk and the hygienic means of production including guidance and requirements for: - The means of production, farm specifications etc. - Processes, milk handling - Animal health - Milk composition related to quality - especially antibiotics, cell count and bacterial content (Hillerton & Berry, 2004) Unpasteurized milk and the associated food products (yogurt, butter, cheese etc.) made from raw milk is safely produced and consumed in many areas around the world. Cheese making is a major industry worldwide, while most of the production comes from large scale industrial producers; a large part is still practiced on a relatively small scale which accounts for the rich diversity of cheeses available. Cheese is made from milk, which contains milk fat and coagulated proteins and preservation is largely achieved by controlling the pH and water activity (Little et al., 2008). In the European Union the production of both fresh and raw milk cheeses is allowed. Between 1995 and 2004 cheese production increased by nearly 15 %, with per capita consumption growing at an average rate of 1.5 % per year. Nearly 40 % of EU milk is consumed as cheese. Four Member States (Germany, France, Italy and the Netherlands) produce more than 75 % of the cheese in the EU. The European dairy industry processes approximately 135 million tons of raw milk into a broad range of products, both for consumption and for use in the production of many food, feed and pharmaceutical products. The raw milk delivered by the EU-25’s 1.6 million dairy farmers, processed by the dairy industry, plays a vital role in rural areas (EU, 2006). About 700 000 tons of raw milk cheeses are produced annually in Europe, particularly in France, Italy and Switzerland (Beuvier & Grappin, 1997). The effort to preserve raw milk cheese production in certain countries of the European Union began in the early 1990’s. Between1990 and1992 the European Union debated the safety of raw milk cheese and was considering the mandatory pasteurization of all dairy products. Some of the northern European countries wanted to forbid the production of raw milk cheeses for sanitary reasons, pointing toward the reduced health risk from pasteurized milk cheeses. They were considering the mandatory pasteurization of all dairy products (Dixon, 2000). Cheese made from raw milk represents an important proportion of the traditional cheeses, particularly in South European countries (Beuvier & Grappin, 1997). In these arias a large variety of traditional cheeses are still produced using raw milk

Importance of supplying dairy cows with essential fatty acids (EFA) and conjugated linoleic acids (CLA) during the transition period on metabolism and health

Ketogenesis and steatosis are common sequelae of the severe mobilization of fat reserves necessary at the onset of lactation when the energy expenditures for milk and maintenance exceed the energy provided by voluntary feed intake in dairy cows. This situation along with a common state of systemic inflammation during early lactation makes dairy cow vulnerable to further metabolic and infectious diseases which impact their profitability. A periparturient supplementation with essential fatty acids (EFA) and conjugated linoleic acids (CLA) has been shown to influence different aspects of lipid and energy metabolisms as well as markers of immune homeostasis in dairy cows. This study aimed at investigating the synergistic effects of EFA, particularly a-linolenic acid (ALA, n-3 fatty acid (FA), and CLA on the liver and plasma proteome profile of dairy cows during the time spanning 3 weeks before to 9 weeks after parturition. Late-gestation Holstein cows were infused from 9 wk ante partum (AP) to 9 wk post partum (PP) into the abomasum with either coconut oil (CTRL, n = 8, 38 & 76 g/d during dry period and lactation, respectively) or a mixture of EFA (Linseed + safflower oil) and CLA (Lutalin, BASF) (EFA+CLA, n = 8, 60 and 120 g/d during dry period and lactation, respectively). An untargeted shotgun proteomics approach based on liquid chromatography coupled with tandem mass spectrometry was performed in liver biopsies and plasma samples collected at days -21, +1, +28, and +63 relative to calving. At each timepoint, differentially abundant proteins (DAP) between treatment groups were identified as the proteins at the intersection between a multivariate supervised Partial Least Squares Discriminant Analysis (PLS-DA) and an univariate analysis (proteins with P-value <0.05 according to a t-test and fold change = 1.3). In liver a total of 1680 proteins was identified at each timepoint, of which 100 DAP between groups were assigned to the metabolism of xenobiotics by cytochrome P450, drug metabolism - cytochrome P450, drug metabolism - other enzymes, arachidonic acid metabolism, cholesterol metabolism and bile secretion, pyruvate metabolism, steroid hormone biosynthesis, glycolysis/gluconeogenesis, glutathione metabolism, and citrate cycle at all timepoints. At each timepoint, cytochrome P450, as a central hub to these annotated pathways, was related to specific CYP enzymes comprising CYP51A1, CYP1A1, CYP4F2, and CYP4V2. A total of 241 unique proteins was identified in the serum, in which a cluster of apolipoproteins (APO) containing APOC3, APOA1, APOA4, and APOC4 was increased in response to EFA+CLA according to the stage of lactation. Overabundant APO were annotated by GO (gene ontology) terms related to triglyceride (TAG) homeostasis, cholesterol and lipoprotein metabolisms, inflammation, and innate immune response. Altogether, these findings provided novel insights into the molecular mechanisms involved in cytochrome P450, cholesterol, and TAG metabolism by which dietary supplemented EFA+CLA influences metabolic and immune adaptation around parturition. Our results suggest that EFA+CLA could synergistically attenuate hepatic TAG accumulation and marginally mitigate inflammation. Nevertheless, further research is necessary to confirm our results at the other (metabolome) levels

Effects of Essential Fatty Acids and Conjugated Linoleic Acid Supplementation on Fatty Acid Pattern in Blood Plasma and Milk and on the Inflammatory Response in Dairy Cows from Late Gestation to Early Lactation

This study investigated the effects of abomasal infusion of essential fatty acids, especially alpha-linolenic acid, and conjugated linoleic acid on their distribution in milk fat and blood plasma and on the plasma inflammatory response in dairy cows from late to early lactation. The most important essential fatty acids for ruminants are alpha-linolenic acid and linoleic acid. They are abundant in pasture which is nowadays reduced in the ration of dairy cows due to the replacement of fresh feeds by preserved diets. Conjugated linoleic acid is formed as a by-product during ruminal biohydrogenation of essential fatty acids and has been associated with positive effects on the energy metabolism and immune system. Forty rumen-cannulated Holstein Friesian cows were assigned to four treatment groups in their late second lactation. Prior to supplementation, cows were fed a total mixed rations with a low-fat content. In late gestation cows were abomasally treated with coconut oil, linseed and safflower oil, conjugated linoleic acid, or both. Performance data, milk composition and fatty acid pattern in milk and plasma as well as inflammatory response parameters in plasma were measured regularly. Furthermore, liver tissue was tested for the abundance of genes related to the inflammatory response.:TABLE OF CONTENTS ABBREVIATIONS 1. INTRODUCTION 2. LITERATURE OVERVIEW 2.1 Essential Fatty Acids and Conjugated Linoleic Acid 2.1.1 Essential Fatty Acids (EFA) 2.1.2 Conjugated Linoleic Acid (CLA) 2.1.3 EFA in Dairy Cow Nutrition 2.2 Fatty Acid Distribution in Blood, Erythrocyte Membranes, and Milk Fat 2.2.1 Plasma Lipids 2.2.2 EFA and CLA in Plasma Lipids 2.2.3 EFA and CLA in Erythrocyte Membranes 2.2.4 EFA and CLA in Milk Fat 2.3 Effects of EFA and CLA on Inflammatory Processes during the Transition Period 2.3.1 Metabolic and Immunological Challenges during the Transition Period 2.3.2 Effects of EFA on the Metabolism, Inflammatory- and Immune Response 2.3.3 Effects of CLA on the Metabolism, Inflammatory- and Immune Response 2.4 Scope of the Thesis 3. PUBLICATION 4. GENERAL DISCUSSION 4.1 Abomasal Infusion 4.2 Animal Performance 4.3 Distribution of EFA and CLA in Blood and Milk Fat 4.4 Effects of EFA and CLA on Plasma and Hepatic Acute Phase and Inflammatory Response 4.5 Conclusion and Practical Considerations 4.5.1 Summary of EFA effects 4.5.2 Summary of CLA effects 4.5.3 Summary of synergistic effects of EFA and CLA 4.5.4 Summary of Observations apart from Treatments and Practical Considerations 5. SUMMARY 6. ZUSAMMENFASSUNG 7. REFERENCES APPENDI

Denovo synthesized fatty acids as regulators of milk fat synthesis

The objectives of the dissertation research were to determine the role of denovo synthesized fatty acids (DNFA) in the regulation of milk fat synthesis. Milk fat responses to increasing amounts of short- and medium-chain fatty acids (SMCFA), added in the proportion as synthesized denovo, were studied in lactating dairy cows. The results showed a significant linear increase in milk fat concentration with SMCFA supplementation. However, milk fat yield was similar for all treatments. A subsequent study was aimed at increasing the availability of SMCFA during trans-10, cis-12 CLA-induced milk fat depression (MFD) in lactating dairy cows to determine whether SMCFA can rescue part of CLA-induced MFD. Post-ruminal infusion of butterfat (BF) was used as a source of SMCFA. The BF treatment was compared to a mixture of fats containing only the long-chain FA (LCFA) with or without trans-10, cis-12 CLA infusion. Milk fat content and yield were significantly reduced with trans-10, cis-12 CLA. However, increased availability of SMCFA with BF infusion had no effects on milk fat yield and concentration. Trans-10, cis-12 CLA significantly reduced the mRNA expression of transcription factor SREBP-1c along with its downstream targets including ACC,FASN, LPL, SCD and AGPAT. The increased availability of SMCFA had no effect on either lipogenic gene or protein expression suggesting that nutritional manipulation was not sufficient to rescue trans-10, cis-12 CLA-induced MFD. Finally, the effects of combination of a Rosiglitazone (ROSI), a PPAR-γ agonist, and trans-10, cis-12 CLA were examined on mammary and hepatic lipogenesis in lactating mice. Mammary lipogenesis was significantly reduced with trans-10, cis-12 CLA, reducing the milk fat content and mRNA expression of lipogenic transcription factors SREBP1-c and PPAR- γ. Trans-10, cis-12 CLA significantly increased hepatic lipid accumulation, while the mRNA expression of SREBP1-c and PPAR- γ were not altered. On the contrary, ROSI had no effects on mammary lipogenesis. However, ROSI significantly rescued trans-10, cis-12 CLA-induced hepatic steatosis

Efeitos da suplementação de ácidos graxos em vacas leiteiras durante períodos de desafio fisiológico : transição e estresse calórico

Orientador: Prof. Dr. Rodrigo de AlmeidaCoorientador: Prof. Dr. João Alberto NegrãoTese (doutorado) - Universidade Federal do Paraná, Setor de Ciências Agrárias, Programa de Pós-Graduação em Zootecnia. Defesa : Curitiba, 30/04/2024Inclui referênciasResumo: Este estudo teve como objetivo avaliar os efeitos da suplementação lipídica durante períodos de desafio fisiológico em vacas leiteiras, focando nos ácidos graxos de cadeia média (AGCM) e nos ácidos graxos ômega-3 e ômega-6. O estudo buscou compreender as mudanças metabólicas, inflamação e estresse oxidativo, assim como a regulação de mediadores lipídicos em vacas submetidas a estresse calórico. No primeiro experimento, conduzido no Paraná, 168 vacas Holandesas foram divididas em dois grupos: Controle (sem suplementação de AGCM) e Suplemento lipídico (50 g/dia de óleo de coco e palma, 0,065% AGCM na MS). Os experimentos duraram 35 dias, abrangendo 21 dias pré-parto e 14 dias pós-parto. Foram medidas a produção e composição do leite, perfil de ácidos graxos do leite, ruminação, peso e escore de condição corporal (ECC), além dos metabólitos sanguíneos (cálcio total, cálcio iônico, glicose, GGT, AST, colesterol, bilirrubina, albumina, ácidos graxos não esterificados e beta-hidroxibutirato) em diferentes dias (-7, 0, 3, 7 e 14 d) e análise de expressão gênica (HPRT1, IL-6, SAA3, GPx3, CASP8, TLR4, MyD88, STAT1, STAT5, LPK, ACACA, FASN, LPL, SCD, SREBF1). A suplementação com AGCM não afetou significativamente a produção de leite ou sua composição (P > 0,05), mas influenciou a expressão gênica e o perfil de ácidos graxos no leite (P 0,05). A dose de AGCM pode ter sido insuficiente para maiores efeitos no desempenho produtivo e metabólico. O segundo experimento, realizado no Centre de Recherche en Sciences Animales de Deschambault (CRSAD), em Quebec, Canadá, investigou os efeitos da infusão abomasal de ácidos graxos ômega-3 e ômega-6 na regulação de mediadores lipídicos durante o estresse calórico. Vinte vacas Holandesas multíparas foram distribuídas aleatoriamente em um delineamento de quadrado latino incompleto com períodos de 10 dias. Os tratamentos incluíram: Termoneutralidade com alimentação em pares + óleo de milho (TNPF/n6), Estresse térmico + óleo de milho (HS/n6) e Estresse térmico + óleo de peixe (HS/n3). Os óleos (159 g/dia) foram infundidos no abomaso em 11 dois bolus. Amostras de sangue foram coletadas nos dias 0, 5 e 10 para análise lipidômica por cromatografia LC-MS/MS. Os ácidos graxos ácido eicosapentaenoico (EPA), ácido docosahexaenoico (DHA), ácido docosapentaenoico (DPA) e ácido araquidônico (AA), juntamente com os oxilipídeos da via da lipoxigenase 5-HETE, 5- oxoETE, 15-HETE, 15-oxoETE, 17-HDoHe e oxilipídeos do citocromo 450: 19,20-EpDPE e 19,20-DiHDPA, aumentaram em HS/n3 em comparação com HS/n6 (P 0.05), but it influenced gene expression and the milk fatty acid profile (P 0.05). The dose of MCFA may have been insufficient for greater effects on productive and metabolic performance. The second experiment carried out at the Centre de recherche en sciences animales de Deschambault (CRSAD), in Quebec, Canada, investigated the effects of abomasal infusion of omega-3 and omega-6 fatty acids on the regulation of lipid mediators during heat stress. Twenty multiparous Holstein cows were randomly distributed in an incomplete Latin square design with 10-day periods. Treatments included: Thermoneutrality with pair feeding + corn oil (TNPF/n6), Heat stress + corn oil (HS/n6), and Heat stress + fish oil (HS/n3). The oils (159 g/day) were infused into the abomasum in two boluses. Blood samples were collected on days 0, 5, and 10 for lipidomic analysis by LC-MS/MS chromatography. The fatty acids eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), and arachidonic acid 13 (AA), together with the oxylipids of the lipoxygenase pathway 5-HETE, 5-oxoETE, 15- HETE, 15-oxoETE, 17-HDoHe and cytochrome 450 oxylipids: 19,20-EpDPE and 19,20- DiHDPA, increased in HS/n3 compared to HS/n6 (P < 0.10). 9-oxoODE was reduced in HS/n6 compared to TNPF/n6. HS/n3 treatment led to elevated concentrations of antiinflammatory oxylipids compared to HS/n6 (P < 0.10). During heat stress, substrate availability, route of synthesis and duration of exposure indicate the potential for exogenous modulation of lipid mediators by omega-3. In summary, lipid supplementation, especially with MCFA and omega-3 fatty acids, can influence metabolism and immune response in dairy cows. Although MCFA supplementation did not show significant effects on milk production and blood metabolites, it did affect gene expression and milk fatty acid profile. Omega-3 beneficially modulated lipid mediators during heat stress, highlighting its potential to improve the health and productivity of dairy cows