Gene expression and fatty acid profile of subcutaneous adipose tissue in lambs supplemented with stearic acid

Gene expression and fatty acid profile of subcutaneous adipose tissue in lambs supplemented with stearic acid | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Gene expression and fatty acid profile of subcutaneous adipose tissue in lambs supplemented with stearic acid Charline Godinho Padilha, Rafaella Horstmann, Rayllana Larsen, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6631087/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This study evaluated the effect of stearic acid (SA) supplementation on the fatty acid (FA) profile and the expression of genes involved in lipid synthesis in the subcutaneous adipose tissue of ewe lambs. Eight ewe lambs (50% Lacaune × 50% Texel), five months old, were used in a completely randomized design with two treatments: Control and SA (1.5% SA in dry matter intake). The experiment lasted 70 days, and the animals underwent subcutaneous adipose tissue biopsy on day 0 (initial period) and day 70 (end period) of supplementation. Samples from initial and end period were analyzed for FA profile, while gene expression analysis was conducted only on samples from end period. Statistical analysis was performed using SAS (OnDemand for Academics), with FA profile data analyzed using PROC MIXED, considering treatments, period, and their interactions as fixed effects, and animal as a random effect. Gene expression data were analyzed using PROC TTEST, considering treatments as fixed effects and animal as a random effect. SA supplementation increased lauric acid in subcutaneous adipose tissue by 68.8% over time, despite causing its own reduction (82%) and the decrease of C18:1 trans -11 (44.4%). SA supplementation increased the relative mRNA abundance of the GPAT gene (P = 0.0103), which is involved in triglyceride synthesis in subcutaneous adipose tissue. In conclusion, SA partially modified the FA profile and the expression of genes involved in subcutaneous adipose tissue synthesis in ewe lambs. Lipids sheep nutrigenomics genes Figures Figure 1 Figure 2 Figure 3 HIGHLIGHTS Stearic acid supplementation causes its own decrease in subcutaneous adipose tissue in ewe lambs. Ewe lambs supplemented with stearic acid have a higher concentration of lauric acid in subcutaneous adipose tissue. Ewe lambs supplemented with stearic acid demonstrate a higher relative mRNA abundance of the GPAT gene, which is involved in the synthesis of subcutaneous adipose tissue triglycerides. INTRODUCTION Lipid supplementation is used to enhance energy intake during specific life stages of animals according to their nutritional requirements. However, over time, additional benefits have been associated with the inclusion of fat in animal diets, such as increased milk production, higher fat yield (Jenkins, 1993 ; Loften, 2014), alterations in the milk fatty acid profile (Boerman et al., 2017 ), and changes in ruminal BH (Jenkins, 1993 ). Among the main lipid supplements, stearic acid (SA) stands out (Loften et al., 2014 ; Boerman et al., 2017 ; Souza et al., 2018 ), demonstrating positive effects when included in ruminant diets, particularly concerning fat content and milk production (Steele & Moore, 1968 ; Piantoni et al., 2015 ). Due to this potential effect, some studies have investigated whether this fatty acid (FA) could mitigate milk fat depression (MFD) (Toral et al., 2016; Aguiar et al., 2021 ), while others have reported that SA does not influence milk production or milk fat yield (Boerman et al., 2017 ; Horstmann et al., 2021). Compared to other FAs, SA appears to be more specifically directed toward adipose tissue, contributing to improved energy balance in animals, as well as participating in metabolic regulation and gene transcription in ruminants (Bionaz et al., 2011 ). It has also been shown to alter the expression of genes involved in lipid synthesis (Horstmann et al., 2021; Aguiar et al., 2021 ; Bessani et al., 2022). However, the mechanisms underlying the effects of SA on lipogenic metabolism in ruminants are not yet fully understood. Studies evaluating the influence of fat supplementation on FA profiles and the expression of genes involved in lipid synthesis are typically conducted in lactating animals through mammary glands (MG) studies. Moreover, research focusing on subcutaneous adipose tissue remains limited, as most studies in this field aim to assess animals bred for meat production. Additionally, studies using sheep as a developmental model are scarce. Thus, the objective of this study was to evaluate the ability of SA to alter the FA profile of subcutaneous adipose tissue and the expression of genes involved in lipid synthesis in ewe lambs. MATERIALS AND METHODS Experimental design, animals and treatments The study was conducted at the Center for Agroveterinary Sciences of the State University of Santa Catarina (latitude: 27º47’32”S and longitude: 50º18’23”W), and all procedures were approved by the Ethics Committee under protocol no. 8029060422. The experiment lasted 70 days and eight ewe lambs (50% Lacaune × 50% Texel), aged five months and weighing 35.1 ± 2.8 kg of body weight (BW), were used in a completely randomized design with two groups: (1) Control and (2) SA (1.5% of stearic acid in dry matter intake). The ewe lambs were nulliparous and housed in collective pens according to their treatment group. The composition of the SA-rich supplement is presented in Supplementary Table 1. Diet and management All animals had ad libitum access to water, and the diet was formulated according to SRNS (2010) (Table 1 ). All treatments were isoenergetic and isoproteic. The concentrate was provided individually, and SA was mixed into a portion of the concentrate to ensure total supplement consumption by the animals. Afterward, the remaining concentrate for that feeding period was provided. Table 1 Ingredients and composition of the experimental diet. Treatments CON SA Ingredientes, % de MS Corn silage 59.8 55.6 Ground corn 17.6 17.7 Soybean meal 17.6 17.6 Kaolim 3.5 7.1 Mineral/ vitamin nucleus 1.5 1.2 Chemical composition, % de MS DM¹ 88.5 90.1 CP 2 14.4 14.1 NDF 3 44.4 41.3 NSC 4 29.5 28.6 Mineral matter 9.2 12.4 Ether extract 3.6 4.6 Fa total 5 2.27 3.26 C18:0, % FA total 5.59 6.64 EM 6 (Mcal/kg) 2.35 2.34 DMI estimated 7 (kg/d) 1.42 1.44 ¹ Dry matter; ² Crude protein; ³ Fiber in neutral detergent; 4 Non-structural carbohydrates; 5 Total fatty acids estimated according to NASEM (2021); 6 Metabolizable energy; 7 Dry matter intake estimated; * The diet was calculated individually according to the SRNS (TEDESCHI et al., 2010). Silage was offered by treatment pen, and the diet was adjusted every 21 days according to the animals' growth. Feed was provided twice daily at a rate of 110% of voluntary dry matter intake. Samples of silage and concentrate were collected every 21 days throughout the experimental period, along with their respective refusals, and a composite sample of each material was subsequently prepared for composition analysis. (Table 1 near here). The animals were weighed, and their body condition score (BCS) was assessed every 21 days through the lumbar region. The BCS evaluation was performed by a single evaluator using a scale from 1 to 5, where 1 represents very thin animals and 5 represents excessively fat animals. Biopsy of subcutaneous adipose tissue Subcutaneous adipose tissue biopsies were performed on all animals on days 0 and 70 of the experimental period. Before the procedure, the biopsy site was shaved and disinfected with a 10% iodized alcohol solution. Local anesthesia was administered with 8 mL of lidocaine hydrochloride, applied in a circular pattern around the incision site. After 10 minutes, the biopsy was performed through an incision in the predetermined region (lateroventral to the sacroiliac area), collecting approximately 130 mg of tissue per animal. The samples were rinsed with saline solution, stored in duplicate in cryotubes containing phosphate-buffered saline (PBS), and placed in a nitrogen tank at -196°C. They were then stored at -20°C and − 80°C for subsequent fatty acid profile and gene expression analyses, respectively. The incision was closed using surgical adhesive, and the animals received intramuscular injections of meloxicam 2% (2.5 mL/100 kg) and oxytetracycline (1 mL/10 kg). The animals were monitored, and the incision site was cleaned daily for three days. RNA extraction, complementary DNA (cDNA) synthesis and quantitative Real-Time PCR (RT-qPCR) RNA extraction and complementary DNA (cDNA) synthesis was carried out weighing the samples (~ 80 mg) and immersing them in Eppendorf tubes containing QiAzol Lysis Reagent (Qiagen Sciences, Germantown, MD, USA). The tissues were then homogenized using a portable tissue homogenizer and RNA extraction was performed using the RNeasy Lipid Tissue Kit (Qiagen Sciences, Germantown, MD, USA), following the manufacturer’s protocol, with DNase treatment (RNase-free DNase set, Qiagen Sciences, Germantown, MD, USA) applied directly to the extraction column to prevent DNA contamination. RNA purity was verified using a NanoDrop ND-2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA), and the quality determined by A260/280 ratio (~ 2.1 ± 0.05). Total RNA was transcribed to complementary DNA (cDNA) using GoScript™ Reverse Transcription Mix and random primers (Promega Corporation, Madison, WI, USA). RT-qPCR was performed on a 48-well plate (Micro Amp, Applied Biosystems), using 5 µL of cDNA from the standard curve obtained by serial dilution of a pooled sample, along with 10 µL of a mix containing RNase-free H₂O, primers for the target gene, and SyberGreen (GoTaq qPCR Master Mix, Promega, Madison, WI, USA). Each sample was analyzed in triplicate of cDNA and mix. Quantification was performed using the StepOne Real-Time PCR System (Applied Biosystems), and data analysis was conducted with StepOne software version 2.1 (Applied Biosystems), generating dissociation curves to confirm the presence of a single amplification product. A regression equation was generated based on the RT-qPCR cycle values to assess reaction efficiency. The primers used in the analysis are listed in Supplementary Table 2. Fatty acid profile analysis The subcutaneou adipose tissue samples were weighed (50 mg), and fatty acids were methylated following the method described by Kramer et al. (1997), using sodium methoxide and acetyl chloride as basic and acidic catalysts, respectively. The resulting fatty acid methyl esters (FAMEs) were analyzed by gas chromatography (Focus GC model; Thermo Scientific, Milan, Italy) equipped with a flame ionization detector and a fused silica capillary column SP-2560 (100 m × 25 mm × 0.2 μm film thickness; Supelco, Bellefonte, Pennsylvania, USA). Hydrogen was used as the carrier gas (1 mL/min), and nitrogen was used as the auxiliary gas. The detector and injector temperatures were set at 250°C, with a split ratio of 15:1. The oven temperature program was set to 7°C for 4 min, followed by an increase of 13°C/min to 175°C, held for 27 min, then increased by 4°C/min to 215°C, and maintained for 31 min (Kramer et al., 1997). The FAMEs were identified by comparing them with three reference standards (Supelco FAME Mix # C4-C24, CLA trans -9, cis-11 #16413, and CLA trans -10 , cis -12 #04397; Sigma-Aldrich). The cis/trans -18:1 isomers were identified based on their elution order under the same chromatographic conditions (Kramer et al., 2002). Statistical analysis Statistical analysis was performed using SAS (OnDemand for Academics), considering significance at P ≤ 0.05 and a trend at P > 0.05 and < 0.10. Fatty acid profile data were analyzed using the PROC MIXED procedure, with treatment, period, and their interaction classified as fixed effects, while the animal was considered a random effect. For gene expression analysis, the PROC TTEST procedure was used, with treatment as a fixed effect and animal as a random effect. The analysis was based on the normalized gene values, which were obtained using the geometric mean of housekeeping genes (ribosomal protein S18 and β-actin). Data are presented as relative RNA abundance, with the Control group set at 100%. Studentized residuals beyond ± 3.0 were considered outliers and excluded from the statistical analysis. Data normality was confirmed using the Shapiro-Wilk test. RESULTS Dry matter intake and animal performance Total dry matter intake (DMI) did not differ between treatments. However, silage intake was 8% lower, while concentrate intake was 12.7% higher in animals supplemented with stearic acid (SA). Regarding total estimated metabolizable energy (ME) intake, a difference was observed between treatments (P = 0.0001), with SA-supplemented animals consuming 5.6% more ME (2.24 vs. 2.13 Mcal/day) and showing an 11% increase in ME intake from concentrate (1.61 vs. 1.45 Mcal/day; P = 0.0001). Body condition score (BCS) also differed between treatments (P = 0.004), with a mean score of 3.8 in the SA group and 3.4 in the Control group. However, no significant differences were observed in live weight (LW) or average daily gain (ADG). Fatty acid profile of subcutaneous adipose tissue The fatty acid (FA) profile of subcutaneous adipose tissue is presented in Table 2 . The interactions between period and treatment were observed for C12:0 (P = 0.01), C18:0 (P = 0.001), C18:1 trans -11 (P = 0.06) and CLA cis -9 trans -11 (P = 0.08). (Table 2 near here) Table 2 Effect of stearic acid (SA) supplementation on the fatty acid profile in adipose tissue of ewe lambs. Treatment CON SA CON SA P – value 3 Inicial End SEM 2 Treatment Period 4 Treat x Per. 5 C10:0 0.21 0.22 0.20 0.26 0.01 0.61 0.60 0.26 C12:0 0.21 0.22 0.16 0.27 0.001 0.006 0.91 0.009 C14:0 2.9 3.2 3.0 2.9 0.07 0.45 0.60 0.28 C14:1 0.20 0.22 0.19 0.19 0.01 0.45 0.34 0.40 C16:0 26.2 26.4 26.0 24.7 0.35 0.23 0.17 0.30 C16:1 1.8 1.8 1.9 2.5 0.04 0.02 0.002 0.44 C18:0 22.0 24.2 19.5 13.3 0.10 0.09 < 0.001 0.001 C18:1 trans -9 0.49 0.50 0.43 0.49 0.01 0.16 0.16 0.34 C18:1 trans -11 1.1 1.3 1.2 0.9 0.07 0.59 0.16 0.06 C18:1 cis -9 35.0 32.3 36.3 38.8 0.13 0.98 0.07 0.27 C18:1 cis- 11 0.68 0.72 0.70 0.79 0.02 0.003 0.18 0.43 C18:2 n-6 1.86 1.78 1.70 2.2 0.01 0.36 0.67 0.22 C18:3 n-3 0.42 0.43 0.37 0.54 0.03 0.66 0.49 0.11 CLA cis -9 trans -11 0.18 0.22 0.25 0.21 0.01 0.19 0.24 0.08 C20:4 n- 6 0.11 0.12 0.13 0.13 0.005 0.04 0.02 0.55 ¹The treatments were CON (control) and 1.5% SA (stearic acid) in dry matter intake; ²Standard error of the mean; ³Significance level (P < 0.05); ⁴Significance of the treatment administration period; ⁵Interaction between treatment and time. The C12:0 levels were 68.8% (P = 0.001) higher in the SA group during the end period compared to the animals in the CON group (Fig. 1 A). The C18:0 value in the subcutaneous adipose tissue of the animals of group SA during the initial period was 10% lower (P = 0.06) compared to animals in the CON group in the same period, while in the comparison between the animals in the SA group and the CON group in the final period, this value was 46.6% lower (P = 0.0013; Fig. 1 B). (Fig. 1 near here) In the SA-treated group, C12:0 levels were 22.7% higher at the end of the experimental period compared to the initial period (P = 0.03: Fig. 2 A). Conversely, the C18:0 content in subcutaneous adipose tissue was 82% lower at the end of the period compared to the initial period in SA-supplemented animals (P = 0.001; Fig. 2 B). The levels of C18:1 trans -11 in subcutaneous adipose tissue did not differ between treatments at either the initial (P = 0.4507) or final (P = 0.7573) time points (Fig. 1 C). However, within the SA treatment group, a significant change was observed between the beginning and end of the supplementation period (P = 0.03); the SA supplementation led to a 44.4% reduction in C18:1 trans -11 over time (Fig. 2 C). The only FA showing a treatment effect between periods in the Control group was CLA cis -9 trans -11 (P = 0.08), which increased by 38.9% in the final period. Gene Expression SA supplementation increased the relative mRNA abundance of the GPAT gene by 67.9% compared to the Control group (P = 0.01) (Fig. 3 ). (Fig. 3 near here). There was no effect of SA supplementation (P > 0.05) on the expression of genes involved in de novo fatty acid synthesis (ACACAα and FASN), fatty acid uptake and transport (LPL, FABP4, and CD36), gene regulation (PPARγ), or triglyceride synthesis (DGAT and AGPAT), as shown in Table 3 . (Table 3 near here). Table 3 Relative abundance of mRNA of genes involved in lipid synthesis in adipose tissue of ewe lambs supplemented with stearic acid (SA) compared to Control (CON). Treatment Relative gene mRNA abundance (%) CON SA SEM P - value CD36 1.00 1.06 0.02 0.5737 ACACAα 1.00 1.34 0.04 0.3911 FASN 1.00 1.26 0.03 0.2980 GPAT 1.00 1.68 0.02 0.0103 DGAT 1.00 1.13 0.03 0.5671 AGPAT 1.00 1.03 0.01 0.7189 PPARγ 1.00 1.07 0.02 0.6472 LPL 1.00 1.07 0.02 0.6682 FABP4 1.00 1.12 0.02 0.4472 Values are presented as the mean ± SEM; ACACAα: Acetyl-CoA carboxylase alpha; FASN: Fatty acid synthase; CD36: CD36 molecule; LPL: Lipoprotein lipase; FABP4: Fatty acid-binding protein 4; GPAT: Glycerol-3-phosphate acyltransferase 1; AGPAT: Acylglycerolphosphate acyltransferase 6; DGAT: Diacylglycerol acyltransferase 1; PPARγ: Peroxisome proliferator-activated receptors. DISCUSSION Several studies have evaluated supplementation with sources rich in SA on productive and performance responses in cows (Boerman et al., 2017 ) and dairy sheep (Toral et al., 2015 ). However, to the best of the author's knowledge, no studies have evaluated the effect of SA in developing animals. In this study, it was observed that animals in the SA group exhibited an increase of 68.8% in the lauric acid (C12:0) content in subcutaneous adipose tissue. The mechanisms underlying this occurrence remain unclear; however, this finding may be considered beneficial, as studies indicate that medium-chain fatty acids, such as C12:0, contribute to milk fat synthesis in cows (Vyas et al., 2012 ) and have been associated with certain health benefits for consumers (Dayrit, 2015 ). Conversely, the SA content in the subcutaneous adipose tissue of animals receiving a supplement rich in this FA was 82% lower at the end of the 70-day supplementation period. Also, the concentration of SA in the subcutaneous adipose tissue of animals in the SA group was 10% lower compared to animals in the CON group in the same period, while in the comparison between the animals in the SA group and the CON group this difference increased to 46.6% in the final period. Although this was not tested in the present study, it may be explained by the digestibility of SA, which decreases as its inclusion in the diet increases (Ferlay et al., 1993; Boerman et al., 2015), consequently reducing its absorption and availability in adipose tissue. The addition of SA to the diet reduces the levels of de novo synthesized FAs, which may explain the absence of short-chain and most medium-chain FAs in adipose tissue (Hansen & Knudsen, 1987 ). It was also observed that SA was not capable of altering the expression of genes involved in FA synthesis (ACACAα and FASN), FA uptake and transport (LPL, FABP4, and CD36), gene regulation (PPARγ), or some genes involved in triglyceride synthesis (DGAT and AGPAT). In our study, the only gene that showed a difference in abundance in the subcutaneous adipose tissue of ewe lambs supplemented with SA compared to the Control was the GPAT gene. According to Bernard et al. ( 2006 ), this gene plays a crucial role in the final step of fat synthesis, as FAs are esterified to glycerol via glycerol-3-phosphate acyltransferase (GPAT), which acetylates glycerol-3-phosphate, considered the first step in triglyceride synthesis (Gonzalez-Baro et al., 2006). Given that SA was expected to increase the relative abundance of other genes involved in lipid synthesis in adipose tissue, it is necessary to consider the characteristics of the supplement and its effects on animal metabolism. A primary factor could be the potential hypophagic effects that may occur with saturated FA supplementation, although such effects are more pronounced with the use of unsaturated fat sources (Harvatine et al., 2005). Additionally, supplement digestibility may have influenced the observed results, as an increased influx of SA into the duodenum leads to a linear decrease in the digestibility of this FA (Boerman et al., 2015). Studies suggest that SA may have low solubility, impairing its incorporation into micelles and thus reducing absorption (Pantoja et al., 1995 ). In addition, Piantoni et al. ( 2015 ) calculated the digestibility of SA supplements and reported a digestibility of only 28.4%, emphasizing the low transfer of SA to milk fat compared to the supplementation of other FAs, such as C16:0, and a similar phenomenon could potentially occur in adipose tissue. Additionally, studies suggest that the absence of effects from FA supplementation in sheep may be related to the supplementation duration (Horstmann et al., 2021). However, studies with an average duration of four to seven weeks have reported changes in FA profile composition and gene expression (Toral et al., 2010 ; Gómez-Cortés et al., 2011 ; Castro-Carrera et al., 2015 ). Therefore, it is expected that changes in the mRNA abundance of genes involved in lipid metabolism would be more easily observed after relatively long supplementation periods. However, in our study, the duration of SA supplementation was longer than in previous studies, yet the effects of SA were not significant. Besides the potential reduced digestibility of SA, it is plausible to consider that the absence of effects on the expression of most lipogenic genes may also be related to the animals' positive energy balance. Previous data in cows indicate that a high availability of exogenous FAs (through duodenal infusion of rapeseed oil) did not affect adipose tissue lipogenic activities during early lactation but caused a reduction after peak lactation, when activity increased due to energy balance variations (Chilliard et al., 1991 ). CONCLUSION SA partially modifies the FA profile of subcutaneous adipose tissue in ewe lambs. Furthermore, SA does not alter the expression of most genes involved in FA synthesis and transport, except for the GPAT gene, which is crucial in triglyceride synthesis. Declarations Author contributions The authors' contributions to this work are as follows: C. G. P. and D. E. O. conceived the research; C.G.P., R.H. and R.L. carried out the adipose tissue biopsy procedure; C.G.P., R.H. and R.L. performed gene expression analysis; C.V.D.M.R. carried out analyzes of the fatty acid profile of adipose tissue; C.G.P. and D.E.O. conducted the statistical analysis of the data; and C.G.P. wrote the article. All authors involved in this article read, reviewed, and approved the final version of the manuscript. Conflict of interest We confirm that there are no known conflicts of interest associated with this publication, and that there has been no significant financial support for this work that could have influenced its outcome. We confirm that the manuscript has been read and approved by all named authors and that no other persons satisfied the criteria for authorship. Acknowledgments We thank the study group on nutrigenomics of ruminants and non-ruminants (NUTRIGER) and the study, research, and extension group with sheep (GEPEO) at the Universidade do Estado de Santa Catarina/CAV for providing the animals, materials, and equipment used in this study. We also thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) and the Fundo Estadual de Apoio a Manutenção e ao Desenvolvimento da Educação Superior (FUMDES) for the scholarship granted to Charline Godinho Padilha. We also would like to thank CNPq (Process 407240/2021-7) and UDESC/FAPESC (Process 853/2023) via the Programa Institucional de Apoio à Pesquisa (PAP), for financial assistance for this study. Statement of animal rights All procedures involving animals were conducted in accordance with the ethical standards outlined in the journal’s author guidelines. The study was approved by the Ethics Committee of the State University of Santa Catarina (protocol no. 8029060422), where the research was carried out at the Center for Agricultural and Veterinary Sciences (latitude: 27º47’32” S; longitude: 50º18’23” W). The authors further confirm that all experimental protocols complied with the European Union directives for the protection of animals used for scientific purposes. Data Availability Statement All data generated or analyzed during this study are available from the corresponding author upon request. We recognize the importance of transparency and reproducibility in scientific research and are committed to facilitating data sharing to support verification of results and further scientific inquiry. Funding This research was funded by the Coordination for the Improvement of Higher Education Personnel - Brazil (CAPES) and by the State Fund to Support the Maintenance and Development of Higher Education (FUMDES), through a scholarship granted to Charline Godinho Padilha. In addition to the additional financial support for carrying out the research provided by CNPq (Process 407240/2021-7) and UDESC/FAPESC (Process 853/2023) through the Institutional Research Support Program (PAP). References Bernard L., Leroux C., Chilliard Y., 2006. Characterisation and nutritional regulation of the main lipogenic genes in the ruminant lactating mammary gland. In: Sejrsen K, Hvelplund T, Nielsen MO (Eds.) Ruminant Physiology. Academic, Wageningen, pp. 295-326. Bionaz M., Thering BJ., Loor JJ., 2011. Fine metabolic regulation in ruminants via nutrient-gene interactions: saturated long-chain fatty acids increase expression of genes involved in lipid metabolism and immune response partly through PPAR-α activation. Br. J. Nutr., 107(2):179-191. doi: 10.1017/s0007114511002777. Boerman JP., et. 2015. Intestinal digestibility of long-chain fatty acids in lactating dairy cows: A meta-analysis and meta-regression. J. Dairy Sci., 98:8889-8903. Boerman JP., de Souza J., Lock AL., 2017. Milk production and nutrient digestibility responses to increasing levels of stearic acid supplementation of dairy cows. J. Dairy Sci., 100(4):2729-2738. Castro-Carrera T, et al. 2015. Dietary sunflower oil modulates milk fatty acid composition without major changes in adipose and mammary tissue fatty acid profile or related gene mRNA abundance in sheep. Animal, 9(4):582-591. Chilliard Y., et al. 1991. Duodenal rapeseed oil infusion in early and midlactation cows. 5. Milk fatty acids and adipose tissue lipogenic activities. J. Dairy Sci., 74(6):1844-1854. doi: 10.3168/jds.s0022-0302(91)78349-5. Dayrit FM., 2015. The properties of lauric acid and their significance in coconut oil. J. Am. Oil Chem. Soc., 92:1-15. de Aguiar GC., et al. 2021. Stearic acid does not overcome conjugated linoleic acid trans-10, cis-12-induced milk fat depression in lactating ewes. Br. J. Nutr., Oct 27:1-7. doi: 10.1017/S000711452100430X. de Souza J., Preseault CL., Lock AL., 2018. Altering the ratio of dietary palmitic, stearic, and oleic acids in diets with or without whole cottonseed affects nutrient digestibility, energy partitioning, and production responses of dairy cows. J. Dairy Sci., 101(1):172-185. doi: 10.3168/jds.2017-13460. Gómez-Cortés P, et al. 2011. Effects of different forage:concentrate ratios in dairy ewe diets supplemented with sunflower oil on animal performance and milk fatty acid profile. J. Dairy Sci., 94(9):4578-4588. doi: 10.3168/jds.2010-3803. Gonzalez-Baró MR., et al. 2006. Regulation of triglyceride metabolism II. Function of mitochondrial GPAT1 in the regulation of triacylglycerol biosynthesis and insulin action. AJP: Gastrointestinal and Liver Physiology, 292(5):1195-1199. Hansen HO., Knudsen J., 1987. Effect of exogenous long-chain fatty acids on lipid biosynthesis in dispersed ruminant mammary gland epithelial cells: Esterification of long-chain exogenous fatty acids. J. Dairy Sci., 70(7):1344-1349. Harvatine KJ., Allen MS., 2005. The effect of production level on feed intake, milk yield, and endocrine responses to two fatty acid supplements in lactating cows. J. Dairy Sci., 88(11):4018-4027. doi: 10.3168/jds.s0022-0302(05)73088-5. Horstmann R., at el. 2021. Stearic acid (C18:0) reduces the expression of lipogenic genes and productivity in late-lactating dairy ewes. Small Rumin. Res., 195:1-5. doi: 10.1016/j.smallrumres.2020.106296. Hussein M., et al. 2013 Conjugated linoleic acid-induced milk fat depression in lactating ewes is accompanied by reduced expression of mammary genes involved in lipid synthesis. J. Dairy Sci., 96(6):3825–3834. doi: 10.3168/jds.2013-6576. Jenkins TC., 1993. Lipid metabolism in the rumen. J. Dairy Sci., 76(12):3851-3863. Kramer JK., Blackadar CB., Zhou J., 2002. Evaluation of two GC columns (60-m SUPELCOWAX 10 and 100-m CP Sil 88) for analysis of milkfat with emphasis on CLA, 18:1, 18:2 and 18:3 isomers, and short- and long-chain FA. Lipids, 37(8). Kramer JK., et al. 1997. Evaluating acid and base catalysts in the methylation of milk and rumen fatty acids with special emphasis on conjugated dienes and total trans fatty acids. Lipids, 32:1219–1228. Loften JR, et al. 2014. Invited review: Palmitic and stearic acid metabolism in lactating dairy cows. J. Dairy Sci., 97(8):4661-4674. doi: 10.3168/jds.2014-7919. Pantoja J., Firkins JL., Eastridge ML., 1995. Site of digestion and milk production by cows fed fats differing in saturation, esterification, and chain length. J. Dairy Sci., 78:2247–2258. Piantoni P., Lock AL., Allen MS., 2015. Milk production responses to dietary stearic acid vary by production level in dairy cattle. J. Dairy Sci., 98(3):1938-1949. Steele W., Moore JH., 1968. The effects of a series of saturated fatty acids in the diet on milk-fat secretion in the cow. J. Dairy Res., 35:361–370. Ticiani E., et al. 2016. Transcriptional regulation of acetyl-CoA carboxylase α isoforms in dairy ewes during conjugated linoleic acid-induced milk fat depression. Animal, 10:1677-1683. Toral PG., et al. 2010. Changes in milk fatty acid profile and animal performance in response to fish oil supplementation, alone or in combination with sunflower oil, in dairy ewes. J. Dairy Sci., 93(4):1604-1615. Toral PG., et al. 2015. Comparison of the nutritional regulation of milk fat secretion and composition in cows and goats. J. Dairy Sci., 98(10):7277-7297. doi: 10.3168/jds.2015-9649. Toral PG., et al. 2010. Does supplemental 18:0 alleviate fish oil-induced milk fat depression in dairy ewes? J. Dairy Sci., 99(2):1-13. Vyas D., Teter BB., Erdman RA., 2012. Milk fat responses to dietary supplementation of short- and medium-chain fatty acids in lactating dairy cows. J. Dairy Sci., 95:5194–5202. doi: 10.3168/jds.2011-5277. Supplementary Files SupplementaryfilesCharlineGodinhoPadilha.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6631087","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":464299254,"identity":"d740bdde-be51-48a8-8461-fdddc45f776f","order_by":0,"name":"Charline Godinho Padilha","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABA0lEQVRIiWNgGAWjYDACCSB+YIBgy4EYBx4Q0pKA0GJgDNaSQFALgm2Q2MCAJIIN8M9ufvYhoeCOHIPY4YO3eSr+pM8PO/wQaIudnG4DDkvuHDOekWDwzJhBOi3ZmueMQe7G22kGQC3JxmYHsGsxkEgwBvrlcGKDdI6Z5Mw2oJbZCSAtBxK34dSS/hmkpb5BOv+b5Mx/BumGs9M/ENCSA7YlgUE6h03iY4NBgrx0Dn5bJG7kFIO0GLZJpxlbfDhmbLhBOqfgQIIBbr/wz0jfzPDhz2F5funkhzcSauTk5Wenb/7wocJODpcWOGCDOxWs0oCAchQg30CK6lEwCkbBKBgJAABu513nNolcgwAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0003-0371-241X","institution":"UDESC: Universidade do Estado de Santa Catarina","correspondingAuthor":true,"prefix":"","firstName":"Charline","middleName":"Godinho","lastName":"Padilha","suffix":""},{"id":464299255,"identity":"cfaa0319-6873-48eb-8b4f-c34d83f0ef6f","order_by":1,"name":"Rafaella Horstmann","email":"","orcid":"","institution":"UDESC: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Rafaella","middleName":"","lastName":"Horstmann","suffix":""},{"id":464299256,"identity":"b099cf22-dd6b-4d32-940a-805fdc8161e9","order_by":2,"name":"Rayllana Larsen","email":"","orcid":"","institution":"UDESC: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Rayllana","middleName":"","lastName":"Larsen","suffix":""},{"id":464299257,"identity":"e53c4cbd-a87c-4372-a63f-2b4ea5f2cb99","order_by":3,"name":"Cláudio Vaz Di Mambro Ribeiro","email":"","orcid":"","institution":"UFBA: Universidade Federal da Bahia","correspondingAuthor":false,"prefix":"","firstName":"Cláudio","middleName":"Vaz Di Mambro","lastName":"Ribeiro","suffix":""},{"id":464299258,"identity":"28743a60-5050-4579-88fb-13a0563884b0","order_by":4,"name":"Dimas Estrasulas de Oliveira","email":"","orcid":"","institution":"UDESC: Universidade do Estado de Santa Catarina","correspondingAuthor":false,"prefix":"","firstName":"Dimas","middleName":"Estrasulas","lastName":"de Oliveira","suffix":""}],"badges":[],"createdAt":"2025-05-09 19:24:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6631087/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6631087/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":83853970,"identity":"b829b394-b231-4e6b-9cfc-1e0a8e823e45","added_by":"auto","created_at":"2025-06-03 17:00:24","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":23388,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of the interaction between stearic acid (SA) treatment and the control (CON) and the experimental period (initial and final period) on the concentration of C12:0 (A), C18:0 (B) and C18:1 \u003cem\u003etrans-\u003c/em\u003e11 (C) in the adipose tissue of lambs.\u003c/p\u003e\n\u003cp\u003eValues are presented as the mean ± standard error of the mean (SEM). Different letters demonstrate interactions between stearic acid (SA) and Control (CON) treatment and period\u003c/p\u003e\n\u003cp\u003e(P\u0026lt;0.05).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6631087/v1/fdc476ec26d7cd58029a0725.png"},{"id":83854414,"identity":"11178718-9d53-461f-a6b2-1fbd5166ce30","added_by":"auto","created_at":"2025-06-03 17:08:24","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":27408,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of stearic acid (SA) supplementation over time on the concentration of C12:0 (A), C18:0 (B), and C18:1 \u003cem\u003etrans\u003c/em\u003e-11 (C) in the adipose tissue of lambs.\u003c/p\u003e\n\u003cp\u003eValues are presented as the mean ± standard error of the mean (SEM). Different letters demonstrate differences between the initial and final period in stearic acid (SA) treatment (P\u0026lt;0.05).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6631087/v1/8b6a70d0a3c7dee7319e70f5.png"},{"id":83853981,"identity":"2e96c31f-8cdc-45e1-9362-8772a33f83c9","added_by":"auto","created_at":"2025-06-03 17:00:24","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":19903,"visible":true,"origin":"","legend":"\u003cp\u003eAbundance of the GPAT gene in adipose tissue of ewe lambs supplemented with stearic acid (SA) compared to Control (CON)\u003c/p\u003e\n\u003cp\u003eValues are presented as the mean ± standard error of the mean (SEM). Different letters demonstrate important differences between stearic acid (SA) and Control (CON) treatment (P\u0026lt;0.05).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6631087/v1/9c0120eecb86e595769961a8.png"},{"id":85753020,"identity":"01b85ea1-34d2-4578-8a52-e101b01543c6","added_by":"auto","created_at":"2025-07-01 10:29:45","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":795026,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6631087/v1/660b7d63-ee75-4d8d-9c74-2a19f3abfa0b.pdf"},{"id":83854417,"identity":"f1e3381b-eaef-4919-ac9f-db2d2e5da677","added_by":"auto","created_at":"2025-06-03 17:08:24","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":19273,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryfilesCharlineGodinhoPadilha.docx","url":"https://assets-eu.researchsquare.com/files/rs-6631087/v1/259c1f9423ec02cd01e05201.docx"}],"financialInterests":"","formattedTitle":"Gene expression and fatty acid profile of subcutaneous adipose tissue in lambs supplemented with stearic acid","fulltext":[{"header":"HIGHLIGHTS","content":"\u003cul\u003e\n \u003cli\u003eStearic acid supplementation causes its own decrease in subcutaneous adipose tissue in ewe lambs.\u003c/li\u003e\n \u003cli\u003eEwe lambs supplemented with stearic acid have a higher concentration of lauric acid in subcutaneous adipose tissue.\u003c/li\u003e\n \u003cli\u003eEwe lambs supplemented with stearic acid demonstrate a higher relative mRNA abundance of the GPAT gene, which is involved in the synthesis of subcutaneous adipose tissue triglycerides.\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"INTRODUCTION","content":"\u003cp\u003eLipid supplementation is used to enhance energy intake during specific life stages of animals according to their nutritional requirements. However, over time, additional benefits have been associated with the inclusion of fat in animal diets, such as increased milk production, higher fat yield (Jenkins, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Loften, 2014), alterations in the milk fatty acid profile (Boerman et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), and changes in ruminal BH (Jenkins, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1993\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmong the main lipid supplements, stearic acid (SA) stands out (Loften et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Boerman et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Souza et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), demonstrating positive effects when included in ruminant diets, particularly concerning fat content and milk production (Steele \u0026amp; Moore, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1968\u003c/span\u003e; Piantoni et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Due to this potential effect, some studies have investigated whether this fatty acid (FA) could mitigate milk fat depression (MFD) (Toral et al., 2016; Aguiar et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), while others have reported that SA does not influence milk production or milk fat yield (Boerman et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Horstmann et al., 2021). Compared to other FAs, SA appears to be more specifically directed toward adipose tissue, contributing to improved energy balance in animals, as well as participating in metabolic regulation and gene transcription in ruminants (Bionaz et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). It has also been shown to alter the expression of genes involved in lipid synthesis (Horstmann et al., 2021; Aguiar et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Bessani et al., 2022).\u003c/p\u003e \u003cp\u003eHowever, the mechanisms underlying the effects of SA on lipogenic metabolism in ruminants are not yet fully understood. Studies evaluating the influence of fat supplementation on FA profiles and the expression of genes involved in lipid synthesis are typically conducted in lactating animals through mammary glands (MG) studies. Moreover, research focusing on subcutaneous adipose tissue remains limited, as most studies in this field aim to assess animals bred for meat production. Additionally, studies using sheep as a developmental model are scarce. Thus, the objective of this study was to evaluate the ability of SA to alter the FA profile of subcutaneous adipose tissue and the expression of genes involved in lipid synthesis in ewe lambs.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eExperimental design, animals and treatments\u003c/h2\u003e \u003cp\u003e The study was conducted at the Center for Agroveterinary Sciences of the State University of Santa Catarina (latitude: 27\u0026ordm;47\u0026rsquo;32\u0026rdquo;S and longitude: 50\u0026ordm;18\u0026rsquo;23\u0026rdquo;W), and all procedures were approved by the Ethics Committee under protocol no. 8029060422. The experiment lasted 70 days and eight ewe lambs (50% Lacaune \u0026times; 50% Texel), aged five months and weighing 35.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8 kg of body weight (BW), were used in a completely randomized design with two groups: (1) Control and (2) SA (1.5% of stearic acid in dry matter intake). The ewe lambs were nulliparous and housed in collective pens according to their treatment group. The composition of the SA-rich supplement is presented in Supplementary Table\u0026nbsp;1.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eDiet and management\u003c/h3\u003e\n\u003cp\u003eAll animals had ad libitum access to water, and the diet was formulated according to SRNS (2010) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). All treatments were isoenergetic and isoproteic. The concentrate was provided individually, and SA was mixed into a portion of the concentrate to ensure total supplement consumption by the animals. Afterward, the remaining concentrate for that feeding period was provided.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eIngredients and composition of the experimental diet.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCON\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSA\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIngredientes, % de MS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCorn silage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e59.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e55.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGround corn\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSoybean meal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKaolim\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMineral/ vitamin nucleus\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemical composition, % de MS\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDM\u0026sup1;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e88.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e90.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCP\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNDF\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e44.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e41.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNSC\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e28.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMineral matter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e12.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEther extract\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFa total\u003csup\u003e5\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18:0, % FA total\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.64\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEM\u003csup\u003e6\u003c/sup\u003e (Mcal/kg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDMI estimated\u003csup\u003e7\u003c/sup\u003e (kg/d)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"3\"\u003e\u0026sup1; Dry matter; \u0026sup2; Crude protein; \u0026sup3; Fiber in neutral detergent; \u003csup\u003e4\u003c/sup\u003e Non-structural carbohydrates; \u003csup\u003e5\u003c/sup\u003e Total fatty acids estimated according to NASEM (2021); \u003csup\u003e6\u003c/sup\u003e Metabolizable energy; \u003csup\u003e7\u003c/sup\u003e Dry matter intake estimated; * The diet was calculated individually according to the SRNS (TEDESCHI et al., 2010).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e Silage was offered by treatment pen, and the diet was adjusted every 21 days according to the animals' growth. Feed was provided twice daily at a rate of 110% of voluntary dry matter intake. Samples of silage and concentrate were collected every 21 days throughout the experimental period, along with their respective refusals, and a composite sample of each material was subsequently prepared for composition analysis. (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e near here).\u003c/p\u003e \u003cp\u003eThe animals were weighed, and their body condition score (BCS) was assessed every 21 days through the lumbar region. The BCS evaluation was performed by a single evaluator using a scale from 1 to 5, where 1 represents very thin animals and 5 represents excessively fat animals.\u003c/p\u003e\n\u003ch3\u003eBiopsy of subcutaneous adipose tissue\u003c/h3\u003e\n\u003cp\u003eSubcutaneous adipose tissue biopsies were performed on all animals on days 0 and 70 of the experimental period. Before the procedure, the biopsy site was shaved and disinfected with a 10% iodized alcohol solution. Local anesthesia was administered with 8 mL of lidocaine hydrochloride, applied in a circular pattern around the incision site. After 10 minutes, the biopsy was performed through an incision in the predetermined region (lateroventral to the sacroiliac area), collecting approximately 130 mg of tissue per animal.\u003c/p\u003e \u003cp\u003eThe samples were rinsed with saline solution, stored in duplicate in cryotubes containing phosphate-buffered saline (PBS), and placed in a nitrogen tank at -196\u0026deg;C. They were then stored at -20\u0026deg;C and \u0026minus;\u0026thinsp;80\u0026deg;C for subsequent fatty acid profile and gene expression analyses, respectively. The incision was closed using surgical adhesive, and the animals received intramuscular injections of meloxicam 2% (2.5 mL/100 kg) and oxytetracycline (1 mL/10 kg). The animals were monitored, and the incision site was cleaned daily for three days.\u003c/p\u003e\n\u003ch3\u003eRNA extraction, complementary DNA (cDNA) synthesis and quantitative Real-Time PCR (RT-qPCR)\u003c/h3\u003e\n\u003cp\u003eRNA extraction and complementary DNA (cDNA) synthesis was carried out weighing the samples (~\u0026thinsp;80 mg) and immersing them in Eppendorf tubes containing QiAzol Lysis Reagent (Qiagen Sciences, Germantown, MD, USA). The tissues were then homogenized using a portable tissue homogenizer and RNA extraction was performed using the RNeasy Lipid Tissue Kit (Qiagen Sciences, Germantown, MD, USA), following the manufacturer\u0026rsquo;s protocol, with DNase treatment (RNase-free DNase set, Qiagen Sciences, Germantown, MD, USA) applied directly to the extraction column to prevent DNA contamination.\u003c/p\u003e \u003cp\u003eRNA purity was verified using a NanoDrop ND-2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA), and the quality determined by A260/280 ratio (~\u0026thinsp;2.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05). Total RNA was transcribed to complementary DNA (cDNA) using GoScript\u0026trade; Reverse Transcription Mix and random primers (Promega Corporation, Madison, WI, USA).\u003c/p\u003e \u003cp\u003eRT-qPCR was performed on a 48-well plate (Micro Amp, Applied Biosystems), using 5 \u0026micro;L of cDNA from the standard curve obtained by serial dilution of a pooled sample, along with 10 \u0026micro;L of a mix containing RNase-free H₂O, primers for the target gene, and SyberGreen (GoTaq qPCR Master Mix, Promega, Madison, WI, USA). Each sample was analyzed in triplicate of cDNA and mix. Quantification was performed using the StepOne Real-Time PCR System (Applied Biosystems), and data analysis was conducted with StepOne software version 2.1 (Applied Biosystems), generating dissociation curves to confirm the presence of a single amplification product. A regression equation was generated based on the RT-qPCR cycle values to assess reaction efficiency. The primers used in the analysis are listed in Supplementary Table\u0026nbsp;2.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFatty acid profile analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe subcutaneou adipose tissue samples were weighed (50 mg), and fatty acids were methylated following the method described by Kramer et al. (1997), using sodium methoxide and acetyl chloride as basic and acidic catalysts, respectively. The resulting fatty acid methyl esters (FAMEs) were analyzed by gas chromatography (Focus GC model; Thermo Scientific, Milan, Italy) equipped with a flame ionization detector and a fused silica capillary column SP-2560 (100 m \u0026times; 25 mm \u0026times; 0.2 \u0026mu;m film thickness; Supelco, Bellefonte, Pennsylvania, USA). Hydrogen was used as the carrier gas (1 mL/min), and nitrogen was used as the auxiliary gas. The detector and injector temperatures were set at 250\u0026deg;C, with a split ratio of 15:1.\u003c/p\u003e\n\u003cp\u003eThe oven temperature program was set to 7\u0026deg;C for 4 min, followed by an increase of 13\u0026deg;C/min to 175\u0026deg;C, held for 27 min, then increased by 4\u0026deg;C/min to 215\u0026deg;C, and maintained for 31 min (Kramer et al., 1997). The FAMEs were identified by comparing them with three reference standards (Supelco FAME Mix # C4-C24, CLA \u003cem\u003etrans\u003c/em\u003e-9, cis-11 #16413, and CLA \u003cem\u003etrans\u003c/em\u003e-10\u003cem\u003e, cis\u003c/em\u003e-12 #04397; Sigma-Aldrich). The \u003cem\u003ecis/trans\u003c/em\u003e-18:1 isomers were identified based on their elution order under the same chromatographic conditions (Kramer et al., 2002).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis was performed using SAS (OnDemand for Academics), considering significance at P \u0026le; 0.05 and a trend at P \u0026gt; 0.05 and \u0026lt; 0.10. Fatty acid profile data were analyzed using the PROC MIXED procedure, with treatment, period, and their interaction classified as fixed effects, while the animal was considered a random effect.\u003c/p\u003e\n\u003cp\u003eFor gene expression analysis, the PROC TTEST procedure was used, with treatment as a fixed effect and animal as a random effect. The analysis was based on the normalized gene values, which were obtained using the geometric mean of housekeeping genes (ribosomal protein S18 and \u0026beta;-actin). Data are presented as relative RNA abundance, with the Control group set at 100%. Studentized residuals beyond \u0026plusmn; 3.0 were considered outliers and excluded from the statistical analysis. Data normality was confirmed using the Shapiro-Wilk test.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eDry matter intake and animal performance\u003c/h2\u003e \u003cp\u003eTotal dry matter intake (DMI) did not differ between treatments. However, silage intake was 8% lower, while concentrate intake was 12.7% higher in animals supplemented with stearic acid (SA). Regarding total estimated metabolizable energy (ME) intake, a difference was observed between treatments (P\u0026thinsp;=\u0026thinsp;0.0001), with SA-supplemented animals consuming 5.6% more ME (2.24 vs. 2.13 Mcal/day) and showing an 11% increase in ME intake from concentrate (1.61 vs. 1.45 Mcal/day; P\u0026thinsp;=\u0026thinsp;0.0001).\u003c/p\u003e \u003cp\u003eBody condition score (BCS) also differed between treatments (P\u0026thinsp;=\u0026thinsp;0.004), with a mean score of 3.8 in the SA group and 3.4 in the Control group. However, no significant differences were observed in live weight (LW) or average daily gain (ADG).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003eFatty acid profile of subcutaneous adipose tissue\u003c/b\u003e\u003c/h2\u003e \u003cp\u003eThe fatty acid (FA) profile of subcutaneous adipose tissue is presented in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The interactions between period and treatment were observed for C12:0 (P\u0026thinsp;=\u0026thinsp;0.01), C18:0 (P\u0026thinsp;=\u0026thinsp;0.001), C18:1 \u003cem\u003etrans\u003c/em\u003e-11 (P\u0026thinsp;=\u0026thinsp;0.06) and CLA \u003cem\u003ecis\u003c/em\u003e-9 \u003cem\u003etrans\u003c/em\u003e-11 (P\u0026thinsp;=\u0026thinsp;0.08). (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e near here)\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of stearic acid (SA) supplementation on the fatty acid profile in adipose tissue of ewe lambs.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCON\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eCON\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eSA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c9\" namest=\"c7\"\u003e \u003cp\u003eP \u0026ndash; value\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eInicial\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eEnd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSEM\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ePeriod\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eTreat x Per.\u003csup\u003e5\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC10:0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC12:0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.006\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.009\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC14:0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC14:1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC16:0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e26.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e26.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC16:1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.002\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18:0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e22.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e24.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e19.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e13.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.001\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18:1 \u003cem\u003etrans\u003c/em\u003e-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18:1 \u003cem\u003etrans\u003c/em\u003e-11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18:1 \u003cem\u003ecis\u003c/em\u003e-9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e35.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e32.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e36.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e38.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18:1 \u003cem\u003ecis-\u003c/em\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.43\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18:2 n-6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC18:3 n-3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCLA \u003cem\u003ecis\u003c/em\u003e-9 \u003cem\u003etrans\u003c/em\u003e-11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC20:4 n- 6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.55\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"9\"\u003e\u0026sup1;The treatments were CON (control) and 1.5% SA (stearic acid) in dry matter intake; \u0026sup2;Standard error of the mean; \u0026sup3;Significance level (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05); ⁴Significance of the treatment administration period; ⁵Interaction between treatment and time.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe C12:0 levels were 68.8% (P\u0026thinsp;=\u0026thinsp;0.001) higher in the SA group during the end period compared to the animals in the CON group (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). The C18:0 value in the subcutaneous adipose tissue of the animals of group SA during the initial period was 10% lower (P\u0026thinsp;=\u0026thinsp;0.06) compared to animals in the CON group in the same period, while in the comparison between the animals in the SA group and the CON group in the final period, this value was 46.6% lower (P\u0026thinsp;=\u0026thinsp;0.0013; Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e near here)\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn the SA-treated group, C12:0 levels were 22.7% higher at the end of the experimental period compared to the initial period (P\u0026thinsp;=\u0026thinsp;0.03: Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). Conversely, the C18:0 content in subcutaneous adipose tissue was 82% lower at the end of the period compared to the initial period in SA-supplemented animals (P\u0026thinsp;=\u0026thinsp;0.001; Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe levels of C18:1 \u003cem\u003etrans\u003c/em\u003e-11 in subcutaneous adipose tissue did not differ between treatments at either the initial (P\u0026thinsp;=\u0026thinsp;0.4507) or final (P\u0026thinsp;=\u0026thinsp;0.7573) time points (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC). However, within the SA treatment group, a significant change was observed between the beginning and end of the supplementation period (P\u0026thinsp;=\u0026thinsp;0.03); the SA supplementation led to a 44.4% reduction in C18:1 \u003cem\u003etrans\u003c/em\u003e-11 over time (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003eThe only FA showing a treatment effect between periods in the Control group was CLA \u003cem\u003ecis\u003c/em\u003e-9 \u003cem\u003etrans\u003c/em\u003e-11 (P\u0026thinsp;=\u0026thinsp;0.08), which increased by 38.9% in the final period.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eGene Expression\u003c/h2\u003e \u003cp\u003eSA supplementation increased the relative mRNA abundance of the GPAT gene by 67.9% compared to the Control group (P\u0026thinsp;=\u0026thinsp;0.01) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e near here).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThere was no effect of SA supplementation (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) on the expression of genes involved in \u003cem\u003ede novo\u003c/em\u003e fatty acid synthesis (ACACAα and FASN), fatty acid uptake and transport (LPL, FABP4, and CD36), gene regulation (PPARγ), or triglyceride synthesis (DGAT and AGPAT), as shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e near here).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eRelative abundance of mRNA of genes involved in lipid synthesis in adipose tissue of ewe lambs supplemented with stearic acid (SA) compared to Control (CON).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"1\" nameend=\"c8\" namest=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRelative gene mRNA abundance (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCON\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eSA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eSEM\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eP - value\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCD36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.5737\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eACACAα\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.3911\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFASN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.2980\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGPAT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.0103\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDGAT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.5671\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAGPAT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.7189\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePPARγ\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.6472\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLPL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.6682\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFABP4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e1.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e0.4472\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eValues are presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM; ACACAα: Acetyl-CoA carboxylase alpha; FASN: Fatty acid synthase; CD36: CD36 molecule; LPL: Lipoprotein lipase; FABP4: Fatty acid-binding protein 4; GPAT: Glycerol-3-phosphate acyltransferase 1; AGPAT: Acylglycerolphosphate acyltransferase 6; DGAT: Diacylglycerol acyltransferase 1; PPARγ: Peroxisome proliferator-activated receptors.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eSeveral studies have evaluated supplementation with sources rich in SA on productive and performance responses in cows (Boerman et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and dairy sheep (Toral et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). However, to the best of the author's knowledge, no studies have evaluated the effect of SA in developing animals. In this study, it was observed that animals in the SA group exhibited an increase of 68.8% in the lauric acid (C12:0) content in subcutaneous adipose tissue. The mechanisms underlying this occurrence remain unclear; however, this finding may be considered beneficial, as studies indicate that medium-chain fatty acids, such as C12:0, contribute to milk fat synthesis in cows (Vyas et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) and have been associated with certain health benefits for consumers (Dayrit, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eConversely, the SA content in the subcutaneous adipose tissue of animals receiving a supplement rich in this FA was 82% lower at the end of the 70-day supplementation period. Also, the concentration of SA in the subcutaneous adipose tissue of animals in the SA group was 10% lower compared to animals in the CON group in the same period, while in the comparison between the animals in the SA group and the CON group this difference increased to 46.6% in the final period. Although this was not tested in the present study, it may be explained by the digestibility of SA, which decreases as its inclusion in the diet increases (Ferlay et al., 1993; Boerman et al., 2015), consequently reducing its absorption and availability in adipose tissue.\u003c/p\u003e \u003cp\u003eThe addition of SA to the diet reduces the levels of \u003cem\u003ede novo\u003c/em\u003e synthesized FAs, which may explain the absence of short-chain and most medium-chain FAs in adipose tissue (Hansen \u0026amp; Knudsen, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1987\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIt was also observed that SA was not capable of altering the expression of genes involved in FA synthesis (ACACAα and FASN), FA uptake and transport (LPL, FABP4, and CD36), gene regulation (PPARγ), or some genes involved in triglyceride synthesis (DGAT and AGPAT). In our study, the only gene that showed a difference in abundance in the subcutaneous adipose tissue of ewe lambs supplemented with SA compared to the Control was the GPAT gene. According to Bernard et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2006\u003c/span\u003e), this gene plays a crucial role in the final step of fat synthesis, as FAs are esterified to glycerol via glycerol-3-phosphate acyltransferase (GPAT), which acetylates glycerol-3-phosphate, considered the first step in triglyceride synthesis (Gonzalez-Baro et al., 2006).\u003c/p\u003e \u003cp\u003eGiven that SA was expected to increase the relative abundance of other genes involved in lipid synthesis in adipose tissue, it is necessary to consider the characteristics of the supplement and its effects on animal metabolism. A primary factor could be the potential hypophagic effects that may occur with saturated FA supplementation, although such effects are more pronounced with the use of unsaturated fat sources (Harvatine et al., 2005). Additionally, supplement digestibility may have influenced the observed results, as an increased influx of SA into the duodenum leads to a linear decrease in the digestibility of this FA (Boerman et al., 2015). Studies suggest that SA may have low solubility, impairing its incorporation into micelles and thus reducing absorption (Pantoja et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e1995\u003c/span\u003e). In addition, Piantoni et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) calculated the digestibility of SA supplements and reported a digestibility of only 28.4%, emphasizing the low transfer of SA to milk fat compared to the supplementation of other FAs, such as C16:0, and a similar phenomenon could potentially occur in adipose tissue.\u003c/p\u003e \u003cp\u003eAdditionally, studies suggest that the absence of effects from FA supplementation in sheep may be related to the supplementation duration (Horstmann et al., 2021). However, studies with an average duration of four to seven weeks have reported changes in FA profile composition and gene expression (Toral et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; G\u0026oacute;mez-Cort\u0026eacute;s et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Castro-Carrera et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Therefore, it is expected that changes in the mRNA abundance of genes involved in lipid metabolism would be more easily observed after relatively long supplementation periods. However, in our study, the duration of SA supplementation was longer than in previous studies, yet the effects of SA were not significant.\u003c/p\u003e \u003cp\u003eBesides the potential reduced digestibility of SA, it is plausible to consider that the absence of effects on the expression of most lipogenic genes may also be related to the animals' positive energy balance. Previous data in cows indicate that a high availability of exogenous FAs (through duodenal infusion of rapeseed oil) did not affect adipose tissue lipogenic activities during early lactation but caused a reduction after peak lactation, when activity increased due to energy balance variations (Chilliard et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1991\u003c/span\u003e).\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eSA partially modifies the FA profile of subcutaneous adipose tissue in ewe lambs. Furthermore, SA does not alter the expression of most genes involved in FA synthesis and transport, except for the GPAT gene, which is crucial in triglyceride synthesis.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors' contributions to this work are as follows: C. G. P. and D. E. O. conceived the research; C.G.P., R.H. and R.L. carried out the adipose tissue biopsy procedure; C.G.P., R.H. and R.L. performed gene expression analysis; C.V.D.M.R. carried out analyzes of the fatty acid profile of adipose tissue; C.G.P. and D.E.O. conducted the statistical analysis of the data; and C.G.P. wrote the article. All authors involved in this article read, reviewed, and approved the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe confirm that there are no known conflicts of interest associated with this publication, and that there has been no significant financial support for this work that could have influenced its outcome. We confirm that the manuscript has been read and approved by all named authors and that no other persons satisfied the criteria for authorship.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank the study group on nutrigenomics of ruminants and non-ruminants (NUTRIGER) and the study, research, and extension group with sheep (GEPEO) at the Universidade do Estado de Santa Catarina/CAV for providing the animals, materials, and equipment used in this study. We also thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) and the Fundo Estadual de Apoio a Manutenção e ao Desenvolvimento da Educação Superior (FUMDES) for the scholarship granted to Charline Godinho Padilha. We also would like to thank CNPq (Process 407240/2021-7) and UDESC/FAPESC (Process 853/2023) via the Programa Institucional de Apoio à Pesquisa (PAP), for financial assistance for this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatement of animal rights\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll procedures involving animals were conducted in accordance with the ethical standards outlined in the journal’s author guidelines. The study was approved by the Ethics Committee of the State University of Santa Catarina (protocol no. 8029060422), where the research was carried out at the Center for Agricultural and Veterinary Sciences (latitude: 27º47’32” S; longitude: 50º18’23” W). The authors further confirm that all experimental protocols complied with the European Union directives for the protection of animals used for scientific purposes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are available from the corresponding author upon request. We recognize the importance of transparency and reproducibility in scientific research and are committed to facilitating data sharing to support verification of results and further scientific inquiry.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was funded by the Coordination for the Improvement of Higher Education Personnel - Brazil (CAPES) and by the State Fund to Support the Maintenance and Development of Higher Education (FUMDES), through a scholarship granted to Charline Godinho Padilha. In addition to the additional financial support for carrying out the research provided by CNPq (Process 407240/2021-7) and UDESC/FAPESC (Process 853/2023) through the Institutional Research Support Program (PAP).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eBernard L., Leroux C., Chilliard Y., 2006. Characterisation and nutritional regulation of the main lipogenic genes in the ruminant lactating mammary gland. In: Sejrsen K, Hvelplund T, Nielsen MO (Eds.) Ruminant Physiology. Academic, Wageningen, pp. 295-326.\u003c/li\u003e\n\u003cli\u003eBionaz M., Thering BJ., Loor JJ., 2011. Fine metabolic regulation in ruminants via nutrient-gene interactions: saturated long-chain fatty acids increase expression of genes involved in lipid metabolism and immune response partly through PPAR-\u0026alpha; activation. Br. J. Nutr., 107(2):179-191. doi: 10.1017/s0007114511002777.\u003c/li\u003e\n\u003cli\u003eBoerman JP., et. 2015. Intestinal digestibility of long-chain fatty acids in lactating dairy cows: A meta-analysis and meta-regression. J. Dairy Sci., 98:8889-8903.\u003c/li\u003e\n\u003cli\u003eBoerman JP., de Souza J., Lock AL., 2017. Milk production and nutrient digestibility responses to increasing levels of stearic acid supplementation of dairy cows. J. Dairy Sci., 100(4):2729-2738.\u003c/li\u003e\n\u003cli\u003eCastro-Carrera T, et al. 2015. Dietary sunflower oil modulates milk fatty acid composition without major changes in adipose and mammary tissue fatty acid profile or related gene mRNA abundance in sheep. Animal, 9(4):582-591.\u003c/li\u003e\n\u003cli\u003eChilliard Y., et al. 1991. Duodenal rapeseed oil infusion in early and midlactation cows. 5. Milk fatty acids and adipose tissue lipogenic activities. J. Dairy Sci., 74(6):1844-1854. doi: 10.3168/jds.s0022-0302(91)78349-5.\u003c/li\u003e\n\u003cli\u003eDayrit FM., 2015. The properties of lauric acid and their significance in coconut oil. J. Am. Oil Chem. Soc., 92:1-15.\u003c/li\u003e\n\u003cli\u003ede Aguiar GC., et al. 2021. Stearic acid does not overcome conjugated linoleic acid trans-10, cis-12-induced milk fat depression in lactating ewes. Br. J. Nutr., Oct 27:1-7. doi: 10.1017/S000711452100430X.\u003c/li\u003e\n\u003cli\u003ede Souza J., Preseault CL., Lock AL., 2018. Altering the ratio of dietary palmitic, stearic, and oleic acids in diets with or without whole cottonseed affects nutrient digestibility, energy partitioning, and production responses of dairy cows. J. Dairy Sci., 101(1):172-185. doi: 10.3168/jds.2017-13460.\u003c/li\u003e\n\u003cli\u003eG\u0026oacute;mez-Cort\u0026eacute;s P, et al. 2011. Effects of different forage:concentrate ratios in dairy ewe diets supplemented with sunflower oil on animal performance and milk fatty acid profile. J. Dairy Sci., 94(9):4578-4588. doi: 10.3168/jds.2010-3803.\u003c/li\u003e\n\u003cli\u003eGonzalez-Bar\u0026oacute; MR., et al. 2006. Regulation of triglyceride metabolism II. Function of mitochondrial GPAT1 in the regulation of triacylglycerol biosynthesis and insulin action. AJP: Gastrointestinal and Liver Physiology, 292(5):1195-1199.\u003c/li\u003e\n\u003cli\u003eHansen HO., Knudsen J., 1987. Effect of exogenous long-chain fatty acids on lipid biosynthesis in dispersed ruminant mammary gland epithelial cells: Esterification of long-chain exogenous fatty acids. J. Dairy Sci., 70(7):1344-1349.\u003c/li\u003e\n\u003cli\u003eHarvatine KJ., Allen MS., 2005. The effect of production level on feed intake, milk yield, and endocrine responses to two fatty acid supplements in lactating cows. J. Dairy Sci., 88(11):4018-4027. doi: 10.3168/jds.s0022-0302(05)73088-5.\u003c/li\u003e\n\u003cli\u003eHorstmann R., at el. 2021. Stearic acid (C18:0) reduces the expression of lipogenic genes and productivity in late-lactating dairy ewes. Small Rumin. Res., 195:1-5. doi: 10.1016/j.smallrumres.2020.106296.\u003c/li\u003e\n\u003cli\u003eHussein M., et al. 2013 Conjugated linoleic acid-induced milk fat depression in lactating ewes is accompanied by reduced expression of mammary genes involved in lipid synthesis. J. Dairy Sci., 96(6):3825\u0026ndash;3834. doi: 10.3168/jds.2013-6576.\u003c/li\u003e\n\u003cli\u003eJenkins TC., 1993. Lipid metabolism in the rumen. J. Dairy Sci., 76(12):3851-3863.\u003c/li\u003e\n\u003cli\u003eKramer JK., Blackadar CB., Zhou J., 2002. Evaluation of two GC columns (60-m SUPELCOWAX 10 and 100-m CP Sil 88) for analysis of milkfat with emphasis on CLA, 18:1, 18:2 and 18:3 isomers, and short- and long-chain FA. Lipids, 37(8).\u003c/li\u003e\n\u003cli\u003eKramer JK., et al. 1997. Evaluating acid and base catalysts in the methylation of milk and rumen fatty acids with special emphasis on conjugated dienes and total \u003cem\u003etrans\u003c/em\u003e fatty acids. Lipids, 32:1219\u0026ndash;1228.\u003c/li\u003e\n\u003cli\u003eLoften JR, et al. 2014. Invited review: Palmitic and stearic acid metabolism in lactating dairy cows. J. Dairy Sci., 97(8):4661-4674. doi: 10.3168/jds.2014-7919.\u003c/li\u003e\n\u003cli\u003ePantoja J., Firkins JL., Eastridge ML., 1995. Site of digestion and milk production by cows fed fats differing in saturation, esterification, and chain length. J. Dairy Sci., 78:2247\u0026ndash;2258.\u003c/li\u003e\n\u003cli\u003ePiantoni P., Lock AL., Allen MS., 2015. Milk production responses to dietary stearic acid vary by production level in dairy cattle. J. Dairy Sci., 98(3):1938-1949.\u003c/li\u003e\n\u003cli\u003eSteele W., Moore JH., 1968. The effects of a series of saturated fatty acids in the diet on milk-fat secretion in the cow. J. Dairy Res., 35:361\u0026ndash;370.\u003c/li\u003e\n\u003cli\u003eTiciani E., et al. 2016. Transcriptional regulation of acetyl-CoA carboxylase \u0026alpha; isoforms in dairy ewes during conjugated linoleic acid-induced milk fat depression. Animal, 10:1677-1683.\u003c/li\u003e\n\u003cli\u003eToral PG., et al. 2010. Changes in milk fatty acid profile and animal performance in response to fish oil supplementation, alone or in combination with sunflower oil, in dairy ewes. J. Dairy Sci., 93(4):1604-1615.\u003c/li\u003e\n\u003cli\u003eToral PG., et al. 2015. Comparison of the nutritional regulation of milk fat secretion and composition in cows and goats. J. Dairy Sci., 98(10):7277-7297. doi: 10.3168/jds.2015-9649.\u003c/li\u003e\n\u003cli\u003eToral PG., et al. 2010. Does supplemental 18:0 alleviate fish oil-induced milk fat depression in dairy ewes? J. Dairy Sci., 99(2):1-13.\u003c/li\u003e\n\u003cli\u003eVyas D., Teter BB., Erdman RA., 2012. Milk fat responses to dietary supplementation of short- and medium-chain fatty acids in lactating dairy cows. J. Dairy Sci., 95:5194\u0026ndash;5202. doi: 10.3168/jds.2011-5277.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Lipids, sheep, nutrigenomics, genes","lastPublishedDoi":"10.21203/rs.3.rs-6631087/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6631087/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study evaluated the effect of stearic acid (SA) supplementation on the fatty acid (FA) profile and the expression of genes involved in lipid synthesis in the subcutaneous adipose tissue of ewe lambs. Eight ewe lambs (50% Lacaune × 50% Texel), five months old, were used in a completely randomized design with two treatments: Control and SA (1.5% SA in dry matter intake). The experiment lasted 70 days, and the animals underwent subcutaneous adipose tissue biopsy on day 0 (initial period) and day 70 (end period) of supplementation. Samples from initial and end period were analyzed for FA profile, while gene expression analysis was conducted only on samples from end period. Statistical analysis was performed using SAS (OnDemand for Academics), with FA profile data analyzed using PROC MIXED, considering treatments, period, and their interactions as fixed effects, and animal as a random effect. Gene expression data were analyzed using PROC TTEST, considering treatments as fixed effects and animal as a random effect. SA supplementation increased lauric acid in subcutaneous adipose tissue by 68.8% over time, despite causing its own reduction (82%) and the decrease of C18:1 \u003cem\u003etrans\u003c/em\u003e-11 (44.4%). SA supplementation increased the relative mRNA abundance of the GPAT gene (P = 0.0103), which is involved in triglyceride synthesis in subcutaneous adipose tissue. In conclusion, SA partially modified the FA profile and the expression of genes involved in subcutaneous adipose tissue synthesis in ewe lambs.\u003c/p\u003e","manuscriptTitle":"Gene expression and fatty acid profile of subcutaneous adipose tissue in lambs supplemented with stearic acid","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-03 17:00:19","doi":"10.21203/rs.3.rs-6631087/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b2645e44-1c35-4811-8e3c-b08716c4e71c","owner":[],"postedDate":"June 3rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-07-01T10:21:38+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-03 17:00:19","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6631087","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6631087","identity":"rs-6631087","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}