Intramuscular Fatty Acid Research Articles

Gene co-expression network analysis for porcine intramuscular fatty acid composition

In pigs, meat quality depends markedly on the fatty acid (FA) content and composition of the intramuscular fat, which is partly determined by the gene expression in this tissue. The aim of this work was to identify the link between muscle gene expression and its FA composition. In an (Iberian × Duroc) × Duroc backcrossed pig population, we identified modules of co-expressed genes, and correlation analyses were performed for each of them versus the phenotypes, finding four relevant modules. Two of the modules were positively correlated with saturated FAs (SFAs) and monounsaturated FAs (MUFAs), while negatively correlated with polyunsaturated FAs (PUFAs) and the omega-6/omega-3 ratio. The gene-enrichment analysis showed that these modules had over-representation of pathways related with the biosynthesis of unsaturated FAs, the Peroxisome proliferator-activated receptor signalling pathway and FA elongation. The two other relevant modules were positively correlated with PUFA and the n-6/n-3 ratio, but negatively correlated with SFA and MUFA. In this case, they had an over-representation of pathways related with fatty and amino acid degradation, and with oxidative phosphorylation. Using a graphical Gaussian model, we inferred a network of connections between the genes within each module. The first module had 52 genes with 87 connections, and the most connected genes were ADIPOQ, which is related with FA oxidation, and ELOVL6 and FABP4, both involved in FA metabolism. The second module showed 196 genes connected by 263 edges, being FN1 and MAP3K11 the most connected genes. On the other hand, the third module had 161 genes connected by 251 edges and ATG13 was the top neighbouring gene, while the fourth module had 224 genes and 655 connections, and its most connected genes were related with mitochondrial pathways. Overall, this work successfully identified relevant muscle gene networks and modules linked with FA composition, providing further insights on how the physiology of the pigs influences FA composition.

Lauric acid content in intramuscular fat is a reliable indicator of black soldier fly larvae meal consumption in Muscovy ducks

The present research investigated if intramuscular fatty acid (FA) profile could distinguish meat from ducks fed with black soldier fly larvae meal (BSFLM) during fattening. By using stepwise linear discriminant analysis on FA profiles of 96 meat samples, lauric acid (C12:0) was found to be the best predictor, accurately differentiating samples with only two misclassifications. The Fisher classification functions indicated that breast samples with lauric acid content above 0.222 % in intramuscular fat would be classified as BSFLM-fed. The Fisher classification functions were also effective in other poultry species fed BSFLM, with only two misclassified samples out of 42 samples from a dataset compiled from published papers. Misclassifications were linked to an unexpected lauric acid content in the intramuscular fat. In conclusion, given that BSFLM is the only relevant source of lauric acid in the diet, this fatty acid could identify meat from BSFLM-fed ducks and possibly other poultry species, warranting further exploration of other FA as potential identifiers of BSFLM-fed poultry.

Effects of dietary supplementation of polysaccharide from Agaricus blazei Murr on productive performance, egg quality, blood metabolites, intestinal morphology and microbiota of Korean quail.

This study aimed to investigate the effects of dietary supplementation with Agaricus blazei polysaccharide (ABP) at varying concentrations on the performance, egg quality, blood biochemistry, intestinal morphology, and microflora of quail. The study involved a total of 2,700 Korean quails, which were randomly divided into three groups. The measured variables encompassed productive performance, egg parameters, carcass parameters, serum metabolites, immune response parameters, antioxidative properties, and gut microbiome. The addition of ABP did not have a significant effect on average daily feed intake. However, it was found to increase the average daily egg weight and egg production rate, reduce the feed-egg ratio. There were no significant impacts on egg quality measures such as egg shape index, egg yolk index and color, egg yolk and protein content. However, ABP supplementation significantly increased the Hough unit (p<0.01) and decreased the rate of unqualified eggs (p<0.01). Regarding serum parameters, the inclusion led to an increase in total protein concentration (p<0.05) and a reduction in low-density lipoprotein cholesterol (p<0.05). There were no significant effects observed on immune indicators such as immunoglobulin A (IgA) and IgM. ABP supplementation increased the levels of serum antioxidant indicators, including glutathione peroxidase, total superoxide dismutase (p<0.05), and total antioxidant capacity colorimeter (p<0.05). Furthermore, ABP supplementation significantly elevated the intramuscular fatty acid content in quail meat. Additionally, ABP supplementation demonstrated a significant improvement in the diversity of gut microbiota and induced alterations in the composition of the gut microbiota. The findings of this study indicate that dietary supplementation of ABP enhanced production performance and antioxidant capacity while increasing the levels of polyunsaturated fatty acids in quail muscle.

Identification of candidate regulatory genes for intramuscular fatty acid composition in pigs by transcriptome analysis

BackgroundIntramuscular fat (IMF) content and its fatty acid (FA) composition are typically controlled by several genes, each with a small effect. In the current study, to pinpoint candidate genes and putative regulators involved in FA composition, we performed a multivariate integrative analysis between intramuscular FA and transcriptome profiles of porcine longissimus dorsi (LD) muscle. We also carried out a combination of network, regulatory impact factor (RIF), in silico prediction of putative target genes, and functional analyses to better support the biological relevance of our findings.ResultsFor this purpose, we used LD RNA-Seq and intramuscular FA composition profiles of 129 Iberian × Duroc backcrossed pigs. We identified 378 correlated variables (13 FA and 365 genes), including six FA (C20:4n-6, C18:2n-6, C20:3n-6, C18:1n-9, C18:0, and C16:1n-7) that were among the most interconnected variables in the predicted network. The detected FA-correlated genes include genes involved in lipid and/or carbohydrate metabolism or in regulation of IMF deposition (e.g., ADIPOQ, CHUK, CYCS, CYP4B1, DLD, ELOVL6, FBP1, G0S2, GCLC, HMGCR, IDH3A, LEP, LGALS12, LPIN1, PLIN1, PNPLA8, PPP1R1B, SDR16C5, SFRP5, SOD3, SNW1, and TFRC), meat quality (GALNT15, GOT1, MDH1, NEU3, PDHA1, SDHD, and UNC93A), and transport (e.g., EXOC7 and SLC44A2). Functional analysis highlighted 54 over-represented gene ontology terms, including well-known biological processes and pathways that regulate lipid and carbohydrate metabolism. RIF analysis suggested a pivotal role for six transcription factors (CARHSP1, LBX1, MAFA, PAX7, SIX5, and TADA2A) as putative regulators of gene expression and intramuscular FA composition. Based on in silico prediction, we identified putative target genes for these six regulators. Among these, TADA2A and CARHSP1 had extreme RIF scores and present novel regulators in pigs. In addition, the expression of TADA2A correlated (either positively or negatively) with C20:4n-6, C18:2n-6, C20:3n-6, C18:1n-9, and that of CARHSP1 correlated (positively) with the C16:1n-7 lipokine. We also found that these two transcription factors share target genes that are involved in lipid metabolism (e.g., GOT1, PLIN1, and TFRC).ConclusionsThis integrative analysis of muscle transcriptome and intramuscular FA profile revealed valuable information about key candidate genes and potential regulators for FA and lipid metabolism in pigs, among which some transcription factors are proposed to control gene expression and modulate FA composition differences.

STC2 Inhibits Hepatic Lipid Synthesis and Correlates with Intramuscular Fatty Acid Composition, Body Weight and Carcass Traits in Chickens

Stanniocalcin 2 (STC2) is a secreted glycoprotein involved in multiple biological processes. To systemically study the biological role of STC2 in chickens, phylogenetic tree analysis and conservation analysis were conducted. Association analysis between variations in the STC2 gene and the economic traits of Gushi-Anka F2 was conducted. The tissue expression patterns of STC2 expression in different chicken tissues and liver at different stages were detected. The biological role of STC2 in chicken liver was investigated through overexpression and interfering methods in the LMH cell line. Correlation analyses between STC2 expression and lipid components were conducted. (1) The phylogenetic tree displayed that chicken STC2 is most closely related with Japanese quail and most distantly related with Xenopus tropicalis. STC2 has the same identical conserved motifs as other species. (2) rs9949205 (T &gt; C) found in STC2 intron was highly significantly correlated with chicken body weight at 0, 2, 4, 6, 8, 10 and 12 weeks (p &lt; 0.01). Extremely significant correlations of rs9949205 with semi-evisceration weight (SEW), evisceration weight (EW), breast muscle weight (BMW), leg muscle weight (LMW), liver weight and abdominal fat weight (AFW) were revealed (p &lt; 0.01). Significant associations between rs9949205 and abdominal fat percentage, liver weight rate, breast muscle weight rate and leg muscle weight rate were also found (p &lt; 0.05). Individuals with TT or TC genotypes had significantly lower abdominal fat percentage and liver weight rate compared to those with the CC genotype, while their body weight and other carcass traits were higher. (3) STC2 showed a high expression level in chicken liver tissue, which significantly increased with the progression of age (p &lt; 0.05). STC2 was observed to inhibit the content of lipid droplets, triglycerides (TG) and cholesterol (TC), as well the expression level of genes related to lipid metabolism in LMH cells. (4) Correlation analysis showed that the STC2 gene was significantly correlated with 176 lipids in the breast muscle (p &lt; 0.05) and mainly enriched in omega-3 and omega-6 unsaturated fatty acids. In conclusion, the STC2 gene in chicken might potentially play a crucial role in chicken growth and development, as well as liver lipid metabolism and muscle lipid deposition. This study provides a scientific foundation for further investigation into the regulatory mechanism of the STC2 gene on lipid metabolism and deposition in chicken liver.

Short- and Long-Term Effects of Split-Suckling in Pigs According to Birth Weight.

Forty-eight litters were used, with a total number of 645 piglets involved in the study. The split-suckling technique was applied to half of the litters at the end of farrowing by removing the heaviest piglets over three periods of 1 h. The piglets were individually weighed at 0, 1 d, and at weaning. Piglet losses were recorded daily. Traceability was maintained until the carcass splitting and meat analysis took place. Carcasses were eviscerated and weighed individually. Total mortality at weaning was affected by body weight, where the low-body-weight piglets showed a mortality rate almost four times higher than that of the normal-weight piglets. Mortality was highest in the first days of life, especially in the low-body-weight piglets. At weaning, split-suckling treatment caused a slight increase in mortality compared to the control group piglets (25% vs. 17.1%). Split-suckling had a positive effect on weight gain during the first 24 h of life (p = 0.014), and there was an interaction between treatment and parity (p = 0.007), with split-suckling being more effective in the primiparous sows compared to the multiparous sows. The piglets from litters receiving the split-suckling treatment had a lower average daily gain during the lactation period (p < 0.001) than the piglets from the control group. Weight gain during the first 24 h of life of the piglets subjected to split-suckling was higher than those of the control group. A lower IgG and α-tocopherol in plasma in the heavier piglets subjected to split-suckling treatment was observed in comparison to their respective control. The piglets from litters receiving the treatment showed a lower average daily gain during the lactation period (p < 0.001) than the piglets from the control group. No difference in slaughter weight was observed according to treatment. The pigs which received split-suckling treatment showed lower subcutaneous fat thickness (p < 0.0013) and higher lean meat yield (p < 0.0027), this effect being more marked in pigs from primiparous sows. Intramuscular fat concentration was higher in the Longissimus Dorsi muscle of the low-body-weight piglets. In the pigs that received split-suckling treatment, a higher concentration of C18:3n-3 (p = 0.036) and a tendency towards a higher concentration of C18:2n-6 (p = 0.107) and unsaturation index (p = 0.113) was observed in intramuscular fatty acids at slaughter, together with a lower concentration of C16:0 (p = 0.053) and SFA (p = 0.064). In conclusion, long-term response to split-suckling, particularly in low-birth-weight piglets, suggests an alteration in adiposity and metabolic regulation in these piglets that receive high levels of colostrum.

Identifying key DNA methylation sites and their cis-methylation quantitative loci for intramuscular fatty acid traits using genome and methylome data in Yorkshire pigs

Single-nucleotide polymorphisms (SNPs) that are located near CpG sites have the potential to influence DNA methylation levels, leading to distinct patterns of DNA methylation. These methylation quantitative trait loci (meQTLs) can influence methylation levels across the entire genome, thereby impacting phenotypic traits. In this study, we utilised reduced representation bisulphite sequencing (RRBS) to analyse DNA methylation in muscle tissue samples collected from 140 Yorkshire pigs. Additionally, we assessed SNPs using the Porcine 50K chip, examined 17 fatty acid traits, and conducted epigenome-wide association studies (EWAS) and genome-wide association studies (GWAS). Our investigation revealed numerous associations between DNA methylation and fatty acid traits, including C16:0, C18:0, C20:3n-3, and C22:6n-3. Notably, a substantial portion of these associations was specifically identified by EWAS and not GWAS. Subsequently, we performed GWAS to identify meQTLs using methylation levels of significant CpGs from the EWAS as traits. The results revealed the predominance of trans-meQTLs; however, we also discovered a cis-meQTL on SSC12. Finally, we reported a crucial CpG site (SSC12:3735523) and its cis-meQTL on SSC12, which are located in close proximity to the FASN gene. Our findings contribute to the identification of CpG sites and their cis-meQTLs associated with fatty acid traits, utilising comprehensive genome and methylome data. Moreover, our study provides novel insights into the molecular mechanisms underlying fatty acid traits in pigs.

Effect of male pig immunocastration on physical and chemical characteristics of Teruel dry-cured hams

Aim of study: To evaluate the effect of the type of male castration (surgical vs. immunological) on the characteristics of Teruel dry-cured hams. Area of study: Teruel and Zaragoza (Spain). Material and methods: A total of 14 dry-cured hams from Duroc × (Landrace × Large White) male pigs intended for the Spanish Protected Designation of Origin ˈTeruel hamˈ were used. Half of them belonged to surgical castrated males (SCM) and the other half to immunocastrated males (IM). Ham weight losses during processing, physical and chemical characteristics were analyzed. Main results: There were no differences (p&gt;0.05) due to the type of castration in ham weight losses throughout processing, thickness of subcutaneous fat, marbling, maximum stress and intramuscular fatty acid profile. However, hams from IM presented lower (p&lt;0.05) chroma than those from SCM. The type of castration had limited influence on chemical composition; only potassium nitrate and retinol contents were affected (p&lt;0.05), being lower in IM than in SCM. For volatile compounds, IM showed lower (p&lt;0.05) percentages of total alcohols and sulfur compounds and higher (p=0.012) proportion of total acids than SCM. All the hams had negligible androstenone content but IM presented higher (p&lt;0.05) skatole and indole concentrations than SCM, being these levels low. Research highlights: The type of castration in male pigs seems to have scarce influence on the quality of Teruel dry-cured hams, and therefore, immunocastration could be considered as a possible alternative to surgical castration.

Cannabidiol improves muscular lipid profile by affecting the expression of fatty acid transporters and inhibiting de novo lipogenesis

Obesity is one of the principal public health concerns leading to disturbances in glucose and lipid metabolism, which is a risk factor for several chronic diseases, including insulin resistance, type 2 diabetes mellitus, and cardiovascular diseases. In recent years, it turned out that cannabidiol (CBD) is a potential therapeutic agent in the treatment of obesity and its complications. Therefore, in the present study, we used CBD therapy (intraperitoneal injections in a dose of 10 mg/kg of body mass for 14 days) in a rat model of obesity induced by a high-fat diet (HFD). Gas–liquid chromatography and Western blotting were applied in order to determine the intramuscular lipid content and total expression of selected proteins in the white and red gastrocnemius muscle, respectively. Based on fatty acid composition, we calculated de novo lipogenesis ratio (16:0/18:2n-6), desaturation ratio (18:1n-9/18:0), and elongation ratios (18:0/16:0, 20:0/18:0, 22:0/20:0 and 24:0/22:0), in the selected lipid fractions. Two-week CBD administration significantly reduced the intramuscular fatty acids (FAs) accumulation and inhibited de novo lipogenesis in different lipid pools (in the free fatty acid, diacylglycerol, and triacylglycerol fractions) in both muscle types, which coincided with a decrease in the expression of membrane fatty acid transporters (fatty acid translocase, membrane-associated fatty acid binding protein, and fatty acid transport proteins 1 and 4). Moreover, CBD application profoundly improved the elongation and desaturation ratios, which was in line with downregulated expression of enzymes from the family of elongases and desaturases regardless of the metabolism presented by the muscle type. To our knowledge, this study is the first that outlines the novel effects of CBD action on skeletal muscle with different types of metabolism (oxidative vs. glycolytic).

Global analysis of the association between pig muscle fatty acid composition and gene expression using RNA-Seq

Fatty acids (FAs) play an essential role as mediators of cell signaling and signal transduction, affecting metabolic homeostasis and determining meat quality in pigs. However, FAs are transformed by the action of several genes, such as those encoding desaturases and elongases of FAs in lipogenic tissues. The aim of the current work was to identify candidate genes, biological processes, and pathways involved in the modulation of intramuscular FA profile from longissimus dorsi muscle. FA profile by gas chromatography of methyl esters and gene expression by RNA-Seq were determined in 129 Iberian × Duroc backcrossed pigs. An association analysis between the muscle transcriptome and its FA profile was performed, followed by a concordance and functional analysis. Overall, a list of well-known (e.g., PLIN1, LEP, ELOVL6, SC5D, NCOA2, ACSL1, MDH1, LPL, LGALS12, TFRC, GOT1, and FBP1) and novel (e.g., TRARG1, TANK, ENSSSCG00000011196, and ENSSSCG00000038429) candidate genes was identified, either in association with specific or several FA traits. Likewise, several of these genes belong to biological processes and pathways linked to energy, lipid, and carbohydrate metabolism, which seem determinants in the modulation of FA compositions. This study can contribute to elucidate the complex relationship between gene expression and FA profile in pig muscle.

Supplementation with rumen-inert fat in the growing phase altered adipogenic gene expression and the size and number of adipocytes in Hanwoo steers.

We hypothesized that the provision of rumen-inert fat (RIF) to growing cattle (9 to 13 mo of age) would affect the expression of genes involved in lipid metabolism and thereby affect the size and number of adipocytes of steers slaughtered at 30 mo of age. Thirty steers with an average initial body weight (BW) of 239 ± 25 kg were allocated to six pens, balanced for BW and genetic merit for marbling, and assigned to one of two treatment groups: control (only basal diet) or test diet (basal diet with 200 g of RIF per day, on an as-fed basis) for 5 mo. Biopsy samples of longissimus lumborum (LM) muscle were then collected for analysis of fatty acid composition and gene expression. Both groups were then fed the same basal diets during the early and late fattening phases, without RIF, until slaughter (average shrunk BW = 759 kg). Supplementation with RIF increased the longissimus thoracis (LT) intramuscular fatty acid concentration at slaughter (P = 0.087) and numerically increased the quality grade score (P = 0.106). The LM intramuscular relative mRNA expression of genes such as PPARα, ZFP423 and SREBP1, FASN, SCD, FABP4, GPAT1, and DGAT2 were downregulated (P < 0.1) following RIF supplementation. Supplementation of RIF decreased (P < 0.1) diameter and concomitantly increased intramuscular adipocytes per viewing section at slaughter. This likely was caused by promotion of triacylglycerol hydrolysis during the growing phase. Another possible explanation is that the relative mRNA expression of gene ATGL was upregulated by RIF supplementation during the growing (P < 0.1) and the fattening phases (P < 0.05), while the genes associated with fatty acid uptake (FABP4) and esterification (DGAT2) were downregulated during the growing phase and upregulated (P < 0.1) during the fattening phase. This implies that the lipid turnover rate was higher for steers during the growing than fattening phase. This study demonstrated that RIF supplementation during the growing phase induced a carryover effect on the lipogenic transcriptional regulation involved in adipocyte lipid content of intramuscular adipose tissue; increased triacylglycerol hydrolysis during the growing phase subsequently was followed by increased lipid accumulation during the fattening phases.

Supplementation with Sunflower/Fish Oil-Containing Concentrates in a Grass-Based Beef Production System: Influence on Fatty Acid Composition, Gene Expression, Lipid and Colour Stability and Sensory Characteristics of Longissimus Muscle.

Beef contains an array of conjugated linoleic acid (CLA) isomers for which positive effects have been reported in animal models of human disease. The objectives were to develop a CLA-enriched beef production system and to assess its quality. Sixty Spring-born heifers were housed in Autumn and offered unwilted grass silage and a barley/soyabean concentrate or wilted grass silage and a concentrate containing sunflower oil and fish oil. In May, both groups were offered either pasture for 22 weeks, restricted pasture and sunflower oil and fish oil for 22 weeks, or pasture for 11 weeks and restricted pasture and sunflower oil and fish oil for the final 11 weeks. The predominant CLA isomer in beef was cis9, trans11 representing on average, 80% total CLA. The modified winter diet followed by supplementation for 22 weeks resulted in beef that had a CLA concentration that was higher, at a comparable intramuscular fatty acid concentration, than previously reported. The lipid and colour stability (over 10 days in modified atmosphere packaging) and sensory characteristics were generally not negatively affected. There were minor effects on the expression of candidate genes involved in lipid metabolism. Consumption of this beef would make a substantial contribution to the quantity of CLA suggested to have a positive effect on consumer health.

ELOVL gene family plays a virtual role in response to breeding selection and lipid deposition in different tissues in chicken (Gallus gallus)

BackgroundElongases of very long chain fatty acids (ELOVLs), a family of first rate-limiting enzymes in the synthesis of long-chain fatty acids, play an essential role in the biosynthesis of complex lipids. Disrupting any of ELOVLs affects normal growth and development in mammals. Genetic variations in ELOVLs are associated with backfat or intramuscular fatty acid composition in livestock. However, the effects of ELOVL gene family on breeding selection and lipid deposition in different tissues are still unknown in chickens.ResultsGenetic variation patterns and genetic associations analysis showed that the genetic variations of ELOVL genes were contributed to breeding selection of commercial varieties in chicken, and 14 SNPs in ELOVL2-6 were associated with body weight, carcass or fat deposition traits. Especially, one SNP rs17631638T > C in the promoter of ELOVL3 was associated with intramuscular fat content (IMF), and its allele frequency was significantly higher in native and layer breeds compared to that in commercial broiler breeds. Quantitative real-time PCR (qRT-PCR) determined that the ELOVL3 expressions in pectoralis were affected by the genotypes of rs17631638T > C. In addition, the transcription levels of ELOVL genes except ELOVL5 were regulated by estrogen in chicken liver and hypothalamus with different regulatory pathways. The expression levels of ELOVL1-6 in hypothalamus, liver, abdominal fat and pectoralis were correlated with abdominal fat weight, abdominal fat percentage, liver lipid content and IMF. Noteworthily, expression of ELOVL3 in pectoralis was highly positively correlated with IMF and glycerophospholipid molecules, including phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol and phospholipids inositol, rich in ω-3 and ω-6 long-chain unsaturated fatty acids, suggesting ELOVL3 could contribute to intramuscular fat deposition by increasing the proportion of long-chain unsaturated glycerophospholipid molecules in pectoralis.ConclusionsIn summary, we demonstrated the genetic contribution of ELOVL gene family to breeding selection for specialized varieties, and revealed the expression regulation of ELOVL genes and their potential roles in regulating lipid deposition in different tissues. This study provides new insights into understanding the functions of ELOVL family on avian growth and lipid deposition in different tissues and the genetic variation in ELOVL3 may aid the marker-assisted selection of meat quality in chicken.

Performance, meat quality, intramuscular fatty acid profile, rumen characteristics and serum parameters of lambs fed microencapsulated or conventional linseed oil

Microencapsulated linseed oil (MLO) has been used in diets to enhance the omega-3 fatty acid concentration of meat in beef cattle without negatively affecting ruminal microbials and digestion; however, the growth performance, meat quality, intramuscular fatty acid profile, and rumen characteristics in response to MLO have not been examined in sheep. Thus, the aim of the present study was to evaluate the effects of microencapsulated or conventional linseed oil supplement on growth performance, meat quality, intramuscular fatty acid profile, rumen characteristics and serum parameters in Small-tailed Han sheep. Eighteen 5-months-old male sheep (25.31 ± 1.31 kg) were allocated to three groups. After seven days of adaptation to a basal diet, fed for 80 days, the treatments allocated were (1) basal diet (CON); (2) basal diet with added 2% linseed oil (LO); (3) basal diet with added 4% MLO. The results showed that treatments had no effects on growth performance (P ≥ 0.10). Compared to CON and MLO group, the pH&lt;sub&gt;24 h &lt;/sub&gt;of the longissimus dorsi (LD) muscle in LO group was lower (P = 0.07), while the shear force was higher (P = 0.01). Compared to CON group, the addition of linseed oil increased proportions of C17:1, C18:2 n-6c, total polyunsaturated fatty acids (PUFA), PUFA/saturated fatty acids and total n-6, while it decreased the proportion of C17:0, C16:1 and C18:1 c-9 in LD muscle (P &amp;lt; 0.10). The lambs in LO and MLO group had higher proportions of C20:1, C18:3 n-3 and total n-3 in LD muscle than those in CON group (P &amp;lt; 0.10). Compared to CON group, the ruminal pH value of MLO group and the ruminal NH&lt;sub&gt;3&lt;/sub&gt;–N content of LO group were lower (P &amp;lt; 0.10). The total volatile fatty acid, proportion of acetate, and acetate/propionate were deceased in LO and MLO groups (P &amp;lt; 0.05). Furthermore, the two supplements significantly increased the proportions of propionate, butyrate and isovalerate (P &amp;lt; 0.05). Circulating cholesterol and high-density lipoprotein cholesterol concentrations were increased by linseed oil supplementation (P = 0.04). In conclusion, these results indicate that microencapsulated linseed oil did not exhibit any superior effects on muscle fatty acid composition and rumen fermentation of lambs.

Mstn knockdown promotes intramuscular fatty acid metabolism by β oxidation via the up-regulation of Cpt1b

Myostatin (Mstn) is known as growth/differentiation factor-8 (GDF-8). Knockout or knockdown of Mstn gene promotes muscle development and reduces fat content. Here we prepared Mstn knockdown mice by RNA interference, then the morphology of the skeletal muscle, the content of triglyceride (TG), the content and composition of fatty acids in the skeletal muscle were detected. The expression of Mstn reduced in muscle of Mstn knockdown mice compared to the controls. The cross sectional areas of the skeletal muscle myofibers were significantly larger while the content of TG was less than that of the controls, and the ratios of n-3/n-6 and unsat/sat in the knockdown mice increased significantly. Subsequently, we detected the expression of genes associated with fatty acid metabolism. The expression of the genes associated with lipolysis and fatty acid transportation were up-regulated, while the genes associated with fatty acid synthesis were down-regulated. Of these genes, the up-regulation of a gene associated with β oxidation, Cpt1b, was up-regulated remarkably. We further detected the enzyme activity of CPT1 in skeletal muscle and obtained the same results with gene expression. Moreover, chromatin immunoprecipitation assay was performed and we found that SMAD3, a transcription factor downstream of Mstn, directly binds to the promoter of Cpt1b gene. These results showed that knockdown of Mstn up-regulated the expression of Cpt1b through the binding of SMAD3 to the promoter of Cpt1b, then promoted the β oxidation metabolism of intramuscular fatty acids.

Impact of cooking on the content of nutritionally important fatty acids in muscle from lambs raised to have divergent intramuscular fatty acid compositions

The effect of sous vide cooking or grilling longissimus thoracis et lumborum (LTL) on the intramuscular fatty acid concentrations of lambs raised at pasture (Pasture), fed a cereal concentrate-based diet (Concentrate) or fed a cereal concentrate-based diet with a protected linseed oil supplement (Linseed) was assessed using GC-FID. Sous vide and grilling led to significant increases in fatty acid concentrations, particularly in the Pasture group. The apparent retention values were generally higher in the Pasture group. Concentrate, Linseed and Pasture groups had 16.7–19.9 mg, 25.8–28.7 mg, and 38.8–42.9 mg eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) in 100 g cooked meat, respectively. The EPA+DHA in the Pasture group met the requirements for a nutrient claim as a “source of n-3 fatty acids” set by the European Commission and as a “good source of n-3 fatty acids” set by the Food Standards Australia and New Zealand.

PPARGC1A Is a Moderator of Skeletal Muscle Development Regulated by miR-193b-3p.

Meat production performance is one of the most important factors in determining the economic value of poultry. Myofiber is the basic unit of skeletal muscle, and its physical and chemical properties determine the meat quality of livestock and poultry to a certain extent. Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A) as a transcriptional coactivator has been found to be widely involved in a series of biological processes. However, PPARGC1A is still poorly understood in chickens. In this manuscript, we reported that PPARGC1A was highly expressed in slow-twitch myofibers. PPARGC1A facilitated mitochondrial biogenesis and regulated skeletal muscle metabolism by mediating the flux of glycolysis and the TCA cycle. Gain- and loss-of-function analyses revealed that PPARGC1A promoted intramuscular fatty acid oxidation, drove the transformation of fast-twitch to slow-twitch myofibers, and increased chicken skeletal muscle mass. Mechanistically, the expression level of PPARGC1A is regulated by miR-193b-3p. Our findings help to understand the genetic regulation of skeletal muscle development and provide a molecular basis for further research on the antagonism of skeletal muscle development and fat deposition in chickens.

Influence of Dietary Algae Meal on Lipid Oxidation and Volatile Profile of Meat from Lambs with Competent Reticular Groove Reflex.

Dietary lipid sources influence intramuscular fatty acid composition, which in turn may affect the volatile profile of meat. The aim of this work was to investigate the effects of marine algae supplementation (Aurantiochytrium limacinum) on volatile compounds of cooked lamb meat. Forty-eight lambs with 42 days of age were divided into three groups: lambs fed a conventional diet without algae meal supplementation (NOALG), lambs with competent reticular groove reflex (RGR) fed the same diet supplemented with 2.5% marine algae meal mixed in the concentrate (ALGCON), and lambs with competent RGR, receiving the same diet and fed with 2.5% marine algae meal in a milk replacer to bypass the rumen (ALGMILK). Lipid and protein oxidation in raw meat was assessed and volatile compounds in grilled meat were determined. The highest and lowest lipid oxidations were observed in the ALGMILK and NOALG groups, respectively. Protein oxidation was unaffected. Out of 56 identified compounds, 12 volatiles significantly increased in both algae groups and 6 of them exclusively in the ALGCON treatment. Algae meal supplementation and its form of administration, either protected or not from rumen degradation, are important factors to consider in lipid oxidation and the aromatic profile of lamb meat.

Introducing Mediterranean Lupins in Lamb Diets: Effects on Carcass Composition, Meat Quality, and Intramuscular Fatty Acid Profile.

Simple SummaryThe main aim of this preliminary study was to evaluate the effects of replacing soybean meal with lupins on carcass traits, meat characteristics, meat characteristics, and meat fatty acid profile in lambs. Two trials were conducted: In trial 1, the soybean meal was partially replaced by Lupinus albus or Lupinus luteus; in trial 2, lambs were fed four diets with graded levels of Lupinus luteus, ranging from 0 to 200 g/kg. The lambs were slaughtered to evaluate carcass characteristics, meat composition, and fatty acids profile. Carcass composition was not affected (p > 0.05) by diet in both trials. Meat quality attributes did not vary (p < 0.05) between trials 1 and 2. Overall, fatty acid content was not affected by diet (p > 0.05) in both trials. Soybean meal produced the same results as lupins in this study, indicating the latter as a potential alternative protein source, although research should focus on meat palatability.The objective of this preliminary study was to evaluate the effects of partial replacement of soybean meal by lupins on lambs’ diets, on the carcass traits, meat characteristics, and meat fatty acid profile. Two trials were conducted: In trial 1, the soybean meal (control; C) was partially replaced by Lupinus albus or Lupinus luteus (50 g/kg; LA5 and LL5, respectively); in trial 2, lambs were fed four diets with graded levels of Lupinus luteus (0, 100, 150 and 200 g/kg; C, LL10, LL15, LL20, respectively). At the end of the feeding trials, animals were slaughtered to evaluate carcass characteristics and meat composition, including fatty acids. Carcass composition in tissues was not affected (p > 0.05) by diet in both trials. Additionally, no significant (p < 0.05) differences were observed in meat quality attributes between diets in trials 1 and 2. Overall, the Longissimus muscle’s fatty acid content was not affected by diet (p > 0.05) in both trials. Carcass and meat quality was overall comparable between lambs fed with soybean meal and lupins, indicating the latter as a potential alternative protein source. However, the lack of significant differences could also be attributed to the small sample size.

Comprehensive Transcriptome Analysis Reveals the Role of lncRNA in Fatty Acid Metabolism in the Longissimus Thoracis Muscle of Tibetan Sheep at Different Ages.

Long noncoding RNA (lncRNA) plays an important regulatory role in mammalian adipogenesis and lipid metabolism. However, their function in the longissimus thoracis (LT) muscle of fatty acid metabolism of Tibetan sheep remains undefined. In this study, fatty acid and fat content in LT muscle of Tibetan sheep were determined, and RNA sequencing was performed to reveal the temporal regularity of lncRNA expression and the effect of lncRNA-miRNA-mRNA ceRNA regulatory network on lipid metabolism of LT muscle in Tibetan sheep at four growth stages (4-month-old, 4 m; 1.5-year-old, 1.5 y; 3.5-year-old, 3.5 y; 6-year-old, 6 y). The results indicated that the intramuscular fat (IMF) content was highest at 1.5 y. Moreover, the monounsaturated fatty acid (MUFA) content in 1.5 y of Tibetan sheep is significantly higher than those of the other groups (P < 0.05), and it was also rich in a variety of polyunsaturated fatty acids (PUFA). A total of 360 differentially expressed lncRNAs (DE lncRNAs) were identified from contiguous period transcriptome comparative groups of 4 m vs. 1.5 y, 1.5 y vs. 3.5 y, 3.5 y vs. 6 y, and 4 m vs. 6 y, respectively. Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis found that the target genes in lncRNA trans-mRNA were significantly related to the protein digestion, absorption, and fatty acid biosynthesis pathways (P < 0.05), which demonstrated that DE lncRNA trans-regulated the target genes, and further regulated the growth and development of the LT muscle and intramuscular fatty acid metabolism in Tibetan sheep. We further analyzed the role of the lncRNA-miRNA-mRNA regulatory network in the lipid metabolism of Tibetan sheep. Additionally, GPD2, LIPE (lipase E hormone-sensitive enzyme), TFDP2, CPT1A, ACACB, ADIPOQ, and other mRNA related to fatty acid and lipid metabolism and the corresponding lncRNA-miRNA regulatory pairs were identified. The enrichment analysis of mRNA in the regulatory network found that the AMPK signaling pathway was the most significantly enriched (P = 0.0000112361). Comprehensive transcriptome analysis found that the LIPE, ADIPOQ, ACACB, and CPT1A that were regulated by lncRNA might change the formation of energy metabolism in Tibetan sheep muscle through the AMPK signaling pathway, and oxidized muscle fibers are transformed into glycolytic muscle fibers, reduced IMF content, and the fatty acid profile also changed.