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Antioxidant supplementation in critical periods may be useful for improvement of piglet early viability and development. We have evaluated the effects of maternal perinatal diet inclusion of a high vitamin E level (VE, 100 mg all-rac-α-tocopheryl acetate /kg), hydroxytyrosol (HT, 1.5 mg/kg), or their combination (VEHT), in comparison to a control diet (C, 30 mg all-rac-α-tocopheryl acetate /kg), on the offspring homeostasis and metabolism, analysing the weaned piglets' adipose tissue transcriptome and adipocyte morphology. Diets were provided to pregnant Iberian sows (n = 48, 12 per treatment) from gestation day 85 to weaning (28 days postpartum) and 48 piglets (n = 12 per treatment) were sampled 5 days postweaning for dorsal subcutaneous adipose tissue analyses. RNA obtained from 6 animals for each diet was used for paired-end RNA sequencing. Results show that supplementation of sows' diet with either vitamin E or hydroxytyrosol had substantial effects on weaned piglet adipose transcriptome, with 664 and 587 genes being differentially expressed, in comparison to C, respectively (q-value<0.10, Fold Change>1.5). Genes upregulated in C were mainly involved in inflammatory and immune response, as well as oxidative stress, and relevant canonical pathways and upstream regulators involved in these processes were predicted as activated, such as TNF, IFNB or NFKB. Vitamin E, when supplemented alone at high dose, activated lipid biosynthesis functions, pathways and regulators, this finding being accompanied by increased adipocyte size. Results suggest an improved metabolic and antioxidant status of adipose tissue in animals born from sows supplemented with individual antioxidants, while the combined supplementation barely affected gene expression, with VEHT showing a prooxidant/proinflamatory functional profile similar to C animals. Different hypothesis are proposed to explain this unexpected result. Findings allow a deeper understanding of the processes taking place in adipose tissue of genetically fat animals and the role of antioxidants in the regulation of fat cells function., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Laviano et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Chronic inflammatory enteropathies (CIEs) are classified based on treatment trials, and new methods are being sought for earlier differentiation and characterization. Giardia infection (GIA) is one of the first differential diagnoses and may be present in CIE-affected dogs. The aim of this study was to evaluate the faecal characteristics and faecal fatty acid profile (short, medium, long, and branched-chain fatty acids) in dogs with food-responsive enteropathy (FRE), immunosuppressant-responsive enteropathy (IRE), and dogs infected with Giardia compared to healthy control (HC) animals as a potential non-invasive indicator of intestinal health that helps in the differentiation of CIEs. The C16:1n-7 percentage ( p = 0.0001) and C16:1n-7/C16:0 ratio ( p = 0.0001) served to differentiate between HC, FRE, and IRE. IRE dogs presented lower levels of short-chain fatty acids (∑SCFAs) ( p = 0.0008) and acetic acid (C2) ( p = 0.0007) compared to the other three groups and lower propionic acid (C3) ( p = 0.0022) compared to HCs. IRE and GIA presented higher faecal fat content ( p = 0.0080) and ratio of iso/anteiso branched-chain fatty acids (BCFAs) compared to HC and FRE. Correlations between some fatty acids and desaturation indices with the canine inflammatory bowel disease activity index and faecal characteristics were observed, suggesting that these compounds could play an important role in the pathogenesis of these diseases., Competing Interests: The authors declare no conflicts of interest.

Pig feeding prior to the extensive fattening phase might affect the final lipid profile and product quality. This study evaluates how maternal supplementation with vitamin E (VITE) (100 mg/kg), hydroxytyrosol (HXT) (1.5 mg/kg), or combined administration (VE + HXT) affects the piglet's plasma and tissues' fatty acid profiles and lipid stability according to the sow's parity number (PN), as well as the possible changes to the lipid profile after extensive feeding. The sows' PN affected the total fatty acid profile of plasma, muscle, and liver of piglets, with lower Δ-9 and Δ-6 desaturase indices but higher Δ-5 in those from primiparous (P) than multiparous (M) sows. Dietary VITE was more effective at decreasing C16:0 and saturated fatty acids in the muscle of piglets born from M than P sows, and modified the liver phospholipids in a different way. Sows' supplementation with HXT increased C18:2n-6 in triglycerides and polyunsaturated fatty acids (PUFA) in muscle phospholipids. In the liver, HXT supplementation also increased free-PUFA and free-n-3 fatty acids. However, lipid oxidation of piglets' tissues was not affected by the antioxidant supplementation, and it was higher in the livers of piglets born from M sows. The fatty acid profile in the muscle of pigs after extensive feeding was not affected by the PN, but it was by the sows' antioxidant supplementation, with positive effects on quality by both compounds.

An adequate intestinal environment before weaning may contribute to diarrhea predisposition and piglet development. This study evaluates how the dietary supplementation of vitamin E (VE) (100 mg/kg), hydroxytyrosol (HXT) (1.5 mg/kg) or the combined administration (VE + HXT) given to Iberian sows from gestation affects the piglet's faecal characteristics, short chain fatty acids (SCFAs), fatty acid profile or intestinal morphology as indicators of gut health; and quantify the contribution of the oxidative status and colostrum/milk composition to the piglet's SCFAs content and intestinal health. Dietary VE increased isobutyric acid (iC4), butyric acid (C4), isovaleric acid (iC5), and ∑SCFAs, whereas HXT increased iC4 and tended to decrease ∑SCFAs of faeces. Piglets from HXT-supplemented sows also tended to have higher faecal C20:4n-6/C20:2 ratio C22:6 proportion and showed lower occludin gene expression in the duodenum. The combination of both antioxidants had a positive effect on iC4 and iC5 levels. Correlation analyses and regression equations indicate that faecal SCFAs were related to oxidative status (mainly plasma VE) and colostrum and milk composition (mainly C20:2, C20:3, C20:4 n-6). This study would confirm the superiority of VE over HXT supplementation to improve intestinal homeostasis, gut health, and, consequently piglet growth.

Different feeding strategies are being applied to sows in order to obtain homogeneous piglets' weights and improved health status. This study evaluated how the dietary supplementation of vitamin E (VE) (100 mg/kg), hydroxytyrosol (HXT) (1.5 mg/kg) or the combined administration (VE + HXT) given to Iberian sows from day 85 of gestation affected the growth pattern of the piglets and their oxidative status; and quantified what these effects were due to. Dietary VE and HXT improved the oxidative status of sows and piglets. Both VE and HXT modified the growth pattern at birth and performances of the piglets in a different way according to the growing period. Piglets' performances were positively correlated with plasma VE and negatively with plasma malondialdehyde (MDA) of the sow. However, the highest variation in growth patterns was explained by the colostrum composition. Significant linear equations were observed between piglets' performances and colostrum saturated (SAT), n-7 monounsaturated fatty acids (C16:1n-7 and C18:1n-7) and different desaturases indices. This study would confirm that VE supplementation to the sow diet could be more adequate than HXT for the improved development during the first weeks of a piglet's life. The combined administration of both antioxidants would not produce additional positive effects compared to the individual supplementation.

Modifying the composition of a sow's milk could be a strategy to improve the intestinal health and growth of her piglet during the first weeks of life. This study evaluated how dietary supplementation of vitamin E (VE), hydroxytyrosol (HXT) or VE+HXT given to Iberian sows from late gestation affected the colostrum and milk composition, lipid stability and their relationship with the piglet's oxidative status. Colostrum from VE-supplemented sows had greater C18:1n-7 than non-supplemented sows, whereas HXT increased polyunsaturated (∑PUFAs), ∑n-6 and ∑n-3 fatty acids. In 7-day milk, the main effects were induced by VE supplementation that decreased ∑PUFAs, ∑n-6 and ∑n-3 and increased the Δ-6-desaturase activity. The VE+HXT supplementation resulted in lower desaturase capacity in 20-day milk. Positive correlations were observed between the estimated mean milk energy output and the desaturation capacity of sows. The lowest concentration of malondialdehyde (MDA) in milk was observed in VE-supplemented groups, whereas HXT supplementation increased oxidation. Milk lipid oxidation was negatively correlated with the sow's plasma oxidative status and to a great extent with the oxidative status of piglets after weaning. Maternal VE supplementation produced a more beneficial milk composition to improve the oxidative status of piglets, which could promote gut health and piglet growth during the first weeks, but more research is needed to clarify this.

Dogs suffering from food-responsive enteropathy (FRE) respond to an elimination diet based on hydrolysed protein or novel protein; however, studies regarding the amino acid profile in FRE dogs are lacking. The aim of this pilot study was to evaluate whether the plasma and faecal amino acid profiles differed between control and FRE dogs and whether these could serve as indicators of severity of illness. Blood, faecal samples, body condition score, and severity of clinical signs based on the canine inflammatory bowel disease activity index were collected before starting the elimination diet. FRE dogs had lower proportions of plasma Asparagine, Histidine, Glycine, Cystine, Leucine, and branched-chain/aromatic amino acids; however, Phenylalanine increased. In faecal samples, Cystine was greater whereas Phenylalanine was lesser in sick dogs compared to control. Leucine correlated negatively with faecal humidity (r = -0.66), and Leucine and Phenylalanine with faecal fat (r = -0.57 and r = -0.62, respectively). Faecal Phenylalanine (r = 0.80), Isoleucine (r = 0.75), and Leucine (r = 0.92) also correlated positively with total short-chain fatty acids, whereas a negative correlation was found with Glycine (r = -0.85) and Cystine (r = -0.61). This study demonstrates the importance of Leucine and Phenylalanine amino acids as indicators of the disease severity in FRE dogs.

To meet the demand for chicken meat production, new additives that promote growth and health without adverse effects on meat quality are being investigated. This study was conducted to investigate the effect of protected sodium butyrate (PSB) (0 vs. 2 g/kg), an olive leaf and grape-based by-product (OLG-mix), or a combined supplementation of PSB and OLG-mix on productive performance, antioxidant status, carcass, and meat quality in broilers. PSB improved performance parameters with greater effect in the initial phase. Both, PSB and OLG-mix increased the plasma superoxide dismutase (SOD); however, PSB supplementation was more effective to delay the lipid oxidation of meat from the initial day of storage. OLG-mix produced meat with greater color intensity, b* value and lesser drip losses than PSB. The combination of PSB + OLG-mix did not produce more marked effects that the individual administration; except to control the oxidation of meat. Linear and positive correlations between antioxidant enzymes and weight gain were observed. Significant linear and negative relationships were quantified between plasma SOD and meat lipid oxidation according to dietary treatment. Therefore, the present study would be a first approximation to the possibilities for predicting growth range and meat quality through the evaluation of the blood oxidative status.

Iberian pigs and its crosses are produced to obtain high-quality meat products. The objective of this work was to evaluate a wide panel of DNA markers, selected by biological and functional criteria, for association with traits related to muscle growth, fatness, meat quality and metabolism. We used 18 crossbred Iberian pigs with divergent postnatal growth patterns for whole genome sequencing and SNP discovery, with over 13 million variants being detected. We selected 1023 missense SNPs located on annotated genes and showing different allele frequencies between pigs with makerdly different growth patterns. We complemented this panel with 192 candidate SNPs obtained from literature mining and from muscle RNAseq data. The selected markers were genotyped in 480 Iberian × Duroc pigs from a commercial population, in which phenotypes were obtained, and an association study was performed for the 1005 successfully genotyped SNPs showing segregation. The results confirmed the effects of several known SNPs in candidate genes (such as LEPR, ACACA, FTO, LIPE or SCD on fatness, growth and fatty acid composition) and also disclosed interesting effects of new SNPs in less known genes such as LRIG3, DENND1B, SOWAHB, EPHX1 or NFE2L2 affecting body weight, average daily gain and adiposity at different ages, or KRT10, NLE1, KCNH2 or AHNAK affecting fatness and FA composition. The results provide a valuable basis for future implementation of marker-assisted selection strategies in swine and contribute to a better understanding of the genetic architecture of relevant traits., (© 2022. The Author(s).)

The aim of this study was to evaluate differences in short-chain fatty acids (SCFAs) and the total fatty acid profile of faeces or plasma as possible indicators of FRE in comparison with healthy dogs. FRE dogs had a lower concentration ( p = 0.026) of plasma α-tocopherol as an indicator of the oxidative status of the animal, and lower C20:5n-3 ( p = 0.033), C22:5n-3 ( p = 0.005), polyunsaturated fatty acids (PUFA) ( p = 0.021) and n-6 ( p = 0.041) when compared with the control dogs; furthermore, sick dogs had higher proportions of plasma C20:3n-6 ( p = 0.0056). The dogs with FRE showed a decrease in the production of faecal levels of SCFAs, mainly propionic acid (C3) ( p = 0.0001) and isovaleric acid (iC5) ( p = 0.014). FRE dogs also had a lower proportion of C15:0 ( p = 0.0003), C16:1n-9 ( p = 0.0095), C16:1n-7 ( p = 0.0001), C20:5n-3 ( p = 0.0034) and monounsaturated fatty acids ( p = 0.0315), and tended to have lower n-3 ( p = 0.058) and a reduced desaturase activity index in the stool when compared with the control group. However, the dogs with chronic enteropathy tended to have greater C20:4n-6 ( p = 0.065) in their faeces as signs of damage at the intestinal level. The faecal parameters were better predictors than plasma. The highest correlations between faecal odd-chain, medium- or long-chain fatty acids and SCFAs were observed for C15:0 that correlated positively with faecal acetic acid (C2) (r = 0.72, p = 0.004), propionic acid (r = 0.95, p = 0.0001), isobutyric acid (iC4) (r = 0.59, p = 0.027) and isovaleric acid (r = 0.64, p = 0.0136), as well as with total SCFAs (r = 0.61, p = 0.02). Conversely, faecal C20:4n-6 showed a high inverse correlation (r = -0.83, p = 0.0002) with C2 and C3 (r = -0.59, p = 0.027). Canine inflammatory bowel disease (IBD) activity (CIBDAI) index correlated negatively mainly with faecal measurements, such as C3 (r = -0.869, p = 0.0005) and C15:0 (r = -0.825, p = 0.0018), followed by C16:1/C16:0 (r = -0.66, p = 0.0374) and iC5 (r = -0.648, p = 0.0310), which would indicate that these fatty acids could be good non-invasive indicators of the chronic inflammatory status, specifically FRE.

The nutritional value of fat consumption depends on both the fatty acid composition and the positional distribution of fatty acids within the triglyceride molecule. This research studies the effect of feeding with three different diets (4% lard-enriched; 11.5% high-oleic sunflower-enriched; and extensive feeding mainly with acorns) on the composition of fatty acids in the sn-2 position (and sn-1,3 ) of triglycerides and the textural properties of subcutaneous fat in heavy Iberian pigs ( n = 210 castrated males). A moderate dietary enrichment with oleic acid in mixed diets did not alter the regulation of the sn-2 position of triglyceride (69.9% and 13.9% of palmitic and oleic acids, respectively), but the extremely high intake of oleic acid in pigs fed mainly on acorns changed the proportions of palmitic and oleic acids at the sn-2 position in the subcutaneous fat of pigs (55.0% and 27.2%, respectively). Hardness, adhesiveness, cohesiveness, gumminess, and chewiness showed the least values in EXT pigs, and the greatest values in LARD-fed barrows. SUN cohesiveness and gumminess did not differ from those fed LARD. In addition, Iberian pigs raised in free-range conditions had a more favorable nutritional lipid profile for human health compared to pigs fed conventional diets.

The aim of the study was to investigate the effect of short-term dietary spirulina supplementation (1% and 3%) and the strain of laying hens (White Leghorn: WL and Rhode Island Red: RIR) on color, nutritional value, and stability of yolk. Egg weight was not affected by any of the studied effects. Yolks from 3%-spirulina supplemented hens had higher retinol and lower α-tocopherol content ( p = 0.0001) when compared to control. The supplementation with 1%-spirulina markedly decreased luminosity and increased redness ( p = 0.0001) and yellowness ( p = 0.0103). Short-term spirulina supplementation slightly modified the fatty acid composition of yolk. The C16-desaturase index increased with the algae, whereas other egg quality indices (hypocholesterolemic, thrombombogenic, n-6/n-3) were not modified. Hen strain mainly affected to the lipid profile. The RIR hens accumulated greater yolk retinol with supplementation doses of 3% ( p < 0.05), while the WL hardly suffered changes in the accumulation. Also, yolks from RIR hens had lower C16:0 ( p = 0.0001), C18:0 ( p = 0.0001), saturated (SAT) ( p = 0.0001), and thrombogenic index ( p = 0.0001), C20:3n-6 ( p = 0.0001), n-6/n-3 ratio ( p = 0.003), Δ-6+5-desaturase ( p = 0.0005), total elongase indices ( p = 0.0001) when compared to WL. Moreover, RIR had higher monounsaturated (MUFA), Δ-9-desaturase and hypocholesterolemic indices ( p < 0.05) than WL. A different response to spirulina supplementation was observed for C18:1n-9, MUFA, Δ-9-desaturase and thiesterase indices ( p < 0.05) according to hen strain. Yolks from RIR had higher MUFA and Δ-9-desaturase indices than WL at 1%-spirulina supplementation, whereas these parameters were less affected in RIR supplemented with 3%. SAT and Δ-9-desaturase were significantly correlated (r = -0.38 and 0.47, respectively) with retinol content according to a linear adjustment ( p < 0.05). Lipid oxidation of yolk was slightly modified by the dietary treatment or hen strain. It was detected a relationship between TBARS and α-tocopherol, C22:5n-3 or C22:6n-3 ( p < 0.05). L* and a* were also inversely or positively related with yolk retinol content according to a linear response ( p < 0.05). The administration of 1% of spirulina in diets of red hens would be an interesting alternative to get healthier eggs from the nutritional point of view, obtaining an adequate color and without modifications in other yolk quality traits.

Grape seeds (GS) and grape skins (GK) are natural sources of polyphenols with featured antioxidant capacity. An experiment was conducted to investigate the effect of these polyphenol sources in diets formulated to contain the same total extractable grape polyphenol content on growth performance, protein and extractable polyphenol digestibility, plasma and meat α- and γ-tocopherol and thigh meat oxidation in broiler chickens. Five experimental diets were formulated: control, control + vitamin E (200 mg/kg), 30 g/kg GS diet, 110 g/kg GK diet, GS + GK diet (a mixture of 24.4 g/kg GS and 13.1 g/kg GK designed to simulate a reconstituted grape pomace). Feeding chickens with 110 g/kg GK reduced ( p < 0.001) daily weight gain, worsened ( p < 0.001) feed conversion ratio, increased ( p < 0.001) non-extractable polyphenol content in the ileum and in the excreta and decreased ( p < 0.05) ileal protein digestibility. Regardless of the grape polyphenol source used, the inclusion of grape byproducts in the diets led to an increase of total extractable polyphenol contents in the ileum ( p < 0.01) and the excreta ( p < 0.001), which resulted ( p < 0.001) in a decrease of extractable polyphenol digestibilities. Alpha- and gamma-tocopherol concentrations increased ( p < 0.001) in plasma and in seven-day stored meat in birds fed the diet combining GS and GK with respect to the control group. As it happened with the vitamin E supplementation, feeding the combination of GS and GK also reduced ( p < 0.001) the concentration of the lipid peroxidation marker (malondialdehyde) in the stored meat of chickens.

Background: Olive-derived antioxidants have been shown to affect the oxidative status of meat and have also been associated with greater consumption of glucose, which might affect glycogen stores and muscle characteristics. This study evaluated the effect of oleuropein extract supplementation (OLE) versus vitamin E + Se (VE), and their combination (VEOLE), in pig diets, on pH, drip loss, the proportion of free fatty acids, and meat stability, and their prediction by blood oxidative status markers., Results: The drip loss of muscle was lower in antioxidant-supplemented groups when compared with controls. α-Tocopherol concentration and total fatty acids profile were not affected by dietary oleuropein supplementation. However, OLE and VEOLE had lower free n-3 polyunsaturated fatty acid (PUFA) levels when compared with VE and tended to have higher free monounsaturated fatty acid (MUFA) levels. Furthermore, the VEOLE group had lower free n-6 PUFA levels when compared with controls or VE, whereas the OLE group had intermediated values. Muscle samples from pigs subjected to the antioxidant-mixed supplementation (VEOLE) had lower malondialdehyde concentration when compared with the others. The VE and OLE groups showed intermediate malondialdehyde values. Chilled meat stability was highly correlated with antioxidant status in vivo., Conclusion: The administration of 96 mg oleuropein kg -1 feed produced similar meat quality characteristics as the use of 100 mg kg -1 α-tocopheryl acetate +0.26 mg kg -1 sodium selenite and it would be an interesting alternative in Mediterranean countries. The VEOLE group was the most effective for reducing lipid oxidation and for the production of polyunsaturated free fatty acids in meat, which would result in lower rancidity formation and better aroma development in products. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

This study evaluates the effect of the dietary combination of oleuropein extract (1200 mg/kg) and betaine (1000 mg/kg), magnesium oxide (600 mg/kg), and α-tocopheryl acetate (400 mg/kg), or a half-dose of these compounds, on pigs' performance, oxidative status, and meat quality characteristics (drip loss, TBARS, and texture and fatty acid profile of intramuscular fat). Sixty-six barrows and females were slaughtered at 120 kg of BW. Performance and carcass yield were not changed by treatments. The high-dose mixture resulted in higher serum ferric reducing/antioxidant power ( p = 0.0026), lower glucose ( p = 0.03) and a tendency to have lower serum TBARS ( p = 0.07) when compared to control. Percentage of drip loss, moisture content, intramuscular fat, or texture parameters were not modified by dietary treatments. Pigs supplemented with the high-dose mixture had higher PUFA ( p = 0.0001), n-6 ( p = 0.0001), n-3 ( p = 0.0095) and lower MUFA ( p = 0.0064) in the neutral lipid fraction of intramuscular fat. Free PUFA, mainly n-3 fatty acids ( p = 0.0009), were also higher in the meat of pigs fed the high-dose mixture compared with the others. A higher mobilization (neutral to free fatty acids hydrolysis) of n-3 and MUFA fatty acids in the muscle from pigs fed the high-dose mixture was observed. However, dietary mixture supplementation tended to increase MUFA ( p = 0.056) and decrease the total PUFA ( p = 0.0074) proportions in muscle polar lipids. This specific fatty acid composition of meat from pigs supplemented with the high-dose mixture could be responsible for the higher meat lipid oxidation observed in this group when compared to the other groups. Consequently, the low-dose mixture would be more adequate for maintaining the oxidative status of pigs and, meat lipid stability.

The aims of this study were to investigate the effect of vitamin D 3 administration in drinking water during lairage time prior to slaughter on physiological stress, oxidative status, and pork quality characteristics. Two experiments were carried out. The first one was performed to examine the effect of vitamin D 3 supplementation in drinking water, and the second one to check the effect of supplementation dose (500,000 IU/L vs. 700,000 IU/L). Serum calcium concentration was greater in pigs receiving vitamin D 3 in water when compared to the control group. In experiment 1, a 40% α-tocopherol increase in meat from the group supplemented with vitamin D 3 (500,000 IU/animal) was observed, that resulted in a tendency of decreased meat malondialdehyde (MDA) values at days 5 and 8 after refrigerated storage. In experiment 2, since water intake was higher (800,000 IU and 1,120,000 IU/animal of vitamin D 3 consumption) effects on oxidative status were more profound and vitamin D 3 supplementation increased serum α-tocopherol and decreased cortisol and serum TBARS. These effects were also observed in meat; TBARS levels were decreased after 3 days of refrigerated storage. In both experiments meat from pigs that received vitamin D 3 in drinking water had a lower proportion of total free-polyunsaturated fatty acids (mainly n-6) when compared to the unsupplemented pigs, and these were positively correlated with TBARS production at day 5 of refrigerated storage ( r = 0.53 and 0.38 for experiments 1 and 2, respectively). Meat from pigs receiving vitamin D 3 in water showed reduced cohesiveness, gumminess, and chewiness values compared to the control group. The magnetic resonance imaging study of muscle confirmed the effects on water retention with lesser transverse relaxation time in pigs supplemented with vitamin D 3 . No vitamin D 3 dose effect was observed, apart from muscle α-tocopherol concentration that was higher in pigs supplemented with 700,000 compared to those supplemented with 500,000 UI/L.

This study evaluates the effect of the dietary glycaemic index (low: L-GI vs. high: H-GI), pre-slaughter fasting times (17 h: f 17 vs. 19 h: f 19 vs. 22 h: f 22 ), and their interaction, on certain meat quality characteristics. Greater duration of fasting resulted in lesser drip loss of muscle, greater LIPE expression (lipolysis) and 10% greater TBARS, with PUFA being the main source of free-fatty acid production. L-GI diets produced a greater drip loss, lipolytic activity and lesser C18:1n-9 relative mobilization, suggesting this group had greater glycolytic potential than H-GI. An interaction was found in meat characteristics related to fat metabolism. The IRS2 gene expression was greater in pigs fed L-GI diets, but only in a low fasting period. LIPE gene expression was greater for L-GI-f 17 group than for L-GI-f 22 . Lesser n-6 PUFA but greater MUFA were found in the free fatty acid fraction of L-GI- f 17 group thus resulting in a faster lipolysis initiation in this group., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Growth is dependent on genotype and diet, even at early developmental stages. In this study, we investigated the effects of genotype, sex, and body weight on the fetal muscle transcriptome of purebred Iberian and crossbred Iberian x Large White pigs sharing the same uterine environment. RNA sequencing was performed on 16 purebred and crossbred fetuses with high body weight (340±14g and 415±14g, respectively) and 16 with low body weight (246±14g and 311±14g, respectively), on gestational day 77. Genotype had the greatest effect on gene expression, with 645 genes identified as differentially expressed (DE) between purebred and crossbred animals. Functional analysis showed differential regulation of pathways involved in energy and lipid metabolism, muscle development, and tissue disorders. In purebred animals, fetal body weight was associated with 35 DE genes involved in development, lipid metabolism and adipogenesis. In crossbred animals, fetal body weight was associated with 60 DE genes involved in muscle development, viability, and immunity. Interestingly, the results suggested an interaction genotype*weight for some DE genes. Fetal sex had only a modest effect on gene expression. This study allowed the identification of genes, metabolic pathways, biological functions and regulators related to fetal genotype, weight and sex, in animals sharing the same uterine environment. Our findings contribute to a better understanding of the molecular events that influence prenatal muscle development and highlight the complex interactions affecting transcriptional regulation during development., Competing Interests: Antonio González-Bulnes and Cristina Ovilo are PLOS One Editorial Board members, however, this does not alter the authors’ adherence to all PLOS One policies.

Olive tree leaves are characterized for having not only a potent antioxidant power but also effects on glucose and lipid metabolism. The impact of the individual oleuropein (OLE), vitamin E + Se (VE), or a combined supplementation of oleuropein, vitamin E, and selenium (VEOLE) was evaluated on pig plasma metabolites under fasting prior to slaughter. VEOLE and OLE had lesser n-3 plasma polyunsaturated fatty acids and greater monounsaturated free fatty acids compared to control. The n-3-fatty acid mobilization was directly correlated with greater cysteine and inversely with oxidized glutathione/reduced glutathione (GSSH/GSH) levels. This faster use of n-3 fatty acids might act as an indicator of glutathione synthesis mediated by an increase of cysteine in plasma. Different correlations and linear adjustments were observed between plasma antioxidant power and free cysteine, free glycine, free glutamine, monounsaturated free fatty acids, and total n-3. The best response to stress was found in VEOLE. Cortisol reached the greatest positive correlation with plasma total n-3 fatty acids, which suggests a faster uptake of n-3 for biological functions such as stress control or energy supply in the brain. From a practical point of view, an enhanced oxidative status as well as control of physiological stress prior to slaughter by the combined antioxidants supplementation might have positive effects on pork quality.

1. This work aims to quantify changes in fatty acid profile, melting point, abdominal fat accumulation and 2-thiobarbituric acid-reactive substances production depending on dietary fat source and age at slaughter, and to estimate the optimal date for the change from an unsaturated fat to a saturated fat diet or vice versa . 2. Treatments established were (1) birds fed 8% tallow from 21 to 49 d (TTT); (2) birds fed 8% tallow from 21 to 37 d and 8% sunflower oil from d 38 to 49 (TSS); (3) birds fed 8% sunflower oil from 21 to 37 d and 8% tallow from d 38 to 49 (STT); (4) birds fed 8% sunflower oil from 21 to 41 d and 8% tallow from d 42 to 49 (SST); (5) birds fed 8% sunflower oil from 21 to 49 d (SSS). Birds from each group were slaughtered on d 21, 29, 38, 40, 42, 44, 46 and 49. 3. The polyunsaturated fatty acids (PUFAs) proportion in the SSS group reached maximum values at d 40 and fitted a quadratic response. This group also showed a decrease in saturated fatty acids (SATs) and monounsaturated fatty acids (MUFAs) of lower intensity than the PUFA increase. The highest synthesis of SAT + MUFA was found in the SSS and TSS groups, whereas these had the lowest body-to-dietary PUFA ratio. 4. A high and quadratic increase in the MUFA proportion was observed during the first 10 d of feeding with the tallow-enriched diet at the expenses of the proportion of PUFA that quadratically decreased (minimum values at d 38). 5. Lipogenic and desaturation capacity decreased with age. 6. The TSS group increased tissue PUFA content faster that the SST group decreased PUFA content after the change in diet which indicates that the earlier feeding has to be taken into consideration for obtaining higher or lower changes in quality parameters. 7. The melting point of the SSS group showed a lower response to the dietary treatment in the initial period when compared to the TTT treatment. 8. The TTT, STT, SST and TSS groups showed similar fat accumulation, and changes in lipid oxidation were related to the day of dietary sunflower oil supplementation. 9. Based on the results, it would be possible to determine the most appropriate dietary programme and optimum slaughter age to obtain chicken meat with the desired quality characteristics.

This study evaluates the effect of the source of dietary selenium supplementation (organic versus mineral) and the combined effect of organic selenium and vitamin E on the fatty acid composition and lipolysis in pork intramuscular fat and other meat quality characteristics such as drip loss and lipid stability. Higher vitamin E deposition, lower drip loss, and higher stability against oxidation were detected in muscle from pigs fed organic selenium. Also higher proportion of free fatty acids was observed in intramuscular fat from pigs fed organic selenium than those fed inorganic selenium, being these mainly coming from neutral lipid fraction. In addition, the inclusion of vitamin E in the diet enhanced such effect. Dietary organic selenium also increased Δ 9 -desaturase and elongase indexes and C18:1n-9 concentration. A related decrease of C18:0 concentration was also observed. Described differences in the lipid fraction composition could affect sensory characteristics of meat and meat products.

Iberian pig production includes purebred (IB) and Duroc-crossbred (IBxDU) pigs, which show important differences in growth, fattening and tissue composition. This experiment was conducted to investigate the effects of genetic type and muscle (Longissimus dorsi (LD) vs Biceps femoris (BF)) on gene expression and transcriptional regulation at two developmental stages. Nine IB and 10 IBxDU piglets were slaughtered at birth, and seven IB and 10 IBxDU at four months of age (growing period). Carcass traits and LD intramuscular fat (IMF) content were measured. Muscle transcriptome was analyzed on LD samples with RNA-Seq technology. Carcasses were smaller in IB than in IBxDU neonates (p < 0.001), while growing IB pigs showed greater IMF content (p < 0.05). Gene expression was affected (p < 0.01 and Fold change > 1.5) by the developmental stage (5,812 genes), muscle type (135 genes), and genetic type (261 genes at birth and 113 at growth). Newborns transcriptome reflected a highly proliferative developmental stage, while older pigs showed upregulation of catabolic and muscle functioning processes. Regarding the genetic type effect, IBxDU newborns showed enrichment of gene pathways involved in muscle growth, in agreement with the higher prenatal growth observed in these pigs. However, IB growing pigs showed enrichment of pathways involved in protein deposition and cellular growth, supporting the compensatory gain experienced by IB pigs during this period. Moreover, newborn and growing IB pigs showed more active glucose and lipid metabolism than IBxDU pigs. Moreover, LD muscle seems to have more active muscular and cell growth, while BF points towards lipid metabolism and fat deposition. Several regulators controlling transcriptome changes in both genotypes were identified across muscles and ages (SIM1, PVALB, MEFs, TCF7L2 or FOXO1), being strong candidate genes to drive expression and thus, phenotypic differences between IB and IBxDU pigs. Many of the identified regulators were known to be involved in muscle and adipose tissues development, but others not previously associated with pig muscle growth were also identified, as PVALB, KLF1 or IRF2. The present study discloses potential molecular mechanisms underlying phenotypic differences observed between IB and IBxDU pigs and highlights candidate genes implicated in these molecular mechanisms., Competing Interests: The authors have declared that no competing interests exist, excepting that the coauthors Antonio Gonzalez-Bulnes and Cristina Óvilo are PLOS ONE Editorial Board members. However, this does not alter the authors’ adherence to all the PLOS ONE policies.

This study evaluates the effect of dietary selenium (Se) supplementation source (organic, Se-enriched yeast; SY vs. inorganic, sodium selenite; SS), dose (0.2: L vs. 0.4: H mg/kg) and the combination of Se and vitamin E (VITE+SS) for 26days on drip loss, TBARS, colour changes, myofibrillar protein pattern and proteolysis in pork. The lowest water losses were observed in the SY-H group when compared to the others. SY-H and VITE+SS groups presented lower myofibrillar protein hydrolysis/oxidation. VITE+SS supplementation also resulted in higher PRO, TRP and PHE content at days 2 and 7, whereas the SY group showed increased GLY and CAR and tended to have higher TAU and ANS at day 2. The myofibrillar fragmentation index was not modified by the dietary treatment; however, at day 8, it tended to be higher in groups supplemented with SeY and VITE+SS. The results of the present study might indicate a possible relation between muscle proteolysis and water loss., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study evaluates the effect of organic (Se-enriched yeast; SeY) versus inorganic selenium (sodium selenite; SeS) supplementation and the different response of selenium source according to muscle pH on pork meat quality characteristics. Pigs ( n  = 30) were fed the Se-supplemented diets (0.3 mg/kg) for 65 days. Neither electric conductivity (EC) nor drip loss were affected by the selenium source. The SeY group had lower TBARS in muscle samples after day 7 of refrigerated storage and higher a * values on days 1 and 7 than the SeS group. The effect of dietary selenium source on some meat quality characteristics was affected by muscle pH. Hence, as the muscle pH increases, the drip loss decreases but this effect is more marked with the dietary organic Se enrichment. Muscle pH seems to modulate the action of selenium in pork, especially some meat characteristics such as drip loss.

Iberian ham production includes both purebred (IB) and Duroc-crossbred (IBxDU) Iberian pigs, which show important differences in meat quality and production traits, such as muscle growth and fatness. This experiment was conducted to investigate gene expression differences, transcriptional regulation and genetic polymorphisms that could be associated with the observed phenotypic differences between IB and IBxDU pigs. Nine IB and 10 IBxDU pigs were slaughtered at birth. Morphometric measures and blood samples were obtained and samples from Biceps femoris muscle were employed for compositional and transcriptome analysis by RNA-Seq technology. Phenotypic differences were evident at this early age, including greater body size and weight in IBxDU and greater Biceps femoris intramuscular fat and plasma cholesterol content in IB newborns. We detected 149 differentially expressed genes between IB and IBxDU neonates (p < 0.01 and Fold-Change > 1. 5). Several were related to adipose and muscle tissues development (DLK1, FGF21 or UBC). The functional interpretation of the transcriptomic differences revealed enrichment of functions and pathways related to lipid metabolism in IB and to cellular and muscle growth in IBxDU pigs. Protein catabolism, cholesterol biosynthesis and immune system were functions enriched in both genotypes. We identified transcription factors potentially affecting the observed gene expression differences. Some of them have known functions on adipogenesis (CEBPA, EGRs), lipid metabolism (PPARGC1B) and myogenesis (FOXOs, MEF2D, MYOD1), which suggest a key role in the meat quality differences existing between IB and IBxDU hams. We also identified several polymorphisms showing differential segregation between IB and IBxDU pigs. Among them, non-synonymous variants were detected in several transcription factors as PPARGC1B and TRIM63 genes, which could be associated to altered gene function. Taken together, these results provide information about candidate genes, metabolic pathways and genetic polymorphisms potentially involved in phenotypic differences between IB and IBxDU pigs associated to meat quality and production traits.

The aim of this study was to investigate whether dietary vitamin A level is associated with differences in adipocyte differentiation or lipid accumulation in Iberian pigs at early growing (35.8kg live weight) and at finishing (158kg live weight). Iberian pigs of 16.3kg live weight were allocated to two feeding groups, one group received 10,000IU of vitamin A/kg diet (control); the other group received a diet with 0IU of vitamin A (var) for the whole experimental period. The dietary vitamin A level had no effect on growth performance and carcass traits. The early suppression of vitamin A increased the preadipocyte number in Longissimus thoracis (LT) muscle in the early growth period (P<0.001) and the neutral lipid content and composition (higher MUFA and lower SFA content) at the end of the finishing period (P<0.05). Vitamin A restriction in young pigs increases their lipogenic potential without affecting carcass traits., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This study evaluates the effect of vitamin E supplementation source (micellized natural vs. the synthetic form) and dosage (40, 80, or 120 mg/kg) on α-tocopherol concentration in plasma and muscle, antioxidant capacity, and breast meat quality in turkeys. Three hundred female turkeys were randomly selected at an average live weight 63.2 g±0.5 and distributed into 7 groups. One group (control) was fed a standard diet without vitamin E supplementation and the other 6 were given mixed diets supplemented with the natural (d-α-tocopherol) or synthetic (dl-α-tocopheryl acetate) form of vitamin E in 3 dosages (40, 80, or 120 mg/kg). Following 11 wk feeding, results showed that performance parameters were not modified either by source or dosage of vitamin E supplementation to the turkeys. Plasma and muscle α-tocopherol at d 9 of refrigerated storage were higher when turkeys were supplemented with the natural form at higher doses. Losses in the concentration of α-tocopherol in meat between the beginning and the end of the 9 d refrigerated storage were greater in the groups supplemented with the synthetic form of vitamin E compared to those receiving the natural supplementation. The relationship between plasma α-tocopherol and the Trolox equivalent antioxidant capacity followed a different trend depending on the vitamin E source. Intramuscular fat was not significantly affected by the vitamin E source supplementation; however the slope of the linear regression equation was lower for the natural form than for the synthetic form. Turkeys given the natural form had higher C18:1n-9 but lower C15:1, C17:1, C20:5n-3, and C22:6n-3 in breast muscle. Meat samples from turkeys supplemented with natural vitamin E had higher deoxymyoglobin at d 3, 6, and 9 and lower metmyoglobin at d 9 of refrigerated storage than those receiving the synthetic form. Dietary supplementation with medium doses (80 mg/kg) micellized d-α-tocopherol is an interesting feeding strategy for ensuring antioxidant status and improving meat quality., (© 2015 Poultry Science Association Inc.)

Background: Quantification of γ- and α-tocopherol in dry-cured shoulders of Iberian pigs was evaluated as a tool for differentiating feeding backgrounds or regimens. Samples (n = 115) were obtained over two different seasons from the four categories of pigs described in the Industry Quality Policy, i.e. pigs fed in free-range conditions (FREE-RANGE), pigs fed in free-range conditions and provided feed supplements (FREE-FEED), pigs fed outdoors with feed and with access to grass (FEED-OUT) and pigs fed in intensive conditions with feed (FEED). Linear discriminant functions were calculated and validated., Results: The validation results showed that 20% of the muscle samples were not correctly classified into the four feeding categories, giving an 80% success rate. The FEED group had the lowest proportion of errors, with 100% of samples correctly classified. For the FREE-RANGE group, 87% of samples were assigned to the correct feeding system by cross-validation; however, 13% were considered as FREE-FEED. A higher rate of correct classification can be obtained when using three categories or by calculating the weight gain in free-range conditions using regression equations., Conclusion: Taking into account the high variability of the samples and the high success in classification, these results are of interest and may be applied in practical situations., (© 2014 Society of Chemical Industry.)

Background: The two main genetic types in Iberian pig production show important phenotypic differences in growth, fattening and tissue composition since early developmental stages. The objective of this work was the evaluation of muscle transcriptome profile in piglets of both genetic types, in order to identify genes, pathways and regulatory factors responsible for their phenotypic differences. Contemporary families coming from pure Iberian pigs (IB) or from crossing with Duroc boars (DU×IB) were generated. Piglets (14 from each genetic type) were slaughtered at weaning (28 days) and longissimus dorsi was sampled for composition and gene expression studies. RNA was obtained and hybridized to Affymetrix Porcine Genechip expression arrays., Results: Loin muscle chemical composition showed significant differences between genetic types in intramuscular fat content (6.1% vs. 4.3% in IB and DUxIB animals, respectively, P = 0.009) and in saturated (P = 0.019) and monounsaturated fatty acid proportions (P = 0.044). The statistical analysis of gene expression data allowed the identification of 256 differentially expressed (DE) genes between genetic types (FDR < 0.10), 102 upregulated in IB and 154 upregulated in DU×IB. Transcript differences were validated for a subset of DE genes by qPCR. We observed alteration in biological functions related to extracellular matrix function and organization, cellular adhesion, muscle growth, lipid metabolism and proteolysis. Candidate genes with known effects on muscle growth were found among the DE genes upregulated in DU×IB. Genes related to lipid metabolism and proteolysis were found among those upregulated in IB. Regulatory factors (RF) potentially involved in the expression differences were identified by calculating the regulatory impact factors. Twenty-nine RF were found, some of them with known relationship with tissue development (MSTN, SIX4, IRX3), adipogenesis (CEBPD, PPARGC1B), or extracellular matrix processes (MAX, MXI1). Correlation among the expression of these RF and DE genes show relevant differences between genetic types., Conclusion: These results provide valuable information about genetic mechanisms determining the phenotypic differences on growth and meat quality between the genetic types studied, mainly related to the development and function of the extracellular matrix and also to some metabolic processes as proteolysis and lipid metabolism. Transcription factors and regulatory mechanisms are proposed for these altered biological functions.

The effect of a diet enriched with polyunsaturated n-3 fatty acids (PUFA) on endocrine, reproductive, and productive responses of rabbit females and the litters has been studied. Nulliparous does (n=125) were fed ad libitum from rearing to second weaning two diets supplemented with different fat sources: 7.5g/kg lard for the control diet (group C; n=63) or 15g/kg of a commercial supplement containing a 50% ether extract and 35% of total fatty acids (FAs) as PUFA n-3 (Group P; n=62). Dietary treatments did not affect apparent digestibility coefficients of nutrients, or reproductive variables of does including milk production, mortality and average daily gain of kits over two lactations. However, on Day 5 and 7 post-induction of ovulation, progesterone of Group P tended to increase to a greater extent than in does of Group C. Total PUFAs, n-6 and n-3 and eicosapentanoic (EPA) contents were greater in adipose tissues of does in Group P than in Group C. Docosapentaenoic acid (DPA), EPA, and docosahexaenoic acid (DHA) concentrations were greater in peri-ovarian than in scapular fat with abdominal fat being intermediate in concentration. In PUFA supplemented does, kit mortality at the second parturition tended to be less than in control does. Also, kits born to does of the PUFA-supplemented group weighed more and were of greater length than from does of control group. In conclusion, effectiveness of dietary intervention on reproductive and performance response is greater in the second parity, which suggests an accumulative long-term beneficial effect of n-3 FA supplementation in reproductive rabbit does., (Published by Elsevier B.V.)

This study evaluated the strategy of supplementing oral micellized natural vitamin E (D-α-tocopherol) to either piglets and/or sows on α-tocopherol concentrations in piglets serum and tissues after weaning. One first experiment tested the influence of the vitamin E supplementation source (natural form in water v. the synthetic form in feed) and dose administered to piglets and/or sows on serum α-tocopherol concentration, α-tocopherol stereoisomer accumulation, antioxidant capacity and immune response of weaned piglets. A second experiment studied the effect of sow source and dose vitamin E supplementation on some of these parameters in piglets. Oral supplementation to sows with natural vitamin E as a micellized form (D-α-tocopherol) at the lowest dose produced a similar concentration of α-tocopherol in serum at days 2, 14 and 28 postpartum to those supplemented with threefold higher dose of the synthetic form in feed. At day 39 of age, neither piglet supplementation source nor dose significantly affected α-tocopherol accumulation in the serum, muscle, subcutaneous fat or liver. Those piglets from sows supplemented with the micellized alcohol form had higher RRR-α-tocopherol stereoisomers (P<0.001) and lower (P<0.001) RRS- RSS- and RSR-α-tocopherol, at day 39 of age than those from sows supplemented with the synthetic form. A predominant importance of sow over piglet vitamin E supplementation was observed on stereoisomer distribution in piglets. Low doses of oral natural vitamin E supplementation to sows or piglets did not increase the oxidative stress of piglets when compared with the use of the synthetic form in feed. Immunoglobulin levels in piglet serum at day 39 were not affected by natural vitamin E supplementation at low doses in drinking water of piglets or sows when compared with the synthetic form in feed. IgA tended to be higher (P=0.145) at day 39 in piglets supplemented with natural vitamin E when compared with those supplemented with the synthetic form. Low doses of oral micellized natural vitamin E supplementation to sows is an interesting feeding strategy, when compared with the use of high doses of the synthetic form in feed, because it results in similar α-tocopherol concentrations, allows a predominant -R stereoisomer distribution in piglets and also maintains their oxidative status in vivo.

The use of stereoisomers of α-tocopherol to correctly classify Iberian pig fat samples according to their feeding system was investigated. Samples were obtained over two different seasons in controlled farms from the four categories of pigs described in the Industry Quality Policy (FREE-RANGE: pigs fed exclusively under free-range conditions; FREE-FEED: pigs fed free-range and supplemented with feed; FEED-OUT: pigs fed outdoors with access to grass and a mixed diet; and FEED: pigs fed exclusively a mixed diet). A higher presence of RRR-stereoisomer indicated a greater consumption of the natural form of tocopherol provided by acorns or grass, whereas a higher proportion of S forms were related to a higher mixed diet intake. Validation results showed 90% success in fat sample classification. Analysis of the RRR-stereoisomer together with γ- and α-tocopherol determination can be considered as a potent tool for distinguishing fat from pigs fed under free-range conditions or exclusively with acorns and grass from those receiving a supplemented diet at any time of their fattening phase., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

This study tested the effect of micellized vitamin E (D-α-tocopherol; 1,400 IU/d) administered 12 and 1 h orally before training for 1 d (ST-VitE) or 8 d (LT-VitE) compared with an unsupplemented control (CONTROL) on plasma α-tocopherol, thiobarbithuric acid-reactive substances (TBARS), total glutathione (GSHt), and trolox equivalent antioxidant capacity (TEAC) in 10 race horses. Different sampling times [immediately before training (BEF) and after intense training (END) or 8 h after recovery (+8h)] were investigated. Plasma α-tocopherol concentration was greater in the ST-VitE group than the CONTROL group at +8h (P < 0.05). Natural vitamin E supplementation increased plasma α-tocopherol (P < 0.001) in the LT-VitE group by approximately 1.6-fold at BEF, END, and +8h. In all groups, TBARS tended to be slightly greater (P = 0.087) immediately after training when compared with values BEF or +8h and the lowest TBARS values were observed at +8h in LT-VitE. Vitamin E supplementation did not affect the GSHt concentrations at BEF, END, or +8h. The TEAC values were modified by the vitamin E administration (P = 0.010). The greatest TEAC was found in the LT-VitE group at all sampling times and similar concentrations were reached in the ST-VitE group at +8h. The CONTROL group was not able to maintain TEAC after training (P < 0.001), indicating consumption of antioxidants (mainly vitamin E) and consequently oxidative stress because of the antioxidant system being overwhelmed by a reduced antioxidant supply. In conclusion, micellized natural vitamin E at 1,400 IU/d for 8 d efficiently increased plasma α-tocopherol concentration of race horses undergoing intense training conditions and maintained the general oxidative status.

Twenty-four castrated male (Duroc × Large Withe × Landrace) with an average weight of 69.4 kg were randomly distributed and located in individual cages and given the experimental diets. Diets were provided ad libitum and were formulated containing three levels of palm oil and hydropalm (hydrogenated palm stearine): 4% and 0% (low saturation), 2% and 2% (moderate saturation) and 0% and 4% (high saturation). Pigs were slaughtered at a local slaughterhouse with an average weight of 129.1 kg. The dietary treatment had no significant influence on growth performances and carcass characteristics. The diet saturation led to an increase of C18:0 and ΣSFA (saturated fatty acids) proportions and to a reduction of C18:1 n-9, C18:2 n-6 and ΣPUFA (polyunsaturated fatty acids) proportions and in subcutaneous backfat outer and inner layers. The dietary treatment affected significantly the melting point of subcutaneous backfat outer layer but had no significant influence on the melting point of the subcutaneous backfat inner layer. However, a linear effect of C18:0 proportion of subcutaneous backfat outer and inner layers on the melting point was observed. Dietary treatment had no significant effect on the main fatty acids pattern of intramuscular neutral and polar lipids from Longissimus dorsi muscle. In neutral lipids of liver a dietary influence (p < 0.05), but not a clear dose--response relationship, was observed for C16:0, C18:1 n-9, ΣSFA and ΣPUFA fatty acids, whereas in polar lipids the dietary treatment had no significant effect on the main fatty acids proportions. No effect of diet saturation was found on rheological properties of fat except for the hardness. The hardness was higher in the pigs that received the diet with 0% of palm oil and 4% hydropalm than in those given the diet with 4% of palm oil and 0% hydropalm.

This study aimed to assess the interaction between different dietary vitamin A (dVitA) levels and the same concentration of vitamin E (100 IU all-rac-α-tocopheryl acetate/kg feed) in growing-finishing pigs. In the first experiment, two fat sources × two dVitA levels (0 v. 100 000 IU) were used. The supplementation of 100 000 IU dVitA induced a range of 5.13 to 30.03 μg retinol/g liver, 62.78 to 426.88 μg retinol palmitate/g liver, and 0.60 to 1.96 μg retinol/g fat. Dietary fat did not affect retinol or retinyl palmitate deposition in pigs. The high concentration of dVitA produced lower fat and liver α-tocopherol concentrations, and increased susceptibility of muscle tissue to oxidation. A second experiment was carried out to study the retinol and α-tocopherol retention at different withdrawal times prior to slaughter (two dVitA levels; 0 v. 100 000 IU). A high dose of 100 000 IU vitamin A during a short 2-week period was enough to induce α-tocopherol depletion in liver and fat to a similar extent as when 100 000 IU were administered during the whole fattening. Muscle, fat and liver α-tocopherol concentrations were not affected by dVitA in the 1300-13 000 IU/kg range, but liver α-tocopherol concentration was higher when vitamin A was removed from the vitamin mix 5 weeks prior to slaughter (experiment 3).

The effect of dietary fat saturation (dFat) and dietary vitamin A (dVitA) level (0 IU vs. 100,000 IU) on performance, carcass characteristics and fatty acid composition in the offspring of two terminal sires: Duroc (DU) and Landrace×Large White (LD×LW) was studied. In the inner backfat layer, the DU-sired pigs had higher C16:0 proportion (P<0.05) and tended (P<0.07) to have higher total saturated fatty acids (SFA) and lower C18:1 n-9 proportions than LD×LW-sired pigs. An interaction sire line×dVitA was observed for intramuscular fat (IMF) content (P<0.005). The effect of supplementation with the high dVitA level in DU-sired pigs was associated with a 20% increase in IMF while no effect was observed in LD×LW pigs. Fatty acid pattern was affected by dFat and sire line. In the inner backfat layer, LD×LW-sired pigs receiving the low dVitA level increased C18:0 proportions by 8% in comparison to animals receiving the high dVitA, whereas in DU-sired pigs the effect of vitamin A was opposite. It is concluded that the effect of dFat and dVitA concentration on IMF content and fatty acid profile of subcutaneous backfat and IMF is variable according to pig genotype.

In a first experiment one group of pigs was maintained in free-range conditions according to the traditional way in a Mediterranean forest (exercised-1) and another group was housed individually and received acorns (sedentary-1). In a second experiment two groups of pigs were fed a mixed diet for the whole experimental period. One of these groups was housed individually in 8m(2) pens (sedentary-2). The other group was housed in a corridor and forced to walk daily (exercised-2). The subcutaneous fat and neutral lipids of muscle from the exercised pigs fed acorns had higher C18:1n-9, MUFA, C18:1/C18:0, MUFA/SAT and lower C16:0 and SAT when compared with the fat from the pigs fed acorns in confinement. Those exercised animals fed the mixed diet had also lower C16:0 and SAT in subcutaneous fat and lower SAT and higher C18:2, C18:3, PUFA and MUFA/SAT in neutral lipids when compared with the sedentary pigs, which may indicate that delta-9-desaturase activity was higher in exercised than in sedentary pigs. Exercised pigs had higher acid and neutral esterases and lower neutral lipase activity than sedentary pigs. No differences in the α-tocopherol concentration and TBARS values of meat samples among the pigs that received a mixed diet either exercised or sedentary were observed. The moderate exercise reduced the postprandrial concentrations of triglycerides in plasma, but did not reduce other plasma levels.

This experiment was conducted to study the effect of vitamin A concentration in the diet on fatty acid composition in pigs. One-hundred and twenty eight Duroc × (Large White × Landrace) castrated male pigs were used. Pigs were randomly distributed and located in cages of four pigs each to the following treatment: vitamin A-enriched diet (100,000IU/kg) (n=16) and control diet with a basal level (7500IU/kg) (n=16). The pigs were slaughtered at an average weight of 125.9kg. No effect of diet vitamin A level on performance, carcass traits and intramuscular fat percentage was observed. The retinol concentration in subcutaneous backfat and retinol and retinol palmitate contents in liver were higher in the pigs that were given the vitamin A enriched diet than in those receiving the control diet. However the vitamin A supplementation did not increase the retinol content in the muscle. Pigs receiving the vitamin A enriched diet showed a higher C16:0 and ∑SFA proportions in the outer layer and C18:0 and ∑SFA proportions in the inner layer. Moreover, the diet rich in vitamin A led to a reduction of C18:1 n-9 and ∑MUFA proportions in subcutaneous backfat inner layer. Dietary vitamin A concentration had not influence on main fatty acids composition of intramuscular lipids. Dietary vitamin A enrichment lead to a higher C16:0, ∑SFA concentration in liver lipids.

Ellagitannin-containing foods (strawberries, walnuts, pomegranate, raspberries, oak-aged wine, etc.) have attracted attention due to their cancer chemopreventive, cardioprotective, and antioxidant effects. Ellagitannins (ETs) are not absorbed as such but are metabolized by the intestinal flora to yield urolithins (hydroxydibenzopyran-6-one derivatives). In this study, Iberian pig is used as a model to clarify human ET metabolism. Pigs were fed either cereal fodder or acorns, a rich source of ETs. Plasma, urine, bile, lumen and intestinal tissues (jejunum and colon), feces, liver, kidney, heart, brain, lung, muscle, and subcutaneous fat tissue were analyzed. The results demonstrate that acorn ETs release ellagic acid (EA) in the jejunum, then the intestinal flora metabolizes EA sequentially to yield tetrahydroxy- (urolithin D), trihydroxy- (urolithin C), dihydroxy- (urolithin A), and monohydroxy- (urolithin B) dibenzopyran-6-one metabolites, which were absorbed preferentially when their lipophilicity increased. Thirty-one ET-derived metabolites were detected, including 25 urolithin and 6 EA derivatives. Twenty-six extensively conjugated metabolites were detected in bile, glucuronides and methyl glucuronides of EA and particularly urolithin A, C, and D derivatives, confirming a very active enterohepatic circulation. Urolithins A and B as well as dimethyl-EA-glucuronide were detected in peripheral plasma. The presence of EA metabolites in bile and in urine and its absence in intestinal tissues suggested its absorption in the stomach. Urolithin A was the only metabolite detected in feces and together with its glucuronide was the most abundant metabolite in urine. No metabolites accumulated in any organ analyzed. The whole metabolism of ETs is shown for the first time, confirming previous studies in humans and explaining the long persistency of urolithin metabolites in the body mediated by an active enterohepatic circulation.

This experiment was undertaken to provide information on the effect of feeding system applied during the finishing period (100-150kg) on the quality of Iberian pig meat. Four feeding systems were applied: pigs fed under free-range conditions with acorns and grass fully available during 111 days (FR(111)), pigs fed concentrate diet in confinement during 28 days and free-range with acorns and grass fully available during 83 days (CDC(28)+FR(83)), pigs fed concentrate diet in confinement during 65 days and free-range with acorns and grass fully available during 46 days (CDC(65)+FR(46)) and pigs fed concentrate diet in confinement with a feed average daily amount of 3.1kg during 111 days (CDC(111)). The CDC(111) pigs had more muscular carcasses characteristics than the remaining groups of pigs. However, the higher concentration of C18:1 n-9 in subcutaneous backfat was found in FR(111) pigs. In the neutral lipids from intramuscular fat of Longissimus dorsi only the C18:2 n-6 and C18:3 n-3 proportions were affected by feeding system, and the n-6/n-3 ratio observed was lower in FR(111) and CDC(28)+FR(83) pigs than in CDC(111) pigs. In the neutral lipids from hepatic fat the proportion of n-6 fatty acids was lower in FR(111) pigs than in CDC(65)+FR(46) and CDC(111) pigs.

Sixteen Iberian barrows of the same age with an average initial live weight of 100.1kg were randomly distributed in two groups of eight pigs each. One group was fed healthy acorns and the other group received acorns infested of Curculio sp. The subcutaneous backfat from pigs fed healthy acorns had higher C18:1n-9, MUFA and C20:5n-3 and lower C18:0 and SFA proportions than that from the pigs fed acorns infested with Curculio. The consumption of acorns infested with Curculio sp. led to a reduction of C18:1n-9, MUFA, C18:2n-6, C18:3n-3, C22:5n-3 and PUFA proportions in neutral lipids from Longissimus dorsi muscle with respect to consumption of healthy acorns, whereas in polar lipids it produced a reduction in C18:1n-9, MUFA and C18:4n-3 proportions and an increase in C18:2n-6, C20:4n-6, n-6 and C20:5n-3 proportions and of n-6/n-3 ratio with respect to the healthy acorns consumption. The pigs fed healthy acorns had higher intramuscular fat percentage in Longissimus dorsi than pigs fed with acorns infested with Curculio (9.95 vs 7.09% SEM=0.60).

The experiment was undertaken to provide information on the influence of grass intake in a high fat diet based on acorns, in either free-range or confinement, on the carcass yield and characteristics, and on the accumulation of fatty acids and tocopherols. Groups raised free-range or with acorns and grass in confinement had the highest total backfat. Grass addition to the feed did not significantly modify the carcass weight and cut yield. However, outdoor raising produced lower carcass weights and yield, Longissimus dorsi muscle and ham weights than those fed the formulated diet or acorns and grass in confinement. Neither grass intake nor outdoor rearing did modify significantly the proportion of saturated fatty acids of the backfat. However, a lower proportion of saturated fatty acids was found in the intramuscular neutral lipids from pigs fed outdoors. Pigs reared free-range had significantly (P=0.0001) higher proportions of C18:1n-9 in the inner layer and in the intramuscular neutral lipids than the other groups. The grass intake significantly (P=0.0001) increased the proportion of C18:3n-3 in the inner and outer fat layers and intramuscular neutral lipids, while the outdoor rearing system not only affected on the proportion of linolenic acid but also C22:5 and C22:6 of the inner backfat layer and intramuscular polar lipids. These results suggest that the outdoor rearing allows a higher accumulation of n-3 fatty acids maybe due to an increased activity of the desaturase and elongase enzymes. On the other hand, the accumulation of tocopherols was not significantly modified neither by the grass intake nor the outdoor rearing, while differences were detected by the type of feeding (diet vs. acorns/acorns and grass). It is concluded, that the outdoor rearing system affects the nutritional value of meat, mainly by improving the fatty acid composition.

A group of 11 pigs was fed with 70 g feed per kg of metabolic weight (H pigs) and another group of 11 pigs was fed with 50g feed per kg of metabolic weight (L pigs). In both experimental groups (H and L pigs), it was observed that the higher initial proportion of C16:0, C18:0 and C18:2 (n-6) in backfat at the beginning of the free-range feeding period, the greater decrease rate of these fatty acid proportions regarding weight gain during the free-range fattening period took place. On the other hand, the greater initial proportion of C18:1 (n-9), the smaller increase rate in the concentration of this fatty acid was observed. The intramuscular neutral lipids from L pigs had higher C18:3 (n-3) and lower proportions of monounsaturated fatty acids (MUFA) than those from H pigs, while intramuscular polar lipids from L pigs had significantly higher proportions of C18:0, PUFA, C18:3 (n-3) and (n-3) and significantly lower MUFA and C18:1 (n-9) proportions than those from H pigs. The alpha-tocopherol concentration found in Longissimus dorsi from L pigs was significantly higher (p < 0.012) than those from H pigs.

The effects of partial replacement of dietary polyunsaturated fatty acid (PUFA) with monounsaturated fatty acid (MUFA) on fatty acid profiles of subcutaneous and intramuscular fat and rheological properties of fat were studied in heavy pigs. No effect of dietary fat was observed on total saturated fatty acids proportion in outer and inner backfat layers or intramuscular neutral and polar lipids. The partial substitution of dietary PUFA by MUFA produced a rise in MUFA of intramuscular polar lipids (p < 0.01) but neutral lipids were not affected. Moreover, it also decreased (p < 0.01) the n-6 proportion of outer and inner backfat layers and intramuscular polar lipids, but no effect was observed for the n-6 in the intramuscular neutral lipids. No significant effect of dietary fat treatment was observed on n-3 fatty acids in outer and inner backfat layers, but the partial substitution of PUFA by MUFA increased (p < 0.01) the n-3 proportion of intramuscular lipid fractions and the consequent decreased in the n-6/n-3 ratio in all lipid location. Significant effects of dietary vitamin E were observed on springiness, cohesiveness and adhesiveness of backfat inner layer. However, consistency was not affected by dietary fat. Further research is needed to establish the effects of different ranges of dietary oil combination in the feeding of heavy pigs.

The experiment was undertaken to provide information of the influence of feeding either free-range or in confinement with different dietary MUFA/PUFA ratios and α-tocopheryl acetate supplementation (40 vs. 200 mg/kg) on tocopherol content and susceptibility to lipid oxidation of muscle and microsomes in Iberian pigs. The grass provided to the pigs had a similar α-tocopherol concentration to that observed for diets supplemented with 200 mg/kg α-tocopheryl acetate, and acorns supplied fourfold higher content of γ-tocopherol than the experimental diets. The α- and γ-tocopherol contents of muscle reflected the tocopherol concentration of the diets. Mono and Medium diets produced a similar MUFA/PUFA ratio in neutral and polar lipids of pig muscle to those fed outdoors. The lowest TBARS numbers were found in muscle samples from pigs fed a MUFA-enriched diet in confinement. No significant influence of free-range feeding or dietary fat on drip loss was found. However, α-tocopheryl acetate supplementation reduced (P<0.05) drip loss. Dietary vitamin E supplementation decreased the membrane lipid oxidation by 18% after 120 min. However, free-range feeding decreased the extent of microsome oxidation by 20%, 56% and 82% after 120 min when compared with those groups fed in confinement with high, medium and low MUFA/PUFA ratios, respectively. The hexanal concentration of muscle showed a similar trend to that observed for microsome induced-oxidation, suggesting, that hexanal determination is a more accurate method to measure lipid oxidation in iberian pig muscle than the thiobarbituric acid test.

The objective of this study was to assess the effect of dietary copper and/or vitamin E supplementation on composition and oxidation of M. longissimus in Iberian pigs and to compare it with meat samples from pigs produced extensively and fed with acorn and grass. Grass had the highest alpha-tocopherol content (> 150 mg/kg DM), while acorns had the highest copper concentration (78.1 mg/kg DM). Dietary treatment did not affect copper composition in muscle and no interactions were detected between copper and alpha-tocopherol. The alpha-tocopherol content in muscle from pigs fed diets supplemented with alpha-tocopheryl acetate (100 mg/kg feed) was 1.5 times greater (p < 0.0001) than those from pigs receiving a basel diet. M. longisssimus dorsi from pigs fed extensively had a higher concentration of alpha-tocopherol than those fed in confinement with the basel level of alpha-tocopheryl acetate, but lower values than pigs fed supplemented levels. Total n-3 fatty acids (p < 0.02) and hematin (p < 0.0001) concentrations were significantly higher in muscle from pigs fed extensively than when fed in confinement. Muscle homogenates from Iberian pigs fed in extensive conditions showed significantly (p < 0.02) higher susceptibility to oxidation than those from pigs fed mixed diets. Dietary alpha-tocopheryl acetate supplementation (100 mg/kg feed) significantly (p < 0.05) reduced lipid oxidation of muscle, while dietary copper concentration did not modify susceptibility to lipid oxidation.

The effect of n-3 fatty acid-enriched diets (in the form of 0.5% linseed oil with either 1.5% sunflower oil or 1.5% olive oil) and alpha-tocopheryl acetate supplementation (200 mg/kg feed) on lipid oxidation (thiobarbituric acid-reactive substances, TBARS) and cholesterol oxide products (COPS) in cooked pork was investigated. Longissimus muscle was studied. Meat from pigs fed 0.5% linseed oil-enriched diets had a higher proportion of n-3 fatty acid than meat from pigs in other dietary groups in neutral (P < 0.0001) and polar lipids (P < 0.0001), and a 20% reduction in the n-6:n-3 ratio was observed. Alpha-tocopheryl acetate supplementation increased (P < 0.05) monounsaturated fatty acids in polar lipids and increased (P = 0.0001) alpha-tocopherol levels in muscle. Alpha-tocopherol concentration in muscle was affected by dietary fat (P < 0.05). Groups receiving diets enriched with sunflower oil had significantly higher alpha-tocopherol levels (P < 0.05) in muscle than those groups receiving olive oil-enriched diets. Numbers of TBARS were significantly (P < 0.05) lower in the group fed supplemental olive oil than in those fed sunflower oil. Dietary linseed oil increased (P < 0.05) lipid oxidation principally at the initial period of storage in cooked pork. Overall, dietary alpha-tocopheryl acetate supplementation significantly increased (P < 0.001) lipid stability and decreased (P < 0.05) total COP production across the dietary groups. Alpha-tocopherol was a more effective antioxidant for decreasing TBARS values in cooked meat when adding sunflower oil to the diets instead of olive oil.

Diets containing different ratios of n-3:n-6 polyunsaturated fatty acids, were fed to piglets for a period of 10 days. Diets with n-3:n-6 ratios of 0.2 and 0.3 decreased the group mean activity of plasma butyrylcholinesterase when compared with a diet with a ratio of 0.1.