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Dietary methionine affects protein metabolism, lean gain and growth performance and acts in the control of oxidative stress. When supplied in large excess relative to growth requirements in diets for pigs, positive effects on pork quality traits have been recently reported. This study aimed to decipher the molecular and biochemical mechanisms affected by a dietary methionine supply above growth requirements in the loin muscle of finishing pigs. During the last 14 days before slaughter, crossbred female pigs (n = 15 pigs/diet) were fed a diet supplemented with hydroxy-methionine (Met5; 1.1% of methionine) or not (CONT, 0.22% of methionine). Blood was sampled at slaughter to assess key metabolites. At the same time, free amino acid concentrations and expression or activity levels of genes involved in protein or energy metabolism were measured in the longissimus lumborum muscle (LM). The Met5 pigs exhibited a greater activity of creatine kinase in plasma when compared with CONT pigs. The concentrations of free methionine, alpha-aminobutyric acid, anserine, 3-methyl-histidine, lysine, and proline were greater in the LM of Met5 pigs than in CONT pigs. Expression levels of genes involved in protein synthesis, protein breakdown or autophagy were only scarcely affected by the diet. Among ubiquitin ligases, MURF1, a gene known to target creatine kinase and muscle contractile proteins, and OTUD1 coding for a deubiquitinase protease, were up-regulated in the LM of Met5 pigs. A lower activity of citrate synthase, a reduced expression level of ME1 acting in lipogenesis but a higher expression of PPARD regulating energy metabolism, were also observed in the LM of Met5 pigs compared with CONT pigs. Principal component analysis revealed that expression levels of many studied genes involved in protein and energy metabolism were correlated with meat quality traits across dietary treatments, suggesting that subtle modifications in expression of those genes had cumulative effects on the regulation of processes leading to the muscle transformation into meat. In conclusion, dietary methionine supplementation beyond nutritional requirements in pigs during the last days before slaughter modified the free amino acid profile in muscle and its redox capacities, and slightly affected molecular pathways related to protein breakdown and energy metabolism. These modifications were associated with benefits on pork quality traits.

The process of oxidative stress occurs all over the production chain of animals and food products. This review summarises insights obtained in different farm species (pigs, ruminants, poultry, and fishes) to underpin the most critical periods for the venue of oxidative stress, namely birth/hatching and weaning/start-feeding phase. Common responses between species are also unravelled in periods of high physiological demands when animals are facing dietary deficiencies in specific nutrients, suggesting that nutritional recommendations must consider the modulation of responses to oxidative stress for optimising production performance and quality of food products. These conditions concern challenges such as heat stress, social stress, and inflammation. The magnitude of the responses is partly dependent on the prior experience of the animals before the challenge, reinforcing the importance of nutrition and other management practices during early periods to promote the development of antioxidant reserves in the animal. When these practices also improved the performance and health of the animal, this further confirms the central role played by oxidative stress in physiologically and environmentally induced perturbations. Difficulties in interpreting responses to oxidative stress arise from the fact that the indicators are only partly shared between studies, and their modulations may also be challenge-specific. A consensus about the best indicators to assess pro-oxidative and antioxidant pathways is of huge demand to propose a synthetic index measurable in a non-invasive way and interpretable along the productive life of the animals.

Adipose tissue is a primary sensor for nutrient availability and regulates many functions including feed intake and energy homeostasis. This study was undertaken to determine the molecular responses of adipose tissue to differences in feed intake and feed efficiency. Subcutaneous adipose tissue was collected from two lines of pigs divergently selected for residual feed intake (RFI), a measure of feed efficiency defined as the difference between actual and expected feed intake, and from a subset of high-RFI pigs that were feed-restricted at the level of the voluntary feed intake of low-RFI pigs during the growing-finishing period. Transcriptomics analyses indicated that the number of genes that were differentially expressed (0.01) between low- and high-RFI pigs ( = 8 per group at each stage) in adipose tissue was much lower when pigs were considered at 19 kg (postweaning) than at 115 kg BW (market weight). Extended investigations were performed at 115 kg BW to compare low-RFI ( = 8), high-RFI ( = 8), and feed-restricted high-RFI ( = 8) pigs. They included in silico pathway analyses of the differentially expressed (DE) genes (0.01) and a complementary proteomic investigation to list adipose proteins with a differential abundance (0.10). Only 23% of the DE genes were affected by both RFI and feed restriction. This indicates that the responses of adipose tissue to RFI difference shared only some common mechanisms with feed intake modulation, notably the regulation of cell cycle (including ) and transferase activity pathway. Two carboxylesterase genes (, ) involved in lipolysis, were among the most overexpressed genes in the low-RFI pigs; they were also affected by feed restriction within the high-RFI line. About 60% of the molecular changes between low- and high-RFI pigs were specific to genetic divergence in feed efficiency, independently of feed intake. Different genes and proteins known to be associated with mitochondrial oxidative metabolism were overexpressed in adipose tissue of low-RFI pigs compared with high-RFI pigs; other proteins participating in the generation of energy were also affected by feed restriction within the high-RFI line. Finally, mitochondrial antioxidant genes were upregulated in low-RFI pigs vs. high-RFI pigs. Altogether, increased oxidative and antioxidant processes in adipose tissue might be associated with improved feed efficiency.

Improving feed efficiency is a relevant strategy to reduce feed cost and environmental waste in livestock production. Selection experiments on residual feed intake (RFI), a measure of feed efficiency, previously indicated that low RFI was associated with lower feed intake, similar growth rate, and greater lean meat content compared with high RFI. To gain insights into the molecular mechanisms underlying these differences, 24 Large White females from 2 lines divergently selected for RFI were examined. Pigs from a low-RFI ("efficient") and high-RFI ("inefficient") line were individually fed ad libitum from 67 d of age (27 kg BW) to slaughter at 115 kg BW (n = 8 per group). Additional pigs of the high-RFI line were feed restricted to the daily feed intake of the ad libitum low-RFI pigs (n = 8) to investigate the impact of selection independently of feed intake. Global gene and protein expression profiles were assessed in the LM collected at slaughter. The analyses involved a porcine commercial microarray and 2-dimensional gel electrophoresis. About 1,000 probes were differentially expressed (P0.01) between RFI lines. Only 10% of those probes were also affected by feed restriction. Gene functional classification indicated a greater expression of genes involved in protein synthesis and a lower expression of genes associated with mitochondrial energy metabolism in the low-RFI pigs compared with the high-RFI pigs. At the protein level, 11 unique identified proteins exhibited a differential abundance (P0.05) between RFI lines. Differentially expressed proteins were generally not significantly affected by feed restriction. Mitochondrial oxidative proteins such as aconitase hydratase, ATP synthase subunit α, and creatine kinase S-type had a lower abundance in the low-RFI pigs, whereas fructose-biphosphate aldolase A and glyceraldehyde-3-phosphate dehydrogenase, 2 proteins involved in glycolysis, had a greater abundance in those pigs compared with high-RFI pigs. Antioxidant proteins such as superoxide dismutase and glutathione peroxidase 3 at the mRNA level and peroxiredoxin-6 at the protein level were also less expressed in LM of the most efficient pigs, likely related to lower oxidative molecule production. Collectively, both the transcriptomic and proteomic approaches revealed a lower oxidative metabolism in muscle of the low-RFI pigs and all these modifications were largely independent of differences in feed intake.

Both breed and production systems are responsible for production efficiency and quality traits of pork. Effects of breed and production system within breed on growth, body fatness, and adipose tissues traits were assessed in the pure Basque (B, nonselected, local French) and conventional Large White (LW) breeds, reared either in a conventional (C, slatted floor), alternative (A, indoor straw bedding and outdoor area), or extensive (E, free range) system. A total of 100 castrated males were produced in 2 replicates, each involving 50 pigs distributed in 5 treatments based on breed and production system (i.e., BC, BA, BE, LWC, and LWA [10 pigs/group and per replicate]). From 35 kg BW to slaughter at around 145 kg BW, the BC, BA, LWC, and LWA pigs received the same growing and finishing diets, whereas the BE pigs had free access to the natural resources of the E pen and received a standard growing-finishing diet at restricted allowance according to the farming practices of the B pork chain. The B pigs had lower (P0.001) ADG and G:F than the LW pigs and were much older (P0.001) at slaughter. The LWA pigs had similar ADG but lower (P = 0.03) G:F than the LWC. Within the B breed, the BA had higher (P = 0.04) and the BE lower (P0.001) ADG compared with BC pigs. The B pigs had a higher (P0.001) carcass dressing an exhibited around 2-fold higher (P0.001) back fat proportion, perirenal fat weight and LM lipid content than the LW pigs. Compared with C, the A system decreased (P = 0.04) carcass dressing within LW but did not influence carcass traits within B pigs. The E system decreased (P ≤ 0.05) carcass dressing, back fat proportion, and LM lipid content in BE compared with BC pigs. The B pigs exhibited larger (P0.001) adipocytes in both subcutaneous adipose tissue (SCAT) and LM than the LW pigs. Malic enzyme activity was higher in SCAT of B than LW pigs despite their greater fatness, and was higher (P ≤ 0.01) in BA but lower (P0.001) in BE than in BC pigs. The B pigs had higher (P0.001) MUFA but lower (P ≤ 0.006) SFA and PUFA fatty acid percentages in SCAT than the LW pigs. Compared with C, the A system had scarce influence on FA composition within each breed, whereas the E system led to lower (P = 0.015) SFA and greater (P0.001) PUFA in SCAT of the B pigs. Altogether, the E production system can counteract the genetic potential of B pigs for growth rate but also body fatness.

Ingested dietary nutrients and feed energy are partitioned among tissues to sustain body growth. Based on the respective costs of the various metabolic pathways allowing use and storage of feed energy into cells, it may be theorized that daily meal frequency could affect growth, body composition or feed efficiency. This study aimed to determine the effects of daily meal frequency on nutrient partitioning, tissue metabolism and composition, and performance. Young growing pigs (30 kg BW) were offered a same amount of feed either in 2 (M2, n = 15) or 12 (M12, n = 16) meals per day during a 3-wk interventional period. Animals fed twice a day had an accelerated weight gain (+6.4%, P0.05) and exhibited a greater G:F (+4%, P = 0.03) than animals fed 12 meals per day during this period. Basal plasma concentrations of glucose, lactate, triglyceride, urea, and leptin were lower (P0.001) in M2 pigs than in M12 pigs. Meal frequency also changed (P0.001) the time-course profiles of plasma concentrations of glucose, insulin, and lactate in response to meal ingestion. A greater rise and a sharper fall in plasma glucose and insulin levels were observed in M2 pigs compared with M12 pigs. In both groups, similarities were observed in the postprandial time courses of plasma concentrations of insulin and of α-amino nitrogen (used as a measure of total AA). Despite these metabolic responses, tissue lipids, glycogen content, and enzyme activities participating in energy metabolism in muscle and liver were similar (P0.10) in both groups at the end of the trial. Percentage of perirenal fat in the body and depth of dorsal subcutaneous fat tissue were not affected by meal frequency, but kidney weight was lower (-18%, P0.001) in M2 pigs than in M12 pigs. Altogether, the less frequent daily meal intake improves the conversion of feed into weight gain, without marked modifications of tissue composition in young pigs.

A quantitative trait locus (QTL) affecting fatness in a way opposite to expectations based on breed means was mapped to swine chromosome 7 (SSC7) using crosses between Large White (LW) and Meishan (MS) founders. Defining the molecular fatness trait more explicitly would allow deducing positional candidate genes, for which expression differences must be analysed in experimental populations. First, mRNA levels of genes representing sequential steps in adipogenesis or involved in lipid metabolism were studied in backfat of pigs having homozygous LW(QTL7)/LW(QTL7) or heterozygous LW(QTL7)/MS(QTL7) alleles and considered at two ages. mRNA level of DLK1 expressed in preadipocytes was greater in MS(QTL7)/LW(QTL7) pigs than in homozygous pigs at 28 days. Transcript abundances of CEBPA involved in differentiation, the prolipogenic FASN gene and the adipocyte-specific marker FABP4 were lower in MS(QTL7)/LW(QTL7) pigs compared with LW(QTL7)/LW(QTL7) pigs at 150 days. Because these results suggest a lag time in terminal differentiation associated with the MS allele, seven genes in the QTL interval were deduced as promising candidates for the QTL effect by bioinformatics analysis. Among them, PPARD and CDKN1A had lower expression levels in MS(QTL7)/LW(QTL7) pigs at both ages. Genotype-related differences were observed in mRNA levels of PPARD target genes involved in cell differentiation (FZD7) or fatty acid oxidation (ACADL and ACOX1) at 150 days. These results re-evaluate the potential of PPARD to explain part of variation in pig adiposity.

The possibility that maternal diets during gestation could affect growth and tissue development of offspring and program their later phenotype is an emerging challenge in pig production. The objective of the current study was to investigate the effects of contrasted protein levels in diets of pregnant sows on the proteomic features of subcutaneous adipose tissue (SCAT) of the offspring at birth and its possible persistence later in age. Sows were fed control (Con), low (LP), or high protein (HP) diets throughout gestation. A subset of piglets was killed at 1 d of age for SCAT sampling. The remaining piglets were cross-fostered to nonexperimental sows during lactation. They were fed standard diets during postweaning and fattening periods until 186 d of age. Modifications in SCAT protein abundance shortly after birth were investigated by 2-dimensional gel electrophoresis followed by mass spectrometry. A total of 65 spots were found differentially expressed (Por= 0.10) in SCAT of 1-d-old experimental piglets vs. Con piglets. Proteins with a greater abundance in LP piglets compared with Con piglets were involved in pathways related to glucose and fatty acid metabolisms, lipid transport, and regulation of apoptosis. Upregulation of 5 proteins representative of these biological pathways in LP group vs. Con group were further validated (P0.05) by Western blot analyses. Furthermore, the specific activity of the key lipogenic enzyme fatty acid synthase was found greater (P = 0.06) in SCAT of 1-d-old LP piglets than in Con piglets. The main changes evidenced in SCAT of HP piglets compared with Con animals at 1 d of age rather concerned proteins putatively involved in AA metabolism or in protein turnover. Adipose tissue contents in some proteins that had displayed a greater (Por= 0.10) abundance in experimental pigs compared with Con at d 1 (e.g., transaldolase, annexin II, and apolipoprotein A4) were, however, similar (P0.10) in the 3 groups at d 186 of age. Enolase 1 has less abundance (P0.05) in LP pigs compared with Con pigs at this stage. In conclusion, the proteomics tool has allowed the identification of early changes in various molecular pathways of SCAT in response to the levels of maternal protein supply during gestation.

Pen housing has been proposed in rabbits as an alternative to standard-sized cages. Rabbits reared in pens show greater physical activity. This study investigated whether jump exercise could modify body composition, muscle biochemical and histological characteristics, and some meat quality traits, including the biomechanical properties of tendons. Male weaned rabbits of similar BW (793 +/- 11 g) were either reared in giant collective cages and had to jump over obstacles to get food and water for 35 consecutive days (EXE), or confined in small isolated cages (SEDN). Rabbits were weighed weekly to determine ADG (n = 79 EXE; n = 46 SEDN) and ADFI (n = 9 cages in EXE; n = 46 cages in SEDN). At approximately 10 wk of age, rabbits were slaughtered in 2 series. After overnight chilling, carcasses in the first series (n = 30 EXE; n = 27 SEDN) were divided into fore, intermediate, and hind parts. Color and ultimate pH were recorded in the biceps femoris (BF) and LM. The Achilles tendon and patellar ligament were dissected from the legs and cooked. Muscles [semimembranosus proprius, semimembranosus accessorius (SMA), and BF] were harvested from the legs in a subset of animals from the second series (n = 10 in EXE; n = 9 in SEDN). Both ADG and ADFI were slightly reduced (P0.10) in EXE rabbits compared with SEDN rabbits. Exercised rabbits showed a greater (P = 0.01) proportion of hind parts than SEDN rabbits. Enzyme activities of 3-hydroxyacyl-CoA dehydrogenase and citrate synthase, which play key roles in fatty acid oxidation and the terminal oxidative degradation of nutrients, respectively, were increased in the semimembranosus proprius, SMA (except citrate synthase), and BF muscles of EXE rabbits compared with SEDN rabbits. Only SMA exhibited a decreased (P = 0.05) activity of the glycolytic enzyme, lactate dehydrogenase, in EXE rabbits compared with SEDN animals. Total lipid content, mean diameter of perimysial adipocytes, and activities of core lipogenic enzymes in the SMA and BF muscles did not differ between EXE and SEDN rabbits. Meat color in BF was shifted toward greater a* (red; P = 0.001) and b* (yellow; P = 0.02) values in EXE rabbits compared with SEDN rabbits. Cooked Achilles tendon and patellar ligaments in the legs had greater stiffness (Por = 0.05) in EXE rabbits compared with SEDN rabbits. This experiment demonstrates that rabbit muscles turn to a more oxidative metabolic pattern in response to jump exercise. The quality of attachment of cooked meat to bone is also improved in active rabbits.

Intramuscular fat content is generally associated with improved sensory quality and better acceptability of fresh pork. However, conclusive evidence is still lacking for the biological mechanisms underlying i.m. fat content variability in pigs. The current study aimed to determine whether variations in i.m. fat content of longissimus muscle are related to i.m. adipocyte cellularity, lipid metabolism, or contractile properties of the whole muscle. To this end, crossbred (Large White x Duroc) pigs exhibiting either a high (2.82 +/- 0.38%, HF) or a low (1.15 +/- 0.14%, LF) lipid content in LM biopsies at 70 kg of BW were further studied at 107 +/- 7 kg of BW. Animals grew at the same rate, but HF pigs at slaughter presented fatter carcasses than LF pigs (P = 0.04). The differences in i.m. fat content between the 2 groups were mostly explained by variation in i.m. adipocyte number (+127% in HF compared with LF groups, P = 0.005). Less difference (+13% in HF compared with LF groups, P = 0.057) was noted in adipocyte diameter, and no significant variation was detected in whole-muscle lipogenic enzyme activities (acetyl-CoA carboxylase, P = 0.9; malic enzyme, P = 0.35; glucose-6-phosphate dehydrogenase, P = 0.75), mRNA levels of sterol-regulatory element binding protein-1 (P = 0.6), or diacylglycerol acyltransferase 1 (P = 0.6). Adipocyte fatty acid binding protein (FABP)-4 protein content in whole LM was 2-fold greater in HF pigs than in LF pigs (P = 0.05), and positive correlation coefficients were found between the FABP-4 protein level and adipocyte number (R2 = 0.47, P = 0.02) and lipid content (R2 = 0.58, P = 0.004). Conversely, there was no difference between groups relative to FABP-3 mRNA (P = 0.46) or protein (P = 0.56) levels, oxidative enzymatic activities (citrate synthase, P = 0.9; beta-hydroxyacyl-CoA dehydrogenase, P = 0.7), mitochondrial (P = 0.5) and peroxisomal (P = 0.12) oxidation rates of oleate, mRNA levels of genes involved in fatty acid oxidation (carnitine-palmitoyl-transferase 1, P = 0.98; peroxisome proliferator-activated receptor delta, P = 0.73) or energy expenditure (uncoupling protein 2, P = 0.92; uncoupling protein 3, P = 0.84), or myosin heavy-chain mRNA proportions (P0.49). The current study suggests that FABP-4 protein content may be a valuable marker of lipid accretion in LM and that i.m. fat content and myofiber type composition can be manipulated independently.

Muscle fat stores at slaughter partly determine the dietetic and sensory quality traits of pork meat. Nutritional strategies during the growing-finishing period are able to modify intramuscular fat content; however, the underlying mechanisms remain largely unknown. The objective of this study was to determine some of the cellular, biochemical, and molecular bases of muscle fat content variation in response to feeding regimen in pigs. Crossbred pigs of 30 kg BW were allocated to three feeding groups: free access ([C], n = 10) to a standard diet (3.25 kcal of DE/kg, 9.5 g of lysine/kg), standard diet at 75% of the spontaneous voluntary intake ([FR], n = 10), or both low protein and energy intakes ([PR], n = 10) in order to get the same growth rate as the FR pigs and the same body composition as the C pigs. At slaughter (110 kg BW), FR and PR pigs were 30 d older than C pigs (P0.001). In agreement with the protocol, carcass adiposity was similar in PR and reduced (P0.01) in FR pigs compared with C animals. Lipid content in longissimus lumborum muscle was reduced by 25% in FR pigs and increased by 40% in PR pigs compared to C pigs (P0.001). Commensurate variations in the diameter of muscle adipocytes were observed between the three feeding groups (P0.001). The muscle activities of malic enzyme and glucose-6-phosphate dehydrogenase, generating reduced nicotinamide adenine dinucleotide phosphate for fatty acid synthesis, were depressed (P0.05) in both FR and PR groups, compared to the C group. The expression level of the sterol regulatory element binding protein that was chosen as the putative candidate at the molecular level was not modified by the feeding regimen. No variations in the oxidative enzyme markers were denoted, whereas lactate dehydrogenase activity was reduced by 13% (P0.05) in PR group compared to other groups. In conclusion, moderate long-term feed restriction results in decreased lipogenic capacity of muscle adipocytes and intramuscular fat content. In contrast, the reduction of both protein and energy intakes more likely results in an imbalance between multiple aspects of muscle energy metabolism, in favor of intramuscular fat accretion.