Intramuscular fat content in different muscles,locations, weights and genotype-sexes and its prediction in live pigs with computed tomography

Intramuscular fat (IMF) content depends on sex, genotype and diet and varies with pig growth. The aim of the present work was to determine the evolution of IMF by genotype-sex, muscle and muscle location, to determine relationships between IMF content of different muscles and to predict IMF in live pigs with computed tomography (CT). For this purpose, 155 pigs of seven combinations of genotype-sex were CT scanned and slaughtered at 70, 100 and 120 kg. From the carcasses, fat thickness was measured at several locations along the midline. Loin samples from three anatomical positions (between the eighth and ninth last ribs, between the third and fourth last ribs and between the third and fourth lumbar vertebrae) and three ham muscles (biceps femoris, semimembranosus and gluteus medius) were extracted, weighed and IMF was determined with near-IR equipment. From CT images, the distribution of volume by Hounsfield value (unit related with the density) was obtained for each muscle and anatomical location. Marbling was evaluated in the three loin locations. The effects of genotype-sex and live weight and their interaction were included in the statistical model. For prediction of IMF with CT images, partial least square regression was used. The results show differences in IMF content by genotype-sex and muscle. In general, the most cranial part of the loin presented higher IMF content, as well as the biceps femoris muscle of the ham. Depending on the genotype-sex, IMF content increased during all growth or increased until 100 kg and then became constant. Correlation coefficients between IMF content by muscle/location were between 0.74 and 0.83 within loin locations and between 0.53 and 0.70 for ham muscles. Correlation coefficients between marbling and IMF content evaluated at the same location varied between 0.51 and 0.66. Prediction of IMF content from CT images is not accurate enough (residual predictive deviation statistical values lower than 1.3). Muscle weight increase with animal growth and allometric coefficients varied between 0.89 and 0.97 for the muscles evaluated. The conclusions of the present work are that IMF content differs between and within muscle, during growth and by genotype-sex and that prediction of IMF in CT images of live pigs is not accurate.     

Age-related changes and nutritional regulation of myosin heavy-chain composition in longissimus dorsi of commercial pigs

The objective of this study is to investigate the age-related changes of and the effects of dietary conjugated linoleic acid (CLA) on muscle-fibre types in commercial pigs. We divided 25 crossbred male pigs into five age groups (7, 30, 60, 100 and 180 days) and 30 finishing pigs into two dietary groups (one fed a CLA-enriched diet and the other fed a control diet for 30 days). We analysed the composition (%) of myosin heavy-chain (MyHC) mRNA according to the absolute copies of each MyHC (I, IIa, IIb and IIx) mRNA, and the activities of succinate dehydrogenase (SDH) and malate dehydrogenase (MDH) in the longissimus muscle. From days 7 to 180, the MyHC I mRNA abundance and SDH and MDH activities presented a decreasing trend, the MyHC IIb mRNA abundance presented a steady trend and the MyHC IIa and IIx mRNA abundances presented an increasing trend. On day 30, MyHC I and IIb mRNA abundances were at their lowest (P < 0.05), and the MyHC IIa and IIx mRNA abundances were at their highest (P < 0.05). In the CLA group, the MyHC I mRNA abundance and the activities of SDH and MDH were improved in the longissimus muscle, whereas pressure loss, drip loss and average back fat depth significantly decreased (P < 0.01) and shear force significantly increased (P < 0.01). Loin eye area, feed conversion rate and meat colour showed some tendency to be improved. These results indicated that more oxidative fibres might convert to glycolytic fibres with increasing age or weight, and that the early developmental stage might be a key stage for this conversion. During the finishing stage, the proportion of oxidative fibres might be increased by dietary CLA supplementation, which may contribute to the water-holding capacity of meat. The results would provide an important basis for the application of muscle-fibre types in the improvement of pork quality.     

Detection of differentially expressed genes in the longissimus dorsi of Northeastern Indigenous and Large White pigs

Recent attention in pig breeding programs has focused on the improvement of pork quality in response to increasing consumer demands. Compared to the fatty-type Northeastern Indigenous (Chinese) breed of pigs, the lean-type Large White has lower intramuscular fat and inferior eating quality from the perspective of the Chinese consumer. In order to investigate the molecular basis of differences in pork quality in Chinese indigenous and Western breeds, longissimus dorsi samples were collected from three adult Northeastern Indigenous and three adult Large White pigs. The RNAs were extracted and hybridized to the porcine Affymetrix GeneChip. Microarray analysis demonstrated differential expression of 1134 genes of which 401 have a known function. One hundred and thirty-six genes were up-regulated and 998 down-regulated in Northeastern Indigenous breed compared to Large White pigs. We screened 10 genes as candidate genes associated with pork quality. We investigated a single nucleotide polymorphism in the 5' regulatory region of the gene FABP4 in 65 Songliao black swine, using PCR-single-strand conformational polymorphism. We found this polymorphism to be highly significantly associated with marbling and intra-muscular fat content (P ≤ 0.01). Genotype BB had higher marbling than AB and AA, but there was no significant difference between AB and AA. Genotype BB and AB had higher intra-muscular fat content than AA, but there was no significant difference between BB and AB. These results help to elucidate the genetic mechanisms behind differences in pork quality and provide a theoretical basis for selection and genetic improvement of meat quality traits in pigs.     

Proteome analysis of early post-mortem changes in two bovine muscle types: M. longissimus dorsi and M. semitendinosis

To study early post-mortem changes in muscle tissues from bull calves, cytosole proteins from two muscles: M. longissimus dorsi (LD) and M. semitendinosis (ST) at 0 and 24 h after slaughter were analysed by 2-DE. Principal component analysis (PCA) and rotation testing were used to analyse the protein patterns in the two muscles in order to select protein spots that were significantly different at the two time-points. Selected proteins were identified by MALDI-TOF/TOF. Five proteins, namely cofilin, lactoylglutathione lyase, substrate protein of mitochondrial ATP-dependent proteinase SP-22, HSP 27 and HSP20, were changed in both LD and ST muscles during post-mortem storage. Fifteen additional protein changes were observed in either LD or ST muscles, and some of these changes have not previously been observed to change during post-mortem storage of bovine muscles. Further studies will reveal the relevance of these biomarkers for meat quality.     

Functional study of Villin 2 protein expressed in longissimus dorsi muscle of Korean native cattle in different growth stages

The aim of this study was to investigate protein profiles related to the induction of adipogenesis within the bovine longissimus dorsi muscle (BLDM) by proteomic analysis. We analyzed BLDM proteins at different growth stages to clarify the physiological mechanisms of marbled muscle development in 20 head of Korean native cattle (11 month: 10 head, 17 month: 10 head). BLDM proteins were analyzed by two-dimensional electrophoresis and image analysis. Villin 2 was specifically identified by mass spectrometry and a protein search engine. Villin 2 protein expression in BLDM decreased during the fat development stage in test steers. In a Western blot cell culture study of spontaneously immortal bovine muscle fibroblasts, the abundance of Villin 2 was shown to be down-regulated during differentiation into muscle. In 3T3-L1 mouse embryonic fibroblasts, Villin 2 was decreased during differentiation into adipocytes. The results suggest that Villin 2 may be related to the induction of transdifferentiation and adipogenesis in bovine longissimus dorsi muscle.     

Developmental expression changes of the genes involved in IGFI signaling pathway in longissimus dorsi muscle of Tongcheng and Yorkshire pigs during postnatal growth

IGFI signaling pathway is sufficient to regulate myofibre hypertrophy postnatally, which is associated with muscle mass in economically livestock. In the present study, we drafted the developmental expression pattern of eight genes implicated in IGFI system across six stages of postnatal myofibre growth in Yorkshire and Tongcheng pigs. The results indicated that GRB2 may contribute to increased DNA content in postnatal myofibre hypertrophy via GRB2–Ras–Raf–MEK–ERK sub-pathway; INSR, PDK1, IRS1 and eIF4E may contribute to high growth rate via stimulating the rate of protein synthesis and inhibiting the rate of protein degradation. In addition, the results suggested 60 days maybe a very important stage in postnatal myofibre growth. Moreover, higher mRNA level of IRS1 and GLUT4 maybe associated with inferior meat quality in Yorkshire compared to Tongcheng pig. Therefore, IGFI signaling pathway regulates myofibre hypertrophy postnatally via complicated signal effectors, which may have negative impact on meat quality simultaneously.     

Effects of energy or protein restriction followed by realimentation on the composition of gain and meat quality characteristics of Musculus longissimus dorsi in pigs

The effect of realimentation of fattening pigs on gain composition and intramuscular fat content in the longissimus dorsi muscle and their relationship to meat tenderness was investigated in 60 gilts. From day 90 to 168 of age, Group C (control) was fed according to the requirements, while from day 90 to 118 of age, Group R was fed restrictively (protein and energy) and Group P was fed restrictively with protein alone. This period was followed by adequate feeding. Comparative slaughter techniques were used; animals were slaughtered at the beginning of the study (day 90), at the end of the restriction period (day 118), and after two periods of realimentation (day 146 and day 168). Interactions between experimental treatments indicated that muscle protein gain (p < 0.05), muscle fat gain (p < 0.01), and shear force (p < 0.01) differed depending on period of realimentation. Muscle protein deposition differed only during the second period of realimentation, where Group R deposited more protein compared with Groups C and P (6.0, 4.5 and 5.0 g/d, respectively). Muscle fat deposition during the first period of realimentation was lower in Groups R and P than in Group C (0.73, 0.74 and 1.20 g/d, respectively) while in the second period Group R deposited considerably more fat than Groups C and P (2.39, 1.20 and 1.23 g/d, respectively). At the end of the first period, the shear force of Groups R and P was lower when compared with Group C (2.44, 2.79 and 3.22 kg, respectively); however, at final age it did not differ between treatments.     

Promoter variant-dependent mRNA expression of the MEF2A in longissimus dorsi muscle in cattle

The myocyte enhancer factor 2A (MEF2A) gene encodes a member of the myocyte enhancer factor 2 (MEF2) protein family that is involved in vertebrate skeletal, cardiac, and smooth muscle development and differentiation during myogenesis. According to recent studies, MEF2 genes might be major regulators of postnatal skeletal muscle growth; thus, they are considered to be important, novel candidates for muscle development and body growth in farm animals. The aim of the present study was to search for polymorphisms in the bovine MEF2A gene and analyze their effect on the MEF2A mRNA expression level in the longissimus dorsi muscle of Polish Holstein-Fresian cattle. In total, 4094?bp of the whole coding sequence and the promoter region of MEF2A were re-sequenced in 30 animals, resulting in the detection of 6 novel variants as well as one previously reported SNP. Three linked mutations in the promoter region (-780T/G, g.-768T/G, and g.-222A/G) and only two genotypes were identified in two Polish breeds (TTA/TTA and TTA/GGG). Three SNPs in the coding region [g.1599G/A (421aa), g.1626G/A (429aa), and g.1641G/A (434aa)] appeared to be silent substitutions and segregated as two intragene haplotypes: GGG and AAA. Expression analysis showed that the mutations in the promoter region are highly associated with the MEF2A mRNA level in the longissimus dorsi muscle of bulls carrying two different genotypes. The higher MEF2A mRNA level was estimated in the muscle of bulls carrying the TTA/TTA (p<0.01) genotype as compared with those with TTA/GGG. The results obtained suggest that the nucleotide sequence mutation in MEF2A might be useful marker for body growth traits in cattle.     

Prediction of meat quality from biopsy in live pigs with different RYR1 genotypes

To predict meat quality after slaughter, biopsy samples were taken from musculus longissimus lumborum et thoracis of live pigs at approximately 40 kg and 80 kg of weight. The obtained values from biopsies for pH1 and EC50 (electric conductivity) were compared with measurements after slaughter at a weight of approximately 110 kg. RYR1 genotypes were determined from blood samples using PCR-RFLP. Mating of Nn sows with two nn boars resulted in 72 Nn and 40 nn offspring. Significant differences between the two genotypes were found for pH1 and EC50 values for the three weights. The coefficients of correlation for the Nn genotype of the RYR1 gene between the values after slaughter and both the first and the second biopsy for pH1 and EC50 were very low (r = 0.06, r = 0.14; and r = 0.26, r = 0.26; P ? 0.05). For the nn genotype were r = -0.23, r = -0.15; and r = -0.25, r = -0.11 respectively. The values of pH1 and EC50 were highly correlated (r = -0.52 to -0.84; P < or = 0.001) both within biopsies and after slaughter.     

Genetic heterogeneity and selection signature at the KIT gene in pigs showing different coat colours and patterns

Mutations in the porcine KIT gene (Dominant white locus) have been shown to affect coat colours and colour distribution in pigs. We analysed this gene in several pig breeds and populations (Sicilian black, completely black or with white patches; Cinta Senese; grey local population; Large White; Duroc; Hampshire; Pietrain; wild boar; Meishan) with different coat colours and patterns, genotyping a few polymorphisms. The 21 exons and parts of the intronic regions were sequenced in these pigs and 69 polymorphisms were identified. The grey-roan coat colour observed in a local grey population was completely associated with a 4-bp deletion of intron 18 in a single copy KIT gene, providing evidence that this mutation characterizes the I^d allele described in the early genetic literature. The white patches observed in black Sicilian pigs were not completely associated with the presence of a duplicated KIT allele (I^p), suggesting that genetic heterogeneity is a possible cause of different coat colours in this breed. Selection signature was evident at the KIT gene in two different belted pig breeds, Hampshire and Cinta Senese. The same mutation(s) may cause the belted phenotype in these breeds that originated in the 18th–19th centuries from English pigs (Hampshire) and in Tuscany (Italy) in the 14th century (Cinta Senese). Phylogenetic relationships of 28 inferred KIT haplotypes indicated two clades: one of Asian origin that included Meishan and a few Sicilian black haplotypes and another of European origin.     

Fatigue response of rat medial longissimus muscles induced with electrical stimulation at various work/rest ratios

Muscle fatigue may be a precursor to workplace musculoskeletal disorders, with the low back resulting in the most frequently injured body part. Work/rest ratios have an effect on fatigue due to the amount of rest allowance provided following muscle contraction. This study explored various work/rest ratios by electrically stimulating rat medial longissimus muscles. A 3 V stimulus with 0.2 ms pulse duration was applied at a frequency of 30 Hz. There were four stimulation groups consisting of the following duty cycles (DC) and cycle times (CT): DC25%:CT20s, DC25%:CT280s, DC75%:CT20s, and DC75%:CT180s. Muscle fatigue was measured as a decrease in M-wave amplitude and area, and an increase in M-wave duration. The results indicated that fatigue occurred immediately in each of the groups. The higher duty cycle and shorter cycle time group resulted in significantly greater fatigue than the lower duty cycle and longer cycle time group, as measured by increased M-wave amplitude and area. A longer M-wave duration was observed in the high duty cycle long cycle time group. This suggests that the combination of low duty cycle and long cycle times leads to less fatigue. In high duty cycle scenarios, short cycle times result in less fatigue.     

Estimates of genetic parameters for fatty acid compositions in the longissimus dorsi muscle of Hanwoo cattle

We estimated the heritabilities (h²) and genetic and phenotypic correlations among individual and groups of fatty acids, as well as their correlations with six important carcass and meat-quality traits in Korean Hanwoo cattle. Meat samples were collected from the longissimus dorsi muscles of 1000 Hanwoo steers that were 30-month-old (progeny of 85 proven Hanwoo bulls) to determine intramuscular fatty acid profiles. Phenotypic data on carcass weight (CWT), eye muscle area (EMA), back fat thickness (BFT), marbling score (MS), Warner–Bratzler shear force (WBSF) and intramuscular fat content (IMF) were also investigated using this half-sib population. Variance and covari.ance components were estimated using restricted maximum likelihood procedures under univariate and pairwise bivariate animal models. Oleic acid (C18:1n-9) was the most abundant fatty acid, accounting for 50.69% of all investigated fatty acids, followed by palmitic (C16:0; 27.33%) and stearic acid (C18:0; 10.96%). The contents of saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) were 41.64%, 56.24% and 2.10%, respectively, and the MUFA/SFA ratio, PUFA/SFA ratio, desaturation index (DI) and elongation index (EI) were 1.36, 0.05, 0.59 and 0.66, respectively. The h² estimates for individual fatty acids ranged from very low to high (0.03±0.14 to 0.63±0.14). The h² estimates for SFAs, MUFAs, PUFAs, DI and EI were 0.53±0.14, 0.49±0.14, 0.23±0.10, 0.51±0.13 and 0.53±0.13, respectively. The genetic and phenotypic correlations among individual fatty acids and fatty acid classes varied widely (−0.99 to 0.99). Notably, C18:1n-9 had favourable (negative) genetic correlations with two detrimental fatty acids, C14:0 (−0.76) and C16:0 (−0.92). Genetic correlations of individual and group fatty acids with CWT, EMA, BFT, MS, WBSF and IMF ranged from low to moderate (both positive and negative) with the exception of low-concentration PUFAs. Low or near-zero phenotypic correlations reflected potential non-genetic contributions. This study provides insights on genetic variability and correlations among intramuscular fatty acids as well as correlations between fatty acids and carcass and meat-quality traits, which could be used in Hanwoo breeding programmes to improve fatty acid compositions in meat.