Meat Science and Muscle Biology Symposium: in utero nutrition related to fetal development, postnatal performance, and meat quality of pork.

Intrauterine growth restriction (IUGR) occurs naturally in pigs and leads to low birth weight of piglets due to undernutrition caused by placental insufficiency. For 2 main reasons, low birth weight causes economic loss. First, low birth weight pigs have a greater mortality and increasing the litter size causes more low birth weight piglets within litters. Second, surviving low birth weight piglets have reduced performance (i.e., ADG, feed conversion rate, and percentage meat). To develop dietary strategies for preventing IUGR, knowledge of the biological basis of IUGR is required. Muscle fiber number, formed during myogenesis, is correlated positively with performance traits and has been shown in several studies to be reduced in low birth weight pigs. Postnatal muscle hypertrophy is due to satellite cell number per fiber at birth and their rate of proliferation as well as protein deposition (i.e., protein synthesis and degradation). Previous studies and some recent ones indicate that low birth weight littermates in mice are born with fewer satellite cells and studies on pigs show that the rate of satellite cell proliferation may vary within litters. Proteomics studies show that protein synthesis and degradation is downregulated in IUGR pigs and low birth weight pigs also produce meat with less tenderness. Alternative maternal feeding strategies to prevent IUGR have been examined. Increasing maternal global nutrition had no beneficial effect on performance and muscle growth traits in several studies. Feeding excess maternal dietary protein also did not influence muscle growth traits whereas moderately decreased maternal dietary protein may decrease muscle fiber number and performance. On the other hand, addition of L-carnitine to the maternal gestation or lactation diet may increase birth and weaning weights or the muscle fiber number, respectively, in low birth weight pig offspring. Finally, promising data have been obtained on reproductive traits in pigs after addition of functional AA, such as arginine and glutamine, to the gestational diet. Although much is known about the biological basis of IUGR, we still need to learn much more about the mode of action before maternal dietary strategies can be developed to prevent IUGR.

Advances in research on the prenatal development of skeletal muscle in animals in relation to the quality of muscle-based food. I. Regulation of myogenesis and environmental impact.

Skeletal muscle development in vertebrates - also termed myogenesis - is a highly integrated process. Evidence to date indicates that the processes are very similar across mammals, poultry and fish, although the timings of the various steps differ considerably. Myogenesis is regulated by the myogenic regulatory factors and consists of two to three distinct phases when different fibre populations appear. The critical times when myogenesis is prone to hormonal or environmental influences depend largely on the developmental stage. One of the main mechanisms for both genetic and environmental effects on muscle fibre development is via the direct action of the growth hormone-insulin-like growth factor (GH-IGF) axis. In mammals and poultry, postnatal growth and function of muscles relate mainly to the hypertrophy of the fibres formed during myogenesis and to their fibre-type composition in terms of metabolic and contractile properties, whereas in fish hyperplasia still plays a major role. Candidate genes that are important in skeletal muscle development, for instance, encode for IGFs and IGF-binding proteins, myosin heavy chain isoforms, troponin T, myosin light chain and others have been identified. In mammals, nutritional supply in utero affects myogenesis and the GH-IGF axis may have an indirect action through the partitioning of nutrients towards the gravid uterus. Impaired myogenesis resulting in low skeletal myofibre numbers is considered one of the main reasons for negative long-term consequences of intrauterine growth retardation. Severe undernutrition in utero due to natural variation in litter or twin-bearing species or insufficient maternal nutrient supply may impair myogenesis and adversely affect carcass quality later in terms of reduced lean and increased fat deposition in the progeny. On the other hand, increases in maternal feed intake above standard requirement seem to have no beneficial effects on the growth of the progeny with myogenesis not or only slightly affected. Initial studies on low and high maternal protein feeding are published. Although there are only a few studies, first results also reveal an influence of nutrition on skeletal muscle development in fish and poultry. Finally, environmental temperature has been identified as a critical factor for growth and development of skeletal muscle in both fish and poultry.

Advances in research on the prenatal development of skeletal muscle in animals in relation to the quality of muscle-based food. II--Genetic factors related to animal performance and advances in methodology.

Selective breeding is an effective tool to improve livestock. Several selection experiments have been conducted to study direct selection responses as well as correlated responses in traits of skeletal muscle growth and function. Moreover, comparisons of domestic with wild-type species and of extreme breeds provide information on the genetic background of the skeletal muscle phenotype. Structural muscular components that differed with increasing distance in lean growth or meat quality in mammals were found to be myofibre number, myofibre size, proportions of fibre types as well as the numbers and proportions of secondary and primary fibres. Furthermore, markers of satellite cell proliferation, metabolic enzyme activities, glycogen and fat contents, the expression of myosin heavy chain isoforms, of activated AMPKα and other proteins in skeletal muscle tissue and circulating IGF1 and IGF-binding proteins have been identified to be involved in selection responses observed in pigs, cattle and/or chicken. The use of molecular methods for selective breeding of fish has only recently been adopted in aquaculture and studies of the genetic basis of growth and flesh quality traits are scarce. Some of the molecular markers of muscle structure/metabolism in livestock have also been identified in fish, but so far no studies have linked them with selection response. Genome scans have been applied to identify genomic regions exhibiting quantitative trait loci that control traits of interest, for example, muscle structure and meat quality in pigs and growth rate in chicken. As another approach, polymorphisms in candidate genes reveal the relationship between genetic variation and target traits. Thus, in large-scale studies with pigs' associations of polymorphisms in the HMGA2, CA3, EPOR, NME1 and TTN genes with traits of carcass and meat quality were detected. Other studies revealed the significance of mutations in the IGF2 and RYR1 genes for carcass lean and muscle fibre traits in pigs. Mutations in the myostatin (MSTN) gene in fish were also examined. Advances in research of the genetic and environmental control of traits related to meat quality and growth have been made by the application of holistic 'omics' techniques that studied the whole muscle-specific genome, transcriptome and proteome in relation to muscle and meat traits, the development of new methods for muscle fibre typing and the adaptation of biophysical measures to develop parameters of muscle fibre traits as well as the application of in vitro studies. Finally, future research priorities in the field are defined.

Birth weight and postnatal dietary protein level affect performance, muscle metabolism and meat quality in pigs.

Intrauterine growth restriction (IUGR), resulting in low birth body weight (LBW) occurs naturally in pigs. However, IUGR may also cause persistent changes in physiology and metabolism resulting in poorer performance, organogenesis and meat quality. As IUGR pigs have a lower daily gain from birth to slaughter they may differ in utilization of nutrients and requirements for dietary protein compared with their larger littermates. Thus, the objective in this study was to examine the interaction between birth body weight (BW) and the postnatal dietary protein level, in relation to postnatal performance, organogenesis, muscle metabolism and meat quality. The experiment was carried out with offspring from 16 purebred Danish Landrace gilts mated to Danish Landrace boars. The female and entire male pigs with LBW that survived at weaning were compared with the female and male pigs with the highest/high birth body weight (HBW) within each litter. The offspring were reared individually from weaning and were fed ad libitum a diet containing either a normal level of protein (NP) for optimal growth or an isocaloric diet containing a 30% lower protein content (LP) from 3 weeks to 150 days of age. At slaughter, we found no interactions between birth weight group and dietary protein level for any of the measured traits. The relative crown-rump length (cm/kg) at birth indicates that LBW pigs were thinner than HBW pigs. Daily gain and feed intake were reduced by 14% and 10%, respectively, while the kg feed/kg gain was slightly increased by 3% in LBW pigs compared with HBW pigs. The LP diet reduced daily gain by 27% due to reduced feed intake and increased kg feed/kg gain by 12% and 21%, respectively compared with the NP diet. LBW male pigs produced meat with a higher shear force than male HBW pigs and also LP pigs produced meat with higher shear force than NP pigs. The activity of lactate dehydrogenase in the Longissimus dorsi muscle (LD) was reduced in pigs fed the LP diet. Calpastatin was increased in LD of LBW pigs and decreased in pigs fed the NP diet. In conclusion, these results suggest a rejection of our hypothesis that low birth weight littermates have a lower requirement for dietary protein compared with heavy weight littermates. Furthermore, LBW male pigs and LP fed pigs of both genders produced less tender meat than HBW pigs or NP fed pigs, respectively.

In vitro primary satellite cell growth and differentiation within litters of pigs.

Postnatal muscle growth is dependent on satellite cell (SC) proliferation, differentiation and fusion to increase the DNA content of existing muscle fibres and thereby the capacity to synthesize protein. The purpose of the present study was to examine the ability of isolated SCs from low, medium and high weaning weight litter mates of pigs to proliferate and differentiate, and to affect protein synthesis and degradation after fusion into myotubes. At 6 weeks of age, SCs from the lowest weight (LW), medium weight (MW) and highest weight (HW) female pigs within eight litters were isolated. Thereby, eight cultures of SCs were established for each of the three weight groups within litter, representing three groups of SCs from pigs exhibiting differences in postnatal muscle growth performance. Proliferation was estimated as the number of viable cells at different time points after seeding. SC differentiation was evaluated by measuring the activity of the muscle-specific enzyme, creatine phosphokinase, and protein synthesis and degradation were measured by incorporation and release of 3H-tyrosine, respectively. A tendency towards a difference in proliferation between SC cultures was found (P = 0.09). This was evident as the number of viable cells at day 3 was lower in cultures from LW pigs than from HW (P < 0.05) and MW (P < 0.01) pigs. Differentiation was significantly different between cultures (P < 0.05). There was a significant difference between LW and MW cultures at 72 h (P < 0.05), and a tendency towards a difference between LW and HW cultures at 45 h (P = 0.07). Protein synthesis per µg protein or per µg DNA did not differ among SC cultures from LW, MW and HW pigs. Neither did protein degradation rate differ significantly among SC cultures from LW, MW and HW pigs. Overall, the results show that SCs from LW pigs seem to proliferate and differentiate at a slower rate than SCs from MW and HW pigs. The results found in this study show no difference in the ability of SCs to affect protein synthesis or degradation between SCs from litter mates exhibiting different growth rates in vivo.

Creatine monohydrate and glucose supplementation to slow- and fast-growing chickens changes the postmortem pH in pectoralis major.

The energy status of the chicken at slaughter has a large impact on the development of pH postmortem and thus on color and water-holding capacity (WHC). Supplementation of creatine monohydrate and glucose (CMH+GLU) may increase the creatine content in the muscles before slaughter, thereby delaying the formation of lactic acid and postponing the pH decline. The objective of this study was to examine the impact of supplementing CMH+GLU in the drinking water within the last 48 h before slaughter on the pH decline, meat color, and WHC in the pectoralis major from 2 strains of Ross chickens. Forty Ross 308 (fast-growing) female chickens and 40 Ross 1972 (slow-growing) female chickens had free access to drinking water either supplemented with CMH (15 g/ L) and glucose (50 g/L) within the last 48 h before slaughter or without supplementation. All chickens were slaughtered at 42 or 43 d of age irrespective of weight. Temperature and pH were measured at 1 and 30 min and at 1, 3, 8, and 24 h postmortem. Also, WHC measured as drip loss and color were determined postmortem. The CMH+GLU supplementation decreased pH (P < 0.05) at all time points between 1 min and 8 h postmortem in both strains, whereas at 24 h postmortem only pH in Ross 308 chickens were decreased significantly upon supplementation. Lightness was significantly increased in the meat from Ross 308 but not from Ross 1972 chickens upon supplementation. This interaction was significant (P < 0.05). The redness of the meat was decreased upon supplementation (P < 0.05), although only significantly in Ross 1972. The pH was lower for Ross 1972 chickens at the early time points (P < 0.01) and also a higher drip loss (P < 0.05), lightness (P < 0.01), and redness (P < 0.001) were observed. Thus, there seems to be no beneficial effect of CMH+GLU supplementation on chicken meat quality on the basis of results from this experiment.

Serum from heifer calves treated with bovine growth hormone affects the rate of proliferation of C2C12 myogenic cells dependent on the plane of nutrition: the role of insulin-like growth factor-I and IGF-binding proteins-2 and -3.

The present in vitro experiments were carried out in order to study whether variations in the bovine growth hormone (bGH)/insulin-like growth factor (IGF)-I axis induced by plane of nutrition and bGH treatment of heifer calves caused variations in serum-induced proliferation of C2C12 myoblasts. Serum was obtained from two groups each of six heifer calves (195 +/- 8 kg) before (d -1) and after treatment with 15 mg/day of bGH for 6 days (d 6) fed either a low (GHL) or a high plane (GHH) of nutrition. Preceding the experiment all 12 heifer calves were fed at the low plane of nutrition. At d 6, serum concentrations of insulin and IGF-I were increased while that of IGF-binding proteins (IGFBP)-2 was decreased in GHH, but unchanged in GHL calves. Serum-induced proliferation of C2C12 myoblasts, was elevated at d 6 by GHH treatment. Especially human IGFBP-3 but also bovine IGFBP-2 added to cell cultures inhibited the rate of proliferation of C2C12 myoblasts stimulated by human IGF-I. The present results showed that GH treatment causes changes in the GH/IGF axis, which leads to changes in serum-induced growth of C2C12 muscle cells dependent on the plane of nutrition that mimic in vivo effects of GH treatment, which indicate an endocrine contribution of the IGF system. However, drawbacks of this suggestion are discussed.

The estimated accuracy of the EU reference dissection method for pig carcass classification.

This experiment was designed to describe the accuracy of the EU-reference dissection method, and to describe the types of factors influencing the accuracy and assess their size. The experiment was conducted in four different European countries at two abattoirs within each country. A total of 128 carcasses was selected according to carcass weight, fat class and sex, and 8 butchers from different countries dissected the carcasses. Due to the experimental design of the experiment a variation in pig type was found between countries. The accuracy was expressed by the repeatability and reproducibility standard deviation, which were found to be 0.87 and 1.10, respectively, and by the reliability, found to be 0.87. This indicates a high accuracy, although a significant effect was found on the estimation of lean meat percentage (LMP) of butcher, and also that jointing of the carcass was of overall importance to the accuracy of the EU-reference dissection method.

Within-litter variation in muscle fiber characteristics, pig performance, and meat quality traits.

The objective of this study was to examine the intralitter variation in postnatal growth performance, meat quality, and muscle fiber characteristics when littermates were categorized by carcass weight. Thirty-nine litters were weaned at 4 wk of age and had free access to feed from 2 wk of age until slaughter. They were slaughtered by litter at an average BW of 104 +/- 14 kg, and six pigs per litter were selected for analysis: the heaviest- (HW), middle- (MW), and lightest-weight (LW) pig of each sex. Categorizing littermates in LW, MW, and HW pigs at the same age reflected the differences in postnatal growth rate within a litter; thus ADG, muscle mass, and muscle deposition rate differed across pig weight groups (P < 0.001). Also, the total DNA content was different among pig weight groups (P < 0.001) and reflected differences in muscle growth rate. The difference in muscle growth rate between LW and MW pigs could be explained by a larger (P < 0.05) mean fiber area (MFA) in MW pigs, whereas the number of muscle fibers was similar. Growth rate differences between MW and HW pigs could in part be explained by a higher number (P < 0.01) of equal-sized muscle fibers in HW pigs. The difference in MFA was due to a higher estimated DNA and RNA content per muscle fiber in MW and HW compared with LW pigs (P < 0.05). Pigment content was higher in MW and HW compared with LW pigs (P < 0.01), but no other measured meat quality traits were significantly different across pig weight groups. These results indicate that both the number and the growth rate of muscle fibers contribute to intralitter variation in postnatal growth performance.

Increased maternal nutrition of sows has no beneficial effects on muscle fiber number or postnatal growth and has no impact on the meat quality of the offspring.

The objective of this study was to examine how increased feed intake of the sow during early to mid-gestation affects sow performance and the muscle fiber number, performance, and technological meat quality of the offspring. Thirty-nine pregnant sows (Landrace x Large White sows mated to Landrace or Large White boars) in their fourth parity were assigned to one of three treatments: 1) the sows were either fed restrictively (control = 15 MJ of NE/d from d 1 to 90, then 24 MJ of NE/d from d 91 to 112, and again 15 MJ of NE/d from d 113 to 115 of gestation); 2) fed ad libitum from d 25 to 50 (A25-50); or 3) ad libitum from d 25 to 70 (A25-70) and as control in the remaining periods. The offspring were weaned at 4 wk of age and had free access to feed from 2 wk of age until slaughter. They were slaughtered litterwise at an average body weight of 104 +/- 14 kg. Estimates for total, primary (P-), and secondary (S-) muscle fiber number; muscle fiber area; and DNA and RNA content were analyzed in semitendinosus muscle (ST) samples from the heaviest, middle, and lightest weight (LW) pigs of each sex within litter selected at slaughter. Technological meat quality traits (pH at 24 h postmortem, drip loss, Minolta color, and pigment) were analyzed in longissimus dorsi muscle. Fiber number, fiber area, and concentrations and content of DNA and RNA of the offspring were not significantly affected by increased maternal nutrition. The ST muscle weight was lower in offspring from A25-50 than control sows (P = 0.019). Average daily gain, carcass weight, and the muscle deposition rate also were numerically lower for A25-50 than control and A25-70 pigs. An interaction between treatment and pig weight was found for muscle deposition rate (P = 0.006), in that LW pigs from treatment A25-50 had a lower deposition rate than LW pigs from control. We found no effect of treatment on the meat quality traits in the offspring. Also, barrows had a higher (P < 0.05) number of P-fibers, higher daily gain, and carcass weight than female pigs. No differences were found on any meat quality traits between sexes. Thus, ad libitum feeding of pregnant sows from d 25 to 50 or d 25 to 70 of gestation did not have any beneficial effect on muscle fiber number and area in the offspring. It seems that maternal ad libitum feeding from d 25 to 50 in gestation had a negative effect on postnatal muscle growth, with especially the LW pigs being affected.