Intermittent bolus feeding does not enhance protein synthesis, myonuclear accretion, or lean growth more than continuous feeding in a premature piglet model

Optimizing enteral nutrition for premature infants may help mitigate extrauterine growth restriction and adverse chronic health outcomes. Previously, we showed in neonatal pigs born at term that lean growth is enhanced by intermittent bolus compared to continuous feeding. The objective was to determine if prematurity impacts how body composition, muscle protein synthesis, and myonuclear accretion respond to feeding modality. Following preterm delivery, pigs were fed equivalent amounts of formula delivered either as intermittent boluses (INT; n = 30) or continuously (CONT; n = 14) for 21 days. Body composition was measured by DXA and muscle growth was assessed by morphometry, myonuclear accretion, and satellite cell abundance. Tissue anabolic signaling and fractional protein synthesis rates were determined in INT pigs in postabsorptive (INT-PA) and postprandial (INT-PP) states and in CONT pigs. Body weight gain and composition did not differ between INT and CONT pigs. Longissimus dorsi (LD) protein synthesis was 34% greater in INT-PP than INT-PA pigs (P < 0.05) but was not different between INT-PP and CONT pigs. Phosphorylation of 4EBP1 and S6K1 and eIF4E·eIF4G abundance in LD paralleled changes in LD protein synthesis. Satellite cell abundance, myonuclear accretion, and fiber cross-sectional area in LD did not differ between groups. These results suggest that, unlike pigs born at term, intermittent bolus feeding does not enhance lean growth more than continuous feeding in pigs born preterm. Premature birth attenuates the capacity of skeletal muscle to respond to cyclical surges in insulin and amino acids with intermittent feeding in early postnatal life.

143 Nutritional advances in fetal and neonatal development: amino acid supplementation

Nutrient requirements of grazing ruminants change throughout the production year. Also, pasture quality and quantity change throughout the year and are influenced by environment. Therefore, there commonly are times of the year where nutrient requirements are not met from the forage alone. We have long known the importance of nutrition during key development including the conception, fetal, neonatal, and weaning period. Altering nutrition during these periods can have lasting effects on health and productivity. Much of the research in ruminants examining nutritional effects on development has focused on large changes in diet, such as energy or protein restriction, rather than specific nutrients such as amino acids. Amino acids play an important role to meet requirements for lean growth but also have other physiological functions regulating metabolism and health that have important effects on development. Amino acid deficiencies during the fetal and neonatal period(s) can result in small-for-gestational age births or stunting, respectively. Diets that are deficient in protein or amino acids are also often deficient in dietary energy and other nutrients. Supplying specific amino acids (and other nutrients/energy) for absorption, signaling, and use by the animal could be an approach to help overcome negative developmental outcomes. Specific amino acids are thought to function by regulating protein synthesis, immune function, 1-C metabolism, intestinal development and health, blood flow, cellular proliferation, and behavior, to name a few. Some of the amino acids with potential for influencing developmental outcomes include leucine, arginine, glutamine, lysine, methionine, threonine, and tryptophan. Because of the varied functional roles of specific amino acids, there is great potential to develop targeted supplementation programs using amino acids to improve developmental outcomes.

41 Characterizing the amount and variability of intramuscular fat deposition throughout the loin using barrows and gilts from two genotypes

Pork quality is evaluated by commercial processors along the ventral surface of whole boneless loins. However, in retail settings boneless chops are evaluated individually for quality. The objective was to determine the variability in marbling attributable to chop location, sex, and sire line. The population of pigs evaluated were sired by commercial duroc boars selected for meat quality (MQ; n=96) or lean growth (LG; n=96) and equally split between sexes. Upon slaughter and fabrication, bone-in chops were removed from four locations along the loin (A=6th rib, B=10th rib, C=last rib, D=4th lumbar). Pairs of chops from each location were used for visual color, visual marbling, and subjective firmness, Warner-Bratzler shear force, and determination of moisture and extractable lipid (IMF). Variability across location, sex, and sire line was measured using a Levene’s test and the VARCOMP procedure of SAS was used to evaluate the proportion of variability each contributed to total variance. Barrows produced chops with greater IMF (3.64%) and variability (P&lt; 0.001) than gilts (3.20%; P&lt; 0.001). Chops from MQ pigs exhibited greater IMF (4.02%) and variability (P&lt; 0.001) than LG (2.82%; P&lt; 0.001). Chops from location A (3.80%) and D (3.77%) had greater IMF than B (3.34%; P&lt; 0.001) with all three locations greater than C (2.77%; P&lt; 0.001). Variance (s2) also differed (A=1.44% B=1.59% C=1.05% D=2.18%; P&lt; 0.05) across chop locations. Of the variability in IMF, 33.0% was attributed to sire line, 10.16% to chop location and 4.01% to sex, with 52.83% unaccounted for. Location A chops were the most tender (2.57kg; P&lt; 0.001) and C chops the least tender (2.93kg; P&lt; 0.01). No differences in variability (s2; P=0.40) of tenderness were observed across chop location (A=0.31kg B=0.24kg C=0.24kg D=0.23kg). In conclusion, chop location, sex, and sire line all contribute to the amount and variability of pork loin marbling. Supported by National Pork Board Grant #18-118.

Characterizing the amount and variability of intramuscular fat deposition throughout pork loins using barrows and gilts from two sire lines

The objective was to determine the amount and variability of intramuscular fat (IMF) in a pork loin attributable to anatomical chop location, sex, and sire line. Pigs were sired by commercially available terminal Duroc boars selected for meat quality (MQ; n = 96) or lean growth (LG; n = 96) and equally split between barrows and gilts. After slaughter and fabrication, bone-in chops were removed from four locations of each left-side loin (A = 6th rib, B = 10th rib, C = last rib, and D = 4th lumbar vertebrae). An adjacent pair of chops from each location was collected and evaluated for visual color and marbling, subjective firmness, moisture and extractable lipid (IMF) (anterior chop), and Warner–Bratzler shear force (posterior chop). Data were analyzed using the MIXED procedure of SAS as a split-plot design. Homogeneity of variances was tested on raw data using Levene’s test of the GLM procedure and found to be heterogeneous. Thus, a two-variance model was fit using the REPEATED statement of the MIXED procedure, grouped by pig. The mivque(0) option of the VARCOMP procedure was used to calculate the proportion of variability that each factor contributed to the total variance. Barrows (3.64%) produced chops with greater (P &lt; 0.01) IMF content than gilts (3.20%), and barrows (2.14) had greater (P &lt; 0.01) IMF variability than gilts (1.23). Chops from MQ pigs (4.02%) exhibited greater (P &lt; 0.01) IMF content than LG (2.82%), and MQ (1.76) had greater IMF variability (P &lt; 0.01) than LG pigs (0.97). Chops from locations A (3.80%) and D (3.77%) had greater IMF than B (3.34%; P &lt; 0.01), and A, B, and D had greater IMF than C (2.77%; P &lt; 0.01). Variances of IMF also differed (A = 1.44, B = 1.59, C = 1.05, and D = 2.18; P = 0.01) across chop locations. Of the variability in IMF, 33.0% was attributed to sire line, 10.16% to chop location, and 4.01% to sex, with 52.83% not accounted for by these three factors. Location A chops were the most (P &lt; 0.01) tender (2.57 kg) and C chops the least (P &lt; 0.01) tender (2.93 kg), while B and D chops were intermediate and not different from each other. No differences in variability (P = 0.40) of tenderness were observed among chop locations (A = 0.31, kg B = 0.24 kg, C = 0.24 kg, and D = 0.23 kg). These results demonstrated that variability in tenderness values did not reflect the variability of IMF. In conclusion, chop location, sex, and sire line all contribute to the amount and variability of pork loin marbling.

Continuous Feeding Does Not Blunt Satellite Cell Abundance, Myonuclear Accretion, or Lean Growth in a Neonatal Piglet Model of Prematurity

Objectives Lean growth in preterm neonates is poor and may increase the lifelong risk for adverse health outcomes. Strategies are needed to promote skeletal muscle growth in the postnatal period. Skeletal muscle growth, which accounts for the largest fraction of lean mass accretion, requires the coordinated activation of protein synthesis, satellite cell (SC; muscle stem cell) proliferation, differentiation, and fusion into muscle fibers. The objective of this study was to determine the effect of feeding modality on SC abundance, myonuclear accretion, and lean growth in preterm neonatal pigs. Methods Pigs delivered 10 d preterm by C-section (952 ± 205 g) were fitted with an umbilical artery catheter (later replaced with jugular vein catheter) and an orogastric tube for parenteral and enteral nutrition, respectively. Pigs were assigned to continuous (CONT; n = 14; 7.5 mL/[kg·h]) or intermittent bolus (INT; n = 30; 30 mL/kg every 4 h) feeding for 21 d (210 kcal/kg and 16 g protein/kg per d); pigs were advanced from parenteral to enteral feeding over 6 d. Bromodeoxyuridine (BrdU; 25 mg·kg−1) was administered to pigs every 12 h from days 19 to 20, inclusive, to label proliferating cells. Body composition was measured by DXA on day 21. Satellite cell (Pax7+) abundance, myonuclear accretion, and muscle fiber cross-sectional area (CSA) were quantified in the longissimus dorsi muscle by immunofluorescence. Results Sublaminal Pax7+ SC abundance was similar between CONT and INT groups (60.9 vs. 58.3 ± 6.0 per 1000 fibers). The proportion of sublaminal relative to total Pax7+ SCs was similar between CONT and INT groups (78.8 vs. 78.8 ± 2.2%). The abundance of BrdU + myonuclei, an index of myonuclear accretion, did not differ between CONT and INT groups (26.7 vs. 26.7 ± 3.4 per 1000 fibers). Total myonuclei did not differ between CONT and INT groups (420 vs. 403 ± 16 per 1000 fibers). Muscle fiber CSA did not differ between CONT and INT groups (210 vs. 237 ± 15 µm2). Lean and fat masses were similar between groups. Conclusions Unlike term pigs, CONT feeding does not blunt lean growth in preterm pigs compared to INT feeding. The absence of increased lean growth with INT feeding is consistent with the similar SC abundance, myonuclear accretion, and muscle fiber CSA between feeding modalities. Funding Sources Research was supported by NIH and USDA.

Influence of dietary protein content on the chemico-physical profile of dry-cured hams produced by pigs of two breeds

The use of low-protein (LP) feeds is a good strategy to reduce the environmental release of N compounds, but their influence on the quality of the products must be considered. This study explored the influence of LP diet and two pig breeds (BR) with different lean growth ability on the quality traits of dry-cured hams. We analysed 40 left dry-cured hams from pigs of two BR [Duroc-Danbred crosses (Danbred) and Duroc × Large White crosses (Anas)] fed either conventional (147 to 132 g/kg, crude protein) or LP diet. The LP had a crude protein content reduced by 20% with respect to the conventional. The differences in ham quality resulting from protein reduction were small, with a decrease of the protein and an increase of the lipid content of the ham slice in Anas, but not in Danbred (BR × Diet interaction; P = 0.043). Therefore, the use of LP would be feasible and sustainable, without detrimental effects on products. It was found the pig genotypes with different potentials for lean growth may affect the initial ham weight, fat cover and seasoning losses of hams, but they appear to affect little other chemical, physical and textural quality traits of the dry-cured hams.

356 Meal feeding compared with continuous feeding enhances insulin and amino acid signaling to translation initiation in skeletal muscle of pigs

Objectives: Meal feeding enhances skeletal muscle protein synthesis and lean growth more than continuous feeding in piglets. This enhanced muscle protein synthesis with meal feeding is associated with increased activation of mTORC1-dependent translation initiation. The mechanism underlying this response is unknown. We aimed to identify insulin and amino acid signaling components involved in the enhanced lean growth that results from meal feeding vs. continuous feeding in term-born pigs. Methods: Newborn piglets were fed for 21 d an equal amount of sow milk replacer (12.8 g protein and 155 kcal/(kg BW.d)) by gastrostomy tube either as intermittent bolus meals every 4 h (MEAL) or by continuous infusion (CON). After 21 d, gastrocnemius muscle was collected from CON, and before (MEAL-0) or 60 min after a meal (MEAL-60). Components of the insulin and amino acid signaling pathways up- and downstream of mTORC1 that regulate protein translation were measured. Results: Phosphorylation of AKT and TCS2 was greater in MEAL-60 than in MEAL-0 and CON (P &lt; 0.05). The association of Sestrin2 with GATOR2 was similar in CON and MEAL-0 but was lower in MEAL-60 (P &lt; 0.05). The abundances of RagA-mTOR, RagC-mTOR, and Rheb-mTOR, but not CASTOR1-GATOR2, complexes were higher in MEAL-60 than in CON and MEAL-0 (P &lt; 0.05). The phosphorylation of S6K1 and 4EBP1 was higher in MEAL-60 than CON and MEAL-0 (P &lt; 0.05). The abundances of Sestrin2, GATOR2, CASTOR1, RagA, RagC, and Rheb and the phosphorylation of eIF2alpha, eEF2, ERK1/2 and AMPK were unaffected by treatments. Conclusions: Our results demonstrate that the enhanced rate of skeletal muscle protein synthesis and lean growth with meal feeding compared with continuous feeding are due to the enhanced activation of both insulin and amino acid signaling pathways that result in the greater stimulation of translation initiation. Support: NIH HD085573, USDA CRIS 6250-51000-055, NIH HD072891, USDA NIFA 2013-67015-20438.

PSIII-7 Production trait differences between show pig and commerical genetic lines

The objective was to compare growth and body composition of pigs sired by boars from the show pig industry to those sired by modern commercial genetic lines. Data included one group of 174 male pigs from 16 sires and 50 dams. Pigs were sired by Large White (LW) show pig sires (SHOW) or one of two maternal LW commercial genetic lines (W or P). Dams of all pigs evaluated were from line P. Pigs were weaned at 27 d of age and placed into a mechanically ventilated nursery (0.23 m2 per pig). After 35 d in the nursery, pigs moved to a naturally ventilated finisher (0.74 m2 per pig) with fully slatted floors. Weight, 10th rib backfat, and loin eye area (LEA) were collected at 156 d of age (backfat and LEA adjusted to 114 kg). Data was analyzed using PROC GLM with a fixed effect of genetic line and covariates of weaning age and birth weight. A one day increase in weaning age increased (P < 0.01) 156 d weight by 658 g. Nursery average daily gain (ADG) was greater (P < 0.01) for pigs sired by SHOW and W when compared to P (414 and 397 vs. 351 g, respectively) yet finishing ADG did not differ (P > 0.05) (821 and 817 vs. 797 g, respectively). Backfat was thicker (P < 0.01) for SHOW when compared to W or P (16.7 vs. 14.7 and 14.7 mm, respectively) and LEA was larger (P < 0.05) for SHOW when compared to W or P (47.3 vs. 44.8 and 44.0 cm2, respectively). Finishing lean gain per day was greater (P < 0.05) for SHOW and W when compared to P (323 and 322 vs. 309 g per d, respectively). Results suggest pigs sired by LW SHOW are fatter, heavier muscled, and have comparable lean growth to those sired by commercial LW genetic lines.

Continuous Feeding Does Not Blunt Skeletal Muscle Protein Synthesis and Lean Growth Compared to Intermittent Bolus Feeding in the Preterm Piglet (OR26-06-19)

Objectives Refining early feeding strategies for premature infants is essential for mitigating adverse outcomes of prematurity. In neonatal term piglets, continuous feeding blunts growth compared to intermittent bolus feeding. Our objective was to determine the impact of feeding modality on lean growth in preterm pigs. We hypothesized that intermittent bolus feeding can mitigate low lean growth rates in preterm neonates compared to continuous feeding. Methods Pigs obtained by C-section (105 d gestation; 952 ± 205 g body weight) were fitted with an umbilical artery catheter (later replaced with jugular vein catheter) and an orogastric tube for parenteral and enteral nutrition, respectively. Pigs were assigned to continuous (CONT; 7.5 mL/[kg·h]) or intermittent bolus (INT; 30 mL/kg every 4 h over 15 min) feeding for 21 d. Pigs initially received parenteral nutrition and were advanced to full oral feeds over 6 d (220 kcal/kg and 16 g/kg protein per day). Body composition (by DXA), plasma insulin, and skeletal muscle anabolic signaling and fractional protein synthesis rates (PS; L-[ring-2H5]phenylalanine) were determined in INT pigs in the postabsorptive (before a meal, INT-PA; n = 13) and postprandial (after a meal, INT-PP; n = 16) states and in CONT pigs (n = 14). Results Body weight gain, lean mass, and fat mass did not differ between INT and CONT pigs. Insulin was lower before feeding for INT pigs than CONT pigs (P < 0.05). Insulin increased with feeding for INT pigs and exceeded that of CONT pigs at 30 and 60 min (P < 0.01) before returning to baseline levels at 240 min. In the longissimus dorsi (LD), gastrocnemius, and soleus muscles, the abundance of the eIF4E·eIF4G complex, which is required for translation initiation, was greater in INT-PP and CONT pigs than INT-PA pigs (P < 0.01), but did not differ between INT-PP and CONT pigs. PS in the LD muscle was greater in INT-PP pigs than INT-PA pigs (P < 0.01), but did not differ between INT-PP and CONT pigs. Conclusions Continuous feeding does not blunt translation initiation and protein synthesis in skeletal muscle compared to intermittent bolus feeding in preterm piglets. The resulting absence of enhanced lean growth with intermittent bolus compared to continuous feeding contrasts with term piglets and may be a consequence of prematurity. Funding Sources USDA CRIS 6250-51000-055, NIH HD072891, and USDA NIFA 2013-67015-20438.