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Skeletal muscle growth is largely dependent on the proliferation and differentiation of muscle-specific stem cells known as satellite cells (SC). Previous work has shown that dietary inclusion of the vitamin D3 metabolite, 25-hydroxycholecalciferol (25OHD3), also called calcidiol, can promote skeletal muscle growth in post-hatch broiler chickens. Improving vitamin D status of broiler breeder hens by feeding 25OHD3 in addition to vitamin D3 has also been shown to positively impact progeny. Yet, whether combined pre- and post-hatch supplementation with 25OHD3 produces an additive or synergistic SC-mediated, skeletal muscle growth response remains unanswered. To evaluate the effect of combined maternal and post-hatch dietary 25OHD3 supplementation on the growth and SC mitotic activity of the Pectoralis major (PM) muscles in broiler chickens, a randomized complete block design experiment with the main effects of maternal diet (MDIET) and post-hatch diet (PDIET) arranged in a 2 × 2 factorial treatment structure was conducted. From 25 to 36 wk of age, broiler breeder hens were fed 1 of 2 MDIET formulated to provide 5,000 IU D3 (MCTL) or 2,240 IU of D3 + 2,760 IU of 25OHD3 per kg of feed (M25OHD3). Their male broiler chick offspring (n = 400) hatched from eggs collected from 35 to 36 wk of age were reared in raised floor pens. Broilers were fed 1 of 2 PDIET formulated to provide 5,000 IU of D3 per kg of feed (PCTL) or 2,240 IU of D3 + 2,760 IU of 25OHD3 per kg of feed (P25OHD3). Muscle was collected at days 4, 8, 15, 22, and 29 and stored until immunofluorescence analysis. Data were analyzed as a 2-way ANOVA with SAS GLIMMIX. Dietary 25OHD3 was effectively transferred from hen plasma to egg yolks (P = 0.002) and to broiler progeny plasma (days 4 to 22; P ≤ 0.044). Including 25OHD3 in either MDIET or PDIET altered PM hypertrophic growth prior to day 29 (P ≥ 0.001) and tended to reduce Wooden Breast severity (P ≤ 0.089). Mitotic SC populations were increased in PM of MCTL:P25OHD3 and M25OHD:PCTL-fed broilers at d 4 (P = 0.037). At d 8, the PM mitotic SC populations were increased 33% by P25OHD3 (P = 0.054). The results of this study reveal that combined maternal and post-hatch 25OHD3 supplementation does not produce additive or synergistic effects on SC-mediated broiler muscle growth. However, vitamin D status improvement through dietary 25OHD3 inclusion in either the maternal or post-hatch diet stimulated broiler breast muscle growth by increasing proliferating SC populations., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Large variations in hatching egg incubation temperatures have been previously shown to negatively impact posthatch growth in broiler chickens. The objective was to determine whether small incubation temperature variations owing to incubator tray location (LOC) could alter posthatch female and male broiler growth performance and carcass characteristics. Broiler hatching eggs were obtained from a 40-week-old commercial broiler breeder flock and incubated in trays placed in the bottom (BOT), middle (MID), and top (TOP) thirds of the racks (n = 4 racks per incubator tray LOC) in a single-stage incubator in a commercial hatchery. Chicks hatched from the 3 LOC (n = 720 per LOC) were vent sexed, vaccinated, and separate-sex reared with 12 birds per pen in a floor-pen facility and fed a common corn and soybean meal-based diet for 41 d. At day 41, all birds (n = 720) were processed to determine carcass and carcass part yields and incidence and severity of the meat quality defects wooden breast (WB) and white striping (WS). No LOC × Sex interactions were observed (P > 0.05). Growth performance and incidence and severity of WB and WS were similar among LOC (P > 0.05). However, broilers from BOT trays had heavier tender and breast weights than broilers from warmer MID trays (P < 0.05). Broilers from the BOT trays had higher breast meat yield as a proportion of carcass weight (25.00%) than warmer MID (24.54%) broilers (P < 0.05). However, broilers from warmer MID trays had greater carcass yield than those from cooler TOP trays (P < 0.05). As expected, male broilers had heavier carcass, breast, tender, wings, drumsticks and thighs weights and were more severely affected by WB than females (P < 0.05). Overall, these data indicate that the inherent differences in environmental factors among incubation LOC can impact broiler carcass and breast meat yields., (Copyright © 2020. Published by Elsevier Inc.)

Muscle satellite cells (MSCs) are myogenic stem cells that play a critical role in post-hatch skeletal muscle growth and regeneration. Activation of regeneration pathways to repair muscle fiber damage requires both the proliferation and differentiation of different MSC populations as well as the function of resident phagocytic cells such as anti-inflammatory and pro-inflammatory macrophages. The Wooden Breast (WB) phenotype in broiler chickens is characterized by myofiber degeneration and extensive fibrosis. Previous work indicates that the resident MSC populations expressing the myogenic regulatory factors, Myf-5 and Pax7 are larger and more proliferative in broilers severely affected with WB vs. unaffected broilers. To further characterize the cellular and molecular changes occurring in WB-affected muscles, samples from pectoralis major (PM) muscles with varying severity of WB (WB score 0 = normal; 1 = mildly affected; 2 = severely affected) were collected at 25 and 43 days post-hatch ( n = 8 per score per age) and processed for cryohistological and protein expression analyses. Collagen per field and densities of macrophages and MyoD+, Myf-5+, and Pax7+ MSC populations were quantified on immunofluorescence-stained cryosections. Relative collagen protein expression was quantified by fluorescent Western Blotting. In both 25 and 43-days-old broilers, the proportion of collagen per field ( P ≤ 0.021) and macrophage density ( P ≤ 0.074) were greater in PM exhibiting severe WB compared with normal. At day 43, populations of MyoD+, Myf-5+:MyoD+ MSC were larger and relative collagen protein expression was greater in WB-affected vs. unaffected broilers ( P ≤ 0.05). Pax7+ MSC relative to total cells was also increased as WB severity increased in 43-days-old broilers ( P ≤ 0.05). Densities of Myf-5+ ( P = 0.092), MyoD+ ( P = 0.030), Myf5+:MyoD+ ( P = 0.046), and Myf-5+:MyoD+:Pax7+ ( P = 0.048) MSC were greater in WB score 1 birds compared with WB score 0 and 2 birds. Overall, alterations in the resident MSC and macrophage populations and collagen protein content were observed in WB-affected muscle. Further investigation will be required to determine how these changes in cell population kinetics and local autocrine and paracrine signaling are involved in the apparent dysregulation of muscle maintenance in WB-affected broilers., (Copyright © 2020 Ferreira, Kindlein, Flees, Shortnacy, Vieira, Nascimento, Meloche and Starkey.)

Myogenic stem cells (MSC), also known as satellite cells, play a key role in mediating post-hatch skeletal muscle growth in broiler chickens. Information regarding the heterogeneity of MSC populations and myofiber size distributions in different muscles of modern broiler strains of both sexes is scarce. Thus, myofiber cross-sectional area (CSA) and myogenic regulatory factor expression by MSC populations in 2 functionally different muscles (Biceps femoris, BF and Pectoralis major, PM) from both sexes of broilers from 2 strains, Ross 708 x Ross 708 (ROSS; Aviagen Group, Huntsville, AL) and Red Ranger (RR; S&G Poultry, Clanton, AL) was evaluated. Female and male broilers from both strains (n = 80 birds per strain) were reared for 43 d on a common corn and soybean meal-based diet. At 43 d of age, 8 birds per sex per strain (n = 32 total) were euthanized and PM and BF muscle samples from each bird were analyzed using cryohistology, immunofluorescence (IF) staining, and fluorescence microscopy. Cryosections from each muscle from each bird were IF stained to detect Myf-5, MyoD, and Pax7-expressing MSC. The CSA of every fiber in each digital image was measured and MSC populations (Myf-5+, MyoD+, Pax7+, Myf-5+: MyoD+, Myf-5+: Pax7+, MyoD+: Pax7+, and Myf-5+: MyoD+: Pax7+) were enumerated. Myofiber CSA was similar among the 2 strains and 2 sexes (P > 0.05). As expected, mean myofiber CSA was greater in the predominantly fast-twitch PM than the predominantly slow-twitch BF muscle (P = 0.006). At 43 d of age, ROSS broilers had a 32% larger population of Myf-5+: MyoD+: Pax7+ MSC compared with RR (P = 0.04). The heterogeneity of MSC was similar among the sexes. While the total MSC population was larger in the BF muscle compared with the PM (P < 0.05), the Myf-5+ and Pax7+ MSC densities were similar among muscles (P > 0.05). The major differences in myofiber morphometrics and MSC heterogeneity were not among the different broiler strains or sexes but were instead, between the 2 functionally different muscles., (© 2019 Poultry Science Association Inc.)

The degenerative myopathy known as wooden breast (WB) has been increasingly observed in the breast muscle (PM) of commercial broilers during the last decade. Previous research has demonstrated that WB may be induced or ameliorated by modulating dietary digestible Lys (dLys) concentrations. Two concurrent experiments (Exp) were conducted to verify the effects of feeding 2 diets formulated to 75% and 100% of recommended dLys concentrations from 15 to 25 d of age on production responses and WB incidence (Exp 1), and the characterization of muscle stem cell activity in broilers affected by WB (Exp 2). At 25 and 43 d of age, birds were injected with 5΄-bromo-2΄-deoxyuridine (BrdU) prior to the collection of PM tissue to label mitotically active cells. Muscle samples were processed for cryosectioning and immunofluorescence staining and microscopy in order to determine myofiber cross-sectional area (CSA), to enumerate Myf-5+ and Pax7+ myogenic stem cell populations, and to determine the mitotic activity (BrdU+) of these populations. The reduced dLys diet produced broilers with differing (P < 0.001) incidences of WB within the same flock (Exp 1), with some detrimental effects on performance and processing characteristics. In Exp 2, broilers with severe WB had increased numbers (P = 0.016) and proportions (P = 0.022) of mitotically active, myogenic stem cells, as well as increased proportions (P < 0.05) of large CSA myofibers relative to broilers unaffected by WB at 25 d of age. At 43 d of age, broilers affected by severe WB had a greater (P = 0.011) total population of myogenic stem cell types (Myf-5+, Pax7+, or Myf-5+:Pax7+) and a concurrent increase (P = 0.007) in the mitotic activity (Myf-5+:BrdU+, and Pax7+:BrdU+, and Myf-5+:Pax7+:BrdU+) of these cells. Additionally, a greater (P < 0.05) proportion of small CSA myofibers was observed in broilers with severe WB. These results provide evidence that myofiber CSA, as well as the heterogeneity and mitotic activity of myogenic stem cell populations were altered in the presence of WB.

Quantitative control of nutrient intake may decrease the incidence of wooden breast (WB) and white striping (WS) myopathies with some impairment of live performance. Two experiments (Exp) utilizing Yield Plus × Ross 708 male broilers were conducted to determine if a reduction in myopathies may be obtained through a qualitative approach by reducing digestible lysine (dLys) density. All birds received an identical starter diet until 11 d of age. In Exp 1 (63 pens; 22 birds/pen), each pen was then randomly assigned to 1 of the following 7 dietary treatments (TRT) for a 47 d production period. Seven dietary treatments were provided for the grower 1 (G1; 12 to 18 d of age) and grower 2 (G2; 19 to 26 d of age) phases: 1) 100% of primary breeder recommendations for dLys throughout Exp; 2) 85% of TRT 1 dLys for G1; 3) 85% of TRT 1 dLys for G2; 4) 85% of TRT 1 dLys for G1 and G2; 5) 75% of TRT 1 dLys for G1; 6) 75% of TRT 1 dLys for G2; 7) 75% of TRT 1 for G1 and G2. In Exp 2 (24 pens; 30 birds/pen), birds were randomly assigned to 1 of the following 4 dietary TRT (Table 2) during a 61 d production period. Four dietary treatments were provided for the grower (G; 12 to 28 d) and finisher 1 (F1; 29 to 40 d of age) phases: 1) 100% of primary breeder recommendations for dLys 2) 85% of TRT 1 dLys for G; 3) 85% of TRT 1 dLys for F1; 4) 85% of TRT 1 dLys for G and F1; thereafter, birds received common finisher 1 (Exp 1: 27 to 42 d of age), finisher 2 (Exp 2: 41 to 48 d of age) and withdrawal (Exp 1: 43 to 47 d of age; Exp 2: 49 to 61 d of age) diets. Ideal amino acid ratios were not maintained in reduced dLys diets in either Exp. At 48 (Exp 1; 18 birds/pen) and 62 (Exp 2; 30 birds/pen) d of age, selected birds were processed and fillets were visually scored for WB and WS. No differences (P > 0.05) in cumulative live performance responses between TRT 1 and the remaining TRT were observed in either Exp. In Exp 1, the incidence of severe WB (20.8%) and WS (42.3%) at 48 d of age among birds receiving TRT 7 was reduced (P < 0.01) compared with TRT 1 (WB: 36.6%; WS: 64.3%), at the expense of reduced (P = 0.003) breast weights and yield. In Exp 2, the incidence of severe WB (18.8%) and WS (17.8%) at 62 d of age for birds receiving TRT 4 was reduced (P < 0.05) compared with TRT 1 (WB: 39.3%; WS: 38.3%), without any detrimental effects on processing characteristics. These results indicate that altering dietary dLys during critical periods of the growth trajectory may be a viable strategy for reducing the incidence and severity of WB and WS.

Two experiments (Exp) were conducted to determine if reductions in the incidence and severity of wooden breast (WB) and white striping (WS) may be obtained by reducing dietary nutrient density. In each Exp, Yield Plus × Ross 708 male broiler chicks were placed into 63 pens (22 birds/pen). All birds received an identical prestarter diet until 7 d of age, after which time each pen was randomly assigned to 1 of the following 7 dietary treatments (TRT) for the starter (8 to 14 d), grower (15 to 24 d), finisher 1 (Exp 1: 26 to 35 d; Exp 2: 26 to 42 d), and withdrawal (Exp 2: 43 to 48 d) phases: 1) 100% of primary breeder recommendations for digestible amino acid and metabolizable energy density throughout Exp; 2) 95% of TRT 1 until 14 d of age, then as TRT 1; 3) 95% of TRT 1 until 24 d of age, then as TRT 1; 4) 95% of TRT 1 throughout Exp; 5) 90% of TRT 1 until 14 d of age, then as TRT 1; 6) 90% of TRT 1 until 24 d of age, then as TRT 1; 7) 90% of TRT 1 throughout Exp. At 36 d (Exp 1) and 49 d (Exp 2), 18 birds per pen were processed and evaluated for WS and WB. In Exp 1, reduced dietary density in the starter phase (TRT 2 and TRT 5) resulted in increased (P ≤ 0.05) incidences of severe WB (32.9% and 34.7%) relative to TRT 1 (18.2%). In Exp 2, broilers assigned to TRT 7 had reduced (P < 0.01) incidences of severe WB (20.8%) and WS (42.3%) relative to the control (WB: 36.5%; WS: 64.5%). In both Exp, plasma creatine kinase and lactate dehydrogenase increased (P ≤ 0.05) with increasing scores for WB and WS. Reducing dietary nutrient density from 8 to 14 d may exacerbate fillet myopathies in broilers reared to 35 d of age. Although reducing dietary energy and amino acid density to 90% of recommendations from 1 to 48 d reduced the severity of myopathies, these reductions occurred with compromises in live performance. Altogether, these results indicated that concurrent manipulation of dietary amino acid and energy density is not a viable practical solution for breast myopathies.

An experiment was conducted to determine if myopathies of the Pectoralis major muscles are influenced by differences in growth trajectory achieved through a controlled feeding program. Male Yield Plus × Ross 708 broiler chicks were placed into 28 pens (25 birds/pen) equipped with plastic slats to prevent coprophagy. All birds received identical starter (1 to 10 d), grower (11 to 32 d), finisher (33 to 42 d), and withdrawal (43 to 50 d) diets that were formulated to meet or exceed nutrient recommendations of the primary breeder. Each pen of birds was randomly assigned to one of 4 pair-feeding programs (TRT 1: ad libitum; TRT 2: 95% of TRT 1 intake; TRT 3: 90% of TRT 1 intake; and TRT 4: 85% of TRT 1 intake) with 7 replicate pens per treatment. Feed intake and mortality were recorded daily. Individual BW was recorded at 31, 42, and 49 d of age. Blood samples were collected from 4 birds per pen at 31, 41, and 48 d of age and subsequently analyzed for plasma creatine kinase (CK) and lactate dehydrogenase (LDH). At 32, 43, and 50 d of age, 4 birds per pen were euthanized for necropsy. The right breast fillet of each bird was visually scored for white striping (WS) and wooden breast (WB). Linear decreases (P ≤ 0.01) in feed intake, BW gain, feed conversion ratio, and mortality were observed with decreasing feed allocation. Linear decreases (P ≤ 0.01) in severity were observed for WS and WB at 33, 43, and 50 d with decreasing feed allocation. Severity of WB at 33 and 43 d, as well as that of WS at 43 and 50 d, decreased (P ≤ 0.05) quadratically with decreasing feed allocation. Reduced feed allocation produced quadratic decreases (P ≤ 0.05) in CK and LDH concentrations at 31, 41, and 48 days. These results indicate that the incidence of breast fillet myopathies in broilers may be reduced through controlled feeding programs.

An experiment consisting of 3 nearly identical trials was conducted to determine the AMEn content of distillers dried grains with solubles (DDGS) to validate 4 previously published prediction equations for AMEn of corn DDGS in broilers. In addition, prior research data were used to generate a best-fit equation for AMEn based on proximate analysis. Fifteen samples of DDGS ranging in ether extract (EE) from 4.98 to 14.29% (DM basis) were collected from various dry-grind ethanol plants and were subsequently fed to broiler chicks to determine AMEn content. A corn-soybean meal control diet was formulated to contain 15% dextrose and test diets were created by mixing the control diet with 15% DDGS at the expense of dextrose. In each trial, male Ross × Ross 708 chicks were housed in grower battery cages and received a common starter diet until the experimental period. Each cage was randomly assigned to 1 of the dietary treatments (trial 1 and trial 2: control + 6 test diets, 13 replicates per diet; trial 3: control + 3 test diets, 12 replicates per diet). Experimental diets were fed over a 6-d acclimation period, followed by a 48-h total excreta collection period. On a DM basis, AMEn of the 15 DDGS samples ranged from 1,975 to 3,634 kcal/kg. Analyses were conducted to determine gross energy, CP, EE, DM, starch, total dietary fiber, neutral detergent fiber, crude fiber (CF), acid detergent fiber, and ash content of the DDGS samples. All results were reported on a DM basis. Application of the 4 equations to the validation data resulted in root mean square error (RMSE) values of 335, 381, 488, and 502 kcal/kg, respectively. Least absolute shrinkage and selection operator technique was applied to proximate analysis data for 30 corn coproducts adapted from prior research and resulted in the following best-fit equation: [AMEn (kcal/kg) = 3,673 - (121.35 × CF) + (51.29 × EE) - (121.08 × ash); P < 0.01; R(2) = 0.70; R(2) adj = 0.67; RMSE = 270 kcal/kg]. The RMSE values obtained through validation were not consistent with the expectation of predictive performance based on internal measures of fit for each equation. These results indicated that validation is necessary to quantify the expected error associated with practical application of each individual prediction equation to external data., (Poultry Science Association Inc.)

An experiment consisting of 2 identically designed trials was conducted to determine the nutrient composition and AMEn content of distillers dried grains with solubles (DDGS) to develop prediction equations for AMEn in broilers. Fifteen samples of DDGS ranging in ether extract (EE) from 3.15 to 13.23% (DM basis) were collected from various dry-grind ethanol plants and were subsequently fed to broiler chicks to determine AMEn content. A corn-soybean meal control diet was formulated to contain 15% dextrose, and test diets were created by mixing the control diet with 15% DDGS at the expense of dextrose. In each trial, 672 male Ross × Ross 708 chicks were housed in grower battery cages with 7 birds per cage (0.06 m(2)/bird) and received a common starter diet until 10 d of age. Each cage was randomly assigned to 1 of 16 dietary treatments, with 6 replicate pens per treatment. Experimental diets were fed over a 6-d acclimation period from 10 to 16 d of age, followed by a 48-h total excreta collection period. Gross energy (GE) and CP of the experimental diets and excreta were determined to calculate AMEn for each DDGS sample. On a DM basis, AMEn of the 15 DDGS samples ranged from 1,869 to 2,824 kcal/kg. Analyses were conducted to determine the GE, CP, EE, DM, starch, total dietary fiber (TDF), neutral detergent fiber (NDF), acid detergent fiber (ADF), and ash content of the DDGS samples. Stepwise regression resulted in the following best-fit equation for AMEn (DM basis) based on the adjusted coefficient of determination (R(2)adj), SE, and prediction error sum of squares (PRESS): AMEn, kcal/kg = -12,282 + (2.60 × GE) + (89.75 × CP) + (125.80 × starch) - (40.67 × TDF; R(2)adj = 0.86; SE = 98.76; PRESS = 199,819; P ≤ 0.001). These results indicated that the composition of DDGS with variable EE content may be used to predict AMEn in broiler chicks.