Woody breast myopathy broiler show age-dependent adaptive differential gene expression in Pectoralis major and altered in-vivo triglyceride kinetics in adipogenic tissues.

A study was conducted to understand the differentially expressed genes in Pectoralis (P) major under woody breast (WB) myopathy condition in a high yielding broiler strain using RNA-sequencing at the growing (d 21) and finishing (d 42 and d 56) grow-out ages. Follow-up study was conducted to understand the in vivo triglyceride (TG) synthesis (d 49) occurring in adipogenic tissues using deuterium oxide (2H2O) as a metabolic tracer. Results indicated the top physiological systems affected in myopathy broiler were related to the musculo-skeletal system (d 21, 42, and 56) and cardiovascular system (d 42 and 56). Ubiquitin-specific proteases are expressed higher in myopathy broiler at d 21 (OTUD1) and d 42 (SACS) that potentially indicated higher degradation of muscle protein occurring at those ages. While genes related to transcription factors and muscle cell differentiation (ZNF234, BTG2) and muscle growth (IGF1) were upregulated with myopathy broiler suggesting concurrent muscle fiber regeneration. The downregulation of PYGB and MGAM genes related to carbohydrate transport and metabolism at d 42 potentially indicated nutrient-deficient state of myopathy affected fibers; whereas the nutrient-deficient physiological state of cells seemed to be counteracted by up-regulation of genes related to carbohydrate (ALDOB, GPD1L2) at d 56. There was a reduced (P < 0.05) in vivo TG synthesis in liver of the myopathy broiler (0.123 %/hr) compared to non-myopathy broiler (0.197 %/hr). The majority of TG synthesized in liver with myopathy broiler could conceivably be delivered to P. major (rather than to abdominal fat pad storage) to fulfil the increased energy need of muscle cells (via TG lipolysis and fatty acid [FA] oxidation). The increased utilization of FAs in the WB affected muscle could result in reduced secretion of FAs into blood circulation leading to sub-optimal availability of FAs for re-esterification for TG synthesis in liver. Results indicated that myopathy broiler at later age (d 56) of grow-out period were synchronously going through adaptive physiological processes of feedback responses to adverse cellular states.

Effects of dietary energy levels on Pectoralis major mixed muscle protein turnover and body composition in two broiler lines housed in different grow-out environments.

This study determined the Pectoralis (P) major mixed muscle protein turnover (PT) in two meat broiler lines, Line A and Line B, during the finishing grow-out feeding period (21-42 days) as affected by the dietary metabolizable energy (ME) levels and ambient temperatures. Experimental finishing diets consisted of 80, 90, 100, 110 and 120% ME of recommended nutrient guidelines for energy level. Fractional synthesis rates (FSR) or fractional degradation rates (FDR) were measured in P. major at day 36 and 42. Protein and fat mass gain were measured, and respective energy retention efficiencies as protein and fat (EREp and EREf) were determined. Metabolic heat production (HP) was also reported. Experimental feeding studies were conducted in cool season (24 hr mean: 69.91˚F and 63.98% RH) and in hot season (24 hr mean: 77.55˚F and 86.04% RH). Results showed that FSR or FDR values were not affected by dietary ME levels at day 36, whereas reduced FSR (p < .05) were observed at day 42 fed diets with reduced ME levels (≤100% ME) which could have resulted from greater maintenance energy requirement of maturing broilers at that age. Broilers fed reduced ME diets (≤100% ME) maintained protein mass (equivalent to broilers fed ≥100%-120% ME) by reduced FDR and increased feed intake. Grow-out ambient temperature did not affect FSR or FDR values across ME levels. Line B retained higher protein mass, lower fat mass and greater HP compared to Line A. This was followed by higher feed intake in Line B. Further, Line B exhibited higher EREp and lower EREf across dietary ME levels. In summary, PT homeostasis and body composition changes in broiler lines studied seemed to be regulated by the birds' intent to normalize energy intake as per physiological need by controlling feed intake.

Effects of dietary energy levels on performance and carcass yield of 2 meat-type broiler lines housed in hot and cool ambient temperatures.

Two meat-type broiler lines, line A and line B were fed experimental diets from 22-42 d with objectives to determine the effects of dietary metabolizable energy (ME) levels on feed intake (FI), performance, body composition, and processing yield as affected by environmental grow-out temperatures. Two thousand fifty male chicks from line A and 2,050 male chicks from line B were reared in 90-floor pens, 45 chicks per pen utilizing primary breeder nutrition and husbandry guidelines for starter (1-10 d) and grower (11-21 d) phases. Experimental finisher diets consisted of 5 increasing levels of apparent nitrogen corrected ME (2,800, 2,925, 3,050, 3,175, and 3,300 kcal/kg set at 19.5% crude protein and 1.0% dLys at each level) to represent 80, 90, 100, 110, and 120% ME of Evonik AminoChick energy level giving 2 × 5 factorial design and were fed from 22-42 d. All other amino acid levels in diets were formulated to a fixed ratio of dLys level. There were nine replicate pens for each diet and each line. The experiment was conducted twice-once in hot season (barn averages: 77.55 ˚F and 86.04% RH) and another in cool season (barn averages: 69.91 ˚F and 63.98% RH) of the year. Results showed that FI and feed conversion ratios (FCR) decreased (P < 0.05) linearly (R2 = 0.9) by 61.25 g and 0.073 units for every 10% increase in dietary ME for combined analysis of lines and seasons. The % fat mass of total body mass increased by 0.57%, whereas % protein mass decreased by 0.21% across ME levels (R2 > 0.9). However, there was no difference (P > 0.05) in % weights (of live weight) for wings, breast filet, tenders, or leg quarters across ME levels for both lines except % fat pad that increased (P < 0.05) by 0.20% for each 10% increment in dietary ME level. Line B had higher cumulative FI, BW gain, % lean, and protein mass of body mass than line A in hot season (P < 0.05). Feed intake was not different between lines in cool season (P > 0.05), whereas higher BW and improved FCR were observed for line A. Line A had higher % fat mass in both seasons. In summary, performance and yield results as affected by dietary ME levels were line specific and were affected by grow-out seasons. The optimal dietary ME level for the ME range studied (2,800-3,000 kcal/kg) at a constant recommended amino acid level lies in determining the best performance and profitability indices by taking into account the grow-out production inputs and processing yield outputs.

Effect of digestible amino acids to energy ratios on performance and yield of two broiler lines housed in different grow-out environmental temperatures.

Two broiler lines, Line A and Line B, were fed experimental diets from 22 to 42 d with objectives to determine effects of digestible amino acids (AA) to metabolizable energy ratios on feed intake (FI), performance, and processing yield. Experimental diets were formulated to 3,150 kcal/kg with 5 levels of digestible lysine (dLys)-80, 90, 100, 110, and 120% of recommended AA level giving g dLys/Mcal values of 2.53, 2.85, 3.17, 3.48, and 3.80, respectively. All other AA were formulated to a fixed ratio to dLys. A total of 4,050 chicks were utilized in each trial (9 replicate pens for each AA level and each line; 45 chicks/pen) conducted twice: one in hot environmental temperature (HT) (24 h mean ∼85.3 °F; 80.9% RH) and another in cool environmental temperature (CT) (24 h mean ∼71.6 °F; 61.7% RH). Results showed that FI was not impacted by dietary AA levels in HT for both lines. Higher FI (P < 0.05) was observed in CT for lower dietary AA levels (<100% AA level) for both lines, with overall higher FI occurring in Line B. Higher FI for Line B was also accompanied by higher body weight in HT and CT. Treatment diets had quadratic effects on average daily gain (ADG), feed conversion ratio (FCR), and processing yields (breasts and tenders) in both HT and CT, with broilers in CT performing better (P < 0.05). The optimal response values for ADG in HT and CT were 89.72 g and 113.44 g occurring at 120 and 109.5% AA level, respectively. The optimal response values for FCR in HT and CT were 1.79 and 1.58 occurring at 120 and 117.5% AA level, respectively. The optimal response values for breast meat yield in HT and CT were 575.9 g and 776.5 g occurring at 112.6 and 114.5% AA level, respectively. The optimal response values for tender meat yield in HT and CT were 119.8 g and 154.9 g occurring at 120 and 115% AA level, respectively. Line A had a higher breast and tender yield % (of live weight) for both environmental temperatures which correlated to body composition data with higher % protein mass and % digestible AA retention. In this study, findings indicated that effects of increased digestible AA density on FI, performance, and processing yield are specific to strain and grow-out temperature, but the optimum response was attained for both lines with diets containing 110 to 120% AA levels (3.48-3.80 g dLys/Mcal) during the 22 to 42 d finisher period.

In vivo collagen and mixed muscle protein turnover in 2 meat-type broiler strains in relation to woody breast myopathy.

Two meat-type broiler strains, strain A and strain B, were reared in floor pens (25 birds/pen; 45 pens/strain) for pectoralis (P) major collagen and mixed muscle protein turnover (PT) study from 0-56 D using primary breeder nutrition and husbandry guidelines. Forty broilers (n = 10/strain for collagen PT; n = 10/strain for mixed muscle PT) were selected at each sampling age at day 21, 28, 35, 42, and 56 and infused with 1-13C proline (Pro) and 15N-phenylalanine (Phe) which are used as amino acid tracers for collagen and mixed muscle PT measurements, respectively. Muscle and plasma samples were collected, and enrichments of 1-13C Pro and 15N-Phe were determined using mass spectrometry. Fractional synthesis rate (FSR) and fractional degradation rate (FDR) were measured for collagen and mixed muscle using precursor-product principle. At day 42, after separating the sampled broilers as myopathy (woody breast [WB] score > 1) and nonmyopathy (WB = 0), plasma metabolites were screened for differential 3-methyhistidine (3-MH) expression for both strains. Data were analyzed using one-way ANOVA using t test. Results showed that collagen and mixed muscle FSR and FDR in pectoralis major decreased (P < 0.05) for both strains as the broilers aged. FSR for collagen and FDR for mixed muscle were higher for strain B than those for strain A (P < 0.05). Total collagen was higher (P < 0.05) for strain B. Differentially expressed 3-MH in plasma was higher (P < 0.05) for myopathy-affected broilers indicating greater muscle degradation occurring in myopathy-affected broiler types for both strains. 3-MH Expression in plasma was higher for strain B than for strain A. The research findings showing an increased collagen content per unit muscle weight in pectoralis major in strain B (than in strain A) could be due to higher mixed muscle FDR and increased collagen FSR occurring during the grow-out period. The increased degradation of muscle fibers and probable replacement of muscle-specific protein with connective tissue, mainly collagen, was an evident pathophysiological phenomenon occurring in myopathy-affected broilers.

Effects of dietary amino acid levels and ambient temperature on mixed muscle protein turnover in Pectoralis major during finisher feeding period in two broiler lines.

Two broiler lines A and B were fed experimental diets from 21 to 42 days with an objective to determine Pectoralis major protein turnover (PT) as affected by the dietary amino acid (AA) levels and ambient temperature. Experimental diets (n = 9 replicate pens per diet) were formulated to 3,150 kcal/kg with five levels of digestible lysine (dLys) -80, 90, 100, 110 and 120% of recommended AA level giving g dlys/Mcal values of 2.53, 2.85, 3.17, 3.48 and 3.80 respectively. All other AA was formulated to a fixed ratio to dLys. Fractional synthesis or degradation rates (FSR or FDR) of P. major were measured on day 36 and day 42 for all dietary treatment levels for both broiler lines using stable isotope of AA (15 N-phenylalanine) as metabolic tracer. Experimental feeding studies were conducted once in hot season (24-hr mean ~ 85.3°F; 80.9% RH) and repeated in cool season (24-hr mean ~ 71.6°F; 61.7% RH) of the year. The FSR values increased (p < .05) as digestible AA in diet increased for both broiler lines in hot season until break point FSR occurring at 106.2% AA level. The average FSR values measured were higher for Line B at day 36 (20.98%/D for Line B vs. 20.69%/D for Line A) and at day 42 (16.07%/D for Line B vs. 12.47% D for Line A). FDR values observed at day 36 and day 42 were not different between lines (p > .05). Similar trends but elevated values of FSR and FDR in cool season than in hot season were recorded for both the lines. Line B showed the higher mixed muscle protein accretion (%/D) than Line A by actually increasing the FSR which was correlated by higher lean mass deposition and higher feed intake (p < .05). The overall findings indicated that PT response in P. major due to effects of digestible AA levels and ambient temperature was different and line-specific.

Characterizing Woody Breast Myopathy in a Meat Broiler Line by Heat Production, Microbiota, and Plasma Metabolites.

Selection for quantitative traits in meat broilers such as breast yield and growth rate exert physiological pressure leading to ante mortem histological and biochemical alterations in muscle tissues. The poultry industry has recently witnessed a myopathy condition affecting Pectoralis major (breast muscle) of broilers, called woody breast (WB), an etiology still unclear to scientific community. A study was conducted to characterize the WB myopathy in a meat broiler line at its finishing phase (d 41) in terms of heat production (HP), microbiota and plasma metabolites. Two treatment groups were studied-WB affected (myopathy) and normal (non-myopathy) broiler; n = 20 in each group. Indirect calorimetry was utilized for HP measurement. Furthermore, body composition (BC) analysis was also performed using dual-energy x-ray absorptiometry (DEXA). Microbiota in ileal digesta was studied with PCR amplified 16s rRNA gene. LC-MS targeted metabolomics was performed to understand differential expression of plasma metabolites. Results showed that there was difference in fasting HP (P < 0.05) between these two treatment groups, with non-myopathy broiler producing more heat which was indicative of higher body protein content validated by higher protein: fat ratio by BC results. Less protein content in myopathy bird could be due to probable higher mixed muscle degradation occurring in lean tissue as marked by elevated 3-methylhistidine expression in plasma. Microbiota results showed unclassified Lactobacillus as predominant genus with higher abundance occurring in myopathy group; whereas at species level, L. acidipiscis was predominant bacteria for non-myopathy broiler. Differentially significant metabolites (P < 0.05) identified from plasma metabolome between these two treatment groups were homocysteine, cyclic GMP, trimethylamine N-oxide (TMAO), tyramine, carnitine, and acetylcarnitine, which were all associated to cardiovascular system. The findings suggest that more research in meat broilers could be opted toward delivering reduced vascularity issues to alleviate this myopathy condition.