Broiler Pectoralis Major Muscle Research Articles

Effects of Wooden Breast Myopathy on Meat Quality Characteristics of Broiler Pectoralis Major Muscle and Its Changes with Intramuscular Connective Tissue.

This study aimed to investigate the effect of wooden breast (WB) myopathy on chemical composition, meat quality attributes and physiochemical characteristics of intramuscular connective tissue (IMCT) of broiler pectoralis major (PM) muscle. Thirty-six fillets were classified into varying degrees of WB condition, including normal, moderate and severe. Results show that WB myopathy altered the collagen profile in PM muscle by increasing total collagen content and decreasing collagen solubility. The composition of macromolecules in IMCT, including hydroxylysyl pyridoxine cross-linking, decorin and glycosaminoglycans, were increased with the severity of WB myopathy. Differential scanning calorimetry analysis indicated higher denaturation temperatures and lower denaturation enthalpy of IMCT for WB. Secondary structures of α-helix and β-sheet in the IMCT of WB were changed to β-turn and random coil. In addition, chemical composition and meat quality attributes showed a correlation with collagen profile and IMCT characteristics. Overall, this study emphasizes the effect of WB myopathy on IMCT and their contributions to meat quality variation.

Chronic heat stress inhibits glycogen synthesis through gga-miR-212-5p/GYS1 axis in the breast muscle of broilers

Studies have demonstrated that chronic heat stress can accelerate glycolysis, decrease glycogen content in muscle, and affect muscle quality. However, the consequences of chronic heat stress on glycogen synthesis, miRNA expression in pectoralis major (PM) muscle, and its regulatory functions remain unknown. In this study, high-throughput sequencing and cell experiments were used to explore the effects of chronic heat stress on miRNA expression profiles and the regulatory mechanisms of miRNAs in glycogen synthesis under chronic heat stress. In total, 144 cocks were allocated into 3 groups: the normal control (NC) group, the heat stress (HS) group, and the pair-fed (PF) group. In total, 30 differently expressed (DE) miRNAs were screened after excluding the effect of feed intake, which were mainly related to metabolism, signal transduction, cell growth and death. Furthermore, the gga-miR-212-5p/GYS1 axis was predicted to participate in glycogen synthesis through the miRNA-mRNA analysis, and a dual-luciferase reporter test assay confirmed the target relationship. Mechanistically, chronic heat stress up-regulated gga-miR-212-5p, which could inhibit the expression of GYS1 in the PM muscle. Knocking down gga-miR-212-5p alleviates the reduction of glycogen content caused by chronic heat stress; overexpression of gga-miR-212-5p can reduce glycogen content. This study provided another important mechanism for the decreased glycogen contents within the PM muscle of broilers under heat stress, which might contribute to impaired meat quality.

Shotgun proteomics reveals changes in the pectoralis major muscle of broilers supplemented with passion fruit seed oil under cyclic heat stress conditions

The aim of this study was to characterize the proteins differentially expressed in the pectoralis major muscle of broilers supplemented with passion fruit seed oil (PFSO) under cyclic heat stress conditions. Ninety one-day-old male chicks were housed in cages arranged in a climatic chamber, where they were kept under cyclic heat stress for eight hours a day from the beginning to the end of the experiment. The birds were divided into two experimental groups, one group supplemented with 0.9% PFSO and a control group (CON) without PFSO supplementation. At 36 days of age, 18 birds were slaughtered to collect muscle samples. From pools of breast fillet samples from each group, proteolytic cleavage of the protein extracts was performed, and later, the peptides were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The 0.9% PFSO supplementation revealed the modulation of 57 proteins in the pectoralis major muscle of broilers exposed to cyclic heat stress. Among them, four proteins were upregulated, and 46 proteins were downregulated. In addition, seven proteins were expressed only in the CON group. These results suggest that PFSO may increase heat tolerance, with a possible reduction in oxidative stress, activation of neuroprotective mechanisms, protection against apoptosis, decrease in inflammatory responses, and regulation of energy metabolism.

Research Note: Expression levels of collagen-related genes in PSE conditions and white striping features of broiler pectoralis major muscle

The objective of this study was to compare the meat quality characteristics and collagen-related gene expression levels in the broiler pectoralis major (PM) muscle among the meat quality groups, including normal, pale, soft, and exudative (PSE), and white striping (WS) groups. The group was classified by their WS degree (moderate or severe striping) and quality traits including pH and lightness values at 24 h postmortem (normal group: pH24 h ≥5.7, 48≤L*≤53, without WS features; PSE group: pH24 h <5.7, L* >53, without WS features; WS group: pH24 h ≥5.7, 48≤L*≤53, with moderate or severe striping). The WS group revealed no differences in all measured meat quality traits compared to the normal group (P > 0.05). PM muscles exhibiting PSE conditions without WS indicated lower pH15 min and pH24 h values (P < 0.05). Whereas, lower lightness and cooking loss values were observed in the normal and WS groups compared to the PSE group (P < 0.05). No significant difference was observed in the level of type I collagen among the groups (P > 0.05), whereas a higher type III collagen level was observed in the WS group than in the other groups (P < 0.05). Additionally, the WS group showed a higher type IV collagen level compared to the normal group (P < 0.05) and a level not different from that of the PSE group (P > 0.05). In contrast, the expression levels of matrix metalloproteinase (MMP) 2, involved in type IV collagen degradation, and angiopoietin-like protein 7, associated with collagen accumulation, were higher in the WS group compared to the normal group (P < 0.05). However, no difference was detected in the MMP1 level among the all groups (P > 0.05). These results suggest that the occurrence of WS features in broiler PM muscle, unlike PSE and normal conditions, can be influenced by the expression levels of collagen-related genes associated with abnormalities in extracellular matrix components.

Effects of the Probiotic Enterococcus faecium on Muscle Characteristics of Chickens.

The use of antibiotics in farm animals is one of the main reasons for the development of resistant bacterial strains (e.g., zoonotic pathogens). Therefore, save alternatives are needed. Here, we examined how post-hatch application (day one to seven of life) of the probiotic Enterococcus faecium AL41 (EF) affects the development and tissue properties of the broiler pectoralis major muscle (PM). Expression of regulators, namely IGF-1, PAX7, and MYF5, was also investigated. At day 1 (n = 6), and days 5, 8, and 12 (n = 10), muscle samples were taken from control and EF supplemented chicks. From day 5 on, myonuclei number per fiber was elevated in EF chicks. Improved capillarization (from day 8), larger myofibers, increased body and PM weights (day 12) were found in the EF group. Part of our findings is explainable by higher intramuscular expression of IGF-1 and lower MYF5 expression in EF birds. In both groups IGF-1 expression decreases with age, thereby increasing the cellular myogenic potential. However, a strong increase in PAX7 expression and more PAX7-positive nuclei were found in EF chicks at day 12. We conclude that EF supplementation improves PM growth and health due to positive effects on bioavailability and fusion capacity of SATC progeny and better tissue perfusion.

Research Note: PSE condition is associated with increased apoptotic potential in broiler pectoralis major muscle

This study compared the meat quality and sensory quality characteristics of broiler pectoralis major (PM) muscle between meat quality classes, including reddish, firm, and non-exudative (RFN) and pale, soft, and exudative (PSE) conditions. Additionally, we also investigated the associations between the meat quality classes and expression levels of cytochrome c, initiator (caspase 9), and effector caspases (caspase 3 and 7) at the early postmortem period. A total of 135 PM muscles from broilers were used, and meat quality classes were determined according to the pH24 h and lightness values and classified into the RFN (N = 81) and PSE (N = 54) conditions. The PSE breasts showed lower muscle pH15 min and pH24 h values compared to the RFN breasts (P < 0.05). A lower lightness value was observed in the RFN group compared to the PSE group (P < 0.001), whereas there were no significant differences in redness and yellowness between the groups (P > 0.05). The PSE group exhibited higher extent of water loss, including drip loss and cooking loss, compared to the RFN group (P < 0.05). For these reasons, samples from the RFN group required less force to breakdown the cooked meat with more moisture in the mouth after chewing compared to samples from the PSE group (P < 0.001); however, flavor intensity did not differ between the groups (P > 0.05). At 15 min postmortem, all apoptosis-related molecules, including cytochrome c, caspase 9, caspase 3, and caspase 7, were present at higher levels in the PSE group than in the RFN group (P < 0.05). These results indicated that higher apoptotic potentials were associated with the development of PSE chicken breasts. Therefore, the variation of meat quality in chicken breast can be explained as being affected by the expression levels of apoptosis-related factors at the early postmortem period.

Acute stress deteriorates breast meat quality of Ross 308 broiler chickens by inducing redox imbalance and mitochondrial dysfunction.

This study investigated the effects of acute stress on breast meat quality, redox status, and mitochondrial function in pectoralis major (PM) muscle of broilers. A total of 168 broiler chickens (42-d-old, Ross 308) were randomly divided into control (CON) and preslaughter transport (T) treatments. A broiler was an experimental unit. Each treatment consisted of 84 broilers, and they were put in 12 crates with 7 broilers each. Broilers in the T group were transported according to a designed protocol, and the CON broilers were kept in crates under normal living conditions before slaughtering. Based on the meat quality traits assessed at postmortem 24 h, all PM muscles of the transported broilers were further classified into normal (T-NOR) and pale, soft, and exudative (PSE)-like (T-PSE) groups for the determination of redox status in PM muscle and isolated mitochondria, energy metabolites, mitochondrial electron transport chain complexes activities, as well as mitochondrial function-modulating genes expression. Compared with CON, the extent of lipid peroxidation as well as protein oxidation were significantly increased in both PM muscles and mitochondria in T-PSE (P < 0.05), whereas not in T-NOR. Higher activities of glutathione peroxidase, total superoxide dismutase, and Cu-Zn superoxide dismutase were observed in PM muscle of T-NOR broilers when compared with CON (P < 0.05). Preslaughter transport increased the generation of reactive oxygen species, as well as enhanced antioxidant capacity in PM mitochondria of broilers (P < 0.05). Compared with CON, the ATP content, activities of complexes I and III, as well as relative mitochondrial membrane potential and swelling were significantly decreased in T-PSE (P < 0.05), whereas no significant changes in either ATP content or complex I activity were observed in T-NOR. Preslaughter transport enhanced the mRNA expression of regulators involved in the glutathione system, thioredoxin 2 system, and mitochondrial biosynthesis in PM muscle of broilers (P < 0.05). Moreover, we noticed a more evident enhancement effect in T-NOR than in T-PSE (P < 0.05). Overall, this work indicates that acute stress-induced redox imbalance and mitochondrial dysfunction have significant implications for the development of PSE-like meat.

Association of Serum Glucose, Serotonin, Aspartate Aminotransferase, and Calcium Levels with Meat Quality and Palatability Characteristics of Broiler Pectoralis Major Muscle.

Simple SummaryApoptosis is a critical and highly regulated cell death process after exsanguination, and serum apoptosis-related molecules are associated with apoptotic and glycolytic potentials. Thus, the activities and production of these molecules can directly and/or indirectly affect variation in meat quality characteristics. However, there is limited information on the complex relationship between serum biochemical molecules and their effects on the meat and eating quality of broilers. In the present study, the activity of aspartate aminotransferase, a cytoprotective molecule, was found to be associated with other apoptosis-related molecules, including blood glucose, serotonin, and calcium levels, at exsanguination. All serum parameters analyzed in this study were related to the early postmortem and ultimate muscle pH. Thus, apoptosis-related molecules may affect the glycolytic rate in broiler pectoralis major muscle. Additionally, our results demonstrated that the variations in the sensory quality traits of broilers could be somewhat explained by the levels of apoptosis-related serum parameters at exsanguination.This study investigated the correlations between apoptosis-related blood biochemical parameters measured at exsanguination and the meat and sensory quality characteristics of broiler pectoralis major muscle. The concentration of serotonin showed a positive correlation with aspartate aminotransferase (AST) activity (p < 0.001) and a negative correlation with calcium content (p < 0.01). All serum parameters showed relationships with muscle pH at 15 min and/or 24 h postmortem (p < 0.05). Serum AST activity, which had a negative correlation with calcium content (p < 0.01), was positively related with muscle pH and negatively correlated with Warner–Bratzler shear force values (WBS, p < 0.05). Principal component analysis results revealed the associations between AST activity and meat quality traits, including pH24h, lightness, and WBS. Furthermore, cooked breast with higher AST activity and lower calcium level tended to exhibit higher scores of tenderness and overall acceptability than that with lower AST activity and higher calcium level (p < 0.05).

Development of Imaging System for Online Detection of Chicken Meat with Wooden Breast Condition.

In recent years, the wooden breast condition has emerged as a major meat quality defect in the poultry industry worldwide. Broiler pectoralis major muscle with the wooden breast condition is characterized by hardness upon human palpation, which can lead to decrease in meat value or even reduced consumer acceptance. The current method of wooden breast detection involves a visual and/or tactile evaluation. In this paper, we present a sideview imaging system for online detection of chicken breast fillets affected by the wooden breast condition. The system can measure a physical deformation (bending) of an individual chicken-breast fillet through high-speed imaging at about 200 frames per second and custom image processing techniques. The developed image processing algorithm shows the over 95% classification performance in detecting wooden breast fillets.

529 Late-Breaking: Effects of in Ovo Injection of High Yield Broilers with Nicotinamide Riboside on Pectoralis Major Morphometrics, Muscle Fiber Density, and mRNA Expression

Abstract The objective of this study was to determine the effects of in ovo injection of high yield broilers with nicotinamide riboside (NR) on broiler pectoralis major muscle (PMM) development, growth, and mRNA expression. Fertilized Cobb 700 broiler eggs were randomly assigned to 1 of 4 treatments within a 2×2 factorial design with NR dose (DOS; 0 or 2.5 mM) and injection location (LOC; albumen or yolk) as main effects. Eggs were injected with their assigned treatment at d 10 of incubation. Day-15 and -18 embryos and hatched chicks were euthanized, right PMM and muscle fiber morphometric measurements were collected, and left PMM were utilized for mRNA expression analysis of sirtuin-1 and cyclin D1, 2 and 3. There were no LOC×DOS interactions or main effects for d-15 and -18 PMM measurements (P &amp;gt; 0.23). There were LOC×DOS interactions for hatched chick PMM weight and length, and muscle fiber density (P &amp;lt; 0.04). When NR was injected into the albumen, PMM weight decreased (P = 0.02) and length and fiber density were unaffected (P &amp;gt; 0.33), while PMM weight and fiber density increased when NR was injected into the yolk (P &amp;lt; 0.05). There were no LOC×DOS interactions for all genes analyzed for d-15 and 18-embryos (P &amp;gt; 0.23), but NR embryos had greater expression of all genes (P &amp;lt; 0.03) except d-18 cyclin-D2 (P = 0.08). There were LOC×DOS interactions for hatched chick expression of all genes (P &amp;lt; 0.04). There was no difference in sirturin-1 expression when chicks were injected with 0 mM NR, but 2.5-mM yolk chicks had greater expression than 2.5-mM albumen chicks. When NR was injected into the albumen, expression of all cyclins was unaffected (P &amp;gt; 0.08), while expression increased when NR was injected into the yolk (P &amp;lt; 0.01). Increased expression of sirturin-1 and the cyclins may be responsible for NR catalyzed increase in PMM weight and fiber density.

Effects of feed restriction on the occurrence of white striping and wooden breast myopathies, performance, carcass characteristics and some blood parameters in broiler chickens

This study was planned to determine the effects of feed restriction applied at different times and levels on the occurrence of white striping and wooden breast myopathies, performance, carcass characteristics and some blood parameters in broilers. In total, 600 male broilers were randomly placed to 5 groups with eight replicates. Control group was fed ad libitum throughout experimental period. T1 and T2 groups received feed as 80% and 70% of control group from 11 to 24 days of age, respectively. T3 and T4 groups received feed as 80% and 70% of control group from 25 to 39 days of age, respectively. T2, T3 and T4 groups had the lower mean values for feed intake and body weight gain from 0 to 49 days of age, final body weight, carcass weight, serum alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels, compared to control group. T3 and T4 groups had lower dressing percentage, breast meat yield, serum aspartate aminotransferase (AST) and creatine kinase (CK) levels than control. T2, T3 and T4 groups had lower incidence for white striping and wooden breast myopathies than control. T3 and T4 groups had also the lower incidence for severe degree white striping and wooden breast myopathies than control. In conclusion, feed restriction to 70% of ad libitum intake between 11 and 24 days of age and feed restriction to 80% and 70% of ad libitum intake between 25 and 39 days of age may reduce the incidence and severeness of white striping and wooden breast myopathies, although there is some loss in performance traits. Feed restrictions to 80% and 70% levels of ad libitum intake between 25 and 39 days of age may be more effective in reducing these myopathies on pectoralis major muscle of broilers.

Influence of Woody Breast Myopathy on Sarcomere Length and Tensile Strength in Commercial Broiler Pectoralis major Muscle

Woody breast syndrome is characterized by degenerative changes at the muscle fiber level and accumulation of connective tissue between the fibers. This study explored effects of the syndrome on muscle properties by focusing on a comparison of the sarcomere lengths between normal and woody breast muscles, including cranial and middle parts, surface and deeper layers, electrically stimulated and nonstimulated muscles, and their combinations. Tensile strengths longitudinally and transversally to the muscle fiber direction in the cranial and middle parts of the muscles were determined. The overall sarcomere lengths of woody breasts were longer than normal muscles (1.93 μ m vs. 1.88 μm; P &amp;lt; 0.05). The surface layer had overall longer sarcomere lengths than the deeper layer (cranial surface vs. cranial deeper layer: 1.97 vs. 1.89 μ m; middle surface vs. middle deeper layer: 1.93 vs.1.84 μ m; P &amp;lt; 0.05). Compared with normal breast muscles, woody breast muscles had longer sarcomeres in the surface layer; however, sarcomere length did not differ in the deeper layer. Electrically stimulated chicken breasts generally had longer sarcomere lengths (2.00 vs. 1.82 μm; P &amp;lt; 0.001). There was no significant difference in tensile strength between normal and focal woody breast (mild local lesion, usually in the cranial end) samples when fiber direction or sample location was studied (P &amp;gt; 0.05), but there was a clear difference between normal and focal versus diffuse (severe woody breast lesion throughout the muscle) samples (P &amp;lt; 0.001). Tensile strength was much greater in diffuse woody breast muscles when extended longitudinally or transversely to the fiber direction. In conclusion, although this study did not show sarcomere lengths in living muscle, it suggests an imbalance in sarcomere lengths in different parts of the breast muscle, which may induce a reduction in the functionality and strength of the muscle.

Physical, Chemical and Histological Characterization of Pectoralis major Muscle of Broilers Affected by Wooden Breast Myopathy.

Simple SummaryWooden breast myopathy is a muscle abnormality resulting from the genetic improvement practiced by the poultry industry and which is widespread throughout the planet. Recent research has tried to unravel this myopathy for years and how the quality of the meat is affected, in search of its etiology and the solution to this problem. Unfortunately, the problems related to the myopathies of the affected birds will only be solved when the cause of the abnormality is in fact unveiled and controlled; in the meantime, it is up to us researchers to contribute scientifically to research on this type of meat that is fit for consumption but that, for often presenting non-standard appearance for sale in the form of whole fillets, is discarded.This study aimed to characterize the effects of wooden breast myopathy (WBM) on quality of broiler chicken breast meat. Normal samples (absence of myopathy), moderate-degree samples (hardness only in one area of the breast fillet) and severe-degree samples (hardness throughout the breast fillet) were classified. In macroscopic analysis, the pectoral muscle affected by the WBM showed, in general, pale color with stiff, irregular and reddish regions (suffusions and petechiae), with the presence of white striations. In microscopic analysis, the myopathy was characterized by loss of the polygonal aspect of the muscle fibers. Samples with moderate degree of the myopathy showed greater (p = 0.0266) water retention capacity. There was an increase (p = 0.004) in total collagen concentration in samples from the severe-degree group 0.29% in normal samples to 0.43% and 0.48% in samples from moderate- and severe-degree groups, respectively. Samples of chicken breast affected by the severe-degree WBM showed lower (p < 0.0001) myofibrillar fragmentation index (64.51) and lower (p = 0.0002) fat concentration (2.17%) than normal chicken samples (80.45 and 3.79%, respectively). Samples affected by WBM are larger and heavier and present poorer physical quality when compared to normal chicken meat. Histologically it is possible to observe loss of the polygonal aspect of muscle fibers.

Research Note: Evaluation of the incidence of white striping and underlying myopathic abnormalities affected by fast weight gain in commercially fed broiler chickens

Significant improvements in genetics, nutrition, and food efficiency have had a great impact on the rapid growth of broilers, notably with increases in muscle mass. However, with rapid growth, the broiler industry has been negatively impacted by the increased incidence of myopathies, including white striping. White striping affects the pectoralis major muscle of broilers, particularly the larger breasts of rapidly growing modern commercial broiler lines. In this study, we documented the growth process of commercial broiler chickens from hatching to market weight at 6 wk. Gross pathology and histopathology analyses were performed on pectoralis major muscle collected weekly from birds culled from 1 to 6 wk. The severity of both gross and histologic pathologies in the breast muscle increased over time. White striping was initially observed at week 2, with a rise in the incidence and severity through the sixth week. Mild histopathology was noted in week 2, characterized by macrophage infiltration and limited phagocytosis of the muscle. Muscle condition deteriorated with age and weight gain, with more prevalent macrophages, phagocytosis, and interstitial fibroblasts. By week 5 and 6, there was severe myopathy including regions of obliterated muscle tissue. Linear regression models show a positive correlation between white striping, gross pathology, and histopathology relative to weight and age.

The unique physiological features of the broiler pectoralis major muscle as suggested by the three-dimensional ultrastructural study of mitochondria in type IIb muscle fibers.

Typical skeletal muscles are composed of mixed muscle fiber types, which are classified as slow-twitch (type I) and fast-twitch (type II) fibers, whereas pectoralis major muscles (PMs) in broiler chickens are 100% composed of type IIb fast-twitch fibers. Since metabolic properties differ among muscle fiber types, the combination of muscle fiber types is involved in physiological functions and pathological conditions in skeletal muscles. In this study, using serial block-face scanning electron microscopy, we compared three-dimensional (3D) mitochondrial properties in type IIb fibers in broiler PMs and those in type I fibers of broiler gastrocnemius muscles (GMs) heterogeneously composed of slow- and fast-twitch muscle fibers. In type I fibers in the GMs, elongated mitochondria with numerous interconnections to form a substantial network among myofibrils were observed. Along with lipid droplets sandwiched by mitochondria, these features are an adaptation to effective oxidative respiration and constant oxidative damage in slow-twitch muscle fibers. In contrast, type IIb fibers in the PMs showed small and ellipsoid-shaped mitochondria with few interconnections and no lipid droplets, forming a sparse network. The mitochondrial spatial network comprises of active mitochondrial dynamics to reduce mitochondrial damage; therefore, type IIb fibers possess physiologically low capacity to maintain mitochondrial wellness due to static mitochondrial dynamics. Based on 3D mitochondrial properties, we discussed the contrasting physiological functions between type I and IIb fibers and proposed a high contractile power and low stress resistance as unique physiological properties of broiler PMs.

High leucine levels affecting valine and isoleucine recommendations in low-protein diets for broiler chickens

Four experiments were conducted to estimate the optimal standardized ileal digestible (SID) level of branched-chain amino acids in low-protein diets during the starter, grower, and finisher periods, using the response surface methodology, and to study their effects on performance and mRNA expression of genes involved in the mechanistic target of rapamycin (mTOR) pathway of broiler chickens from 8 to 21 D of age. In experiments 1, 2, and 3, a total of 1,500 Cobb male broiler chickens were assigned to 15 diets of a central composite rotatable design (CCD) of response surface methodology containing 5 levels of SID Leu, Val, and Ile with 5 replicate pens of 20 birds each. A 3-factor, 5-level CCD platform was used to fit the second-order polynomial equation of broiler performance. In experiment 4, a total of 540 8-day-old Cobb male broiler chickens were distributed in a completely randomized 2 x 3 x 3 factorial arrangement with 2 SID Leu levels (1.28 or 1.83%), 3 SID Val levels (0.65, 0.90, or 1.20%), and 3 SID Ile levels (0.54, 0.79, or 1.09%) for a total of 18 treatments with 5 replicate cages of 6 birds each. High Leu levels impaired (P < 0.05) gain:feed when birds were fed marginal Val or Ile diets. However, gain:feed was restored when both Val and Ile were supplemented to reach adequate or high levels. High Leu levels increased (P < 0.05) mRNA expression of S6K1 and eEF2 genes only in birds fed high Ile levels. Dietary SID Leu, Val, and Ile levels required for gain:feed optimization in low-protein diets were estimated at 1.37, 0.94, and 0.87% during the starter period; 1.23, 0.82, and 0.75% during the grower period; and 1.15, 0.77, and 0.70% during the finisher phase, respectively. Higher Val and Ile levels are required to optimize the effect of Leu supplementation on mRNA expression of mTOR pathway genes in the pectoralis major muscle of broilers from day 1 to 21 after hatch.

Physiological and Pathological Mitochondrial Clearance Is Related to Pectoralis Major Muscle Pathogenesis in Broilers With Wooden Breast Syndrome.

Wooden breast syndrome (WB) constitutes an emerging myopathy in the pectoralis major muscle (PM) of broiler chickens, characterized by myofiber hypertrophy and degeneration along with severe fibrosis. WB pathogenesis has been considered to involve hypoxia induced by rapid growth of the PM. In this study, we focused on mitochondrial morphology and dynamics in the myofibers, as these organelles are sensitive to damage by hypoxia, and examined the effects on WB pathogenesis. Specifically, the PMs of a flock of 35 broilers at 50 days of age were evaluated. First, the severity of disease in each bird was determined by measuring histopathological indices including the fibrotic area (FA) in the muscle and circularity of myofibers (CM). These values were 29.4 ± 9.6% and 0.70 ± 0.042, respectively, showing variety among the flock. Myofiber vacuolization was observed in all birds including numerous small- or large-rimmed vacuoles, with the former consisting of ultrastructurally autophagosome-like vacuoles engulfing degenerated mitochondria. The large-rimmed vacuoles frequently occurred in the PMs with more severe FA and CM, indicating a relationship between altered autophagy/mitophagy and WB severity. Next, the expression levels of hypoxia-adaptive and mitochondrial dynamics-related genes were analyzed, and their correlations with the histopathological indices were examined. The histopathological indices were negatively correlated with the expression of vascular endothelial growth factor A (VEGFA), indicating that less angiogenesis owing to weakened hypoxia-inducible factor signaling induces more severe WB pathology. In addition, the observed negative correlation with mitochondrial dynamics-related genes implied that WB pathology deteriorates concomitant with reduced mitochondrial dynamics. Furthermore, the expression of mitochondrial dynamics-related genes showed strong positive correlation with that of VEGFA and autophagy-/mitophagy-related genes. These results revealed that the PMs of broilers possess the mechanism of physiological clearance of mitochondria damaged by the hypoxia resulting from the continuous mitochondrial dynamics and autophagy/mitophagy accompanying rapid PM growth. In turn, the altered mitochondrial clearance induced by chronic hypoxia and the accumulation of damaged mitochondria likely underly the severe pathological features of WB.

Exploring the Factors Contributing to the High Ultimate pH of Broiler Pectoralis Major Muscles Affected by Wooden Breast Condition.

The elevated ultimate pH (pHu) found in wooden breast (WB) meat suggests an altered muscular energetic status in WB but also could be related to a prematurely terminated post-mortem pH decline. The aims of this study were to explore the factors contributing to the elevated pHu and establish whether the occurrence of WB defect alters muscle post-mortem carbohydrate metabolism and determine if the contractile apparatus reflects such changes. A total of 24 carcasses from Ross 308 male chickens were obtained from a commercial producer and harvested using commercial processing procedures. Carcasses were categorized into unaffected (NORM) and WB groups (n = 12 each), and samples were collected from cranial bone-in pectoralis major (PM) muscles at 15 min and 24 h post-mortem for the determination of pH, glycolytic metabolites, adenonucleotides, buffering capacity, phosphofructokinase (PFK) activity, and in vitro pH decline. Twenty-four additional deboned PM samples (12 NORM and 12 WB) were collected from the same processing plant to assess muscle histology and sarcomere length at four different locations throughout the PM muscle. Data show that the reduced glycolytic potential of WB muscles only partially explains the higher (P < 0.001) pHu of WB meat, as residual glycogen along with unaltered PFK activity suggests that neither glycogen nor a deficiency of PFK is responsible for arresting glycolysis prematurely. The dramatic reduction in ATP concentrations in the early post-mortem period suggests a defective ATP-generating pathway that might be responsible for the reduced pH decline in WB samples. Further, the addition of excess of ATPase extended post-mortem glycolysis of WB meat in an in vitro glycolytic system. WB-affected samples have longer (P < 0.001) sarcomeres compared to NORM, indicating the existence of compromised energy-generating pathways in myopathic muscles that may have had consequences on the muscle contraction and tension development, as in vivo, also during the post-mortem period. Considering the overall reduced glycolytic potential and the myodegenerative processes associated with WB condition, we speculate that the higher pHu of WB meat might be the outcome of a drastically impaired energy-generating pathway combined with a deficiency and/or a dysfunction of muscle ATPases, having consequences also on muscle fiber contraction degree.

Phosphoproteome analysis of sarcoplasmic and myofibrillar proteins in stress-induced dysfunctional broiler pectoralis major muscle

Postmortem changes of sarcoplasmic and myofibrillar protein phosphorylation in pectoralis major (PM) muscle of broilers under pre-slaughter stress were investigated. Broiler chickens were randomly distributed to unstressed control and transport under high environmental temperature groups. PM muscle samples of transport-stressed broilers were classified into normal or pale, soft and exudative (PSE)-like. Sarcoplasmic fraction in PSE-like meat had a higher global phosphorylation level than that in normal meat at the early postmortem stage. The myofibrillar proteins showed diverse phosphorylation patterns at different postmortem times. The stress-induced highly phosphorylated sarcoplasmic proteins were glycometabolic enzymes, which partially contributed to accelerated glycolysis rate. The phosphorylation levels of most sarcomeric proteins identified in the myofibrillar fraction were affected by postmortem time, implying their roles in regulating muscle rigor mortis development. This work contributes to a deeper understanding of the biochemical processes that may lead to stress-induced changes in meat quality.

Stable reference genes for expression studies in breast muscle of normal and white striping-affected chickens.

The normalization with proper reference genes is a crucial step to obtain accurate mRNA expression levels in quantitative PCR (qPCR) studies. Therefore, in this study, 10 reference candidate genes were evaluated to determine their stability in normal pectoralis major muscle of broilers and those counterparts affected with White Striping (WS) myopathy at 42days age. Four different tools were used for ranking the most stable genes: GeNorm, NormFinder, BestKeeper and Comparative Ct (ΔCt), and a general ranking was performed using the RankAggreg tool to select the best reference genes among all tools. From the 10 genes evaluated in the breast muscle of broilers, 8 were amplified. Most of the algorithms/tools indicated the same two genes, RPL30 and RPL5, as the most stable in the broilers breast muscle. In addition, there was agreement among the tools for the least stable genes: MRPS27, GAPDH and RPLP1 in the broilers breast muscle. Therefore, it is interesting to note that even with different tools for evaluating gene expression, there was consensus on the most and least stable genes. These results indicate that the Ribosomal protein L30 (RPL30) and Ribosomal protein L5 (RPL5) can be recommended for accurate normalization in qPCR studies with chicken pectoralis major muscle affected with White Striping and other myopathies.