This review is a summary of a Poultry Science Association symposium addressing myopathies in broilers\' breast meat, focusing on the interactions between genetics, nutrition, husbandry, and meat processing. The Pectoralis major myopathies (woody breast [WB]; white striping [WS]; spaghetti meat [SM]) and Pectoralis minor (\"feathering\") are described, followed by discussing their prevalence, potential causes, current and future ways to mitigate, as well as detection methods (in live birds and meat) as well as ways to utilize affected meat. Overall, breast myopathies remain an important focus across the poultry industry and whilst a lot of data and knowledge has been gathered, it is clear that there is still a lot to understand. As there are multiple factors impacting the occurrence of breast myopathies, their reduction relies on a holistic approach. Ongoing balanced breeding strategies by poultry breeders is targeting the longer-term genetic component but comprehending the significant influence from nongenetic factors (short-term solutions such as nutrition) remains a key area of opportunity. Consequently, understanding the physiology and biological needs of the muscle through the life of the bird is critical to reduce the myopathies (e.g., minimizing oxidative stress) and gain more insight into their etiology.

Relationships between texture measurements and meat water properties were investigated in raw intact broiler breast fillets with the wooden breast (WB) condition. Texture measurements included subjective WB scores and blunt Meullenet-Owens Razor Shear (BMORS). Water properties were determined with low-field nuclear magnetic resonance (LF-NMR). Spearman correlation was used to estimate relationships between WB scores and water properties, while Pearson correlation was used for relationships between BMORS force and water properties. LF-NMR measurements exhibited 3 water components: protein-associated or hydration water T2b, intra-myofibrillar water or immobilized water T21, and extra-myofibrillar water or free water T22 in chicken breast meat. Significant and strong Spearman correlations were found between the WB scores and T21 time constant, the abundance (normalized areas) of T22, and the proportion of T21 and T22 (rs > 0.60, P < 0.001). Strong Pearson correlations (r = 0.72) were noted only between the T21 time constant and BMORS force. These results demonstrate that water may contribute to the specific texture characteristics measured with subjective WB scoring (palpable hardness and rigidity) and BMORS (hardness and share force) in raw broiler breast fillets with the WB condition.

OBJECTIVE: The aim of this study was to investigate the roles of ribonucleotide reductase subunit M2 (RRM2; subunit of ribonucleotide reductase) in severe woody breast (WB) and normal breast muscles.
METHODS: 40 8-week-old male Ross-708 broiler chickens.
METHODS: Quantitative PCR was performed to determine gene expression, and commercial ELISA/assay kits were used to obtain several enzymatic activities.
RESULTS: Results showed that RRM2 activity (P = .0002) and RRM2 (P = .05) and hydroxymethylbilane synthase expression (impaired oxygen transport and metabolism, P = .002) were reduced in WB, while caveolin-3 (defected membrane integrity, P = .09), endoglin (increased fibrosis, P = .06), and secreted protein acidic rich in cysteine (metabolic dysregulation, P = .09) expression tended to increase in WB. WB tended to have increased levels of homocysteine (P = .06), aspartate aminotransferase mitochondria (P = .02), pyruvate kinase (P = .04), DNA damage (P = .06), creatine kinase (P = .05), and triglyceride (P = .002) but decreased ATPase activity (P = .01), all indicating mitochondria dysfunction and tissue damage.
CONCLUSIONS: In this study, differences in various enzyme activities and increased DNA damage suggest that RRM2-mediated mitochondrial abnormalities may play a role in WB myopathy.

Previous work has shown that dietary treatments affect woody breast (WB) incidence differently, which indicates that gut conditions such as gut barrier function, inflammation, and oxidative stress are likely related to WB. In this study, dietary supplementation with antibiotics (bacitracin) or probiotics (Bacillus subtilis) was investigated for their effects on the expression of transcripts related to gut barrier function, inflammation, and oxidative stress in the mucus lining of the jejunum from broilers with or without WB. A split-plot experimental design was used in this study. The dietary treatments served as the main plot factor and the breast muscle condition was the subplot factor. On d 41, jejunum mucus was collected from 1 bird from each of 3 replicate pens in each 3 dietary treatment groups that exhibited WB and an additional bird that contained a normal breast (3 biological replicates/treatment/phenotype; 3 × 3 × 2, total N = 18). Total RNA was extracted using a commercial RNA extraction kit. The expression levels of CLDN1, MUC6, TLR2A, TLR2B, TLR4, IFN-γ, IL-1β, IL-8L1, IL-10, NOS2, and SOD were determined using 2-step RT-qPCR analysis. The gene expression difference in ΔCt values was determined after normalizing with the chicken 18S rRNA gene. When the significant differences occurred between treatments, the relative fold change was calculated using the ΔΔCt method and the significance level was calculated. The PROC GLM procedure of SAS 9.4 was used, and the level of significance was set at P ≤ 0.05. There were no significant interactive effects between diet and the breast muscle condition on the expression of any of the genes tested. However, birds with WB exhibited higher MUC6 (P < 0.0001) gene expression levels than birds with normal breast muscles. In addition, the expression of SOD decreased in birds that were fed the antibiotic diet when compared to birds that were fed the probiotic diet (P = 0.014). In conclusion, WB identified in broilers tested in the current study is attributed to increased expression of mucin, indicating a correlation between WB incidence and gel-forming mucin secretion and pathogen signaling.

This study was conducted on chicken pectoralis major muscle with different wooden breast severity in combination with different sampling locations to investigate the effects of wooden breast syndrome on protein traits and total myofiber area, and their associations. Contents of sarcoplasmic, salt-soluble myofibrillar and salt-insoluble protein and proportion of total myofiber area significantly declined with increasing severity in the superficial part of muscle, whereas the amount of heat-soluble/insoluble collagen and protein denaturation as well as the area of degenerated myofibers, connective tissue and cellular infiltrates increased. Myofibril protein content indicators showed strong positive correlations to total myofiber area. Moreover, PCA results indicated that severe wooden breast is positively linked to muscle collagen content and to protein denaturation. Our results suggest that decrease in sarcoplasmic and myofibrillar proteins is associated with reduction of myofiber area. In turn, the muscle fibers are replaced by connective tissue, accompanied by excessive myofibrillar and sarcoplasmic protein denaturation.

Bioelectrical impedance analysis (BIA) was established to quantify diverse cellular characteristics. This technique has been widely used in various species, such as fish, poultry, and humans for compositional analysis. This technology was limited to offline quality assurance/detection of woody breast (WB); however, inline technology that can be retrofitted on the conveyor belt would be more helpful to processors. Freshly deboned (n = 80) chicken breast fillets were collected from a local processor and analyzed by hand-palpation for different WB severity levels. Data collected from both BIA setups were subjected to supervised and unsupervised learning algorithms. The modified BIA showed better detection ability for regular fillets than the probe BIA setup. In the plate BIA setup, fillets were 80.00% for normal, 66.67% for moderate (data for mild and moderate merged), and 85.00% for severe WB. However, hand-held BIA showed 77.78, 85.71, and 88.89% for normal, moderate, and severe WB, respectively. Plate BIA setup is more effective in detecting WB myopathies and could be installed without slowing the processing line. Breast fillet detection on the processing line can be significantly improved using a modified automated plate BIA.

The cellular events leading to the development of the woody breast myopathy in broiler breast muscle are unclear. Affected woody breast muscle exhibits muscle fiber degeneration/regeneration, connective tissue accumulation, and adverse morphological changes in mitochondria. Ribonucleotide reductase (RNR) is an enzyme for the synthesis of dNTP, which is important for mitochondria DNA content (mtDNA). RNR consists of two subunits: RRM1/RRM2. A decrease in RRM2 is associated with a decrease in mtDNA and mitochondria proteins, leading to impaired ATP production. The objective of this study was to investigate potential RNR differences between woody breast (WB) and normal (N) breast muscle by examining RRM2 expression and associated pathways. Gene expression and enzyme activities were examined by qPCR and commercial kits. Results showed that RRM2 expression reduced for WB (p = 0.01) and genes related to mitochondria, including ATP6 (p = 0.03), COX1 (p = 0.001), CYTB (p = 0.07), ND2 (p = 0.001) and ND4L (p = 0.03). Furthermore, NDUFB7 and COX 14, which are related to mitochondria and ATP synthesis, tended to be reduced in WB. Compared to N, GLUT1 reduced for WB (p = 0.05), which is responsible for glucose transport in cells. Consequently, PDK4 (p = 0.0001) and PPARG (p = 0.008) increased in WB, suggesting increased fatty acid oxidation. Citric synthase activity and the NAD/NADH ratio (p = 0.02) both reduced for WB, while WB increased CHRND expression (p = 0.001), which is a possible indicator of high reactive oxygen species levels. In conclusion, a reduction in RRM2 impaired mitochondria function, potentially ATP synthesis in WB, by increasing fibrosis and the down-regulation of several genes related to mitochondria function.

The woody breast (WB) myopathy is an unintended outcome of fast broiler chicken growth and high breast muscle yields. Myodegeneration and fibrosis in the living tissue are results of hypoxia and oxidative stress driven by lack of blood supply to muscle fibers. The study aim was to titrate a vasodilator ingredient, inositol-stabilized arginine silicate (ASI), as a feed additive to improve blood flow and ultimately, breast meat quality. A total of 1,260 male Ross 708 broilers were assigned to: 1) a control basal diet, or the control diet plus increasing ASI: 2) 0.025% ASI, 3) 0.05% ASI, 4) 0.10% ASI, or 5) 0.15% ASI. At d 14, 28, 42, and 49, growth performance was measured on all broilers and serum from 12 broilers/diet was analyzed for creatine kinase and myoglobin presence. On d 42 and 49, 12 broilers/diet were measured for breast width, then left breast fillets were excised, weighed, palpated for WB severity, and visually scored for degree of white striping (WS). At 1 d postmortem, 12 raw fillets/treatment underwent compression force analysis, and at 2d postmortem, the same fillets were analyzed for water-holding capacity. mRNA was isolated from 6 right breasts/diet at both d 42 and 49 for qPCR quantification of myogenic gene expression. Birds fed the lowest dose of 0.025% ASI had a 5-point/3.25% feed conversion ratio reduction compared to birds fed 0.10% ASI over wk 4 to 6 and reduced serum myoglobin at 6-wk of age compared to the control. Breasts from birds fed 0.025% ASI received 42% greater normal WB scores at d 42 compared to control fillets. At d 49, breasts from broilers fed 0.10 and 0.15% ASI received 33% normal WB scores. At d 49, 0.025% AS-fed broiler breasts showed no severe WS. Increased myogenin expression was observed in 0.05 and 0.10% ASI breast samples on d 42 and myoblast determination protein-1 expression was upregulated in breasts from birds fed 0.10% ASI on d 49 compared to the control. Therefore, a dietary inclusion of 0.025, 0.10, or 0.15% ASI was beneficial in reducing WB and WS severity and promoting muscle growth factor gene expression at age of harvest without diminishing bird growth or breast muscle yields.

Genetic selection for breast yields and fewer days to market has inadvertent effects on broiler meat quality. Woody breast (WB) and white striping (WS) are pectoralis major myopathies prevalent in commercial broilers. Effects of voluntary exercise on these disorders, specifically, are unknown. A second-generation laser enrichment device shown to induce activity in Ross 308 and 708 birds was implemented using 1,360 Ross 708 broilers randomly assigned to laser enrichment or control for 49 d. Laser-enriched birds were exposed to 6-min laser periods 4 times daily. Seventy focal birds were gait and contact dermatitis scored weekly. Blood was collected wk 5 to 7 from 56 broilers for serum corticosterone, myoglobin, and troponin. Seventy broilers were sampled for breast muscle width, fillet dimensions, and WB and WS at wk 6 and 7. One and 2-day postmortem, fillet compression force and water-holding capacity were measured. Serum corticosterone was reduced by up to 21% in laser-enriched birds wk 5 to 7 (P < 0.01). Serum myoglobin was increased in laser-enriched broilers by 5% on wk 5 (P < 0.01) but increased in control birds wk 6 to 7 by up to 13% (P < 0.01). Serum troponin was reduced in laser-enriched broilers by 9% at wk 5 (P < 0.01). Laser exposure increased breast width and fillet weight at d 42 by 1.08 cm (P < 0.05) and 30 g (P < 0.05). At d 49, fillet height was increased 0.42 cm in laser-enriched birds (P < 0.05). Laser enrichment reduced severe WS incidence at d 42 by 24% (P < 0.05) and on d 49 by 15% (P < 0.10). Severe WB score was numerically reduced by 11% in laser enrichment on d 42 and 18% on d 49 (P > 0.05). Water-holding capacity was improved in laser-enriched breasts (P < 0.01) and expression of myostatin and insulin-like growth factor 2 were increased on d 49 (P ≤ 0.01. Laser enrichment reduced markers of stress and muscle damage while improving breast muscle quality and is therefore a potential effective enrichment for commercial broilers.

Starting in approximately 2010, broiler breast meat myopathies, specifically woody breast meat, white striping, spaghetti meat, and gaping have increased in prevalence in the broiler meat industry. Omic methods have been used to elucidate compositional, genetic, and biochemical differences between myopathic and normal breast meat and have provided information on the factors that contribute to these myopathies. This review paper focuses on the genomic, transcriptomic, proteomic, metabolomic, and other omics research that has been conducted to unravel the molecular mechanisms involved in the development of these myopathies and their associated factors and potential causes. SIGNIFICANCE: This review manuscript summarizes poultry meat quality defects, also referred to as myopathies, that have been evaluated using omics methods. Genomics, transcriptomics, proteomics, metabolomics and other methodologies have been used to understand the genetic predisposition, the protein expression, and the biochemical pathways that are associated with the expression of woody breast meat, white striping, and other myopathies. This has allowed researchers and the industry to differentiate between chicken breast meat with and without myopathic muscle as well as the environmental and genetic conditions that contribute to differences in biochemical pathways and lead to the phenotypes associate with these different myopathies.