Gene Expression Essential for Myostatin Signaling and Skeletal Muscle Development Is Associated With Divergent Feed Efficiency in Pedigree Male Broilers

Background: Feed efficiency (FE, gain to feed) is an important genetic trait as 70% of the cost of raising animals is due to feed costs. The objective of this study was to determine mRNA expression of genes involved in muscle development and hypertrophy, and the insulin receptor-signaling pathway in breast muscle associated with the phenotypic expression of FE.Methods: Breast muscle samples were obtained from Pedigree Male (PedM) broilers (8 to 10 week old) that had been individually phenotyped for FE between 6 and 7 week of age. The high FE group gained more weight but consumed the same amount of feed compared to the low FE group. Total RNA was extracted from breast muscle (n = 6 per group) and mRNA expression of target genes was determined by real-time quantitative PCR.Results: Targeted gene expression analysis in breast muscle of the high FE phenotype revealed that muscle development may be fostered in the high FE PedM phenotype by down-regulation several components of the myostatin signaling pathway genes combined with upregulation of genes that enhance muscle formation and growth. There was also evidence of genetic architecture that would foster muscle protein synthesis in the high FE phenotype. A clear indication of differences in insulin signaling between high and low FE phenotypes was not apparent in this study.Conclusion: These findings indicate that a gene expression architecture is present in breast muscle of PedM broilers exhibiting high FE that would support enhanced muscle development-differentiation as well as protein synthesis compared to PedM broilers exhibiting low FE

Enrichment of Autophagy and Proteosome Pathways in Breast Muscle of Feed Efficient Pedigree Male Broilers

Background: Feed efficiency (FE) is an important genetic trait in poultry and livestock. Autophagy (self-eating) and proteosomes are cellular processes that remove damaged cell components (e.g., proteins, organelles). As evidence of extensive protein oxidation was observed in Pedigree Male (PedM) broilers exhibiting a low FE (LFE) phenotype compared to a high FE (HFE) phenotype, the main goal of this study was to assess gene and protein expression of the autophagy and proteosome pathways in breast muscle obtained in PedM broilers exhibiting HFE and LFE phenotypes.Methods: Feed efficiency was calculated as weight gain divided by feed intake gain in individual PedM broilers that were measured between 6 and 7 weeks of age. Targeted gene expression was conducted on breast muscle using quantitative real-time polymerase chain reaction (qPCR) to determine mRNA expression of genes associated with the autophagy pathway; AMP-activated protein kinase alpha 1 (AMPKα1), mammalian target of rapamycin (mTOR), Beclin 1, and autophagy genes (Atg) 3, Atg7, and Atg16L1. Binomial distribution analysis was conducted on transcriptomic and data obtained by RNAseq and shotgun proteomics, respectively on the same set of tissues for genes associated with autophagy, vacuole formation, and proteosome expression.Results: Greater efficiency was attained in the HFE PedM broilers by greater weight gain on the same amount of feed consumed resulting in FEs of 0.65 ± 0.01 and 0.46 ± 0.01 in the HFE and LFE phenotypes, respectively. Targeted mRNA expression analysis revealed significant (P < 0.05) elevations in AMPKa1, mTOR, Atg16L1, and Atg7 and a marginal (P = 0.07) elevation in Beclin1. Binomial distribution analysis transcriptomic and proteomic data revealed significant skews favoring autophagy-, vacuole-, and proteosome-related genes in the HFE phenotype. These results indicate that the autophagy and proteosome expression is enhanced in the HFE compared to the LFE pedigree male broiler phenotype suggesting that protein and organelle quality control may be enhanced in high feed efficiency

Potential Role of Ribonucleotide Reductase Enzyme in Mitochondria Function and Woody Breast Condition in Broiler Chickens

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

Nutritional Strategies to Improve Meat Quality and Composition in the Challenging Conditions of Broiler Production: A Review

Poultry meat is becoming one of the most important animal protein sources for human beings in terms of health benefits, cost, and production efficiency. Effective genetic selection and nutritional programs have dramatically increased meat yield and broiler production efficiency. However, modern practices in broiler production result in unfavorable meat quality and body composition due to a diverse range of challenging conditions, including bacterial and parasitic infection, heat stress, and the consumption of mycotoxin and oxidized oils. Numerous studies have demonstrated that appropriate nutritional interventions have improved the meat quality and body composition of broiler chickens. Modulating nutritional composition [e.g., energy and crude protein (CP) levels] and amino acids (AA) levels has altered the meat quality and body composition of broiler chickens. The supplementation of bioactive compounds, such as vitamins, probiotics, prebiotics, exogenous enzymes, plant polyphenol compounds, and organic acids, has improved meat quality and changed the body composition of broiler chickens

Whole Genome Resequencing of Arkansas Progressor and Regressor Line Chickens to Identify SNPs Associated with Tumor Regression

Arkansas Regressor (AR) chickens, unlike Arkansas Progressor (AP) chickens, regress tumors induced by the v-src oncogene. To better understand the genetic factors responsible for this tumor regression property, whole genome resequencing was conducted using Illumina Hi-Seq 2 × 100 bp paired-end read method (San Diego, CA, USA) with AR (confirmed tumor regression property) and AP chickens. Sequence reads were aligned to the chicken reference genome (galgal5) and produced coverage of 11× and 14× in AR and AP, respectively. A total of 7.1 and 7.3 million single nucleotide polymorphisms (SNPs) were present in AR and AP genomes, respectively. Through a series of filtration processes, a total of 12,242 SNPs were identified in AR chickens that were associated with non-synonymous, frameshift, nonsense, no-start and no-stop mutations. Further filtering of SNPs based on read depth ≥ 10, SNP% ≥ 0.75, and non-synonymous mutations identified 63 reliable marker SNPs which were chosen for gene network analysis. The network analysis revealed that the candidate genes identified in AR chickens play roles in networks centered to ubiquitin C (UBC), phosphoinositide 3-kinases (PI3K), and nuclear factor kappa B (NF-kB) complexes suggesting that the tumor regression property in AR chickens might be associated with ubiquitylation, PI3K, and NF-kB signaling pathways. This study provides an insight into genetic factors that could be responsible for the tumor regression property

Water properties in intact wooden breast fillets during refrigerated storage

ABSTRACT: The wooden breast (WB) condition notably alters moisture content and water holding capacity (WHC) in broiler breast fillets. The purpose of this study was to investigate water properties during refrigerated storage from 4 h to 168 h postmortem using time domain nuclear magnetic resonance (TD-NMR). Water properties measured included mobility (T), proportion (P), and abundance per 100 g of meat (A). Changes in meat quality indicators including compression force, color, pH, cumulative purge loss, and proximate composition were also measured. Compression force and energy of the WB fillets were higher than normal fillets (P < 0.05). Slopes of changes in lightness of the WB and normal fillets were different in skin and bone side (P < 0.05). The slope of the purge loss from the WB fillets was higher than the normal fillets (P < 0.05). Time domain nuclear magnetic resonance analysis showed 4 water populations in intact broiler fillets with transverse relaxation time (T2) constants at approximately 4 to 5 milliseconds (ms) (designated as 2b, corresponding to hydration water or bound water), 40 to 60 ms (designated as 21, corresponding to intra-myofibrillar water or immobilized water), 80 to 210 ms (designated as 22a, corresponding to extra-myofibrillar water or free water with lower mobility) and 210 to 500 ms (designated as 22b, corresponding to extra-myofibrillar water or free water with higher mobility) during early postmortem storage (between 4 h and 72 h postmortem) and only 3 populations (2b, 21, and 22a) after 72 h postmortem. There were interaction effects (P < 0.05) between storage time and WB condition for all water properties except T2b, A2b/100 g, and T22b. The linear change of T21, P21, A21/100 g, T22a, A22a/100 g, P22b, and A22b/100 g in stored WB samples were different from the normal fillets (P < 0.05). During storage, P21 and A21/100 g of the WB fillets exhibited faster linear increases than those of the normal fillets, whereas T21 and T22a of the normal fillets and A22a/100 g, P22b, and A22b/100 g of the WB fillets showed faster linear decreases (P < 0.05). Our data demonstrate that the WB condition affects changes in water properties in broiler fillets during postmortem refrigerated storage

Mitochondrial metabolism: a driver of energy utilisation and product quality?

High feed efficiency is a very desirable production trait as it positively influences resource utilisation, profitability and environmental considerations, albeit at the possible expense of product quality. The modern broiler is arguably the most illustrative model species as it has been transformed over the past half century into an elite feed converter. Some producers are currently reporting that 42-day-old birds gain 1 kg of wet weight for every 1.35 kg of dry weight consumed. Its large breast muscle is exclusively composed of large, low mitochondrial-content Type IIB fibres, which may contribute to low maintenance costs and high efficiency. In an effort to gain a better understanding of individual variation in chicken feed efficiency, our group has been exploring the biology of the mitochondrion at multiple levels of organisation. The mitochondrion is the organelle where much biochemical energy transformation occurs in the cell. Using Cobb-Vantress industrial birds as our primary experimental resource, we have explored the tissue content, structure and function of the mitochondrion and its relationship to growth, development, efficiency and genetic background. While much remains to be understood, recent highlights include (1) variation in muscle mitochondrial content that is associated with performance phenotypes, (2) altered muscle mitochondrial gene and protein expression in birds differing in feed efficiency, (3) variation in isolated mitochondrial function in birds differing in feed efficiency and (4) evidence for an unexpected role for the mitochondrially localised progesterone receptor in altering bird muscle metabolism. Mitochondrial function is largely conserved across the vertebrates, so the same metabolic principles appear to apply to the major production species, whether monogastric or ruminant. A speculative role for the mitochondria in aspects of meat quality and in influencing postmortem anaerobic metabolism will conclude the manuscript

Copy number variation study in Japanese quail associated with stress related traits using whole genome re-sequencing data.

Copy number variation (CNV) is a major driving factor for genetic variation and phenotypic diversity in animals. To detect CNVs and understand genetic components underlying stress related traits, we performed whole genome re-sequencing of pooled DNA samples of 20 birds each from High Stress (HS) and Low Stress (LS) Japanese quail lines using Illumina HiSeq 2×150 bp paired end method. Sequencing data were aligned to the quail genome and CNVnator was used to detect CNVs in the aligned data sets. The depth of coverage for the data reached to 41.4x and 42.6x for HS and LS birds, respectively. We identified 262 and 168 CNV regions affecting 1.6 and 1.9% of the reference genome that completely overlapped 454 and 493 unique genes in HS and LS birds, respectively. Ingenuity pathway analysis showed that the CNV genes were significantly enriched to phospholipase C signaling, neuregulin signaling, reelin signaling in neurons, endocrine and nervous development, humoral immune response, and carbohydrate and amino acid metabolisms in HS birds, whereas CNV genes in LS birds were enriched in cell-mediated immune response, and protein and lipid metabolisms. These findings suggest CNV genes identified in HS and LS birds could be candidate markers responsible for stress responses in birds

Proteomic analyses on chicken breast meat with white striping myopathy

ABSTRACT: White striping (WS) is an emerging myopathy that results in significant economic losses as high as $1 billion (combined with losses derived from other breast myopathies including woody breast and spaghetti meat) to the global poultry industry. White striping is detected as the occurrence of white lines on raw poultry meat. The exact etiologies for WS are still unclear. Proteomic analyses of co-expressed WS and woody breast phenotypes previously demonstrated dysfunctions in carbohydrate metabolism, protein synthesis, and calcium buffering capabilities in muscle cells. In this study, we conducted shotgun proteomics on chicken breast fillets exhibiting only WS that were collected at approximately 6 h postmortem. After determining WS severity, protein extractions were conducted from severe WS meat with no woody breast (WB) condition (n = 5) and normal non-affected (no WS) control meat (n = 5). Shotgun proteomics was conducted by Orbitrap Lumos, tandem mass tag (TMT) analysis. As results, 148 differentially abundant proteins (|fold change|>1.4; p-value < 0.05) were identified in the WS meats compared with controls. The significant canonical pathways included BAG2 signaling pathway, glycogen degradation II, isoleucine degradation I, aldosterone signaling in epithelial cells, and valine degradation I. The potential upstream regulators include LIPE, UCP1, ATP5IF1, and DMD. The results of this study provide additional insights into the cellular mechanisms on the WS myopathy and meat quality

MicroRNA profiling associated with muscle growth in modern broilers compared to an unselected chicken breed

Background: Genetically selected modern broiler chickens have acquired outstanding production efficiency through rapid growth and improved feed efficiency compared to unselected chicken breeds. Recently, we analyzed the transcriptome of breast muscle tissues obtained from modern pedigree male (PeM) broilers (rapid growth and higher efficiency) and foundational Barred Plymouth Rock (BPR) chickens (slow growth and poorer efficiency). This study was designed to investigate microRNAs that play role in rapid growth of the breast muscles in modern broiler chickens. Results: In this study, differential abundance of microRNA (miRNA) was analyzed in breast muscle of PeM and BPR chickens and the results were integrated with differentially expressed (DE) mRNA in the same tissues. A total of 994 miRNA were identified in PeM and BPR chicken lines from the initial analysis of small RNA sequencing data. After filtering and statistical analyses, the results showed miR-2131-5p, miR-221-5p, miR-126-3p, miR-146b-5p, miR-10a-5p, let-7b, miR-125b-5p, and miR-146c-5p up-regulated whereas miR-206 down-regulated in PeM compared to BPR breast muscle. Based on inhibitory regulations of miRNAs on the mRNA abundance, our computational analysis using miRDB, an online software, predicated that 118 down-regulated mRNAs may be targeted by the up-regulated miRNAs, while 35 up-regulated mRNAs appear to be due to a down-regulated miRNA (i.e., miR-206). Functional network analyses of target genes of DE miRNAs showed their involvement in calcium signaling, axonal guidance signaling, and NRF2-mediated oxidative stress response pathways suggesting their involvement in breast muscle growth in chickens. Conclusion: From the integrated analyses of differentially expressed miRNA-mRNA data, we were able to identify breast muscle specific miRNAs and their target genes whose concerted actions can contribute to rapid growth and higher feed efficiency in modern broiler chickens. This study provides foundation data for elucidating molecular mechanisms that govern muscle growth in chickens