Blood Plasma Biomarkers for Woody Breast Disease in Commercial Broilers

Abstract

Woody breast (WB) myopathy results in poor muscle quality. The increasing incidence of WB over the last several years indicates a need for improved prediction or early diagnosis. We hypothesized that the use of body fluids, including blood, may be more suitable than breast muscle tissue in developing a minimally invasive diagnostic tool for WB detection. To identify potential early-age-biomarkers that may represent the potential onset of WB, blood samples were collected from 100, 4 wks old commercial male broilers. At 8 wks of age, WB conditions were scored by manual palpation. A total of 32 blood plasma samples (eight for each group of WB and non-WB control birds at two time points, 4 wks and 8 wks) were subjected to shotgun proteomics and untargeted metabolomics to identify differentially abundant plasma proteins and metabolites in WB broilers compared to non-WB control (Con) broilers. From the proteomics assay, 25 and 16 plasma proteins were differentially abundant (p < 0.05) in the 4 and 8 wks old samples, respectively, in WB compared with Con broilers. Of those, FRA10A associated CGG repeat 1 (FRAG10AC1) showed >2-fold higher abundance in WB compared with controls. In the 8 wks old broilers, 4 and 12 plasma proteins displayed higher and lower abundances, respectively, in WB compared with controls. Myosin heavy chain 9 (MYH9) and lipopolysaccharide binding protein (LBP) showed more than 2-fold higher abundances in WB compared with controls, while transferrin (TF) and complement C1s (C1S) showed more than 2-fold lower abundances compared with controls. From the untargeted metabolomics assay, 33 and 19 plasma metabolites were differentially abundant in birds at 4 and 8 wks of age, respectively, in WB compared with controls. In 4 wks old broilers, plasma 3-hydroxybutyric acid (3-HB) and raffinose concentrations showed the highest and lowest fold changes, respectively, in WB compared with controls. The blood plasma 3-HB and raffinose concentrations were confirmed with targeted biochemical assays. Blood biomarkers, such as 3-HB and raffinose, may be suitable candidate targets in the prediction of WB onset at early ages.

Identification of circulating metabolites associated with wooden breast and white striping

“…For example, increased expression of the LPL gene in wooden breast- affected pectoralis major suggests an increased uptake of fatty acids from circulating lipoproteins, as LPL encodes the rate-limiting enzyme in lipoprotein metabolism [ 31 , 39 ]. Evidence supporting altered lipid metabolism in wooden breast has been demonstrated in numerous studies including a recent metabolomics study by Kong et al (2021) who reported elevated levels of 3-hydroxybutyric acid in blood plasm from 4-week-old WB affected chickens [ 53 ].…”

Molecular and gene expression analyses of chicken oncomodulin and their association with breast myopathies in broilers

“…Briefly, live birds underwent physical palpation for WB prediction and at less than 10 min postslaughter, WB and/or WS conditions of the breast muscles (pectoralis major) were evaluated by visual examination. Based on a scoring scale of 0 to 3 ( Kang et al, 2020 ; Kong et al, 2021 ), they were classified into control (Con, n = 6; WB and WS scores = 0), WB (n = 7; WB score ≥ 2, WS score ≤ 1), and WB/WS (n = 7; both WB and WS scores ≥ 2) categories. Breast muscle samples were collected at less than 10 minutes post-mortem, immediately snap-frozen in dry ice, and stored at -80°C until further use.…”

Breast muscle white striping and serum corticosterone reduced in broilers exposed to laser environmental enrichment

“… Maharjan et al (2020) observed elevated plasma metabolites indicative of vascularity problems, localized hypoxia, and muscle atrophy. Kong et al (2021) identified multiple plasma proteins and metabolites that differed in WB-affected commercial broilers, possible WB predictors, as early as 4 wk of age. Santos et al (2021) detected increased plasma creatine kinase, aspartate transaminase, and lactate dehydrogenase, believed to be indicative of muscle damage, in conventional broilers with the greatest occurrence of breast myopathies.…”