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.ArticleFrontiers in Pharmacology.11:579(2020)journal articl

Two Novel Glycoside Hydrolases Responsible for the Catabolism of Cyclobis-(1→6)-α-nigerosyl

The actinobacterium Kribbella flavida NBRC 14399(T) produces cyclobis-(1 -> 6)-alpha-nigerosyl (CNN), a cyclic glucotetraose with alternate alpha-(1 -> 6)- and alpha-(1 -> 3)-glucosidic linkages, from starch in the culture medium. We identified gene clusters associated with the production and intracellular catabolism of CNN in the K. flavida genome. One cluster encodes 6-alpha-glucosyl-transferase and 3-alpha-isomaltosyltransferase, which are known to coproduce CNN from starch. The other cluster contains four genes annotated as a transcriptional regulator, sugar transporter, glycoside hydrolase family (GH) 31 protein (Kfla1895), and GH15 protein (Kfla1896). Kfla1895 hydrolyzed the alpha-(1 -> 3)-glucosidic linkages of CNN and produced isomaltose via a possible linear tetrasaccharide. The initial rate of hydrolysis of CNN (11.6 s(-1)) was much higher than that of panose (0.242 s(-1)), and hydrolysis of isomaltotriose and nigerose was extremely low. Because Kfla1895 has a strong preference for the alpha-(1 -> 3)-isomaltosyl moiety and effectively hydrolyzes the alpha-(1 -> 3)-glucosidic linkage, it should be termed 1,3-alpha-isomaltosidase. Kfla1896 effectively hydrolyzed isomaltose with liberation of beta-glucose, but displayed low or no activity toward CNN and the general GH15 enzyme substrates such as maltose, soluble starch, or dextran. The k(cat)/K-m for isomaltose (4.81 +/- 0.18 s(-1) mM(-1)) was 6.9- and 19-fold higher than those for panose and isomaltotriose, respectively. These results indicate that Kfla1896 is a new GH15 enzyme with high substrate specificity for isomaltose, suggesting the enzyme should be designated an isomaltose glucohydrolase. This is the first report to identify a starch-utilization pathway that proceeds via CNN

Rapid growth rate results in remarkably hardened breast in broilers during the middle stage of rearing: A biochemical and histopathological study.

The high incidence of meat of impaired quality poses a serious problem in the poultry industry. In recent years, the incidence of the pectoralis major muscle that appeared pale colored, remarkably hardened, and exudative, called "wooden breast" or "woody breast" has increased in slaughter houses. In the present study, 19-day-old Ross 308 broiler chickens affected (n = 10) and unaffected (n = 10) with remarkably hardened breast were selected from a commercial broiler farm, and reared to 55 days of age under a controlled environment. Among the affected birds, 5 of 10 birds appeared exhausted with markedly suppressed weight gain and 4 of 10 birds died during the rearing period. In contrast, all unaffected birds survived and most gained weight. Four of 10 unaffected birds lost the ability of back-to-back wing contact by the late stage of rearing. The biochemical analysis of blood plasma samples of 20-day-old birds revealed that creatine kinase and L-aspartate aminotransferase values in most affected birds were higher than those in unaffected birds; however, these values in unaffected birds increased rapidly with lost wing contactability and increasing age. Postmortem examinations revealed that the mean diameter of myofibers in affected birds was smaller than that in unaffected birds. Moreover, symptoms of degenerative and regenerative muscles were observed in most birds in both groups. Among them, a decrease in, or defect of, the characteristic polygonal shape of myofibers was the most common change within the pectoralis major muscles in both groups. The present study demonstrated that broilers affected with remarkably hardened breast during the middle stage of rearing would have suppressed physical status and weight gain, or would die. It was suggested that rapid growth in broilers might be a cause of remarkably hardened breast

Increased serum malondialdehyde concentration in cows with subclinical ketosis

The purpose of this study is to compare the assessment of pre- and postpartum oxidative stress-related causal indicators and other metabolites in cows with postpartum subclinical ketosis (SCK). The prepartum serum malondialdehyde concentration and body condition score (BCS) were elevated in the SCK cows (n=17) compared to healthy controls (n=12), while the insulin sensitivity check index was lower in the SCK cows than in the controls. Oxidative stress is enhanced in cows with prepartum higher BCS, causing decreased insulin sensitivity, and may be associated with onset of postpartum SCK. However, paraoxonase alone might be insufficient to assess the antioxidant state because of no difference in pre- and postpartum activities between the two groups

Rapid growth rate results in remarkably hardened breast in broilers during the middle stage of rearing : A biochemical and histopathological study

The high incidence of meat of impaired quality poses a serious problem in the poultry industry. In recent years, the incidence of the pectoralis major muscle that appeared pale colored, remarkably hardened, and exudative, called “wooden breast” or “woody breast” has increased in slaughter houses. In the present study, 19-day-old Ross 308 broiler chickens affected (n = 10) and unaffected (n = 10) with remarkably hardened breast were selected from a commercial broiler farm, and reared to 55 days of age under a controlled environment. Among the affected birds, 5 of 10 birds appeared exhausted with markedly suppressed weight gain and 4 of 10 birds died during the rearing period. In contrast, all unaffected birds survived and most gained weight. Four of 10 unaffected birds lost the ability of back-to-back wing contact by the late stage of rearing. The biochemical analysis of blood plasma samples of 20-day-old birds revealed that creatine kinase and L-aspartate aminotransferase values in most affected birds were higher than those in unaffected birds; however, these values in unaffected birds increased rapidly with lost wing contactability and increasing age. Postmortem examinations revealed that the mean diameter of myofibers in affected birds was smaller than that in unaffected birds. Moreover, symptoms of degenerative and regenerative muscles were observed in most birds in both groups. Among them, a decrease in, or defect of, the characteristic polygonal shape of myofibers was the most common change within the pectoralis major muscles in both groups. The present study demonstrated that broilers affected with remarkably hardened breast during the middle stage of rearing would have suppressed physical status and weight gain, or would die. It was suggested that rapid growth in broilers might be a cause of remarkably hardened breast

Nutrition During the Early Rearing Period Affects the Incidence of Wooden Breasts in Broilers

This study aimed to evaluate the relationship between early nutrition and the incidence of wooden breasts (WB) in broilers. Sixteen male and twenty female neonatal ROSS 308 broiler chicks were divided equally into four flocks. From 0–12 days of age, starter diet H, composed of 22.4% crude protein (CP), 6.6% crude fat (CF), 1.25% lysine, 0.48% methionine, and ≥3,070 kcal/kg metabolizable energy (ME), was fed to two flocks, and starter diet L, composed of 19.9% CP, 2.5% CF, 1.04% lysine, 0.38% methionine, and ≥2,930 kcal/kg ME, was fed to the remaining two flocks. All the flocks were fed the same commercial finisher diet, composed of 20.3% CP, 7.5% CF, 1.18% lysine, 0.44% methionine, and ≥3,300 kcal/kg ME, from 12–47 days of age. The birds were weighed every 2–5 days, subjected to a wing-lift test, and histology was conducted on the pectoralis major muscle tissue samples from all the birds necropsied at 47 days of age. Significant differences in the mean body weight between groups H and L were observed during 6–16 days and 24–26 days of age in males and during 6–26 days of age in females. Regarding the score evaluation of the individual lesions reflecting wooden breast, the birds in which back-to-back wing contact was not possible had higher lesion scores than those in which back-to-back wing contact was possible. The absence of back-to-back wing contact appeared more frequently in flocks fed the starter diet L, particularly in males. These results indicate that inappropriate nutrition levels in the starter diet increase the incidence of WB. Therefore, avoiding early nutrition deficits is a cost-effective feeding strategy

Nutrition During the Early Rearing Period Affects the Incidence of Wooden Breasts in Broilers

This study aimed to evaluate the relationship between early nutrition and the incidence of wooden breasts (WB) in broilers. Sixteen male and twenty female neonatal ROSS 308 broiler chicks were divided equally into four flocks. From 0–12 days of age, starter diet H, composed of 22.4% crude protein (CP), 6.6% crude fat (CF), 1.25% lysine, 0.48% methionine, and ≥3,070 kcal/kg metabolizable energy (ME), was fed to two flocks, and starter diet L, composed of 19.9% CP, 2.5% CF, 1.04% lysine, 0.38% methionine, and ≥2,930 kcal/kg ME, was fed to the remaining two flocks. All the flocks were fed the same commercial finisher diet, composed of 20.3% CP, 7.5% CF, 1.18% lysine, 0.44% methionine, and ≥3,300 kcal/kg ME, from 12–47 days of age. The birds were weighed every 2–5 days, subjected to a wing-lift test, and histology was conducted on the pectoralis major muscle tissue samples from all the birds necropsied at 47 days of age. Significant differences in the mean body weight between groups H and L were observed during 6–16 days and 24–26 days of age in males and during 6–26 days of age in females. Regarding the score evaluation of the individual lesions reflecting wooden breast, the birds in which back-to-back wing contact was not possible had higher lesion scores than those in which back-to-back wing contact was possible. The absence of back-to-back wing contact appeared more frequently in flocks fed the starter diet L, particularly in males. These results indicate that inappropriate nutrition levels in the starter diet increase the incidence of WB. Therefore, avoiding early nutrition deficits is a cost-effective feeding strategy