Nest substrate and tool shape significantly affect the mechanics and energy requirements of avian eggshell puncture.

Some host species of avian obligate brood parasites reject parasitic eggs from their nest whereas others accept them, even though they recognize them as foreign. One hypothesis to explain this seemingly maladaptive behavior is that acceptors are unable to pierce and remove the parasitic eggshell. Previous studies reporting on the force and energy required to break brood parasites' eggshells were typically static tests performed against hard substrate surfaces. Here, we considered host nest as a substrate to simulate this potentially critical aspect of the natural context for egg puncture while testing the energy required to break avian eggshells. Specifically, as a proof of concept, we punctured domestic chicken eggs under a series of conditions: varying tool shape (sharp versus blunt), tool dynamics (static versus dynamic) and the presence of natural bird nests (of three host species). The results show a complex set of statistically significant interactions between tool shapes, puncture dynamics and nest substrates. Specifically, the energy required to break eggs was greater for the static tests than for the dynamic tests, but only when using a nest substrate and a blunt tool. In turn, in the static tests, the addition of a nest significantly increased energy requirements for both tool types, whereas during dynamic tests, the increase in energy associated with the nest presence was significant only when using the sharp tool. Characterizing the process of eggshell puncture in increasingly naturalistic contexts will help in understanding whether and how hosts of brood parasites evolve to reject foreign eggs.

Effect of vitamin E and alpha lipoic acid on intestinal development associated with wooden breast myopathy in broilers.

Intestinal development is closely associated with inflammatory wooden breast (WB) myopathy. Vitamin E (VE) and alpha lipoic acid (ALA) with antioxidant and anti-inflammatory effects were used independently and in combination to evaluate their effects on intestinal developmental changes in ileal morphology and expression of genes related with gut nutrient transport, structure, and inflammation in broilers during the first 3 wk posthatch. A total of 160 newly hatched Ross 708 broiler chicks were randomly assigned into a control and 3 dietary treatments with 10 replicates of 4 birds each. Supplementation of VE (160 mg/kg) and ALA (500 mg/kg) independently and in combination were fed during the first 3 wk. At 1, 2, and 3 wk of age, one chick from each pen was harvested. Plasma VE concentration and ileal morphology were determined. Gene expression was measured by real-time quantitative PCR. Broilers in VE and combination of ALA and VE group had higher plasma VE concentration than the control and ALA group at 1, 2, and 3 wk of age (P < 0.01). All dietary treatments increased ileal villus height at 1 wk of age (P < 0.01) and decreased intraepithelial lymphocytes at 3 wk of age compared to the control (P ≤ 0.05). Combination of VE and ALA increased collagen type IV alpha 1 chain expression (P ≤ 0.05) and improved basement membrane structure indicating increased gut basement membrane integrity at 2 and 3 wk of age compared to the control. Expression of lipopolysaccharide-induced tumor necrosis factor-alpha factor associated with inflammation was decreased in all dietary treatments at 3 wk of age compared to the control (P < 0.01). Ileal morphology and gene expression were closely correlated with breast muscle morphology and gene expression. These results suggest that VE and ALA especially when they were combined in the diet had positive effects on mitigating intestinal inflammation and improving nutrient transport beginning at 1 wk of age, which is likely critical in reducing the severity of WB.

Supplementation of vitamin E and omega-3 fatty acids during the early posthatch period on intestinal morphology and gene expression differentiation in broilers.

Early posthatch nutrition is important for gut health. Vitamin E (VE) and omega-3 (n-3) fatty acids can improve gut health through antioxidative and anti-inflammatory effects. The objective of this study was to identify the effects of VE, n-3 fatty acids, and combination of both during the starter phase (0-10 d) or grower phase (11-24 d) on intestinal morphology and expression of genes associated with gut health. A total of 210 Ross 708 broilers were randomly assigned into 7 treatments with 10 replicates of 3 birds each. The control group was fed a corn-soybean meal-basal diet during the entire study (0-58 d). Supplementation of VE (200 IU/kg), n-3 fatty acids (n-6/n-3 ratio of 3.2:1), and combination of both were fed during the starter phase (0-10 d) or grower phase (11-24 d). All of the broilers were harvested at 58 d of age. Villus height, crypt depth, villus width, distance between villi, and number of intraepithelial lymphocytes were obtained. Expression of 21 genes was measured using NanoString analysis. Expression of solute carrier family 15 member 1 (P = 0.01) associated with peptide transport and mucin 2 (P = 0.03) related with intestinal mucus barrier was increased in the broilers supplemented with n-3 fatty acids in the grower diet compared with the control. Expression of solute carrier family 7 member 1 associated with amino acid transport was decreased in the group supplemented with n-3 fatty acids during the starter phase compared with the group supplemented with n-3 fatty acids (P = 0.01) or VE and n-3 fatty acids during the grower phase (P = 0.03). These data suggest that VE and n-3 fatty acids supplemented during the grower phase have a positive effect on improving nutrient transport with n-3 fatty acids supplementation in the grower diet showing the most beneficial effect. These findings can be used in the development of nutritional management strategies to improve broiler growth performance and meat quality.

Impacts of shade on cattle well-being in the beef supply chain.

Shade is a mechanism to reduce heat load providing cattle with an environment supportive of their welfare needs. Although heat stress has been extensively reviewed, researched, and addressed in dairy production systems, it has not been investigated in the same manner in the beef cattle supply chain. Like all animals, beef cattle are susceptible to heat stress if they are unable to dissipate heat during times of elevated ambient temperatures. There are many factors that impact heat stress susceptibility in beef cattle throughout the different supply chain sectors, many of which relate to the production system, that is, availability of shade, microclimate of environment, and nutrition management. The results from studies evaluating the effects of shade on production and welfare are difficult to compare due to variation in structural design, construction materials used, height, shape, and area of shade provided. Additionally, depending on operation location, shade may or may not be beneficial during all times of the year, which can influence the decision to make shade a permanent part of management systems. Shade has been shown to lessen the physiologic response of cattle to heat stress. Shaded cattle exhibit lower respiration rates, body temperatures, and panting scores compared with unshaded cattle in weather that increases the risk of heat stress. Results from studies investigating the provision of shade indicate that cattle seek shade in hot weather. The impact of shade on behavioral patterns is inconsistent in the current body of research, with some studies indicating that shade provision impacts behavior and other studies reporting no difference between shaded and unshaded groups. Analysis of performance and carcass characteristics across feedlot studies demonstrated that shaded cattle had increased ADG, improved feed efficiency, HCW, and dressing percentage when compared with cattle without shade. Despite the documented benefits of shade, current industry statistics, although severely limited in scope, indicate low shade implementation rates in feedlots and data in other supply chain sectors do not exist. Industry guidelines and third-party on-farm certification programs articulate the critical need for protection from extreme weather but are not consistent in providing specific recommendations and requirements. Future efforts should include: updated economic analyses of cost vs. benefit of shade implementation, exploration of producer perspectives and needs relative to shade, consideration of shade impacts in the cow-calf and slaughter plant segments of the supply chain, and integration of indicators of affective (mental) state and preference in research studies to enhance the holistic assessment of cattle welfare.

Effect of early posthatch supplementation of vitamin E and omega-3 fatty acids on the severity of wooden breast, breast muscle morphological structure, and gene expression in the broiler breast muscle.

Wooden breast (WB) has arisen primarily in the breast muscle of commercial broilers. It is characterized by palpation of a rigid pectoralis major (p. major) muscle and is under severe oxidative stress and inflammation. Previous studies have shown that vitamin E (VE) has antioxidant properties and omega-3 (n-3) fatty acids have an anti-inflammatory effect. The objectives of this study were to identify the effects of VE and n-3 fatty acids on the severity of WB, morphological structure of the p. major muscle, expression of genes likely associated with WB and to determine the most beneficial supplementation period. A total of 210 Ross 708 broilers were randomly assigned into 7 treatments with 10 replicates of 3 birds each. The control group received a corn-soybean meal basal diet during the entire study (0-58 d). Supplementation of VE (200 IU/kg), n-3 fatty acids (n-6/n-3 ratio of 3.2:1), or combination of both were fed during the starter phase (0-10 d) or grower phase (11-24 d). All broilers were harvested at 58 d of age. Morphological assessment of the p. major muscle included myofiber width, perimysial and endomysial connective tissue space, overall morphological structure, and scoring of WB microscopically. Gene expression was measured using nanostring analysis. Genes associated with muscle development and growth factors, inflammation, extracellular matrix, and glucose metabolism were differentially expressed in the p. major muscle of the broilers supplemented with VE in the grower diet. Greater than 2 times more giant myofibers (≥70 µm) were found in the group supplemented with VE and n-3 fatty acids in the starter diet compared with the group fed VE in the grower diet (P = 0.02). Microscopic evaluation showed that VE supplementation in the grower diet had a 16.19% increase in muscle with no WB compared with the control group (P = 0.05). These data suggest that supplementation of VE during the grower phase may reduce the severity of WB in broilers.

Research Note: The effect of selection for 16-week body weight on turkey serum metabolome.

The phenotype of modern commercial turkeys is substantially different than that of unselected, heritage turkey lines. These phenotypic changes have arisen from alterations in the genome/transcriptome, as well as the influence of many external factors on growth performance including nutrition, environment, and management. To investigate the phenotypic changes resulting from genetic selection for increased body weight, The Ohio State University maintains 2 unique genetic turkey lines: the randombred control (RBC2) line, which is comprised of genetics from 1960 era commercial turkeys and has been maintained without conscious selection for any trait; and the F line, which was originally selected from the RBC2 line and has been selected for increased 16 wk body weight for over 50 generations. This study used broad-spectrum mass-spectrometry profiling techniques to identify and quantify differences in the metabolome of the serum of F and RBC2 turkey lines. Serum samples from both F and RBC2 turkeys were subject to quantitative time of flight liquid chromatography tandem mass spectrometry analyses. Principle component analyses showed distinct populations of metabolites in the F vs. RBC2 serum, suggesting that increased body weight is associated with the accumulation of several metabolites. Comparing the spectral features to online databases resulted in the selection of 104 features with potentially identifiable chemical structures. Of these 104 features, 25 were found at higher levels in the serum of the RBC2 line turkeys, while 79 were found at a greater abundance in the F line turkeys. A more detailed analysis of these 104 features allowed for the putative identification of 49 compounds, which were clustered into 6 functional groups: 1) energy metabolism; 2) vitamins; 3) hormones and signaling molecules; 4) lipid derivatives, fatty acid metabolites, and membrane components; 5) amino acid/protein metabolism; and 6) microbial metabolites. Further validation and experimentation is needed to confirm the identity of these metabolites and understand their biological relevance and association with selection for increased body weight.

Effect of vitamin E and omega-3 fatty acids early posthatch supplementation on reducing the severity of wooden breast myopathy in broilers.

The wooden breast (WB) myopathy is identified by the palpation of a rigid pectoralis major (P. major) muscle and is characterized as a fibrotic, necrotic P. major muscle disorder in broilers resulting in reduced breast meat quality. Breast muscle affected with WB is under severe oxidative stress and inflammation. The objectives were to identify the effects of dietary vitamin E (VE) and omega-3 (n-3) fatty acids independently or in combination when fed during the starter phase (0-10 D) or grower phase (11-24 D) on growth performance, meat yield, meat quality, and severity of WB myopathy and to determine the most beneficial dietary supplementation period. A total of 210 Ross 708 broiler chicks were randomly assigned into 7 experimental groups with 10 replicates of 3 birds each. The control group was fed with corn-soybean meal basal diet with VE (10 IU/kg) and n-3 fatty acids (n-6/n-3 ratio of 30:1) at a standard level during the entire study (0-58 D). Supplementation of VE (200 IU/kg), n-3 fatty acids (n-6/n-3 ratio of 3:1), or combination of both was performed during the starter phase or grower phase. Growth performance, meat yield, meat quality, and WB scores were obtained. There was no significant difference in final body weight and meat yield when VE was increased (P > 0.05). In contrast, n-3 fatty acids supplementation in starter diets significantly decreased final body weight, hot carcass weight, and chilled carcass weight of broilers (P ≤ 0.05). The P. major muscle from broilers supplemented with VE in starter diets had lower shear force than in grower diets (P ≤ 0.05). Supplemental VE reduced the severity of WB and in starter diets showed a more beneficial effect than those fed VE in the grower diets. These data are suggestive that additional supplementation of dietary VE may reduce the severity of WB and promote breast meat quality without adversely affecting growth performance and meat yield.

The Effect of the Wooden Breast Fibrotic Myopathy in Broilers on Fibrillar Collagen Organization and Decorin-Collagen Binding.

The wooden breast myopathy is identified by the palpation of a rigid pectoralis major muscle and results in myofiber necrosis and fibrosis in fast-growing, meat-type broilers. The fibrosis in wooden breast-affected muscle is characterized by the replacement of myofibers with extracellular matrix proteins, especially fibril-forming collagens. Studies have shown differences in collagen organization in fast-growing broiler lines, with tightly packed and highly aligned collagen organizations having a higher phenotypic incidence of wooden breast. The objective of the current study was to analyze collagen fibril organization further in two fast-growing broiler lines (Lines A and B) with incidence of wooden breast compared with a slower growing broiler Line C with no phenotypically detectable wooden breast. The small leucine-rich proteoglycan decorin was also studied for its interaction with collagen by immunogold detection. Decorin binds to fibrillar collagens and organizes their alignment and crosslinking, both of which will affect collagen functional properties. Key findings from the study showed that collagen shifts to larger diameter collagen fibril bundles with the wooden breast myopathy. Specifically, broilers affected with wooden breast from Line A had a more dramatic shift toward larger collagen fibril bundles compared with those affected from Line B. Wooden breast-affected Line A had collagen fibril bundles up to 8.4 µm, whereas Line B maximum size was 5.1 µm. Although decorin-collagen binding was not different overall in the wooden breast myopathy or broiler line, for small-diameter collagen fibril bundles, wooden breast-affected Line A had more decorin-collagen binding than wooden breast-affected Line B. Taken together, these data provide further evidence that multiple fibrotic myopathies are likely in fast-growing meat-type broilers.

Efecto de la miopatía fibrótica de pechuga de madera en pollos de engorde en la organización del colágeno fibrilar y en la unión entre decorina y colágeno. La miopatía de madera de la pechuga se identifica por la palpación de un músculo pectoralis major rígido y da como resultado necrosis y fibrosis de fibras musculares en pollos de engorde de rápido crecimiento. La fibrosis en el músculo afectado por pechuga de madera se caracteriza por la sustitución de las fibras musculares con proteínas de la matriz extracelular, especialmente colágeno que forma fibrillas. Los estudios han demostrado diferencias en la organización del colágeno en las líneas de pollos de engorde de rápido crecimiento, con organizaciones de colágeno altamente alineadas y altamente empacadas que tienen una mayor incidencia fenotípica para la pechuga de madera. El objetivo del presente estudio fue analizar la organización de las fibrillas de colágeno en dos líneas de pollos de engorde de rápido crecimiento (Líneas A y B) con una incidencia de pechos de madera en comparación con la Línea C de pollos de engorde de crecimiento más lento, sin pecho de madera detectable fenotípicamente. El proteoglicano pequeño decorina rica en leucina también se estudió por su interacción con el colágeno mediante detección por el método inmunogold. La decorina se une a los colágenos fibrilares y organiza su alineación y sus enlaces cruzados, los cuales afectarán las propiedades funcionales del colágeno. Los hallazgos más importantes del estudio demostraron que el colágeno se organiza en fibrillas de mayor diámetro en la miopatía de pechuga de madera. Específicamente, los pollos de engorde afectados con la pechuga de madera de la Línea A mostraron una tendencia mayor para mostrar paquetes de fibrillas de colágeno más grandes en comparación con los afectados de la Línea B. La Línea A afectada por la pechuga de madera tuvo paquetes de fibrillas de colágeno de hasta 8.4 µm, mientras que el tamaño máximo de la Línea B fue de 5.1 µm. Aunque en general, el enlace entre la decorina y colágeno no fue diferente en la miopatía de madera de la pechuga o en la línea de pollos de engorde con haces de fibrillas de colágeno de diámetro menor, la línea A afectada por pechuga de madera tuvo más uniones de decorina-colágeno en comparación con la línea B afectada por pechuga de madera. En general, los datos proporcionan evidencia adicional de que es más probable la presentación de miopatías fibróticas múltiples en pollos de engorde de rápido crecimiento.

Potassium carbonate improves fresh pork quality characteristics.

Meat enhancement strategies like sodium tripolyphosphate (STP) are used to improve fresh meat quality attributes like color, water-holding capacity, and tenderness. However, alternatives are necessary because of reduced consumer acceptance of STP. One alternative is potassium carbonate (K2CO3). A study was conducted to evaluate K2CO3's impact on fresh, boneless, center-cut pork loins enhanced with one of five treatments: a negative control, positive control (0.3% STP), and three concentrations of K2CO3 (0.1, 0.3, and 0.5%). Loins were cut into chops, stored under simulated retail display, and analyzed for color (L*, a*, b*), pH, cook loss, and tenderness. For each quality characteristic measured, the 0.3% and 0.5% K2CO3 maintained redness (a*), decreased yellowness (b*), reduced cooking loss, and maintained tenderness compared to STP. SDS-PAGE analysis further determined that both K2CO3 and STP extracted myosin heavy chain. Combined, these data suggest that K2CO3 may function as an alternative to STP in the fresh pork industry provided microbial safety and shelf-life are appropriately controlled.

The effect of nutrient restriction and syndecan-4 or glypican-1 knockdown on the differentiation of turkey pectoralis major satellite cells differing in age and growth selection.

Skeletal muscle growth is mediated by the proliferation and differentiation of satellite cells, whose activity is affected by both nutrition and the expression of syndecan-4 and glypican-1. Previous research has not addressed if there is an interactive effect of nutrition with the expression of syndecan-4 and glypican-1. Thus, the objective of the current study was to determine if the response of satellite cells to nutrient restriction was altered by syndecan-4 or glypican-1 knockdown and if age and growth selection are factors. Satellite cells were isolated from pectoralis major muscle of 1-day, 7-wk, and 16-wk-old turkeys selected for increased 16-wk body weight (F line) and the randombred control (RBC2) line from which the F line was selected. Syndecan-4 or glypican-1 expression was knocked down in both lines using small interfering RNAs along with nutrient restriction of 0 or 20% of the standard cell culture medium either applied during proliferation with subsequent normal differentiation medium (RN) or during differentiation with preceding normal proliferation medium (NR). For both lines, nutrient restriction and syndecan-4 or glypican-1 knockdown had an independent and additive effect on satellite cell differentiation at 72 h of differentiation except for 1 d satellite cells. The 1 d satellite cell differentiation was increased by RN treatment, but when combined with syndecan-4 or glypican-1 knockdown, the increase in differentiation was negated. At 48 h of differentiation, syndecan-4 knockdown in 7 and 16 wk satellite cells and glypican-1 knockdown in 7 wk cells cancelled the effect of the RN treatment, but enhanced the effect of NR treatment at 24 h of differentiation. Growth selection had little effect on the interaction between nutrient restriction and syndecan-4 or glypican-1 knockdown. Taken together, these data demonstrate that the satellite cell response to nutrition is dependent on the expression of syndecan-4 and glypican-1 in an age-dependent manner with growth selection having little impact.

The effect of nutrient restriction on the proliferation and differentiation of turkey pectoralis major satellite cells differing in age and growth rate.

Myogenic satellite cells are critical for posthatch muscle growth, and their activity is sensitive to nutritional regime during the immediate posthatch period. The objective of the current study was to determine if the response of satellite cells to nutrient restriction was dependent on bird age and/or growth rate. Satellite cells were isolated from the pectoralis major (p. major) muscle of 1-d, 7-wk, and 16-wk-old turkeys selected for increased body weight at 16 wk of age (F line) and the randombred control (RBC2) line from which the F line was selected. Nutrient restriction of 0, 5, 10, 20, and 40% of the standard cell culture medium was applied during proliferation with subsequent normal differentiation medium (RN) or during differentiation with preceding normal proliferation medium (NR). Satellite cell proliferation and differentiation decreased with nutrient restriction for all treatment regimens and ages, except for 1-d cell differentiation with the RN treatment, which increased with nutrient restriction. Interestingly, after 24 h of a 5% nutrient restriction during the RN treatment, proliferation increased for 1-d and 7-wk cells. Additionally, after 24 h of 5, 10, and 20% nutrient restriction during the NR treatment, differentiation increased for 1-d and 7-wk cells. The 16-wk cells did not exhibit this response to any treatment regimen. Growth rate had little effect on satellite cell response to nutrient restriction. In this study, satellite cells differentially responded to nutrient restriction depending on age, as well as duration and timing of the nutrient restriction. These data suggest that it is necessary to optimize diets throughout a bird's life to maximize satellite cell activity and p. major muscle growth.

The effect of syndecan-4 and glypican-1 knockdown on the proliferation and differentiation of turkey satellite cells differing in age and growth rates.

Posthatch skeletal muscle growth requires myogenic satellite cells and the dynamic expression of cell membrane-associated proteins. The membrane associated heparan sulfate proteoglycans, syndecan-4 and glypican-1, link the satellite cell niche to the intracellular environment. Sydnecan-4 and glypican-1 are differentially expressed with age in turkey satellite cells and their over-expression impacts both satellite cell proliferation and differentiation, but their effect on satellite cells from lines with different growth potentials is not known. The objective of the current study was to determine if syndecan-4 and glypican-1 regulation of satellite cell proliferation and differentiation is affected by age and growth selection. Pectoralis major satellite cells isolated at 1 d, 7 and 16-wk of age from a Randombred Control 2 (RBC2) line and a 16-wk body weight (F) line selected from the RBC2 line turkeys were studied. Syndecan-4 and glypican-1 expression was knocked down in both lines. The F-line cells proliferated faster than RBC2 line cells regardless of age, while differentiation tended to be greater in RBC2 line cells than F-line cells at each age. Syndecan-4 knockdown decreased proliferation at 7- and 16-wk but not 1 d cells, and increased differentiation at 1 d and 7 wk but not 16 wk cells. Glypican-1 knockdown differentially affected proliferation depending on cell age, whereas differentiation was decreased for 7- and 16-wk but not 1 d cells. These data suggest syndecan-4 and glypican-1 differentially affected satellite cell function in an age-dependent manner, but had little impact on differences in proliferation and differentiation due to growth selection.

The Effect of the Wooden Breast Myopathy on Sarcomere Structure and Organization.

The wooden breast (WB) has been classically identified by the phenotypic presence of a wood-like pectoralis major (p. major) muscle. The WB-affected p. major muscle is characterized by necrotic muscle fibers and the replacement of muscle with connective tissue, water, and fat. The objective of the current study was to determine the effect of the WB myopathy on sarcomere organization by transmission electron microscopy. Sarcomere structure and organization were examined in two broiler lines with a high incidence of WB (Lines A and B) and another broiler line without WB (Line C). Affected muscle had an increase in smaller myofibers with diameters of 20 µm or less. Sarcomere organization decreased with fiber diameter in both Lines A and B. The structure and organization of sarcomeres in Line C were similar to WB-unaffected muscle in Lines A and B. Taken together, these data demonstrate that the WB myopathy detrimentally affects sarcomere organization in a broiler line-specific manner. Disorganization of sarcomere structure will affect the function of the p. major muscle as well as meat quality.

Effect of incubation temperature on neuropeptide Y and neuropeptide Y receptors in turkey and chicken satellite cells.

Neuropeptide Y (NPY) is an appetite stimulating peptide released from the central nervous system and impacts the function of many different cell types. A recent transcriptome study showed that NPY expression was altered when turkey breast muscle satellite cells were incubated at low or high temperatures, suggesting NPY may mediate temperature effects on satellite cells. However, to date minimal information exists describing the expression and function of NPY in satellite cells. The objective of this study was to determine how temperature impacts NPY and NPY receptor gene expression in satellite cells isolated from turkeys and chickens with differing genetic lineages. Two broiler and two turkey breast muscle satellite cell lines were incubated at 35, 38 or 41 °C during proliferation and differentiation. In both turkey lines, NPY, and receptors NPY2R and NPY5R expression increased at elevated temperatures after 72 h of proliferation. During differentiation NPY and NPY5R expression increased in both turkey lines with higher temperatures, whereas NPY2R was minimally affected by temperature. In contrast, in both chicken cell lines there were few significant differences for NPY and NPY receptor expression across temperature during proliferation. During differentiation, the temperature effect was different in the two chicken cell lines. In the BPM8 chicken line, there were few differences in NPY and NPY receptors across temperature; whereas elevated temperatures increased NPY, NPY2R, and NPY5R expression in the 708 line. The differences between turkey and chicken lines suggest NPY has species specific satellite cell functions in response to heat stress.

Fibrillar Collagen Organization Associated with Broiler Wooden Breast Fibrotic Myopathy.

Wooden breast (WB) is a fibrotic myopathy affecting the pectoralis major (p. major) muscle in fast-growing commercial broiler lines. Birds with WB are phenotypically detected by the palpation of a hard p. major muscle. A primary feature of WB is the fibrosis of muscle with the replacement of muscle fibers with extracellular matrix proteins, such as collagen. The ability of a tissue to be pliable and stretch is associated with the organization of collagen fibrils in the connective tissue areas surrounding muscle fiber bundles (perimysium) and around individual muscle fibers (endomysium). The objective of this study was to compare the structure and organization of fibrillar collagen by using transmission electron microscopy in two fast-growing broiler lines (Lines A and B) with incidence of WB to a slower growing broiler Line C with no phenotypically detectable WB. In Line A, the collagen fibrils were tightly packed in a parallel organization, whereas in Line B, the collagen fibrils were randomly aligned. Tightly packed collagen fibrils arranged in parallel are associated with nonpliable collagen that is highly cross-linked. This will lead to a phenotypically hard p. major muscle. In Line C, the fibrillar collagen was sparse in its distribution. Furthermore, the average collagen fibril diameter and banding D-period length were altered in Line A p. major muscles affected with WB. Taken together, these data are suggestive of different fibrotic myopathies beyond just what is classified as WB in fast-growing broiler lines.

Histopathologic and Myogenic Gene Expression Changes Associated with Wooden Breast in Broiler Breast Muscles.

The wooden breast condition is a myopathy affecting the pectoralis major (p. major) muscle in fast-growing commercial broiler lines. Currently, wooden breast-affected birds are phenotypically detected by palpation of the breast area, with affected birds having a very hard p. major muscle that is of lower value. The objective of this study was to compare the wooden breast myopathy in two fast-growing broiler lines (Lines A and B) with incidence of wooden breast to a slower growing broiler Line C with no phenotypically observable wooden breast. One of the characteristics of the wooden breast condition is fibrosis of the p. major muscle. Morphologic assessment of Lines A and B showed significant fibrosis in both lines, but the collagen distribution and arrangement of the collagen fibrils was different. In Line A, the collagen fibrils were tightly packed, whereas in Line B the collagen fibrils were diffuse. This difference in collagen organization may be due to the expression of the extracellular matrix proteoglycan decorin. Decorin is a regulator of collagen crosslinking and is expressed at significantly higher levels in Line A wooden breast-affected p. major muscle, which would lead to tightly packed collagen fibers due to high levels of collagen crosslinking. Furthermore, expression of the muscle-specific transcriptional regulatory factors for proliferation and differentiation of muscle cells leading to the regeneration of muscle in response to muscle damage was significantly elevated in Line A, and only the factor for differentiation, myogenin, was increased in Line B. The results from this study provide initial evidence that the etiology of the wooden breast myopathy may vary between fast-growing commercial broiler lines.

The effect of temperature on apoptosis and adipogenesis on skeletal muscle satellite cells derived from different muscle types.

Satellite cells are multipotential stem cells that mediate postnatal muscle growth and respond differently to temperature based upon aerobic versus anaerobic fiber-type origin. The objective of this study was to determine how temperatures below and above the control, 38°C, affect the fate of satellite cells isolated from the anaerobic pectoralis major (p. major) or mixed fiber biceps femoris (b. femoris). At all sampling times, p. major and b. femoris cells accumulated less lipid when incubated at low temperatures and more lipid at elevated temperatures compared to the control. Satellite cells isolated from the p. major were more sensitive to temperature as they accumulated more lipid at elevated temperatures compared to b. femoris cells. Expression of adipogenic genes, CCAAT/enhancer-binding protein ß (C/EBPß) and proliferator-activated receptor gamma (PPARγ) were different within satellite cells isolated from the p. major or b. femoris. At 72 h of proliferation, C/EBPß expression increased with increasing temperature in both cell types, while PPARγ expression decreased with increasing temperature in p. major satellite cells. At 48 h of differentiation, both C/EBPß and PPARγ expression increased in the p. major and decreased in the b. femoris, with increasing temperature. Flow cytometry measured apoptotic markers for early apoptosis (Annexin-V-PE) or late apoptosis (7-AAD), showing less than 1% of apoptotic satellite cells throughout all experimental conditions, therefore, apoptosis was considered biologically not significant. The results support that anaerobic p. major satellite cells are more predisposed to adipogenic conversion than aerobic b. femoris cells when thermally challenged.

Elevated insulin-like growth factor 2 expression may contribute to the hypermuscular phenotype of myostatin null mice.

OBJECTIVES: Myostatin (Mstn) inhibits while insulin-like growth factors 1 and 2 (Igf1 and Igf2) increase skeletal muscle growth. However, there is little known regarding Mstn regulation of Igf1 and Igf2 expression. Therefore, the objective of this study was to quantify the expression of IGF family members in skeletal muscle and liver throughout the growth phase of Mstn null (MN) mice. Further, differences between male and female mice were investigated. METHODS: Male and female wild type (WT) and MN mice were euthanized at birth (0 d), 7 days (7 d), weaning (21 d), sexual maturity (42 d), and 70 d. For the neonatal periods, 0 d and 7 d, all muscles from the hind limbs were compiled for RNA extraction. At 21 d, 42 d, and 70 d, biceps femoris (BF), tibialis anterior, triceps brachii (TB), and gastrocnemius-soleus complex were collected. RESULTS: As expected, muscle weights were up to 90% greater in MN mice compared with WT mice at 21 d, 42 d and 70 d. However, Igf1 expression was reduced (P ≤ 0.04) at 7d and 21 d in MN mice compared to WT mice. Expression of Igf2 did not differ between genotypes at 0 d and 7d, but, at 21 d, 42 d and 70 d in BF and TB muscles, Igf2 expression was 1.9-2.9 fold greater (P<0.01) in MN compared to WT mice. Hepatic Igf1 and Igf2 levels were minimally affected by genotype; with the exception of a 1.4-fold reduction (P=0.04) in Igf1 expression in 21 d MN mice compared with WT mice. Though male mice were heavier than females starting at 21 d of age, expression differences in Igf1, Igf2, their receptors and binding proteins do not account for growth differences. In every case, when expression was different between sexes, female expression was increased despite increased growth in male mice. CONCLUSION: This study is the first to provide evidence that Mstn may negatively regulate Igf2 expression to control postnatal skeletal muscle growth, however differences in growth between male and female mice are not readily explained by changes in expression of Igf family members.

Sex differences in proximal and distal nephron function contribute to the mechanism of idiopathic hypercalcuria in calcium stone formers.

Idiopathic hypercalciuria (IH) is a common familial trait among patients with calcium nephrolithiasis. Previously, we have demonstrated that hypercalciuria is primarily due to reduced renal proximal and distal tubule calcium reabsorption. Here, using measurements of the clearances of sodium, calcium, and endogenous lithium taken from the General Clinical Research Center, we test the hypothesis that patterns of segmental nephron tubule calcium reabsorption differ between the sexes in IH and normal subjects. When the sexes are compared, we reconfirm the reduced proximal and distal calcium reabsorption. In IH women, distal nephron calcium reabsorption is decreased compared to normal women. In IH men, proximal tubule calcium reabsorption falls significantly, with a more modest reduction in distal calcium reabsorption compared to normal men. Additionally, we demonstrate that male IH patients have lower systolic blood pressures than normal males. We conclude that women and men differ in the way they produce the hypercalciuria of IH, with females reducing distal reabsorption and males primarily reducing proximal tubule function.

Pretreatment with low-energy shock waves reduces the renal oxidative stress and inflammation caused by high-energy shock wave lithotripsy.

The purpose of this study was to determine if pretreatment of porcine kidneys with low-energy shock waves (SWs) prior to delivery of a clinical dose of 2,000 SWs reduces or prevents shock wave lithotripsy (SWL)-induced acute oxidative stress and inflammation in the treated kidney. Pigs (7-8 weeks old) received 2,000 SWs at 24 kV (120 SW/min) with or without pretreatment with 100 SWs at 12 kV/2 Hz to the lower pole calyx of one kidney using the HM3. Four hours post-treatment, selected samples of renal tissue were frozen for analysis of cytokine, interleukin-6 (IL-6), and stress response protein, heme oxygenase-1 (HO-1). Urine samples were taken before and after treatment for analysis of tumor necrosis factor-α (TNF-α). Treatment with 2,000 SWs with or without pretreatment caused a statistically significant elevation of HO-1 and IL-6 in the renal medulla localized to the focal zone of the lithotripter. However, the increase in HO-1 and IL-6 was significantly reduced using the pretreatment protocol compared to no pretreatment. Urinary excretion of TNF-α increased significantly (p < 0.05) from baseline for pigs receiving 2,000 SWs alone; however, this effect was completely abolished with the pretreatment protocol. We conclude that pretreatment of the kidney with a low dose of low-energy SWs prior to delivery of a clinical dose of SWs reduces, but does not completely prevent, SWL-induced acute renal oxidative stress and inflammation.