Development of components of the extracellular matrix, basal lamina and sarcomere in chick quadriceps and pectoralis muscles

1. The purpose of this study was to investigate differences in the development of components of the cell/matrix linkage in two functionally different muscle types: the pectoralis muscle, a major locomotory muscle in birds but not particularly functional in chickens, and the quadriceps muscle, a smaller and more functionally active muscle in the chicken. 2. The development of the extracellular matrix, basal lamina and sarcomere in the pectoralis and quadriceps muscles in chick embryos was examined biochemically to determine differences in the rate of development between these two muscles. Samples of these muscle types were dissected out from chick embryos from embryonic day 10 until 8 weeks post hatch. 3. Using SDS-PAGE electrophoresis and western blotting with antibodies against sarcomeric actin, laminin and collagens I, III and IV, it was apparent that muscle development begins earlier in the quadriceps muscle than in the pectoralis, and that late in the developmental process (d 18) both muscle types were well differentiated. The final concentration of collagens in the mature muscle remained higher in the quadriceps than in the pectoralis muscle. 4. The onset of development of the extracellular matrix, basal lamina and sarcomere was earlier in the quadriceps than the pectoralis, which could have functional implications for these muscles as a whole.     

实验性雏鸭锌中毒症的病理学研究

1日龄天府肉鸭健雏100只随机分为2组，分别喂以止常对照(Zn100mg／kg日粮)和锌中毒(Zn1300mg／kg日粮)日粮4周，进行系统的病理学研究。中毒鸭13d出现症状，18只死亡。尸检主要见腿部肌肉色白或灰白色，外观似蜡样；肌胃平滑肌色淡灰白；消化道充满黑褐色煤焦油样内容物。光镜下，骨骼肌纤维肿胀变性坏死；心肌和肌胃平滑肌局灶性坏死；胸腺、腔上囊、脾脏淋巴细胞减少。电镜观察，免疫器官淋巴细胞线粒体肿胀、嵴断裂。血液病理学变化主要是碱性磷酸酶活性、血清总蛋白与球蛋白含量以及红细胞总数和血红蛋白含量降低，血清谷丙转氨酶活性和锌含量升高。结果表明，高锌对鸭可产生明显的毒害作用。文中还对发病机理进行了探讨。     

Influence of in ovo inoculation with various nutrients and egg size on broiler performance

This study evaluated the influence of inoculation of different nutrients into eggs of 18-d-old broiler embryos. On d 18 of incubation, before transferring, eggs were injected with a solution containing either maltose, a multivitamin supplement, zinc-glycine, glutamine, a mixture containing all these elements, or sodium chloride (control). After hatching, 2,400 male broiler chicks were placed in an experimental broiler house and distributed into 60 floor pens in a completely randomized design comprising a 2 × 6 factorial arrangement of treatments (2 egg weights × 6 solutions), for a total of 12 treatments. Birds and feed consumption were measured weekly. At 42 d of age, 3 birds per replicate pen were randomly selected for processing. Birds derived from heavier eggs had greater hatchability and hatching weight. At 42 d of age, birds from heavier eggs had higher BW, carcass weight, and breast meat weight. The livability of birds derived from heavy eggs was higher at 7 and 14 d of age. The in ovo inoculation of the nutrients to 18-d-old embryos did not influence live performance or carcass traits. The technique of in ovo inoculation of certain nutrients may be used in industrial poultry production, but further studies are required to define the best solutions or mixture of nutrients to be used.     

Pathological and immunological study of goose embryos experimentally infected with duck plague virus

A total of 240 embryonated goose eggs obtained from two susceptible flocks were used. Half of the eggs were inoculated into the allantoic cavity with a virulent strain (7593) of duck plague virus isolated from an acute outbreak, and the other half were inoculated with the attenuated vaccine virus (KAPEVAC). Ten, 100 or 1000 CPU/0.1 ml virus were given on days 12 and 20 of incubation. Embryos that died and surviving embryos killed at 5-day intervals were examined by light and electron microscopy. The yolk and the serum of embryos that survived until hatching were assayed for antibody content. Lymphocytes separated from the blood were used for the immuno-rosette formation and lymphocyte stimulation tests. Pathomorphological changes indicative of virus replication occurred in the liver, kidney, myocardium, gizzard muscle and chorioallantoic membrane (CAM) of the embryos in the case of both virus strains. The time of onset and severity of these changes and the time and rate of embryonic mortality depended on the virulence of the strain used for inoculation, the virus dose and the time of inoculation. Virus-neutralizing (VN) antibodies were demonstrable neither in the yolk nor in the serum of goslings exsanguinated after hatching. The lymphocytes recognized the virus antigen in the in vitro cellular tests and responded to it with blastogenic transformation. As opposed to adult birds, in the embryos duck plague virus infection did not cause damage to the digestive tract mucosa and the lymphoid organs.     

牦牛肌红蛋白的基因克隆测序、分离纯化、含量及其与乳酸脱氢酶和苹果酸脱氢酶活力的关系

为探索牦牛适应高原缺氧环境的分子机制,测定了麦洼牦牛心肌和骨骼肌中肌红蛋白（Mb）含量及乳酸脱氢酶（LDH）、苹果酸脱氢酶（MDH）活力,并对牦牛Mb进行了分离纯化和基因克隆测序。牦牛心肌和骨骼肌中Mb含量显著高于水牛和黄牛（P〈0.01）;牦牛、黄牛和水牛心肌MDH/LDH活力比均显著高于骨骼肌;牦牛心肌MDH/LDH活力比显著高于水牛和黄牛,但在不同牛种骨骼肌之间无显著差异。各组织Mb含量与MDH/LDH活力比呈显著正相关。试验用RT-PCR方法从牦牛心肌中克隆了Mb基因,与普通牛相比,核苷酸序列同源性为99.5%,氨基酸序列完全相同;用盐析、CM-Sephadex阳离子交换层析、Sephadex G-50凝胶层析等方法分离纯化了牦牛Mb,SDS-PAGE显示其分子量约为17 ku。     

Differential Growth in the Supracoracoideus Muscles of Male and Female Turkeys

The growth of supracoracoideus muscles in male and female turkeys was measured at two week intervals for 20 weeks after hatching. Growth in weight of the supracoracoideus was closely related to growth of the adjacent pectoralis and coracobrachialis muscle. Growth curves for muscle length and fascicular length exhibited marked deceleration so that more than 75% of the length at 20 weeks had been reached by 10 weeks. Growth curves for weight, cross sectional area at the muscle midlength and myofibre diameter were approximately linear with 50% or less of the 20 week growth attainment being reached by 10 weeks. Supracoracoideus muscles in most males had a greater cross sectional area and were longer and heavier than those in females. At 20 weeks, the greater mass of supracoracoideus muscles in males was due to radial and not longitudinal myofibre growth.     

Effect of selenium and vitamin E content of the maternal diet on the antioxidant system of the yolk and the developing chick

1. The effects of selenium and vitamin E supplementation of the maternal diet on their transfer to the egg yolk and tissues of the newly hatched chick and on the development of the antioxidant system in the chick liver in early postnatal life were investigated. 2. One hundred Cobb broiler breeder hens were divided into 10 equal groups and housed in pens at 25 weeks of age. Each hen received 1 of the treatment diets which included 0.2 or 0.4 mg/kg selenium, 40, 100, 200 mg/kg vitamin E or their combination. After 6 weeks, the hens were artificially inseminated once per week. From week 8, eggs were collected and placed in an incubator. After hatching, chicks from each group were reared (under standard commercial conditions) to 10 d of age. The chicks were fed on a standard starter commercial broiler diet. At the time of hatching, and at 5 and 10 days old, 4 chicks from each group were sacrificed and blood, liver and brain were collected for the subsequent biochemical analyses. 3. The inclusion of organic selenium or vitamin E in the commercial diet significantly increased their concentration in the egg and in the liver of 1-d-old chicks obtained from the eggs enriched with these substances. A positive effect of such dietary supplementation was seen at d 5 and d 10 of postnatal development. 4. There was a positive effect of selenium supplementation of the maternal diet on glutathione concentration in the liver of 1-d-old and 5-d-old chicks. A combination of a dietary selenium supplementation with high vitamin E doses further increased glutathione concentration in the liver. Dietary selenium supplementation significantly increased selenium-dependent glutathione peroxidase (Se-GSH-Px) activity in the liver of the 1-d-old and 5-d-old chicks and decreased liver susceptibility to peroxidation. 6. It is concluded that the nutritional status of the laying hen determines the efficiency of the antioxidant system throughout embryonic and early postnatal development of the offspring.     

The glygogen content of skeletal muscle,liver and heart in late chick embryos

The liver, heart and skeletal muscle (thigh muscles) contain appreciable quantities of glycogen at the 432nd hr. of incubation. From this time, when pulmonary respiration begins, until “ pipping “ there is a decline in the glycogen content of the liver and heart. Between “ pipping “ and hatching there is evidence of the repletion of the glycogen stores in these tissues. When the shell and shell membrane above the air space are removed at the 432nd hr. there is a progressive increase in the glycogen content of the liver, heart and skeletal muscle until the time of hatching. The depletion of the glycogen stores in normal embryos before “ pipping “ appears to be due to an hypoxic state existing before “ pipping” and relieved when “ pipping “ takes place. When the gas exchange through the air space is reduced by waxing the shell overlying the air space, the fall in the glycogen stores of all three tissues is severe and would appear to be due to accentuation of the hypoxic state prevailing before “ pipping “.

With the exception of a period of about 30 min. after “ pipping”, and for brief periods in the hours before hatching the respiratory quotient remains constant at or around 0–7. This low RQ, is consistent with the synthesis of glycogen from lipid material, derived presumably from the contents of the yolk sac. The evidence is reviewed for believing that synthesis of glycogen proceeds even when depletion of the glycogen stores in the tissues occurs.

The low levels of cardiac glycogen found in those embryos which had restricted access to oxygen (waxed shells) combined with the high mortality in such experiments suggests that survival through the critical period of hypoxia before “ pipping “ depends on the level of the stores of glycogen in the heart at the start of this period of hypoxia.     

Effect of magnesium on reactive oxygen species production in the thigh muscles of broiler chickens

1. The objective of the present study was to investigate the effect of magnesium (Mg) on reactive oxygen species (ROS) production in the thigh muscles of broiler chickens. A total of 96 1-d-old male Arbor Acre broiler chickens were randomly allocated into two groups, fed either on low-Mg or control diets containing about 1.2 g/kg or 2.4 g Mg/kg dry matter. 2. The low-Mg diet significantly increased malondialdehyde (MDA) concentration and decreased glutathione (GSH) in the thigh muscles of broiler chickens. ROS production in the thigh muscle homogenate was significantly higher in the low-Mg group than in the control group. Compared with the control, muscle Mg concentration of broiler chickens from the low-Mg group decreased by 9.5%. 3. Complex II and III activities of the mitochondrial electron transport chain in broilers on low-Mg diet increased by 23 and 35%, respectively. Significant negative correlations between ROS production and the activities of mitochondrial electron transport chain (ETC) complexes were observed. 4. The low-Mg diet did not influence contents of iron (Fe) or calcium (Ca) in the thigh muscles of broiler chickens and did not influence unsaturated fatty acid composition (except C18:2) in the thigh muscles. 5. A low-Mg diet decreased Mg concentration in the thigh muscles of broiler chickens and then induced higher activities of mitochondrial ETC, consequently increasing ROS production. These results suggest that Mg modulates the oxidation-anti-oxidation system of the thigh muscles at least partly through affecting ROS production.     

Macromolecular composition of fast, slow and cardiac muscles of chick during incubation.

Total muscle mass increases about five-fold in fast (anterior latissimus dorsi: ALD) and slow (posterior latissimus dorsi: PLD) muscles and 30-fold in cardiac muscle between the 12th and 20th d of incubation. 2. Increase in muscle protein does not parallel the rise in muscle RNA. 3. The DNA concentration of the ALD muscle decreases while that of the heart and PLD muscle show an increase between 8 and 12 d and a steady decrease thereafter. 4. The increase in glycogen concentration is greater in heart than in ALD and PLD up to the 20th d; 1 d after hatching, ALD has more glycogen than PLD.     

藏绵羊肌红蛋白（Mb）基因克隆及肌肉Mb含量的测定

为探索藏绵羊适应青藏高原缺氧环境的分子机制,克隆了藏绵羊的肌红蛋白（Mb）基因,纯化了心肌Mb,同时比较了心肌和骨骼肌的Mb含量。用RT-PCR克隆了心肌Mb的cDNA,长度为640 bp,编码153个氨基酸。试验采用盐析、CM-Sephadex离子交换层析、Sephadex G-50凝胶层析等方法分离纯化了藏绵羊Mb,SDS-PAGE分析其分子量约为17kD。组织含量测定表明,藏绵羊心肌中Mb含量极显著高于骨骼肌。     

Evaluation of the uterine environment and embryos of prepubertal gilts

A series of three experiments was conducted to test the functional status of the uterus and embryos in prepubertal gilts. In Exp. 1, gilts were induced to ovulate by treating with gonadotropins followed by hCG 72 or 96 h later, and were artificially inseminated 24 h after hCG. Five of the 10 gilts treated at 120 d of age, but none of the gilts treated at 100 of age, maintained pregnancies. We next tested the function of the uterine environment by transferring embryos from postpubertal females into gilts of various ages that had been induced to ovulate but not inseminated (Exp. 2). Pregnancy rate at d 50 of gestation was 44% (4/9) for 100-d-old recipients, 67% (2/3) for 140-d-old recipients, and 60% (3/5) for postpubertal recipients (P > 0.20). Therefore, uteri of 100-d-old gilts are able to maintain pregnancies with conceptuses from postpubertal gilts. In Exp. 3, embryos from 100-d-old and postpubertal gilts were transferred into postpubertal recipients. Uterine horns of recipients were surgically separated before transfer, and embryos from 100-d-old and post-pubertal females were transferred to opposite horns of some recipients (experimental). Other recipients received embryos from postpubertal females in both uterine horns (control). When examined on d 50 to 60 of gestation, three of five control gilts were pregnant and three of seven experimental gilts were pregnant (P > 0.50). In experimental recipients, the survival of embryos from 100-d-old gilts was 38% (8/21) compared to 57% (15/26) for embryos from postpubertal gilts (P > 0.30). Because all uterine horns of pregnant recipients contained fetuses, these results support the hypothesis that embryos from 100-d-old gilts are able to initiate and maintain pregnancies in the uteri of postpubertal gilts. Therefore, the uterine environment of 100-d-old gilts provides an environment that supports development of embryos produced by postpubertal gilts, and the embryos produced by 100-d-old gilts can survive and develop in the uteri of postpubertal gilts. It was only the combination of embryos and uteri of 100-d-old gilts that did not permit pregnancy to be maintained.     

Growth changes of the collagen content and architecture in the pectoralis and iliotibialis lateralis muscles of cockerels

1. Growth changes of the collagen content and architecture in the pectoralis (PT) and iliotibialis lateralis (ITL) muscles were examined using cockerels from 1 to 14 weeks of age. 2. Total collagen content in PT muscle showed little change, but in ITL muscle reached a maximum at 5 weeks and thereafter decreased slightly until 14 weeks. The collagen content was markedly larger in ITL muscle after 5 weeks. Pyridinoline content of collagen increased abruptly from 5 to 14 weeks in both muscles, but no difference between muscle types was detected. 3. The cell size of the endomysial honeycombs increased with the development of myofibres, and the mesh size of the perimysium around the honeycombs enlarged. 4. In both muscles endomysia were an incomplete network of collagen fibrils with many foramina at one week, became a very thin membrane of felt-like fabric in 2 to 5 weeks and thereafter increased in thickness until 11 to 14 weeks. 5. Perimysial width around the secondary fasciculus differed between the muscle types after 5 weeks. In the wider perimysium of ITL muscle, the collagen fibres increased in number and size to make a stack of collagen bands around the fasciculus. In the narrower perimysium of PT muscle, a few platelets of collagen fibres also developed. 6. The perimysial collagen fibre at 1 to 2 weeks had a smooth surface and appeared to be composed of fine collagen fibrils. The fibre at 11 to 14 weeks showed a rugged surface and was composed of coarser collagen bundles that combined with each other into a net-like configuration with very slim meshes. 7. Our results showed that the collagenous components of chicken intramuscular connective tissue changed markedly during the early period of muscle growth in distribution, architecture and quality but with little difference in quantity.     

Granulopoiesis in the pineal gland of chickens

Foci of differentiating heterophilic granulocytes in the pineal gland were studied by light and electron microscopy in chickens, from hatching until 56 weeks of age. Foci of granulopoiesis could be seen in the first 24 hours after hatching. Thereafter, their number and cellular density increased, becoming highest at 2 weeks. From then on, numbers decreased progressively until foci disappeared at 18 weeks. Granulopoietic cells established local associations with fibroblast-like cells. Mature granulocytes reached the bloodstream by the 2 mechanisms described for hematopoietic cells in the bone marrow, either passing between lining cells or through the cytoplasm of lining cells.     

Dietary thyroid hormone improves growth and muscle protein accumulation of black-boned chickens

1. The objective of the study was to examine the effect of dietary thyroid hormone on performance and muscle protein accumulation of black-boned chickens. 2. A total of 720 1-d-old birds was housed in 24 pens and assigned to 4 diets containing triiodothyronine (T3) at 0, 0.1, 0.3 or 0.5 mg/kg. 3. The trial was split into starter (0 to 4 weeks) and grower (5 to 8 weeks) phases. At the beginning of the grower phase, each pen was split into two; one subgroup was fed basal diet (T3-) and the other continued with T3-added diet (T3+). 4. Dietary T3 at 0.1 mg/kg improved average daily body weight gain until week 6 but retarded it thereafter. However, T3 at 0.5 mg/kg depressed growth throughout the whole period. During weeks 7 to 8, T3 retarded growth, whereas growth recovered when T3 was withdrawn from the diet. 5. Birds fed the diet containing 0.1 mg/kg during weeks 0 to 4 and those receiving basal diet during weeks 5 to 8 showed the greatest thigh muscle growth, but this growth was depressed when they were fed the 0.5 mg/kg diet during weeks 0 to 8. 6. Serum T3 concentration was increased, while thyroxine (T4) concentration was decreased by dietary T3 treatment at 2 weeks old. At 4 weeks old, T3 and T4 concentrations decreased. At 6 and 8 weeks old, among T3-treated groups the serum T3 concentration of the T3- group was higher than that of the T3+ group. 7. In conclusion, dietary T3 improved weight gain and muscle growth of black-boned chickens before week 6, whereas supplementation depressed growth after 6 weeks. Optimum dietary T3 concentrations showed a carry-over effect on both body weight gain and thigh muscle growth.     

Total lipid and triacylglycerol contents in the liver of broiler and layer chickens at embryonic stages and hatching

To compare the hepatic function of broiler and layer chickens (Gallus gallus domesticus) at various embryonic stages and hatching, the total lipid and triacylglycerol (TG) content were determined. The chicken embryos accumulated a large amount of lipids in the liver and the total lipid content gradually increased toward hatching, though no significant difference was observed between broilers and layers. The TG contents in the liver increased considerably with developmental stage. At embryonic day 14 the TG content in the liver was similar between broilers and layers; thereafter, it was 1.3 and 2.2 times higher in broilers than in layers at embryonic day 18 and for newly hatched chicks, respectively. Chick embryos accumulate an excessive amount of cholesterol ester in the liver, but cholesterol ester is replaced by TG after hatching. The results of hepatic TG contents in the present study suggest that the development of the hepatic function between broilers and layers may already differ at embryonic stages.     

The zinc-myoglobin relationships in porcine muscles.

Zinc and myoglobin content in muscles from pigs were studied under various conditions. Zinc concentration was considerably higher in red than in white muscles. In muscles, where the metabolic pattern changes from glycolytic to oxidative during the period from birth to weaning, a simultaneous increase in zinc content was seen. A significant positive correlation exists between myoglobin and zinc content under normal conditions. However, while myoglobin concentration decreases due to iron deficiency anaemia no changes occur in zinc content. It is concluded that no functional link seems to exist between zinc metabolism and myoglobin synthesis in porcine muscles.     

Age-related changes in the intramuscular distribution of melanocytes in the Silky fowl

1. The characteristics of melanocyte distribution in skeletal muscles in the Silky fowl were investigated in association with growth. 2. Pectoralis (PT) and iliotibialis lateralis (ITL) muscles from 1-, 3-, 5-, 10-, 20- and 30-week-old Silky males were weighed and collagen type I was detected in frozen sections immunohistochemically. 3. Melanocytes were observed in the collagen type I-immunopositive endomysium and perimysium in both muscles. 4. Image analysis indicated that the total area occupied by melanocytes in histological sections sharply decreased from 0.61% to 0.16% in PT muscle and from 1.67% to 0.33% in ITL muscle at 1 to 3 weeks, and then gradually decreased. The melanocyte area was larger in ITL muscle than in PT muscle until 10 weeks of age. 5. We concluded that the proportion of intramuscular melanocytes in the Silky fowl differs between types of muscles in the early stages of development, and it decreases with growth.     

Nutrient supply enhances both IGF-I and MSTN mRNA levels in chicken skeletal muscle

Nutrient supply may control muscle growth directly and indirectly through its influence on regulatory factors. The present study focuses on its effects on muscle insulin-like growth factors (IGF-I and -II) and myostatin (MSTN). Their mRNA levels were quantified by real time RT-PCR in pectoralis major (PM) and sartorius (SART) muscles from broiler chickens submitted to different feeding regimens (fed or fasted for 48 h) between hatch and 2 days of age and at 4 weeks of age. In the PM of 4 weeks old broilers, mRNA levels were also evaluated after a 16 h-fast and a refeeding period (refed 24 or 48 h after a 48 h-fast). In the PM muscle, both IGF-I and MSTN mRNA levels increased between 0 and 2 days of age in the fed group, while they remained low in the unfed one. A comparable trend was observed in the SART, but with lesser amplitude. In both muscles of 4 weeks old chickens, a 48 h-fast induced a significant reduction in MSTN mRNA levels (20% of fed state). In the PM, this effect required more than 16 h of fasting to occur and was fully reversed by only 24h of refeeding. IGF-I mRNA levels also varied with nutritional state. They decreased significantly with fasting in the SART muscle. By contrast, IGF-II mRNA levels did not vary significantly. Our data shows for the first time that two major paracrine regulators of muscle growth, IGF-I and MSTN, are sensitive to nutrient supply in hatching chicks, and also that fasting reduced IGF-I and MSTN mRNA levels in muscles of older chickens.     

Post mortem muscle metabolism and meat quality in three genetic types of turkey

1. A standard (FG, fast-growing), a black local or 'label', type (SG, slow-growing) turkey line, and the crossbreed between these two lines were compared for muscle post-mortem metabolism and related meat quality traits. 2. Ninety male turkeys (30 of each genetic type) were raised under the same experimental conditions until slaughter at 16 weeks of age. 3. Live weights at 16 weeks of age differed significantly (7.8, 6.0 and 4.2 kg, for the FG, crossbred and SG lines, respectively). Collagen content of Pectoralis superficialis (PS) muscle was higher in SG birds than in the other two types. 4. The rate of post-mortem glycogen depletion and lactate accumulation in PS and Ilio tibialis (IT) muscles were similar in the 3 lines, as were the rate and extent of post-mortem pH fall in PS muscle. In IT muscle, however, SG birds showed a slight but significantly faster pH decline. 5. Colour measurements indicated a paler breast muscle and a higher degree of myoglobin oxidation in SG birds at 24 h post mortem, than in both other lines. But these differences had disappeared after 4 and 7 d post mortem 6. SG birds showed higher drip loss and instrumentally-assessed toughness in breast muscle, compared with crossbred and FG birds. FG birds, however, had the lowest yield of breast meat after curing-cooking. 7. No marked differences in post-mortem metabolism were found between the three lines. However, differences in water-holding capacity of fresh and cured-cooked meat suggest that factors other than the rate and extent of post-mortem pH fall may contribute to the respective characteristics of these lines.