Histological disorders related to the focal disappearance of the epiphyseal growth plate in rats induced by high dose of vitamin A

The histological disorders related to the focal disappearance of the epiphyseal growth plate were examined histochemically in the proximal tibia of rats administered a high dose of vitamin A. Animals were given 100,000 IU/100 g body weight/day of vitamin A for 5 days from 4 weeks after birth (VA rats) or given deionized water as control and sacrificed on Day 12 and 19 of the experiment. Tibiae were examined by immunohistochemistry for type I, II and X collagens, lectin-histochemistry for Helix pomatia and backscattered electron imaging. On Day 12, the abnormally developed calcified cartilage matrix was detected within the epiphyseal growth plate in VA rats. The uncalcified cartilage matrix contained type I collagen but lacked type II collagen. In addition, the eroded regions accompanied with numerous osteoclasts and osteoblasts were detected in the epiphyseal growth plate. On day 19, eroded regions penetrated the epiphyseal growth plate to result in its focal disappearances with the eroded surfaces entirely covered with bone tissue in VA rats. These findings suggested that the cartilage matrix of the epiphyseal growth plate was abnormally calcified and showed the phenotypes like bone matrix. The eroded regions of the epiphyseal growth plate seemed to be caused by the invasion of osteoclasts into the altered cartilage matrix and might develop to the focal disappearances by the modeling or remodeling due to action of osteoclasts and osteoblasts.     

Investigation of the insulin-like growth factor system in the avian epiphyseal growth plate

Components of the insulin-like growth factor (IGF) system were investigated in chondrocytes isolated from the avian growth plate. The genes for IGF-I, IGF-II, type 1 IGF receptor (IGF-R), IGF binding protein-2 (IGFBP-2), IGFBP-3, IGFBP-5 and IGFBP-7 were found to be expressed in both proliferative and hypertrophic chondrocytes. The expression of IGF-II in proliferative chondrocytes was extremely high relative to IGF-I. Although IGF-I expression was significantly increased in hypertrophic chondrocytes, the level was still low relative to IGF-II. In cell culture, IGF-I stimulated proteoglycan synthesis and increased the expression of Indian hedgehog (Ihh) and type X collagen, markers of chondrocyte differentiation. IGF-II was found to be equally efficacious in stimulating proteoglycan biosynthesis. These observations suggest that IGF-II may play a significant role in avian growth plate physiology, which is consistent with several reports on mammalian endochondral bone growth.     

Immunolocalization of basic fibroblast growth factor in porcine epiphyseal growth plate

Recent evidence clearly indicates that basic fibroblast growth factor (bFGF) signal transduction is essential for normal skeletal development. Here, we report that bFGF and its receptor are specifically localized in the terminal hypertrophic chondrocytes of the porcine epiphyseal growth plate, the tissue responsible for longitudinal bone growth. Similar observations were obtained with the chondrocytes immediately adjacent to resorbing blood vessels in the secondary center of ossification of the epiphysis. These results are consistent with a recent proposal that bFGF functions in coupling osteogenesis with chondrogenesis by attracting the vascular invasion of cartilage from adjacent trabecular bone.     

Neurotoxicity induced by a single oral dose of aniline in rats.

The neurotoxicity of aniline and its age-dependent responses were investigated in male rats. Groups of 6 rats, 4-week-old, were treated once with aniline (500, 750 or 1,000 mg/kg) or olive oil by gavage. Additional groups of 6 rats, 7- or 10-week-old, were treated once with 800 mg/kg of aniline or olive oil. Paralytic gait or hindlimb paralysis was observed between post-treatment days 8 and 15 in two out of six rats receiving 1,000 mg/kg of aniline at 4 weeks of age. On post-treatment day 15, spongy change in the white matter of the spinal cord was observed in all rats receiving 750 or 1,000 mg/kg of aniline at 4 weeks of age. The lateral and ventral columns of the thoracic spinal cord were the most severely affected. Spongy change in the facial nerve and spinal trigeminal tracts of pons and medulla oblongata, and mild degeneration of the peripheral nerves was found in 3 out of 6 rats receiving 1,000 mg/kg of aniline. At the ultrastructural level, the spongy change was due to distention of the myelin sheath and splitting of the intraperiod line. Axons were well preserved in the affected nerve fibers. No abnormalities were seen in the neuronal cell bodies. Although transient cyanosis was observed in all rats receiving 800 mg/kg of aniline at 7- or 10-week-old, as well as in rats receiving 750 or 1,000 mg/kg of aniline at 4-week-old, no treatment-related neurobehavioral or morphologic abnormalities were found in the former. These findings demonstrate the neurotoxicity of orally administered aniline for rats, depending upon the age of the animal at the time of administration.     

Severe calcification of mucocutaneous and gastrointestinal tissues induced by high dose administration of vitamin D in a puppy

A three-month-old male Bull Terrier was referred to the Animal Medical Centre, Nihon University with chief complaints of subacute emesis and lethargy. Severe leukocytosis, high CRP, hypercalcemia and hypochloremia were detected. Moreover, severe calcification of gingival mucosa and abdominal skin, and abnormalities of the skeletal system were discerned. Abdominal X-ray and endoscopic examination revealed ulcer and hemorrhage on the mucosal membrane of the stomach. This might have been due to injections of high dose vitamin D at 3 and 2 weeks ago by another practioner, according to the detailed history of medication. After two months, a gastrointestinal and skin disorder disappeared, although calcification of the stomach membranes remained and abnormality of the skeletal system had worsened. Therefore, vitamin D should be carefully administrated to a puppy.     

Immunohistochemical observations on pneumonic lesions caused by Rhodococcus equi in foals.

An immunohistochemical analysis of Rhodococcus equi-induced pneumonia in 10 foals was performed by biotin-streptavidin system. The detection of R. equi was more sensitive in immuno-stain using anti-R. equi serum than in Gram's stain. This bacteria also reacted to anti-BCG serum. Lysozyme and alpha 1-antitrypsin were detectable in macrophages. A particularly intense staining was observed in association with intracellular bacteria. Though a degree of reaction for alpha 1-antichymotrypsin was very low in comparison with lysozyme and alpha 1-antitrypsin, it was also demonstrated in macrophages ingesting R. equi. These bacteria were almost intact under an electron microscope. Therefore, the surface components of R. equi may play important roles of protection from intracellular enzymes of macrophages. The cells containing intracytoplasmic IgM, IgG or IgA were a few in number and scattered predominantly around the pneumonic lesion. It is considered that the bactericidal activity by immunoglobulins may be weak in comparison with phagocytosis by macrophages.     

Effects of perinatal high dose dexamethasone on skeletal muscle development in rats.

Five litters of suckling rats were given either dexamethasone (DEX), 1 mg/kg, subcutaneously, three times daily (n = 4/litter) or vehicle control (n = 4/litter) from day 3 through day 7 after birth. Rats were weighed weekly and were weaned on day 30. On day 60, rats were killed and the soleus (SOL) and extensor digitorum longus (EDL) were removed for the following analyses: 1) wet weight, 2) light microscopic examination of hematoxylin and eosin stained transverse sections, 3) quantitative morphometric analysis of myosin ATPase stained transverse sections (fiber numbers, fiber type percentages and mean fiber diameters), and 4) DNA (total and mg/g wet weight). The following parameters were significantly reduced in treated rats: 1) body weight, 2) wet weight of SOL and EDL, and 3) mean diameter of SOL type I fibers. There was a trend for total DNA of SOL and EDL to be decreased in treated rats but this was not statistically significant. In a second experiment, pregnant rats (n = 4) were given DEX, 1 mg/kg, subcutaneously, twice daily, on days 17 and 18 of gestation. Two rats served as vehicle controls. The prenatally DEX-exposed rats weighed significantly less on weeks 3, 4, 6, 7 and 8. There were significant reductions in the following parameters for treated rats: 1) SOL wet weight, and 2) total number of SOL type I fibers. There was a trend for SOL DNA to be reduced but this was not statistically significant.     

Osteochondrosis and arthrosis in pigs. IV. Effect of overloading on the distal epiphyseal plate of the ulna

The effect of overloading on the distal epiphyseal plate of the ulna is described. The right forelegs of 6 pigs were fixed such that the pigs were forced to walk on 3 legs during 2 periods, the first of 12 days duration at about 50 kg live weight, and the second of 7 days at about 100 kg live weight. Osteochondrosis developed to a more marked degree in the left than in the right epiphyseal plate of the ulna, but this did not lead to reduced lengthwise growth of the bones to any great extent. The overloading led to reversible changes in the distal epiphyseal plate of the ulna. It is conceivable that this was due to disturbances in the metaphyseal blood flow. It is also conceivable that the overloading was neither great enough nor of sufficient duration to affect the epiphyseal blood flow with consequential irreversible damage to, and premature closure of, the epiphyseal plate.     

维生素A和维生素E水平对北京鸭前期生产性能的影响

本试验采用5×2完全随机试验设计,5个维生素A(VA)添加水平(0、2500、5000、10000IU/kg和15000IU/kg),2个维生素E(VE)添加水平10mg/kg和100mg/kg,360只1日龄北京鸭随机分为10个处理组,研究不同水平VA、VE对北京鸭生产性能的影响。结果表明,VA的添加可显著提高北京鸭生长前期的平均日增重和平均日采食量(P0.01);对料肉比的影响差异不显著(P0.05),VE水平对北京鸭前期生产性能无显著影响(P0.05);建议北京鸭生长前期VA的适宜添加量为5000～10000IU/kg,VE为10mg/kg。     

Correction of angular deformities caused by premature closure of the distal ulnar epiphyseal plate in dogs

Surgical correction of angular deformities caused by premature closure of the distal ulnar growth plate in dogs. Correction of angular deformities in 14 cases of premature closure of the distal ulnar growth plate is described. Angular deformities were most frequently seen in large breeds during the most active phase of growth at 3-5 months of age. Preoperative axial deviations measured on radius were from 7 to 38 degrees in valgus and 0 to 35 degrees in antecurvation compared with the contralateral leg. According to the kind of treatment, the dogs were divided into two groups: In group I which consisted of cases with sufficient remaining growth potential of the radius, segmental ostectomy of ulna with insertion of an autologous fat graft in the ostectomy diastasis was performed to allow spontaneous correction by asymmetrical epiphyseal growth. In group II reconstructive osteotomy and stable, internal fixation with a plate was accomplished. The cosmetic result was judged as excellent in 10 cases, good in 3 and less favourable in 1. Postoperative follow up time was in average 33 months. On control 43% of the dogs showed no lameness, 57% were slightly lame after exercise or rest. None of the cases turned out to be permanently lame at the end of the follow up investigation.     

Investigation on the growth of coprophagy-prevented rats with supplemented vitamin B12

The growth of coprophagy-prevented rats was compared under administration of normal levels of vitamin B12 and supplemented amounts. Two experiments in which supplemented amounts of vitamin B12 were administered were conducted under different conditions. Six rats per group were fed under coprophagy-allowed (conventional feeding) and coprophagy-prevented conditions respectively. In the first experiment, coprophagy-prevented rats were fed only feed containing recommended vitamin B12 level and forced fed hydrous faeces, vitamin B12 and folic acid respectively. In the second experiment, coprophagy-prevented rats were fed AIN-93G at the recommended vitamin B12 level (25 microg/kg diet), at 100 times the level and at 1000 times the level respectively. Body weight, feed consumption and amounts of each faeces type were determined in both experiments. In a comparison of body weight gain, we learned that coprophagy prevention reduced the values, but that there was no significant difference in the forced feeding group in the first experiment. Similar results were recognized in the second experiment. Vitamin B12 supplementation was not able to raise feed intake significantly and hence it obviously was not a severely limiting factor under the respective experimental condition which depressed feed intake.     

Morphological changes of epiphyseal plate in the long bone of chondrodysplastic dwarfism in Japanese brown cattle.

A total of 20 cases of disproportionate calves on Japanese Brown cattle were examined morphologically. Investigation of 5 affected calves revealed that the insufficiency of endochondral ossification was confined only to the long bones of the limbs and was not accompanied by other associated changes. On the basis of the histological changes of epiphyseal plate and the affected site, this condition may be called chondrodysplasia of the rhizomelic type. Therefore, this disorder was defined as a bovine dwarfism. From three dimensional image analysis and histological changes of the tibial proximal portion in the other 15 cases, deformity and shortness of the tibia were related to the state of distribution and the degree of damages of the epiphyseal plate. In the affected cartilage matrix, cystoid degeneration, fibrous striation, bone spicule, necrotic foci and rent were found. Inadequate metachromasia suggests the abnormal staining of sulfated glycosaminoglycans and alcian blue stainability may be attributable to the abnormal interactions between proteoglycan and other matrix components. At the interface of cartilage and bone, irregular calcification, fibrotic scar and sealing by osseous tissue with relation to vascularity were found. These changes in this study indicated the failure of modeling on the epiphyseal plate, showing disturbance of chondrocytic differentiation and abnormal formation of the matrix.     

高剂量植酸酶对肉鸡生长性能和消化率的影响

文章旨在研究植酸酶对1~42日龄肉鸡生长发育的影响。采用完全随机试验设计,试验共分为5组,每组6个重复,每个重复24只鸡,各组分别为阳性对照组、负对照、负对照日粮+1000、2000和3000IU/kg植酸酶,试验结束后测定各组对骨功能、骨质量和血液矿物质的影响。随着日粮植酸酶添加水平的升高,显著改善了1~21d肉鸡日增重、采食量和料比(P<0.05),而1~42d肉鸡生长性能表现为显著二次曲线效应(P<0.05),其中负对照组肉鸡日增重和采食量最低(P<0.05),料比最高(P<0.05)。对照组较负对照组显著提高了42d肉鸡胫骨干物质含量(P<0.05)。随着植酸酶添加水平的升高,21d肉鸡血钙浓度表现为显著线性降低(P<0.05),而血磷浓度先升高后降低,负对照日粮+1000IU/kg植酸酶组最高(P<0.05)。42d肉鸡血磷浓度随日粮植酸酶水平的升高表现为二次曲线效应。对照组和负对照组较处理组显著提高了42d肉鸡血钙浓度(P<0.05)。与对照和负对照组相比,负对照添加植酸酶组显著降低了矿物质回肠表观消化系数(P<0.05),而负对照组+3000IU/kg植酸酶较对照组显著降低了能量回肠表观消化系数(P<0.05)。植酸酶在回归分析的基础上提高了肉鸡生长性能,添加高剂量植酸酶3000IU/kg显著提高21d肉鸡日增重。试验为期42d,日粮添加2000IU/kg植酸酶对肉鸡的增重和饲料转化率效果较好。     

Lesions of articular, sternal and growth plate cartilage in rats

Degenerative osteoarthritis was observed in the femur and sternum in specific pathogen free Fisher 344 rats at 13 and 19 months of age. Histological changes consisted of degeneration of the matrix and erosion in the articular cartilage of the femur. In the sternal cartilage, degeneration and necrosis of the matrix, formation of cysts, and cleft-like fractures were observed. In addition, degeneration of the matrix, appearance of eosinophilic streaking, and necrosis and fissure between the growth plate and epiphyseal trabeculae were seen in the femur. The changes in the sternum were essentially the same as the femur.     

Effects of high dietary vitamin A supplementation on tibial dyschondroplasia, skin pigmentation and growth performance in avian broilers

An experiment was conducted to study high dietary vitamin A on tibial dyschondroplasia, growth performance and skin pigmentation in broilers. One hundred and twenty Avian commercial broilers were randomly allotted to three treatments: group C (control group), in which broilers were fed basic diet containing vitamin A 5512IU/kg diet; group A, in which broilers were fed basic diet with addition vitamin A 35512IU/kg; group B, broilers were fed basic diet with supplement vitamin A 65512IU/kg. The experiment lasted 35d and at the end of the trial, broilers were killed and the right proximal tibiotarsi were dissected in longitudinal section for the assessment of TD incidence and TD index, skin from the same area of breast and tibia in broilers were collected to determine pigmentation. The results showed that a high level vitamin A significantly increased the rate of TD incidence and TD index, but middle level vitamin A did not have a significant effect on that. Both low and high retinoic acid decreased growth performance and skin pigmentation in broilers. It suggests that a high dietary vitamin A cause tibial dyschondroplasia in broilers, decreased growth performance and skin pigmentation. It is likely that the effect of vitamin A on TD is mediated through a depression of vitamin D status.     

Effects of vitamin A deficiency on the function of pituitary-gonadal system in male rats

The effects of vitamin A deficiency on the pituitary-gonadal function were examined by measurements of serum and pituitary level of pituitary hormones and serum testosterone concentration, and by investigations of histological changes in the testis and the pituitary gland in vitamin A-deficient (VAD) and supplemented (VAS) rats. The growth of VAD rats was retarded and their body weights were decreased after 9 weeks of experiments and attained about one half of the weight of control animals at 12 weeks. In the VAD rats, serum testosterone concentrations were decreased significantly compared with those in the VAS controls. Serum and pituitary concentrations of GH were significantly lower but those of LH were slightly lower in the VAD rats than those in the controls, while the serum FSH concentration was significantly higher than that in the control rats. The seminiferous tubules in the testes of VAD rats were comprised largely of Sertoli cells and a reduced number of spermatogonia and contained fibrous formation in their lumen. In the pituitary gland, GH cells were significantly reduced in number in the VAD rats, but gonadotropic (GTH) cells were increased remarkably in size and number, showing hypertrophy and vacuolation similar to those in castration cells. The cytological changes in the pituitary gland and the increased discharge of FSH represent a secondary and compensatory change similar to that seen following castration and vitamin E deficiency.