Skeletal muscle fiber size in untrained and endurance-trained horses.

The mean area and minimal diameter of 3 histochemically determined myofiber types (1, 2A, and 2B; myosin ATPase in acid buffer) were calculated in middle gluteal muscle biopsy specimens from 62 stallions, 47 Andalusians and 15 Arabians, ranging in age from 6 to 12 years. Fourteen Andalusians and 7 Arabians were untrained, and the remainder were actively endurance-trained. The 6-month training schedules involved walking, slow trotting, and cantering. Fourteen Andalusians were moderately endurance-trained, whereas the other 19 Andalusians and 8 Arabians were strongly endurance-trained. Significant differences were not recorded between untrained and endurance-trained Arabians with respect to the area (type 1, 3,194 +/- 869 microns 2 and 3,150 +/- 370 microns 2; type 2A, 3,819 +/- 890 microns 2 and 3,380 +/- 356 microns 2; and type 2B, 4,872 +/- 962 microns 2 and 4,417 +/- 646 microns 2) or minimal diameter (type 1, 52.2 +/- 7.4 microns and 52.8 +/- 3.1 microns; type 2A, 58.1 +/- 6.7 microns and 55.0 +/- 2.8 microns; and type 2B, 65.3 +/- 6.4 microns and 63.4 +/- 4.3 microns) of the 3 fiber types, nor between untrained and endurance-trained Andalusians with respect to the area (untrained, 3,990 +/- 690 microns 2; moderately endurance-trained, 3,882 +/- 347 microns 2; and strongly endurance-trained, 3,758 +/- 510 microns 2) and minimal diameter (untrained, 58.1 +/- 4.7 microns; moderately endurance-trained, 59.7 +/- 2.7 microns; and strongly endurance-trained, 58.7 +/- 4.5 microns) of 2A fibers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.     

Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Theileria orientalis: a review

Theileria orientalis (also known historically as T. sergenti and T. buffeli) is responsible for benign or non-transforming theileriosis, and exerts its major effect through erythrocyte destruction. The life cycle of T. orientalis is essentially similar to that of other Theileria species, except that the schizonts do not induce transformation and fatal lymphoproliferation. The pathogenesis of anaemia as a result of infection is not clearly established and may be multifaceted. Clinical signs of weakness, reluctance to walk and abortion are early but non-specific indications of disease, particularly if accompanied by a history of cattle being moved. Physical examination may reveal pallor (pale eyes, vaginal mucosa), pyrexia, and elevated heart and respiratory rates. T. orientalis is an economically important parasite of cattle in New Zealand, Australia and Japan, especially where naïve animals are introduced into an endemic area or in animals under stress. Increased awareness of the risks posed by the parasite is required to enable management practices to be implemented to minimise its impact.     

Morphological and quantitative study of the Leydig cells of pigs fed with anabolic doses of clenbuterol

The effects of clenbuterol administered at anabolic doses on the testicular interstitium were studied in 30 pigs allocated to three experimental groups. The diet of two groups was supplemented with clenbuterol (Clb) (1 ppm), but whereas in the Clb+ group the treatment was given until slaughter (treatment period: 3 months), in the Clb- group the clenbuterol was withdrawn 2 weeks before slaughter (treatment period: 2-5 months); in the control group, the pigs were fed without clenbuterol. For histological procedures, a fractional sampling scheme was applied and routine techniques for light and transmission electron microscopy were used. The results of subjective morphology and morphometrics showed slight differences between the treated and the control groups. Conversely, the stereological results identified a prominent hyperplasia of the Leydig cells and ultrastructural analysis of these cells revealed a conspicuous increase in the organelles related to testosterone production, suggesting a functional activation of the interstitial cells in response to the clenbuterol treatment.     

Quantitative changes in the normal and apoptotic thymocytes of pigs treated with anabolic doses of the beta2 adrenergic agonist clenbuterol

Although the beta2 adrenergic agonists have been seen to have important effects on the mechanisms regulating the development and death of T-cells in the thymus, the side-effects on the immune system of anabolic treatments of these substances have hardly been considered. In order to evaluate the effects exerted by the beta2 adrenergic agonist clenbuterol on the thymocyte population, the thymus of eight pigs treated with anabolic doses of this substance was studied by morphometric methods, regarding apoptotic (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL)-positive) and normal (TUNEL-negative) cells. The thymus of another eight pigs fed without clenbuterol served as a control. The clenbuterol treatment had a clear effect on the thymocyte size, decreasing their mean nuclear area. The T-cell apoptosis index was also affected by the clenbuterol, significantly increasing the apoptosis percentage in the treated group with respect to the control. In the light of our results, the clenbuterol induced thymocyte apoptosis throughout the thymus and caused morphometric changes in the thymocyte population, which was in line with the immunosuppressive role attributed to other beta2 adrenergic agonists.     

Effects of anabolic implants of oestradiol alone or in combination with trenbolone acetate on the ultrastructure of mammary glands in female lambs regarding their interference in prolactin secretion

The side-effects of anabolic steroid implants on mammary gland ultrastructure were evaluated in female lambs treated with oestradiol (n = 10) and with oestradiol plus trenbolone acetate (n = 10). Ten non-implanted lambs were used as controls. Apart from the ultrastructural study of the mammary gland, an assessment of the prolactin pituitary cell population was carried out by immunological methods. Our results showed that oestrogenic implants exert stimulating effects on mammary gland development, both by activating the synthesis process at mammary gland cell levels and by increasing prolactin pituitary production. Nevertheless, there was no evidence of secretory products in the lumen of the gland. Implants containing trenbolone acetate counteracted the mammary stimulus of oestrogens showing ultrastructural images of cell autolysis and necrosis.     

含莫纳可林K红曲降低鸡蛋胆固醇效果的研究

48只68周龄的海兰褐蛋鸡分成三组，按0.5％和10％比例分别添加含0．4％莫纳可林K红曲于各组日粮内，饲喂22d，观测其对鸡蛋胆固醇变化的影响。结果表明，投药后5—6d，10％添加组和5％添加组蛋黄胆固醇开始下降；15—22d的胆固醇含量与1～5d相比，10％添加组和5％添加组分别下降了9．02％和5．10％，而对照组升高了5．22％。