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.     

Effect of ruminal protein degradability on growth and N metabolism in growing beef steers

The objective of two experiments was to correlate plasma levels of urea N (PUN) and the percentage of urine N in the form of urea (UUN) to weight gain in response to different dietary protein regimens for growing Angus steers. In Exp. 1, 60 steers (302 kg BW) were assigned to various levels of dietary N (control plus supplemental N to provide from 100 to 400 g more crude protein daily) within two sources of supplemental N (soybean meal [SBM] or a mixture of two parts corn gluten meal:one part blood meal [CGM:BM]). In Exp. 2, 27 steers (229 kg BW) were fed two levels of SBM, and half of the steers received growth-promoting implants. Steers were housed in groups of 12 and fed individually for 84 d in both experiments. Corn silage was fed at a restricted rate to minimize orts. Jugular blood and urine samples were collected during the experiments. In Exp. 1, maximal ADG of steers fed SBM (1.0 kg) was reached with 671 g/d total crude protein, or 531 g/d metabolizable protein. Maximal ADG of steers fed CGM:BM (0.91 kg) was reached with 589 g/d total crude protein, or 539 g/d metabolizable protein. The DMI was higher (P < 0.07) for steers fed SBM (6.37 kg/d) than for steers fed CGM:BM (6.14 kg/d). Increasing ruminal escape protein from 36% (SBM) to 65% (CGM:BM) of CP decreased (P < 0.05) endogenous production of urea, as evidenced by lower concentrations of urea in blood and lower UUN. In Exp. 2, increasing supplemental protein from 100 to 200 g/d increased (P < 0.05) ADG and PUN. Implants lowered (P < 0.05) UUN, particularly at the higher level of supplemental protein. Protein supplementation of growing steers can be managed to maintain acceptable ADG yet decrease excretion of urea in the urine.     

Effect of tapered normal and interval training on performance of Standardbred pacers

Human athletes taper or reduce their training load before a race to enhance performance, apparently because recovery from the effects of fatigue occurs faster than the loss of fitness from the reduced training. However, there appear to be no previous studies of tapering of equine athletes. Our aim in the present study was, therefore, to investigate the efficacy of tapering with Standardbred pacers. We determined the effect of repeated cycles of tapered training on performance of Standardbred pacers. After 8 weeks of jogging and 3 x 2 week cycles of pace work, 19 horses were randomised to a taper and a control group. The taper group completed 5 consecutive 2 week cycles, each incorporating a 7 day taper; some cycles included high-intensity interval training. The control group continued with 5 more cycles of pace work. All horses completed a 2400 m individual time trial after each cycle. Peak and mean speed of the taper group were faster than those of the control group in all cycles; the differences were clear-cut in all cycles for peak speed (overall 4.4%, 95% confidence interval 1.7 to 7.1%), but only in one of the interval-training cycles for mean speed (2.4%, 0.3 to 4.7%). Four horses in the taper group were injured during interval training. Repeated tapering produces a worthwhile enhancement of performance in Standardbreds, but the addition of interval training appears to increase the risk of injury.     

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％。     

Nutritional stress as a cause of thymic atrophy in broiler chickens

A study was conducted to examine the effect of nutritional stress on the development of the thymus, bursa, and pancreas of 7-to-14-day-old commercial meat-type chickens. One group of 7-day-old chickens was given access to food for only 30 minutes daily for 7 days. The birds were necropsied, and the thymus, pancreas, and bursa were compared with those of the control chickens fed ad libitum and necropsied at 7 and 14 days of age. The thymuses from birds on the restricted diet were atrophied (0.45 +/- 0.08 g) and congested compared with the thymuses from chickens fed ad libitum (1.32 +/- 0.31 g). The bursae from chickens on a restricted diet were also smaller (0.26 +/- 0.08 g) than the bursae from birds with free access to food (0.74 +/- 0.11 g). The restricted diet did not appear to cause any gross or histological pancreatic changes. The thymic lesions produced by nutritional stress were similar to those observed in the runting/stunting syndrome.