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.     

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％。     

Disease of geese caused by a new herpesvirus

A goose flock farmed outdoors in south-eastern Queensland suffered an outbreak of peracute disease with high death rate (97%). Small button ulcers and large plaques overlying lymphocyte aggregates were present on the mucosa of the small intestine of affected birds. Small white foci of necrosis and focal haemorrhages were seen in the livers. Numerous intranuclear inclusion bodies were observed microscopically in hepatocytes and a herpesvirus which grew rapidly in chicken kidney cells was isolated from tissues. Duck virus enteritis (DVE) was suspected but DVE antiserums failed to neutralise the virus. Further serological studies with a limited range of known avian herpesviruses have failed to identify the virus. Experimental transmission resulted in high mortality in geese (100%), lower mortality in ducklings and nil mortality in chickens. Surveillance studies showed no evidence of infection in domestic and wild birds beyond the original farm and the infection appears not to have been established in the area. Wild ducks, which were frequent visitors to the farm dam, were considered the most likely source of the infection.     

Blood supply of the ovine adrenal gland and its relevance in adrenal autotransplantation

SUMMARY The arterial supply and venous drainage of 62 left and 5 right ovine adrenal glands is described, and the contribution of individual arteries to successful adrenal gland autotransplantation was evaluated. Arterial flow was measured by direct collection from the draining adrenal vein. Assessment of function of the transplanted adrenal gland was made from survival of the sheep and by the cortisol response to infusion of ACTH and the aldosterone secretory response to infusion of angiotensin II or potassium. For the left adrenal, the principal arterial supply was from the renal artery in 21 (34%), a lumbar artery in 32 (52%), and the anterior mesenteric artery in 3. The total blood flow was 5.0 ± SEM 0.4 mL/min, the flow from the renal branch 2.3 ± 0.3 mL/min, and the principal lumbar branch 2.6 ± 0.3 mL/min. Venous drainage from the left adrenal was via a major adrenal vein to the left renal vein, but additional tributaries to the renal vein were present in 26% . The arterial supply to the adrenal is regional and omission of a branch at transplantation could result in infarction of portion of the gland. By defining arterial supply and measuring blood flow, selection of the appropriate artery or multiple arteries can achieve an adrenal gland autotransplant survival of 90% .     

Land Cover Type and Fire in Portugal: Do Fires Burn Land Cover Selectively?

The purpose of this study is to investigate if, or under what conditions, fires select given land cover types for burning. If fires burn unselectively then the land cover composition (the proportional area of various land cover types) of individual fires should approximate the land cover composition available in their neighborhood. In this study we test this hypothesis by performing statistical analyses of a data set consisting of paired vectors with the proportions of land cover types present in burned areas and in their respective surroundings. The statistical methods employed (a permutation technique and the C_max statistic) are commonly used in resource selection studies where data is subject to a unit-sum constraint. The results of the analysis of 506 fires that burned in Portugal in 1991 indicate that fires are selective, with small fires exhibiting stronger land cover preferences than large fires. According to the results of a multiple comparison analysis performed for small fires, there is a marked preference for shrubland followed by other forest cover types, while agriculture is clearly avoided. A similar analysis is performed to test if fire selectivity is related to the ecological region where it occurs. The results obtained in this study contribute to the discussion on the relative importance of fuels as a drivers of fire spread.     

Use of high-power ultrasonic shears for laparoscopic ovariectomy in mares

A method for performing laparoscopic ovariectomy with high-power ultrasonic shears in mares is described, along with results in 10 mares. Briefly, after epidural administration of detomidine and local anesthesia with mepivacaine, 3 cannulas were placed in each paralumbar fossa. A laparoscope was placed through the most dorsal cannula, and a grasping forceps was placed through the most ventral cannula and used to grasp and manipulate the ovary. The ultrasonic shears was then placed through the middle cannula. The jaws of the ultrasonic shears were closed across a portion of the ovarian pedicle, and the instrument was discharged until tissue within the jaws was transected; the process was repeated until the entire ovarian pedicle was transected. Following removal of the right ovary, it was passed to the left side of the abdomen and both ovaries were removed through an incision in the left paralumbar fossa. No major complications were identified in any of the 10 mares. However, excessive bleeding necessitating reapplication of the ultrasonic shears (2 ovaries) or application of ligating clips (8) was encountered with 10 of the 20 ovaries. Laparoscopic ovariectomy with a high-power ultrasonic shears appears to be safe in mares.