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.     

A specific DNA probe which identifies Babesia bovis in whole blood.

A genomic library of Babesia bovis DNA from the Mexican strain M was constructed in plasmid pUN121 and cloned in Escherichia coli. Several recombinants which hybridized strongly to radioactively labeled B. bovis genomic DNA in an in situ screening were selected and further analyzed for those which specifically hybridized to B. bovis DNA. It was found that pMU-B1 had the highest sensitivity, detecting 25 pg of purified B. bovis DNA, and 300 parasites in 10 microliters of whole infected blood, or 0.00025% parasitemia. pMU-B1 contained a 6.0 kb B. bovis DNA insert which did not cross-hybridize to Babesia bigemina, Trypanosoma evansi, Plasmodium falciparum, Anaplasma marginale, Boophilus microplus and cow DNA. In the Southern blot analysis of genomic DNA, pMU-B1 could differentiate between two B. bovis geographic isolates, Mexican strain M and Thai isolate TS4. Thus, the pMU-B1 probe will be useful in the diagnosis of Babesia infection in cattle and ticks, and in the differentiation of B. bovis strains.     

Quantitative Aspects of Thyroid Scintigraphy With Pertechnetate (99mTcO4) in Cats

Thyroidal ^99mTcO₄(pertechnetate) uptake percentages were determined in unanesthetized euthyroid (n = 13) and hyperthyroid (n = 18) cats. Maximal uptakes were observed 60 minutes after IV injection of the radionuclide and ranged from 0.3 to 3.9% of the dose in euthyroid cats (median 2.23%) and from 5.2% to 23.9% of the dose in hyperthyroid cats (median 14.8%) ( P < .05). There were no overlaps in pertechnetate uptake percentages during any of the intervals evaluated. It is concluded that the optimal time for visualization of the thyroid by ^99mTcO₄-scanning is 60 minutes after IV injection of the radionuclide. Calculation of the percentage uptake is of additional diagnostic value.     

Principles of fracture fixation in growing animals.

Changes in bone structure and conformation can occur very rapidly in growing animals. Injury to the bones or articular surfaces warrant special consideration because of the potential devastating consequences of secondary growth deformities in the animal. Disturbance of normal growth plate activity, imperfect fixation of an articular fracture, and conformational defects in long bones may create lifelong problems for the animal.     

Comparative virulence of NAD-dependent and NAD-independent Actinobacillus pleuropneumoniae strains.

The virulence of a NAD-independent Actinobacillus pleuropneumoniae serotype 2 strain and NAD-dependent serotype 2, 3 and 9 strains was compared under experimental conditions. Hysterectomy-derived piglets were inoculated endobronchially with 50-500 cfu of these strains. All 23 piglets inoculated with the NAD-dependent strains developed acute disease within 12 hours post inoculation. Twenty-two of these piglets died within 24 hours after the first clinical signs. Three of nine piglets inoculated with the NAD-independent strain did not develop clinical disease. In the other six piglets, disease signs were similar as in the piglets inoculated with the NAD-dependent strains. No differences in clinical disease were observed between colostrum deprived piglets and piglets that obtained colostrum from a SPF sow.     

A Quantitative Study of Ganglion Cells in the German Shepherd Dog Retina

As in the number of mammals, the most prominent feature of the ganglion-cell layer in the retina of the German shepherd dog is the sharp increase in the density of ganglion cells in the central area. There is an area of maximum density and also a ‘cat-like’ visual streak, located dorsal to the optic disc. The isodensity lines of ganglion-cell distribution is roughly concentric. Their values vary from 5300-13 000 cells/mm² in the central area, with the cells densely packed, to 1000 cells/mm² or less in the periphery, where the cells are sparsely distributed. There were some individual differences amongst the animals studied, although all of them were pure-bred dogs. This suggests that the configuration of the retina in the canine species is not only dependent on the breed itself but also on some other parameters such as phylogenetic heritage, environment, aptitude, lifestyle, or even training.     

Efficacy of the morantel sustained release trilaminate matrix against gastrointestinal nematodes in beef calves

The effectiveness of the morantel sustained release trilaminate (MSRT) in controlling gastrointestinal nematodes through a grazing season was evaluated using 60 yearling beef stocker calves randomly divided into 2 groups of 30 animals each. In April 1985, the calves comprising the treatment group each received an MSRT designed to release morantel tartrate continuously for 90 days while those of the control group remained unmedicated. All animals were weighed and samples of rectal feces were taken at 14-day intervals, beginning on Day 0, until trial termination (Day 168). At trial termination, 10 control and 10 treated calves were necropsied for recovery of gastrointestinal nematodes. Three sets of parasite-na?ve tracer calves were utilized to evaluate the initial, interim and final levels of pasture contamination by nematode larvae. Overall, the use of the MSRT resulted in a 75.5% reduction (P less than 0.001) in output of nematode eggs from the principals, an 81.8% reduction (P less than 0.001) in numbers of gastrointestinal nematodes in principals (at trial termination), and a 96.9% reduction (P less than 0.05) of pasture larval nematode contamination (as indirectly indicated by parasite burdens in tracer calves). The mean weight advantage of treated calves was 16.6 kg per head (P less than 0.001).     

Effect of Completion of Vernalization on Generative Development of Winter Rape (Brassica napus L. var. oleifera) Plants

Germinating seeds and young plants of winter rape var. Górczañski were vernalized for 56–63 days under conditions of 9-hour day, at the temperature 2 and 5 °C and in continuous darkness at the temperature 2 °C. After vernalization the plants grew under conditions enabling to complete vernalization: in a glass-house at the temperature day/night 15/10 °C and in semi natural conditions of open vegetation hall in the period from June till August. After sub-optimal vernalization further growth of the plants at lowered temperature increased its effectiveness (completion of vernalization). Depending on the degree of the vernalization of the plants the completion of their vernalization was both obligatory, i.e. conditioning the acquisition of the ability of generative development, and facultative i.e. accelerating this development. It has been demonstrated that the population of plants of the examined variety is strongly differentiated not only with respect of vernalization requirements in the particular plants, but also what regards the effectiveness of vernalization completion. New observations have been made indicating that the mechanisms controlling the successive phases of generative development, i.e. phase of forming flower buds and the flowering phase are not identical which may be interpreted as indicating that the "flowering factor" is polymorphous.