Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Effect of nitrogen fertiliser on temporal and spatial variation of mineral nitrogen and microbial biomass in a silvopastoral system

This paper describes a field study to assess the effect of increasing the frequency of split applications of N fertiliser on the pattern of plant uptake, soil N availability, and microbial biomass C and N. Measurements were taken during the growing season in different positions relative to young trees (Prunus avium L.) in an upland silvopastoral system in its first year after establishment. At fertiliser rates of 72 and 144 kg ha^-1 N applied as NH₄NO₃, increasing the number of split applications increased N uptake by the pasture. Mineral forms of soil N measured 2 weeks after application indicated that residual NH _inf4 ^sup+ -N and total mineral N were also greater in this treatment on certain dates. Soil NO _inf3 ^sup- -N was positively correlated with the soil moisture content, and nitrification reached a maximum in early May and declined rapidly thereafter except within the herbicide-treated areas around the trees where soil moisture had been conserved. Results of the study suggest that high NO _inf3 ^sup- -N in herbicide-treated areas was probably caused by mineralisation of grass residues and low uptake by the tree rather than by preferential urine excretion by sheep sheltering beside the trees. Mean microbial biomass C and N values of 894 and 213 kg ha^-1, respectively, were obtained. Microbial C was slightly increased by the higher frequency of split applications at 144 kg ha^-1 N and was probably related to the greater herbage production with this treatment. Microbial N was not significantly affected by the N treatments. Both microbial biomass C and N increased during the growing season, resulting in the net immobilisation of at least 45 kg ha^-1 N which was later released during the autumn.     

Biochemical analysis of normal articular cartilage in horses

Articular cartilage specimens from the distal articular surface of 32 radiocarpal bones from 24 2- to 5-year-old horses were analyzed. The total collagen content was determined on the basis of the 4-hydroxyproline content, using a colorimetric method. A method for estimating the proportions of types-I and -II collagen by measuring spectrophotometric densities of specific cyanogen bromide peptide bands from mixtures of types-I and -II collagen on sodium dodecyl sulfate-polyacrylamide gels was used. The cyanogen bromide peptides representative of each collagen types-I and -II were identified. The peptide ratios were then computed for each of several standards of type-I and -II mixtures. A standard curve was derived from the correlation between these ratios and the corresponding proportions of type-II collagen in standard mixtures. Galactosamine and glucosamine content (hexosamines) were measured by ion chromatography. The galactosamine-to-glucosamine ratio, chondroitin sulfate and keratan sulfate values, and total glycosaminoglycan content were derived from the measured hexosamine content. The total collagen content averaged 556 mg/g (55.6 mg/100 mg) of tissue (dry weight, [dw]). Type-II collagen was the major collagen type in normal articular cartilage specimens. The ratio of the area under the alpha 1 (II)CB10 peak to the area under the alpha 1 (I)CB 7,8 + alpha 1 (II)CB11 peak was a second-order polynomial function of the proportion of type-II collagen in the specimens. The mean galactosamine and glucosamine content were 20.6 mg/g and 7.9 mg/g (dw), respectively. The mean galactosamine-to-glucosamine ratio was 3.74 +/- 0.62.(ABSTRACT TRUNCATED AT 250 WORDS)     

Abomasal intubation of young calves.

A nonsurgical method for passing a tube into the abomasum of young calves was developed. The tube described may be used for infusion into, or sampling from, the abomasum. The tube was successfully passed into the abomasum of all eight calves used. In two calves two to four days old and in two neonatal calves (greater than 24 h old) reticular groove closure was stimulated by providing a suckling stimulus but in four neonatal calves, passage of the tube was successful without the suckling stimulus. Placement of the tip of the tube was verified by auscultation and, in two calves, by radiography.     

Attenuation of Eimeria species: further characterisation of two lines of Eimeria mitis

The immunogenicity of a 'precocious' and attenuated line (HP10) of Eimeria mitis was studied and the stability of attenuation of two precocious lines was compared with that of an embryo-adapted line. Chicks housed in wire-floored cages and given 1 X 10(5) oocysts of the HP10 line were protected against challenge with the parent Houghton strain and two field strains, but remained partially susceptible to the Durham and one other field strain. However, when chicks were kept in litter pens so that reinfection could occur freely, inoculation with as few as 1 X 10(3) oocysts of the precocious line resulted in complete immunity to both the Houghton and Durham strains for at least five weeks afterwards. The stability of attenuation of the precocious and embyro-adapted lines was examined by relaxing the selection pressures used to produce attenuation and then testing the virulence of the resulting lines. The precocious lines remained attenuated after several consecutive relaxed passages, in contrast to the embyro-adapted line which showed a marked reversion to virulence after six passages in chickens.     

Phenobarbitone-responsive hypersialism in two dogs

Two dogs were examined for hypersialism associated with non-painful, symmetrical enlargement of both mandibular salivary glands. These signs were accompanied by inappetence and weight loss. The underlying cause of the problem could not be determined despite extensive investigations including fine needle aspiration biopsy of affected salivary glands. Hypersialism and mandibular salivary gland enlargement resolved completely during pheno-barbitone administration.