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.     

Arthrographic Analysis of Communication between the Tarsometatarsal and Distal Intertarsal Joints of the Horse

The intra-articular anatomy of 103 equine tarsi was studied by contrast radiography with image intensification and computerized tomography. There was communication between the tarsometatarsal and distal intertarsal joints in 21 of 55 (38%) interpretable tarsometatarsal arthrograms, and in 11 of 48 (23%) interpretable distal intertarsal arthrograms. The difference was not significant. The volume of contrast agent and the pressure of injection did not correlate with communication. Forced injection caused subcutaneous leakage of contrast medium but not communication. Communication occurred via the tarsal canal and the space between the third and the combined first and second tarsal bones. Injection of the distal intertarsal joint from the dorsomedial aspect of the limb, distal to the palpable distal border of the medial branch of the tendon of the tibialis cranialis muscle and between the central, third, and combined first and second tarsal bones, provided reliable access except in the presence of severe periosteal proliferations.     

Specificity of polyclonal antibodies to different antigenic preparations of Pseudomonas syringae pv. pisi strain UQM551 and Pseudomonas syringae pv. syringae strain L

Polyclonal antibodies were produced against sonicated and heat-killed cells of Pseudomonas syringae pv. pisi strain UQM551 and Pseudomonas syringae pv. syringae strain L, and their specificities were compared. Evidence is presented that the serological specificity between these two pathovars lies in surface antigens. Of the surface antigens purified and tested, only flagella and lipopolysaccharide from the cell wall showed no cross-reactivity with heterologous antisera. Antisera to glutaraldehyde-fixed flagella of the two strains showed a high level of specificity. At a species or genus level, antisera prepared from heat-killed cells of P. syringae distinguished this species from all other bacterial species and genera tested, including strains of Pseudomonas fluorescens, Escherichia coli, Agrobacterium and Rhizobium.     

Antagonism of detomidine with tolazoline in isoflurane-anaesthetised ponies

The effects of tolazoline (4.0 mg/kg iv) antagonism of detomidine (0.02 mg/kg iv) were evaluated in isoflurane-anaesthetised, ventilated ponies. Each of 6 ponies received both tolazoline and saline treatment during separate anaesthetic episodes only (no surgery was performed). Detomidine administration produced an increase in blood pressure, decrease in heart rate and decrease in P_aO₂ Tolazoline treatment transiently increased heart rate while blood pressure returned to baseline after both treatments. Arterial oxygenation decreased further after tolazoline treatment while oxgenation recovered towards baseline with saline treatment. No other cardiopulmonary effects were detected. Recovery from anaesthesia tended to be more rapid when detomidine was antagonized. The potential benefit of antagonizing detomidine-induced bradycardia with tolazoline, during isoflurane anaesthesia should be weighed against the potential to produce a decrease in arterial oxygenation. The mechanism for this effect is not clear.     

Silviculture of ash in southern England

In 1992 the Wessex Silvicultural Group held a number of meetingsto study the silviculture of ash, during which 16 sites throughoutcentral southern England were visited. Ash can produce valuabletimber on relatively short rotations, and to take full advantageof this potential stands must be carefully managed. The mainpoints to consider are: (1) site selection: ash is site demandingand grows best on moist calcareous barns of pH 6 to 7. Sitesprone to frost should be avoided. (2) Spacing: if timber productionis an important objective, ash should be established with atleast 2500 stems ha^-1 on bare land and 2000 stems ha^-1 on restockingsites. It is sensitive to exposure when young and requires sideshelter making it unsuitable for pure planting on exposed ground.(3) Weed control: this is essential for successful establishment.It is recommended that an area of at least 1 m² around eachtree should be kept weed free for at least 3 years. (4) Protection:the species is relatively free from squirrel damage but is palatableto voles, rabbits, hares and deer. Protection using appropriateindividual tree protection or fencing is essential. (5) Thinning:once a height of 6–7 m has been reached ash should havefrequent crown thinnings to maintain a live crown over at leastone-third the height of the tree. Under-thinning was by farthe most common fault in the stands visited.     

Effect of pH on rheotaxis of bull sperm using microfluidics

The aim of the present research is to study the effect of pH values on the sperm rheotaxis properties. Semen collected from bulls was diluted with SOF medium (1:10). pH of the medium was adjusted using a digital pH meter to the following pH values: 6.0, 6.2, 6.4, 6.4, 6.8, 7.0. All kinetic parameters of sperm (n = 3,385) were determined through a computer‐assisted sperm analysis (CASA) system using microfluidic devices with controlled flow velocity. The following parameters were determined: total motility (TM%), positive rheotaxis (PR%), straightline velocity (VSL, μm/s), average path velocity (VAP, μm/s), linearity (LIN, as VSL/VCL, %), beat cross‐frequency (BCF, Hz) and curvilinear velocity (VCL, μm/s). Nitric oxide, calcium and potassium were estimated in semen at different pH values. To confirm the effect of nitric oxide and K⁺, we used sodium nitroprusside (an NO donor) and KCL as (a K⁺ donor) to see their effect on sperm PR%. The results showed no difference in TM% at pH (6–7). The PR% was the lowest at pH 6 and 7. The best parameters for the PR% were at pH 6.4–6.6. The concentration of Ca⁺² did not change at different pH values. The mean NO values decreased with the increase of pH; however, the mean values of K⁺ increased with the increase of pH. Addition of high concentration of NO and K⁺ to the semen media at fixed pH level had a negative effect on TM% and PR%. In conclusion, the bull sperm had the best rheotaxis properties at pH 6.4–6.6 and sensitive to the change of seminal NO and K⁺.