Binding Sites for Ca2+-Channel Effectors and Ryanodine in Periplaneta americana—Possible Targets for New Insecticides

The calcium channel and the ‘calcium release channel’ of muscle membrane of the cockroach Periplaneta americana have been characterized. Biological assays with calcium channel blockers and ryanodine on different insects and acari revealed pronounced insecticidal effects with ryanodine, but not with calcium channel blockers, at concentrations between 0·1 and 300 μg ml⁻¹. Skeletal muscle membranes derived either from the tubular network or from the sarcoplasmatic reticulum of P. americana were characterized with respect to the binding of the dihydropyridine (DHP) [³H]isradipine (PN 200-110), the phenyl-alkylamine [³H]verapamil and the alkaloid [³H]ryanodine. Preliminary binding studies with the benzothiazepine [³H]diltiazem suggest a low-affinity binding site with a IC₅₀ value of 3·3 μM . All binding sites tested were sensitive to treatment with proteinase K. Optimal conditions for binding of the radioligand ryanodine revealed the highest specific binding at pH 8 and at calcium chloride concentrations between 100 and 500 μM . EGTA at 10 μM abolished 95% of the ryanodine binding. Binding studies with calcium channel binding sites revealed a pronounced effect of low Ca²⁺ concentrations on specific isradipine binding, whereas verapamil and diltiazem binding were only reduced by the presence of 200 μM EGTA. With respect to high Ca²⁺ concentrations, specific binding of diltiazem, isradipine and verapamil was reduced by 73, 40 and 20%, respectively, at 5 mM Ca²⁺. Radioligand binding experiments showed high-affinity binding sites for ryanodine and isradipine. K_D values of 0·95 nM (B_max=550 fmol mg⁻¹ protein) and 0·75 nM (B_max=213 fmol mg⁻¹ protein) were determined respectively. A lower-affinity binding site was identified in binding studies with verapamil (K_D=7·4 nM and B_max=27 fmol mg⁻¹ protein). [³H]isradipine displacement studies with several dihydropyridines revealed the following ranking of affinity: nitrendipine>isradipine>Bay K8664≪nicardipine. Displacement of [³H]verapamil binding by effectors of the phenylalkylamine binding site showed that bepridil and S(-)verapamil had the highest affinities of the compounds tested followed by (±)verapamil, nor-methylverapamil and R(+)verapamil.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.     

Genetic Analysis of Black Spruce (Picea mariana) Populations from Dry and Wet Areas of a Metal-Contaminated Region in Ontario (Canada)

Genetic variation and genetic structure of black spruce (Picea mariana L.) populations growing in wet land (lowlands) and dry lands (uplands) with different levels of metal contaminations were analyzed using ISSR. Polymorphic loci (P%) ranged from 65% to 90% with a mean of 75%. Nei??s gene diversity (h) varied from 0.264 to 0.359 with a mean of 0.310, and Shannon??s index (I) ranged from 0.381 to 0.524 with a mean of 0.449. The level of genetic variation was higher in populations from wet lands than those from dry lands. Variation within populations accounts for most of total genetic variation. The genetic distance among the black spruce (P. mariana) populations ranged from 0.171 to 0.351. The present study indicates that genetic variation and long-term exposure to metals (more than 30 years) are not associated. Cytological analysis of black spruce seeds from metal-contaminated and -uncontaminated areas showed normal mitotic behavior during prophase, metaphase, anaphase, and telophase.     

Continuous positive airway pressure administered via face mask in tranquilized dogs

Objective – To evaluate the tolerance of a continuous positive airway pressure (CPAP) mask in tranquilized dogs and compare PaO₂ in arterial blood in dogs receiving oxygen with a regular face mask or CPAP mask set to maintain a pressure of 2.5 or 5 cm H₂O. Design – Prospective, randomized clinical study. Setting – University teaching hospital. Animals – Sixteen client‐owned dogs without evidence of cardiopulmonary disease were studied. Interventions – Eight animals were randomly assigned to each of 2 treatment groups: group A received 2.5 cm H₂O CPAP and group B received 5 cm H₂O CPAP after first receiving oxygen (5 L/min) by a regular face mask. Animals were tranquilized with acepromazine 0.05 mg/kg, IV and morphine 0.2 mg/kg, IM. An arterial catheter was then placed to facilitate blood sampling for pHa, PaO₂, and PaCO₂ determinations before and after treatments. Direct mean arterial pressure, heart rate, respiratory rate, and temperature were also recorded after each treatment. Measurements and Main Results – CPAP administration was well tolerated by all animals. The mean arterial pressure, heart rate, respiratory rate, temperature, PaCO₂, and pHa, did not differ at any time point between groups. Differences were seen in oxygenation; in group A, PaO₂ significantly increased from a mean of 288.3±47.5 mm Hg with a standard mask to a mean of 390.3±65.5 mm Hg with the CPAP mask and in group B, PaO₂ increased similarly from 325.0±70.5 to 425.2±63.4 mm Hg (P<0.05); no differences were detected between the 2 CPAP treatments. Conclusions – In healthy tranquilized dogs noninvasive CPAP is well tolerated and increases PaO₂ above values obtained when using a regular face mask.     

The prevalence and clinical relevance of hyperkalaemia in calves with neonatal diarrhoea

One hundred and twenty-four calves with neonatal diarrhoea were investigated in order to assess the prevalence of hyperkalaemia and the associated clinical signs. Hyperkalaemia (potassium concentration >5.8 mmol/L) was recognized in 42 (34%) calves and was more closely associated with dehydration than with decreases in base excess or venous blood pH. In 75 calves with normal blood concentrations of D-lactate (i.e. ?3.96 mmol/L), K concentrations were moderately correlated with base excess values (r = ?0.48, P < 0.001). In contrast, no significant correlation was observed in 49 calves with elevated D-lactate. Only three hyperkalaemic calves had bradycardia and a weak positive correlation was found between heart rate and K concentrations (r = 0.22, P = 0.014). Ten of the 124 calves had cardiac arrhythmia and of these seven had hyperkalaemia indicating that cardiac arrhythmia had a low sensitivity (17%) but a high specificity (96%) as a predictor of hyperkalaemia.In a subset of 34 calves with base excess values ??5 mmol/L and D-lactate concentrations <5 mmol/L (of which 22 had hyperkalaemia), changes in posture/ability to stand could be mainly explained by elevations of K concentrations (P < 0.001) and to a lesser extent by increases in L-lactate concentrations (P = 0.024). Skeletal muscle weakness due to hyperkalaemia alongside hypovolaemia may produce a clinical picture that is similar to that in calves with marked D-lactic acidosis. However, since reductions in the strength of the palpebral reflex are closely correlated with D-lactate concentrations, a prompt palpebral reflex can assist the clinical prediction of hyperkalaemia in calves presenting with a distinct impairment in their ability to stand (specificity 99%, sensitivity 29%).     

The Effect of Sedation on Gastric Emptying of a Liquid Marker in Ponies

OBJECTIVE: The effect of sedation on gastric emptying was evaluated in six ponies by monitoring serum concentrations of acetaminophen (AP) after intragastric administration. EXPERIMENTAL DESIGN: Prospective randomized experimental study. ANIMALS: Six adult ponies, 135 to 275 kg. METHODS: Fifteen minutes after the intravenous administration of xylazine (1 mg/kg), butorphanol (0.05 mg/kg), acepromazine (0.05 mg/kg) or saline, ponies were given AP (20 mg/kg in 350 mL water) by stomach tube. Blood for AP analysis was collected at baseline and 15, 30, 45, 75, 90, 105, and 120 minutes after AP administration. The time (Tmax) to reach peak serum concentration (Cmax), and the area under the AP serum concentration versus time curve (AUC) were determined for each treatment group. RESULTS: Tmax was 31 mins in the control group, and this increased significantly (P<.05) after sedation. Cmax decreased (P<.05) after xylazine administration, and AUC decreased (P<.05) after acepromazine. CONCLUSIONS: This study indicated that sedation has a significant effect on the gastric emptying rate of a liquid in ponies.     

Effects of forest floor planting and stock type on growth and root emergence of Pinus contorta seedlings in a cold northern cutblock

A two-year field trial was conducted to determine the growth response, and root emergence pattern of interior lodgepole pine (Pinus contorta Dougl. var. latifolia Engelm.) seedlings in response to container type and forest floor removal. Seedlings were grown in Styroblocks^TM, Copperblocks^TM, or AirBlocks^TM, and were planted directly into the undisturbed forest floor or into manually prepared planting spots where the forest floor had been scraped away to expose the mineral soil. Seedlings planted into scalped planting spots exhibited marginally but significantly (7%) greater above-ground growth rates (seedling stem volume); whereas seedlings planted into the forest floor produced significantly more (11%) new roots. There were no differences in above- or below-ground biomass. Seedlings grown in Copperblock^TM containers produced a higher proportion of roots near the top of the plug when tested at lifting, however this pattern was not observed in the field. Given that scalping is more costly than forest floor planting, and that the increased shoot growth was relatively small, we recommend that forest floor planting be considered as an alternative to manual spot scalping for sites, such as the site tested here: those with cold, but well-drained soils and where competition from other plants is not a serious problem.     

Case of suspected theobromine poisoning in dairy cattle

High-yielding dairy cows collapsed and died displaying signs of a disturbed central nervous system (muscle tremor, convulsion) and a considerably reduced body condition score. An intense diagnostic screening did not allow to confirm a definite diagnosis. Therefore, further analyses including an evaluation of feeds and feeding were initiated. The herd was fed a total mixed ration (TMR) based on corn and grass silage of moderate nutritive value supplemented with various amounts of chocolate chips. In retrospect, the amount of chocolate chips added to the respective TMR batches could not be quantified. These chips were purchased from a company producing bakery and chocolate products for human consumption and added to the silage and therefore to the TMR in order to increase the energy intake of the animals. Because the TMR, which was fed during the time of the incidence, was no longer available, a sample of a later batch of silage was examined. The amount of chocolate chips was quantified (0.44% per fresh matter), and a theobromine analysis was performed in the chocolate chips of the available batch (69.7 mg/100 g fresh matter). Because of the possible link between chocolate intake and observed signs, an immediate cessation of using the chocolate chips was recommended in addition to an optimisation of the TMR, that is an increase of the energy density. Even though the theobromine intake during the time of the incidence is unknown and information about toxicity of theobromine in ruminants is limited, we suspect that the feeding of chocolate in this case caused all signs including the sudden death of the cows. Further reasons are that no differential diagnoses were established and the problems at the farm stopped after removing the chocolate from the TMR.