Comparative enzyme activities and cytochrome P-450 levels of some rat tissues with respect to their metabolism of several pesticides

Cytochrome P-450, A- and B-esterase, amidase, and glutathione S-aryl transferase were assayed in the postmitochondrial centrifugal fraction, microsomes, and supernatant of rat liver, lungs, kidneys, and testes. Liver microsomes contained the highest P-450 levels and A-esterase activity. B-esterase activity was more generally distributed and higher in the microsomal tissue fractions. Microsomal amidase activity was highest in rat lung and lowest in the liver (per mg protein). Glutathione S-aryl transferase activity was highest in the liver. The in vitro metabolism of carbaryl, phosphamidon, and chlorotoluron by the various centrifugal fractions revealed many differences. Carbaryl metabolism was greater in the liver microsomal fractions than in any other preparation. 1-Naphthol was the major metabolite in all tissue fractions. Although very little metabolism of phosphamidon occurred in the rat, metabolism in the rat liver postmitochondrial fraction was slightly higher with respect to the production of metabolites than in the supernatant and microsomes combined. Chlorotoluron was not metabolized by any of the tissue fractions of the rat. At least a low level of activity toward some compounds was observed in all tissues, but this study confirmed that the liver was the most active metabolizing tissue as well as having the highest levels of enzymatic activity usually associated with pesticide metabolism.     

Characterization of acetylcholinesterases of acaricide resistant and susceptible strains of the cattle tick Boophilus microplus (Can.): II. The substrate specificity and catalytic efficiency of the critical enzyme component

Concentrations and turnover numbers have been determined for the critical acetylcholinesterase component of the organophosphorus susceptible Yeerongpilly and resistant Biarra and Ridgelands strains. The lower specific activity for the substrate acetylthiocholine in the resistant strains was shown to be due to decreased catalytic efficiency and not to any change in the titer of the enzyme. Thioester analogs of acetylthiocholine, with additional methylene groups between the carbonyl carbon and the quaternary nitrogen of the molecule and in the alkyl moiety, have been used to derive K_m and V_max values for the enzymes from each strain. An hypothesis is proposed concerning the alteration of the active site that has occurred to produce the dissimilar aberrant enzymes of the resistant strains and the implications of this hypothesis for inhibitor design are discussed.     

Fetal and maternal lesions in pregnant ewes ingesting locoweed (Astragalus lentiginosus).

Locoweed (Astragalus lentiginosus) was fed to ewes from 70 to 100 days of pregnancy. Ewes were killed at 100, 115, 130, and 145 days, and selected tissues were collected from both the dam and the fetus for histologic examination. Neurovisceral cytoplasmic vacuolation and neuroaxonal dystrophy decreased in dam and fetus with time after the feeding of locoweed was discontinued.     

Characterization of acetylcholinesterases of acaricide-resistant and susceptible strains of the cattle tick Boophilus microplus (Can.): I. Extraction of the critical component and comparison with enzyme from other sources

The critical acetylcholinesterase component, that is responsible for the resistance mechanism of decreased sensitivity to the inhibitor in the cattle tick Boophilus microplus (Can.), has been isolated from the organophosphorus-susceptible Yeerongpilly and resistant Biarra and Ridgelands strains. With the substrate acetylthiocholine this enzyme component gave a pH activity profile similar for all strains. Optimal substrate concentration for Ridgelands was 1.73.10^?2M compared to 2.70.10^?4M for the other strains. Computed Km values were 1.98.10^?5M, 3.65.10^?5M, and 6.43.10^?6M for Yeerongpilly, Biarra, and Ridgelands strains, respectively, while the corresponding V_max figures were 336, 135, and 21 μmoles acetylthiocholine hydrolyzed per gram of larvae per hour. These results are discussed in relation to the resistance mechanism and compared with similar parameters derived for this enzyme from bovine erythrocytes, the sheep blowfly Lucilia cuprina, and the sheep nematode Trichostrongylus colubtiformis.     

Antifungal mode of action of triforine

Rapidly growing mycelia of Aspergillus fumigatus treated with 10 μg/ml triforine (N,N′-bis-(1-formamido-2,2,2-trichloroethyl)-piperazine) showed little or no inhibition in dry weight increase prior to 2 h. By 2.5–3 h, triforine inhibited dry weight increase by 85%. The effects of triforine on protein, DNA, and RNA syntheses corresponded to the effect on dry weight increase both in time of onset and magnitude. Neither glucose nor acetate oxidation were inhibited by triforine.Ergosterol synthesis was almost completely inhibited by triforine even in the first hour after treatment. Inhibition of ergosterol synthesis was accompanied by an accumulation of the ergosterol precursors 24-methylenedihydrolanosterol, obtusifoliol, and 14α-methyl-Δ^{8, 24 (28)}-ergostadienol. Mycelia treated with 5 μg/ml of triarimol (α-(2,4-dichlorophenyl)-α-phenyl-5-pyrimidinemethanol) also accumulated the same sterols as well as a fourth sterol believed to be Δ^{5, 7}-ergostadienol.Identification of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in untreated mycelia indicates that the C-14 methyl group is the first methyl group removed in the biosynthesis of ergosterol by A. fumigatus. The lack of detectable quantities of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in triforine or triarimol-treated mycelia and the accumulation of C-14 methylated sterols in treated mycelia suggests that both fungicides inhibit sterol C-14 demethylation. The accumulation of Δ^{5, 7}-ergostadienol in triarimol-treated mycelia further implies that triarimol also inhibits the introduction of the sterol C-22(23) double bond.Two strains of Cladosporium cucumerinum tolerant to triforine and triarimol were also tolerant to the fungicide S-1358 (N-3-pyridyl-S-n-butyl-S′-p-t-butylbenzyl imidodithiocarbonate).     

Development of an EU protocol for the detection and diagnosis of Potato spindle tuber pospiviroid*

The work described here formed part of the EU SMT DIAGPRO project, to develop diagnostic protocols for 18 regulated pests. The Potato spindle tuber pospiviroid (PSTVd) protocol was developed primarily for testing in vitro‐ and glasshouse‐grown potato plants for the purposes of post‐entry quarantine and the production of pathogen‐tested nuclear stock. After a performance audit of methods used by 12 laboratories in Europe and America by ring testing, four methods were chosen for multilaboratory validation. For most laboratories, the detection limits were 10–20 mg of PSTVd‐infective tissue for R‐PAGE; 0.25–0.5 mg for DIG‐probe; 0.062 mg for RT‐PCR; and 0.0155 mg for TaqMan (this was the lowest weight of infective tissue tested). Some laboratories were able to extend the detection limit to 0.0155 mg for DIG‐probe and RT‐PCR. The DIG‐probe and R‐PAGE are recommended as primary detection methods, with confirmation of viroid presence by any of the four validated detection methods. Specific diagnosis requires the viroid to be sequenced. Other methods may be used for primary detection, providing that they preferably detect all PSTVd isolates and other Pospiviroids that have the potential to infect potato, and detect viroid in at least 1/10 of the tissue weight normally tested per plant.     

Geriatric heart diseases in dogs.

A discussion of the diagnosis and therapy of heart disease in an aged pet does not differ significantly from that in a pet of any age. Mitral regurgitation constitutes by far the most important geriatric heart disease, and the selection of drugs to treat heart disease of aging pets is based on identification of specific pathologic features (eg, atrial fibrillation, left atrial enlargement) for which each aspect of treatment (eg, diuretics, angiotensin-converting enzyme inhibitors, spironolactone) is specific.     

The influence of sampling scheme and interpolation method on the power to detect spatial effects of forest birds in Ontario (Canada)

Spatial ecology is becoming an increasingly important component of resource management, and the general monitoring of how human activities affect the distribution and abundance of wildlife. Yet most work on the reliability of sampling strategies is based on a non-spatial analysis of variance paradigm, and little work has been done assessing the power of alternative spatial methods for creating reliable maps of animal abundance. Such a map forms a critical response variable for multiple scale studies relating landscape structure to biotic function. The power to reconstruct patterns of distribution and abundance is influenced by sample placement strategy and density, the nature of spatial auto-correlation among points, and by the technique used to extrapolate points into an animal abundance map. Faced with uncertainty concerning the influence of these factors, we chose to first synthesize a model reference system of known properties and then evaluate the relative performance of alternative sampling and mapping procedures using it. We used published habitat associations of tree nesting boreal neo-tropical birds, a classified habitat map from the Manitou Lakes area of northwestern Ontario, and point count means and variances determined from field studies in boreal Canada to create 4 simulated models of avian abundance to function as reference maps. Four point sampling strategies were evaluated by 4 spatial mapping methods. We found mixed-cluster sampling to be an effective point sampling strategy, particularly when high habitat fragmentation was avoided by restricting samples to habitat patches >10 ha in size. We also found that of the 4 mapping methods, only stratified ordinary point kriging (OPK) was able to generate maps that reproduced an embedded landscape-scale spatial effect that reduced nesting bird abundance in areas of higher forest age-class fragmentation. Global OPK was effective only for detecting broader, regional-scale differences. This revised version was published online in July 2006 with corrections to the Cover Date.     

Historical range of variability in eastern Cascades forests,Washington, USA

The historical range of variability (HRV) has been suggested as a coarse filter approach to maintain ecosystem sustainability and resiliency. The historical range of variability in forest age structure for the central eastern Cascade Range in Washington State, USA was developed from historical fire return intervals and the manner in which fire acted as both cyclic and stochastic processes. The proportions of seven forest structural stages calculated through these processes were applied to the area of each forest series within the central eastern Cascades landscape. Early successional forest stages were more common in high elevation forest than low elevation forest. The historical proportion of old growth and late successional forest varied from 38 to 63 percent of the forested landscape. These process-based estimates are consistent with those developed from forest structural information. HRV is a valuable planning tool for ecosystem conservation purposes, but must be applied to real landscapes with consideration of both temporal and spatial scale. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001