Selective measurement of lipoprotein lipase and hepatic triglyceride lipase in heparinized plasma from horses.

Affinity chromatography on heparin sepharose was used to identify 2 lipolytic enzymes in heparinized plasma from horses. One enzyme was typical of hepatic triglyceride lipase (HTGL), because it was resistant to inactivation by high concentrations of NaCl, and it did not require the addition of serum for activity. The other enzyme was identified as lipoprotein lipase (LPL), because of its inactivation at NaCl concentrations in excess of 0.2M, and its dependency on addition of serum as a source of apolipoprotein C-II activator. The enzymes were purified by 347-(HTGL) and 442- (LPL) fold, with yields of 54 and 58%, respectively. The partially purified enzymes were used to design incubation conditions that gave optimal activities for each enzyme in vitro. A selective assay was then developed for direct measurement of LPL and HTGL activities in heparinized plasma from horses. Analysis of HTGL took advantage of the almost complete inactivation of LPL when serum cofactor was excluded from the assay at the NaCl concentration that gave optimal HTGL activity. Prior incubation of heparinized plasma with sodium dodecyl sulfate to inhibit HTGL was necessary for measurement of LPL, because HTGL retained 67% of its activity at the NaCl concentration required for optimal LPL activity. Activity of each enzyme was measured in heparinized plasma from 12 Shetland ponies. The mean activity +/- SD for LPL was 3.22 +/- 1.04 mumol of fatty acids/ml of heparinized plasma/h (mumol of FA/ml/h. The mean activity for HTGL was 4.9 +/- 1.56 mumol of FA/ml/h. The performance of the assay was assessed by replicate analysis of pools of each enzyme with high and low activities.(ABSTRACT TRUNCATED AT 250 WORDS)     

Quantitative analysis of canine plasma lipoproteins

A combined ultracentrifugationl/precipitation method for the measurement of lipoprotein cholesterol concentrations was developed and validated for use with canine plasma. Very low density lipoproteins (VLDL) were isolated by flotation ultracentrifugation and low density lipoproteins (LDL) separated from high density lipoproteins (HDL) by precipitation with heparin-manganese chloride. Effective separation of these classes was confirmed by agarose gel electrophoresis of native lipoproteins and by sodium dodecyl sulphate polyacrylamide gel electrophoresis of their apolipoprotein distributions. There was trace contamination of the LDL precipitate with HDL, but this represented less than 4 and 9 per cent of the total plasma HDL in normo- and hypercholesterolaemic dogs, respectively. The intra-assay and interassay coefficients of variation for LDL- and HDL-cholesterol concentrations were between 3·3 and 6·9 per cent, and 7·2 and 9·0 per cent, respectively, for plasma cholesterol concentrations between 2·67 and 8·14 mmoll/litre. The intra-assay coefficient of variation for VLDL-cholesterol was 53·8 and 18·4 per cent at plasma cholesterol concentrations of 2·67 and 8·14 mmol/litre, respectively. The interassay coefficient of variation for VLDL was 22·5 per cent. Storage of plasma at -20°C for between two and eight weeks did not affect VLDL-cholesterol concentrations, but led to an increase in LDL-cholesterol and a decrease in HDL-cholesterol concentrations of approximately 10 per cent. The method described is appropriate for the measurement of lipoprotein concentrations in plasma from normo- and hypercholesterolaemic dogs, but samples should not be subjected to prolonged storage before analysis.     

Efficacy of doramectin against experimental infections by some nematode parasites in cattle in New Zealand

A study was undertaken to determine the efficacy of the novel avermectin, doramectin, against experimental larval and adult infections of three species of nematode parasite important to cattle production in New Zealand. Eighteen worm-free dairy bull beef calves were randomly allocated on live weight to three similar treatment groups. Each calf was given 30,000 Ostertagia ostertagi, 20,000 Cooperia spp. and 10,000 Trichostrongylus axei infective larvae as a single dose. One group was treated with doramectin 6 days after infection while the remaining groups received saline or doramectin 27 days after infection. Given as a single subcutaneous injection behind the ear, doramectin at 200 microg/kg removed 99.9-100% of adult and larval stages of O. ostertagi, Cooperia spp. and T. axei when compared to infections established in untreated controls (p<0.001). No adverse reactions were observed following treatment in the doramectin-treated animals. No injection site lesions were found by palpation following treatment or by injection site examination at necropsy.     

Characterizing historical and modern fire regimes in Michigan (USA): A landscape ecosystem approach

We studied the relationships of landscape ecosystems to historical and contemporary fire regimes across 4.3 million hectares in northern lower Michigan (USA). Changes in fire regimes were documented by comparing historical fire rotations in different landscape ecosystems to those occurring between 1985 and 2000. Previously published data and a synthesis of the literature were used to identify six forest-replacement fire regime categories with fire rotations ranging from very short (<100 years) to very long (>1,000 years). We derived spatially-explicit estimates of the susceptibility of landscape ecosystems to fire disturbance using Landtype Association maps as initial units of investigation. Each Landtype Association polygon was assigned to a fire regime category based on associations of ecological factors known to influence fire regimes. Spatial statistics were used to interpolate fire points recorded by the General Land Office. Historical fire rotations were determined by calculating the area burned for each category of fire regime and dividing this area by fifteen (years) to estimate area burned per annum. Modern fire rotations were estimated using data on fire location and size obtained from federal and state agencies. Landtype Associations networked into fire regime categories exhibited differences in both historical and modern fire rotations. Historical rotations varied by 23-fold across all fire rotation categories, and modern forest fire rotations by 13-fold. Modern fire rotations were an order of magnitude longer than historical rotations. The magnitude of these changes has important implications for forest health and understanding of ecological processes in most of the fire rotation categories that we identified.This revised version was published online in May 2005 with corrections to the Cover Date.     

Effects of mutations in Drosophila nicotinic acetylcholine receptor subunits on sensitivity to insecticides targeting nicotinic acetylcholine receptors

Several strains of Drosophila melanogaster possess mutant alleles in nicotinic acetylcholine receptor (nAChR) subunits, Dα1 and Dβ2 that confer resistance to neonicotinoids such as imidacloprid and nitenpyram, and Dα6, that confers resistance to spinosyns. These mutant strains were bioassayed with a selected set of nAChR active insecticides including neonicotinoids, spinosad, and sulfoxaflor, a new sulfoximine insecticide. All of the neonicotinoids examined, except dinotefuran showed reduced insecticidal efficacy on larvae of the Dα1 mutant, suggesting that this subunit may be important in the action of these insecticides. All of the neonicotinoids, including dinotefuran, showed reduced insecticidal efficacy on larvae possessing the Dβ2 mutation. A similar pattern of broad neonicotinoid resistance to that of Dβ2 alone was also observed for larvae with both the mutations (Dα1 + Dβ2). The Dβ2 mutation exhibited a lower level of cross-resistance to sulfoxaflor (<3-fold) than to any of the neonicotinoids (>13-fold). In contrast, there was no cross-resistance for any of the neonicotinoids or sulfoxaflor in adult flies with the Dα6 mutation, which confers high levels of resistance to spinosad. Thus in the D. melanogaster strains studied, target site resistance observed for the neonicotinoids and the spinosyns does not translate directly to resistance towards sulfoxaflor.