Safety testing of tebufenozide, a new molt-inducing insecticide, for effects on nontarget forest soil invertebrates.

Tebufenozide, a new molt-inducing insecticide that mimics the action of ecdysone, is being considered for use to control defoliating lepidoptera in forests in Canada. Soil microcosms, employing substrates and species from the ecosystems in which spraying is likely to occur, were used to evaluate the effects of this compound on soil invertebrates. The forest earthworm (Dendrobaena octaedra Savigny) and four species of Collembola (Folsomia candida Willem, F.

New approaches to insect tissue culture.

Current methods of insect cell culture have produced a limited variety of cell types in an ever expanding list of insect cell lines. In developing midgut epithelial cell lines, we found that traditional methods in insect cell culture failed to provide healthy cells from mature tissues. Examination of mammalian cell culture literature for this particular cell type provided the insight required to successfully develop a cell-specific line (Baines et al.

A protein complex from Choristoneura fumiferana gut-juice involved in the precipitation of delta-endotoxin from Bacillus thuringiensis subsp. sotto.

A 75 kDa protein from spruce budworm (Choristoneura fumiferana) gut-juice has been isolated and shown to cause a specific precipitation of the delta-endotoxin from Bacillus thuringiensis subsp. sotto. This 75 kDa protein, separated by either column chromatography or SDS-PAGE, caused precipitation of the sotto toxin in both agarose diffusion gels and the PAGE gels.

Effects of a new molt-inducing insecticide, tebufenozide, on zooplankton communities in lake enclosures.

: A potent ecdysone agonist, tebufenozide, has recently been developed as a molt-inducing insecticide to control defoliating lepidopterans. As part of continuing research efforts to assess the effectiveness and environmental safety of this material for insect pest management in Canadian forests, tebufenozide (RH-5992-2F) was applied to large lake enclosures and the effects on zooplankton communities were evaluated. There were significant treatment effects at all test concentrations (0.

Analysis of ecdysteroid action in Malacosoma disstria cells: cloning selected regions of E75- and MHR3-like genes.

IPRI-MD-66 (MD-66) cells respond to 20-hydroxyecdysone (20E, 4 x 10(-6) M) in the medium by producing cytoplasmic extensions, clumping and attaching themselves to the substrate. These morphological changes are at a maximum by 6 days post treatment. Degenerate oligonucleotides, designed on the basis of conserved amino acid sequences in the DNA and ligand binding regions of the members of the steroid hormone receptor superfamily, were used in RNA-PCR to isolate two cDNA fragments, Malacosoma disstria hormone receptor 2 (MdHR2) and Malacosoma disstria hormone receptor 3 (MdHR3) from the MD-66 cells.

Comparison of forest soil microcosm and acute toxicity studies for determining effects of fenitrothion on earthworms.

The effects of the organophosphate pesticide fenitrothion on Eisenia fetida Savigny and Dendrobaena octaedra Savigny were studied using the OECD acute toxicity test and microcosms containing forest organic matter. In the acute toxicity study, both species were relatively susceptible to fenitrothion, although D. octaedra (LC50 = 393.

Variation in tolerance to Bacillus thuringiensis among and within populations of the spruce budworm (Lepidoptera: Tortricidae) in Ontario.

Variation in tolerance to Bacillus thuringiensis Berliner subsp. kurstaki (strain HD-1-S-1980) among and within populations of the spruce budworm, Choristoneura fumiferana (Clemens), was assessed in the laboratory. Force-feeding assays using offspring of females collected as pupae from nine locations throughout Ontario and from a laboratory colony (DCF) demonstrated limited variation in tolerance among populations.

Production of antibodies against Bacillus thuringiensis delta-endotoxin by injecting its plasmids.

Bacillus thuringiensis insecticidal crystalline delta-endotoxin proteins are plasmid encoded. Specific antibodies against the delta-endotoxin were obtained in mice and rabbits by injecting isolated B. thuringiensis DNA plasmids into their muscles.

Effects of lethal and sublethal concentrations of the herbicide, triclopyr butoxyethyl ester, in the diet of zebra finches.

Lethal and sublethal effects of dietary triclopyr butoxyethyl ester (TBEE) on zebra finches (Poephila guttata Gould) were determined in laboratory experiments conducted between 8 January and 1 May 1991. The 8-day median lethal dietary concentration, LC50 (95% confidence interval), of TBEE to zebra finches was 1,923 (1,627 to 2,277) mg/kg. In the sublethal effects experiment, when birds were exposed to 500 mg/kg TBEE in the diet for 29 days, food consumption and body weight were significantly depressed (P < 0.

Toxicity of a new molt-inducing insecticide (RH-5992) to aquatic macroinvertebrates.

The molt-inducing insecticide RH-5992, a potent ecdysone agonist, is being evaluated for potential use in forestry to control defoliating lepidopterans. The possible adverse effects of RH-5992 on nontarget aquatic organisms were studied in two test systems. Acute lethal effects were determined for one aquatic amphipod and 11 species of aquatic insects in laboratory flowthrough toxicity tests.

Detection of Choristoneura fumiferana brush border membrane-binding molecules specific to Bacillus thuringiensis delta-endotoxin by crossed affinity immunoelectrophoresis.

Brush border membrane vesicles (BBMVs) prepared from midguts of Choristoneura fumiferana larvae were pretreated separately with the three CryIA delta-endotoxins from Bacillus thuringiensis subsp. kurstaki HD-1. Crossed affinity immunoelectrophoresis of the solubilized toxin-BBMV complexes, using antibodies raised against the toxins, revealed at least two peaks for each toxin.

Purification and characterization of a trypsin-like digestive enzyme from spruce budworm (Choristoneura fumiferana) responsible for the activation of delta-endotoxin from Bacillus thuringiensis.

A trypsin-like enzyme purified from spruce budworm (Choristoneura fumiferana) gut juice has a molecular mass of 25 kDa and its pH activity profile indicates a pKa of 8. Sequence homology with bovine trypsin of the N-terminus and active site, and the ionization dependence for catalysis, reflect the typical trypsin-like activities measured. The action of this enzyme (designated CFT-1) is compared to the neat gut juice with regard to the proteolytic activation of the delta-endotoxin from Bacillus thuringiensis.

Horseradish peroxidase-labelled probes and enhanced chemiluminescence to detect baculoviruses in gypsy moth and eastern spruce budworm larvae.

Horseradish peroxidase-labelled whole genomic DNA probes and enhanced chemiluminescence procedures were utilized to detect baculovirus in insect macerates blotted on nylon membranes. Detection levels were similar to those found using 32P-labelled probes; 5 x 10(3) occlusion bodies (OBs), 2 x 10(3) OBs and 4 x 10(4) OBs of Lymantria dispar (L.) nuclear polyhedrosis virus (LdNPV), Choristoneura fumiferana (Clem.

Use of synthetic peptides to probe functional domains of a Bacillus thuringiensis toxin.

Two 10-residue peptides exhibiting sequence homology to CryIA(a) toxin were chemically synthesized. One corresponds to a segment from residues 48-57 (peptide A) and is common to all CryIA toxins and the second corresponds to a segment from residues 348-357 (peptide B) and is specific to the CryIA(a) toxin. Antibodies were raised in rabbits against both peptides.

Liquid chromatographic determination of RH-5992, an ecdysone agonist, in some forestry matrixes.

A liquid chromatographic method is described for the analysis of RH-5992 (N'-t-butyl-N'-[3,5-dimethyl-benzoyl]-N- [-ethylbenzoyl]hydrazine) from various terrestrial and aquatic forestry matrixes and stream water. The processed soils, litter, oak foliage, sediment, aquatic plants, and fish were extracted with a mixture of acetone and water; balsam fir needles were extracted with acidified methanol solution; stream water was extracted with dichloromethane. Aliquots of crude extracts were solvent partitioned, concentrated, and cleaned up by using Florisil or Prepsep-NH2 columns.

Effects of the herbicides hexazinone and triclopyr ester on aquatic insects.

Experiments were conducted to measure acute lethal response of aquatic insects to hexazinone (Velpar L) and triclopyr ester (Garlon 4) in flow-through laboratory bioassays, and to determine lethal and behavioral effects of these herbicides on insects in outdoor stream channels. No significant mortality (chi 2 P greater than 0.05) occurred in 13 test species exposed to hexazinone in laboratory flow-through bioassays (1-hr exposure, 48-hr observation) at the maximum test concentration of 80 mg/liter.

Comparative toxicity of the HD-1 and NRD-12 strains of Bacillus thuringiensis subsp. kurstaki to defoliating forest Lepidoptera.

Insecticidal activities of sporulated cultures of the HD-1 and NRD-12 strains of Bacillus thuringiensis subsp. kurstaki were compared against four species of defoliating forest lepidopterans in diet-incorporation assays. There was no difference in LC50 between the two strains to larvae of spruce budworm (Choristoneura fumiferana), gypsy moth (Lymantria dispar), eastern hemlock looper (Lambdina fiscellaria fiscellaria), and whitemarked tussock moth (Orgyia leucostigma), whether expressed as total alkaline soluble protein, activated toxin protein, or International Units as determined by bioassay against Trichoplusia ni.

An interlaboratory comparison of data on brain cholinesterase activity in forest songbirds exposed to aerial application of Zectran.

Zectran (4-dimethylamino-3,5-xylyl N-methyl-carbamate), a carbamate insecticide (active ingredient [AI] mexacarbate), was aerially applied to two 300 ha plots of coniferous forest at dosage rates of 70 and 140 g AI/ha, respectively. The brains of 288 birds collected from the treated areas and 84 birds from untreated areas were sagittally sectioned into approximately equal halves. Each of the laboratories participating in the study, the Forest Pest Management Institute (FPMI) and the Canadian Wildlife Service, Atlantic Region (CWS), assayed one half of each brain for cholinesterase (ChE) activity and the results were compared.

Specificity of Activated CryIA Proteins from Bacillus thuringiensis subsp. kurstaki HD-1 for Defoliating Forest Lepidoptera.

The insecticidal activity of the CryIA(a), CryIA(b), and CryIA(c) toxins from Bacillus thuringiensis subsp. kurstaki HD-1 was determined in force-feeding experiments with larvae of Choristoneura fumiferana, C. occidentalis, C.