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PubMed2005

Ion channels: molecular targets of neuroactive insecticides.

Many of the insecticides in current use act on molecular targets in the insect nervous system. Recently, our understanding of these targets has improved as a result of the complete sequencing of an insect genome, i.e., Drosophila melanogaster. Here we examine the recent work, drawing on genetics, genomics and physiology, which has provided evidence that specific receptors and ion channels are targeted by distinct chemical classes of insect control agents. The examples discussed include, sodium channels (pyrethroids, p,p'-dichlorodiphenyl-trichloroethane (DDT), dihydropyrazoles and oxadiazines); nicotinic acetylcholine receptors (cartap, spinosad, imidacloprid and related nitromethylenes/nitroguanidines); gamma-aminobutyric acid (GABA) receptors (cyclodienes, gamma-BHC and fipronil) and L-glutamate receptors (avermectins). Finally, we have examined the molecular basis of resistance to these molecules, which in some cases involves mutations in the molecular target, and we also consider the future impact of molecular genetic technologies in our understanding of the actions of neuroactive insecticides.

Animals↗

PubMed2006

Oviduct contraction in Drosophila is modulated by a neural network that is both, octopaminergic and glutamatergic.

Fertility is a highly complex and regulated phenomenon essential for the survival of any species. To identify Drosophila fertility-specific neural networks, we used a GAL4/UAS enhancer trap genetic screen that selectively inactivates groups of neurons. We identified a GAL4 line (bwktqs) that has a female sterile phenotype only when it expresses the tetanus toxin light chain (TeTxLC). These flies lack oviduct contraction, lay almost no eggs, sperm accumulate in the oviducts, and fewer than normal are seen in the storage organs. In insects, two neuroactive substances are important for oviduct contraction: octopamine (OA), a monoamine that inhibits oviduct contraction, and glutamate (Glu), a neurotransmitter that induces contraction. It is known that octopaminergic neurons of the thoracic abdominal ganglion (TAG) modulate oviduct contraction, however, the glutamatergic neurons that innervate the oviduct have not been identified yet and the interaction between these two neuroactive substances is not well understood. Immunostaining experiments revealed that the bwktqs line trapped an octopaminergic neural network that innervates the genital tract. We show that wt like oviduct contraction in TeTxLC-inactivated flies can only be rescued by simultaneous application of Glu and OA suggesting that the abdominal bwktqs neurons are both octopaminergic and glutamatergic, the use of an agonist and an antagonist for Glu receptors as well as their direct visualization confirmed its participation in this phenomenon. Our work provides the first evidence that adult abdominal type II visceral innervations co-express Glu and OA and allows us to re-evaluate the previous model of neuronal network controlling insect oviduct contraction.

Animals↗

PubMed2003

Botanical insecticides for controlling agricultural pests: piperamides and the Colorado Potato Beetle Leptinotarsa decemlineata say (Coleoptera: Chrysomelidae).

The efficacy of extracts from two Piperaceae species, Piper nigrum L. and P. tuberculatum Jacq. were evaluated using larvae and adults of the Colorado Potato Beetle Leptinotarsa decemlineata (Say). Young larvae and neonates were the most susceptible; a 24-h LD(50) of 0.064% extract of P. tuberculatum was determined for 4-day-old larvae, while 0.05% extract of P. nigrum reduced larval survival up to 70% within one week after treatment of potato Solanum tuberosum L. (Solanaceae) plants. When an insecticide resistant strain of L. decemlineata larvae was tested with the P. tuberculatum extract, there was less than a 2-fold tolerance ratio compared to the 22-fold tolerance ratio to cypermethrin, a pyrethroid. Older larvae, pre-pupal stage and adults, were less sensitive to the P. nigrum extracts; the 24-h LD(50) was 0.5% (95% C.I. = 0.36, 0.65). However, the same concentration was equally effective under field conditions. In the greenhouse, P. nigrum at 0.5% was as effective at reducing adult L. decemlineata feeding as combinations with 2 separate botanical mixtures, garlic and lemon grass oil. Under field conditions, the residual activity of the P. nigrum extracts was less than 3 h. When adult L. decemlineata were placed on treated plants exposed to full sunlight for 0, 1.5, and 3 h, leaf damage progressively increased as the main active compound, piperine, was found to degrade by 80% after 3 h. An in vitro polysubstrate monoxygenase (PSMO) enzyme assay, using the substrate methoxyresorufin O-demethylation (MROD), determined that the principal P. nigrum active compound, piperine, is responsible for inhibition of that specific enzyme. The results suggest that Piper extracts could be used effectively as contact botanical insect control agents to protect potato plants from developing L. decemlineata larvae at concentrations less than 0.1%. There is also potential for Piper extracts to control insecticide resistant populations in conjunction with other integrated pest management (IPM) strategies used in conventional and organic agriculture.

Agriculture↗

PubMed2004

Induction and transmission of Bacillus thuringiensis tolerance in the flour moth Ephestia kuehniella.

The use of Bacillus thuringiensis (Bt) endotoxins to control insect vectors of human diseases and agricultural pests is threatened by the possible evolution of resistance in major pest species. In addition to high levels of resistance produced by receptor insensitivity (5, 16, 17), several cases of tolerance to low to medium levels of toxin have been reported in laboratory colonies of lepidopteran species (3, 18). Because the molecular basis of some of these cases of tolerance to the toxin are not known, we explored alternative mechanisms. Here, we present evidence that tolerance to a Bt formulation in a laboratory colony of the flour moth Ephestia kuehniella can be induced by preexposure to a low concentration of the Bt formulation and that the tolerance correlates with an elevated immune response. The data also indicate that both immune induction and Bt tolerance can be transmitted to offspring by a maternal effect and that their magnitudes are determined by more than one gene.

Animals↗

PubMed2002

Characterization of mycoviruses and analyses of chitinase secretion in the biocontrol fungus Metarhizium anisopliae.

Metarhizium anisopliae is the best-characterized entomopathogen and is used to control insect pests in sugar cane plantations in Brazil on a commercial scale. We have previously reported the infection of some M. anisopliae strains by dsRNA mycoviruses. Here we describe the purification and characterization of the viruses (MaV-A1, MaV-M5, MaV-RJ) in terms of dsRNA content, capsid proteins, electron microscopy, Western blot, and hybridization patterns. One spontaneous mutant lost some of the high molecular weight dsRNA components and showed significant alterations in colony morphology and spore production, suggesting that viral genes interfere with fungal phenotype. A comparison between dsRNA mycovirus-free and infected M. anisopliae isolates showed that virus-free isolates have increased endochitinase secretion. By comparing the following parameters: the buoyant density in CsCl of the presumed virions; the number and estimated molecular weight of the dsRNA components and the molecular mass of the capsid proteins to other mycoviruses previously described, we suggest the inclusion of MaV-A1 and MaV-M5 in the family Totiviridae and MaV-RJ in the family Partitiviridae.

Antibodies, Viral↗

PubMed1994

Use of malaria prevention measures in Malawian households.

Information on malaria prevention practices in households was obtained in a nation-wide knowledge, attitudes, and practices survey in Malawi. Of the 1,531 heads of household questioned, 55% were able to identify mosquitoes as the cause of malaria. Use of any type of malaria prevention method was reported by 52% of respondents. Among users, 47% used commercial products (insecticide, mosquito coils, bednets), and 64% used natural measures (burning leaves, dung, or wood); 11% used both. The most common commercial measure used was mosquito coils (16%) followed by insecticide spray (11%) and bednets (7%). Increasing household income and educational level of the household head were strongly correlated with use of commercial methods to prevent malaria; households with an income ranked moderate or greater were eight times more likely to have used a purchased product. Use of natural measures was correlated with lower income and educational level. Thirty-six percent of respondents reported having heard or seen information on malaria in the previous year. Use of household malaria preventive measures in Malawi is very low and income-dependent. Educational messages are required to improve understanding and use of affordable measures.

Adult↗

PubMed2004

Perspiration increased human pesticide absorption following surface contact during an indoor scripted activity program.

Homeowners and professional applicators frequently use chemicals to control insect pests in urban environments. The identification and evaluation of determinants of human exposure are critical to conduct reliable and responsible human exposure assessments following indoor residential chemical applications. The effect of sweat on absorbed dose in humans was evaluated with human volunteers who participated in a structured activity program (SAP). Participants (n=20) performed a warm-up exercise to induce light sweating prior to an SAP on chlorpyrifos(cp)-treated nylon carpet. Absorbed daily dosages (ADDs) were calculated using urinary biomonitoring of trichloropyridinol. In two separate exposures, participation in the warm-up exercise prior to the exposure SAP resulted in an increased ADD of CP equivalents by approximately 50%. Measured ADDs averaged 2.8 (SAP 1) and 2.0 (SAP 2) microg CP equivalents/kg/day in volunteers who participated in the warm-up exercise. In participants who rested prior to the exposures, ADDs were significantly lower at 1.9 (SAP 1) and 1.3 (SAP 2) microg CP equivalents/kg/day. Perspiration may also be a determinant of exposure in active children and field workers. Measured ADDs were less than estimates of ADD made from environmental measurements including CP deposition, the California roller, and clothing dosimeters worn by participants.

Adult↗

PubMed2003

Preparation of spray-dried wettable powder formulations of Bacillus thuringiensis-based biopesticides.

Bacillus thuringiensis is the most widely used biopesticide among many methods available to control insects. To make a saleable product, B. thuringiensis must be substantially concentrated by removal of water and formulated to improve longevity, efficacy, and ease of transport of the product. B. thuringiensis subsp. aizawai culture broth as an active ingredient was mixed with various adjuvants and then spray dried. The optimum conditions for spray drying were found to be an outlet temperature of 60-85 degrees C and an inlet temperature of 120-180 degrees C. Various adjuvants had different effects on physical and biological properties of the dried product. Gelatinized tapioca starch and milk powder improved suspensibility but adversely affected wettability of the dried formulated product. Vegetable oil and Tween 20 enhanced wettability but resulted in poor suspensibility. Silica fume was used to enhance flowability because it reduced clumping and caking of the powder resulting from the addition of vegetable oil. Formulation containing 10% wt:wt B. thuringiensis, 10% wt:wt gelatinized tapioca starch, 10% wt:wt sucrose, 38% wt:wt tapioca starch, 20% wt:wt milk powder, 10% wt:wt silica fume, 2% wt:wt polyvinyl alcohol, 5% vol:vol Tween 20, 1% vol:vol refined rice bran oil, and 1% vol:vol antifoam solution was found to be optimum in terms of the physical and biological properties of the dried product. This formulation had 55% suspensibility, 24 s for wetting time, and 5.69 x 10(4) CFU/ml of LC50 value against Spodoptera exigua larvae.

Agrochemicals↗

PubMed1990

Current trends and new directions in crop protection.

Synthetic agricultural chemicals will continue to be the major component in protecting food and fiber crops from attack by insects, fungi, and weeds. This trend will most likely continue through the end of this century and possibly beyond. The discovery and introduction of potent new pesticides such as the sulfonylurea and imidazolinone herbicides and synthetic pyrethroid insecticides have demonstrated that excellent weed and insect control can be achieved with minute amounts of chemical per treated unit area in comparison to older chemicals. In some instances, these reductions approached two orders of magnitude. Further improvements in delivery systems and formulations have also contributed to reductions in the chemical load in the environment. However, increased regulatory demands; environmental concerns; developing resistance in key pests; increasing costs of research, development, manufacturing, and selling; and even a smaller number of basic producers, all in concert impact heavily on future developments in chemical pesticides. We can expect also that, within the next decade, there will be an increasing number of introductions of biological pesticides, including microbials and biochemicals derived from microbiological sources. These new products--including mycoherbicides, mycofungicides, and biological insecticides--are expected to become prominent control measures in the intermediate time span (10-20 years), to be supplemented in the long-term by products and transgenic plants derived from recombinant DNA technology.

Agriculture↗

PubMed2006

Effects of the juvenile hormone analog pyriproxyfen on German cockroach, Blattella germanica (L.), tergal gland development and production of tergal gland secretion proteins.

Male German cockroaches possess secretory glands that secrete fluid into a pair of transverse depressions on the seventh and eighth abdominal tergites. We investigated the effects of altered juvenoid titer during the first part of the last instar on tergal gland secretory tissue development and the production of tergal gland secretion proteins. Male fifth (last) instar nymphs (1-3 days post-emergent) were topically treated with the JH analog pyriproxyfen. Light and transmission electron microscopy demonstrated that treatment with pyriproxyfen produced a visible decrease in the amount of tergal gland tissue present, a deformation of the overall shape of the gland located on tergite seven, and a less orderly arrangement of the secretory cells in the tissue. The protein fraction of tergal gland secretion was examined in pyriproxyfen-treated and control insects 1, 5, and 15 days after the insects molted to the adult stage. Amounts of all tergal secretion proteins were reduced in treated insects.

Animals↗

PubMed1991

Lysozyme in the midgut of Manduca sexta during metamorphosis.

Low levels of lysozyme were found in the midgut epithelium of the tobacco hornworm, Manduca sexta, during the early part of the fifth larval stadium. This was observed in control insects as well as in bacterially challenged insects. No lysozyme was detected in the gut contents of either group of insects which were actively eating or in the early stages of metamorphosis. However, high levels of lysozyme activity were detected in homogenates of midgut tissue collected from insects later in the stadium. Immunocytochemical studies demonstrated that lysozyme accumulates in large apical vacuoles in regenerative cells of the midgut during the larval-pupal molt. These cells, initially scattered basally throughout the larval midgut epithelium, multiply and form a continuous cell layer underneath the larval midgut cells. At the larval/pupal ecdysis the larval midgut epithelium is sloughed off and the regenerative cells, now forming the single cell layer of the midgut, release the contents of their vacuoles into the midgut lumen. This release results in high lysozyme activity in the lumen of the pupal midgut and is thought to confer protection from bacterial infection. This is the first indication that the lysozyme gene may be developmentally regulated in a specific tissue in the absence of a bacterial infection.

Animals↗

PubMed1992

Oral toxicity to flesh flies of a neurotoxic polypeptide.

An insect selective neurotoxic polypeptide from venom of the scorpion Androctonus australis (AaIT, M(r) 8,000) was shown to cross the midgut of the flesh fly Sarcophaga falculata, using assays of oral toxicity, column chromatography, and microscopic autoradiography of the native and radioiodinated toxin. AaIT induced paralysis of flies within 1-2 h after oral administration, with a lethal dose (LD50) of 10 micrograms/100 mg of body weight. Oral toxicity was about 0.14% of toxicity by injection. Hemolymph collection 70-85 min after feeding flies with [125I]AaIT showed that 5% of ingested radioactivity appeared in hemolymph. Most of this represented degradation products, but included about 0.3% of the chromatographically intact toxin. In contrast, hemolymph of identically treated lepidopterous larvae (Manduca, Helioverpa [= Heliothis]) contained degradation products but no intact toxin. [125I]AaIT was shown to cross the midgut of Sarcophaga through a morphologically distinct segment of the midgut previously shown to be permeable to a cytotoxic, positively charged polypeptide of similar molecular weight. These results suggest that Sarcophaga midgut contains a morphologically and functionally distinct segment that transports small peptides, and that employment of neurotoxic polypeptides for insect control may be feasible. Activity might be greatly improved through modification and metabolic stabilization of active peptides.

Animals↗

PubMed1979

[Use of dichlorvos in flour mills].

In the framework of studies on the possible use of dichlorvos (DDVP) in flour mills, the authors investigated the effects of various parameters (such as concentration of the DDVP solution to be used, temperature, location of the samples, different kinds of samples and packages) on the formation of DDVP residues on milling products. It was found that a concentration of 0.6 ml of DDVP-50/m3 and temperatures ranging from 18 degrees to 22 degrees C are particularly suited for insect control. Under these conditions, the DDVP residues on the samples were degraded within a period of 2--4 weeks, according to the kind of product. Uncovered foods shall not be present in rooms where DDVP is being applied. In virtue of the present findings, DDVP is recommended for use in flour mills.

Dichlorvos↗

PubMed2006

Insecticidal, antifeedant and oviposition deterrent effects of the essential oil and individual compounds from leaves of Chloroxylon swietenia DC.

Essential oil from the leaves of Chloroxylon swietenia DC. was obtained by hydrodistillation and cold extraction, and the chemical composition was determined by gas chromatography and gas chromatography-mass spectrometry. The major identified components were limonene, geijerene, pregeijerene, germacrene D and trans-beta-ocimene. Laboratory bioassays of the essential oil and four constituents of essential oil isolates were evaluated for insecticidal, antifeedant and oviposition deterrent effects on tobacco cutworm, Spodoptera litura (F.). Toxicity was determined by topical application of the isolates at varying concentrations. Pure oil, geijerene and pregeijerene were found to be more toxic, with LD50 values of 28.6, 35.4 and 40.7 microg per larva respectively. Maximum feeding deterrence was noted for geijerene and pregeijerene, with DC(50) values of 82.5 and 95.1 microg cm(-2) respectively. Furthermore, pure oil, geijerene and pregeijerene displayed oviposition deterrence, even at low concentrations. These results indicate that these natural products may lead to useful, biodegradable, environmentally safe insect control agents.

Animals↗

PubMed2001

Mode of action of the ecdysone agonist tebufenozide (RH-5992), and an exclusion mechanism to explain resistance to it.

Spruce budworm larvae (Choristoneura fumiferana) upon ingesting tebufenozide (RH-5992) stop feeding and go into a precocious, incomplete molt, leading eventually to death. Like 20-hydroxyecdysone (20E), tebufenozide also acts at the receptor level and transactivates the expression of up-regulated genes but, because of its persistence, the down-regulated genes that are normally expressed in the absence of 20E are not expressed. While tebufenozide is lepidopteran-specific, an analog, RH-5849, is effective on dipterans. This is reflected in the respective effects of the two compounds on Cf-203 (C. fumiferana--203), a lepidopteran cell line and Dm-2 (Drosophila melanogaster--2), a dipteran cell line. Cf-203 cells accumulated [14C]tebufenozide and expressed CHR3 (Choristoneura hormone receptor 3), but Dm-2 cells excluded the material and did not express DHR3 (Drosophila hormone receptor 3). Using yeast ABC (ATP binding cassette) transporter mutants, we determined that PDR5 (pleiotropic drug resistance 5) was responsible for the exclusion. We discovered recently that older instars of the white-marked tussock moth (Orgyia leucostigma) are resistant to tebufenozide, perhaps as a result of such an exclusion system. We are currently cloning PDR5 (pleiotropic drug resistance 5), which is an essential step in studying the resistance mechanism.

Animals↗

PubMed2001

Ryanodine receptor acts as a sensor for redox stress.

Ryanoids have not attained importance as insecticides, but the increasing number of xenobiotic effectors known to influence Ca2+ signalling by interaction with ryanodine receptors (RyRs) may serve to identify new targets for insect control. A historical review of redox control of microsomal Ca2+ transport is given here, followed by recent evidence indicating that hyperactive Cys residues are an essential component of a transmembrane redox sensor. Focus is on the role of sulfhydryl chemistry in RyR regulation; metabolic quinonoid intermediates from pesticides and other environmental contaminants are of interest in this context.

Biological Transport↗

PubMed2002

Fumigant activity of (E)-anethole identified in Illicium verum fruit against Blattella germanica.

The insecticidal activities of materials derived from the fruit of star anise, Illicium verum, against adults of Blattella germanica were examined by direct contact application and fumigation methods, and compared with those of DDVP, deltamethrin and hydramethylnon. The biologically active constituent of the Illicium fruit was characterized as the phenylpropene, (E)-anethole, by spectroscopic analysis. In a filter paper diffusion test with females, (E)-anethole caused 80.3% mortality at 0.159 mg cm-2 at 1 and 3 days after treatment (DAT), whereas 16.7% mortality at 3 DAT was achieved at 0.079 mg cm-2. DDVP and deltamethrin gave > 90% mortality at 0.019 mg cm-2 at 1 DAT. At 0.009 mg cm-2, DDVP and deltamethrin showed 73.3 and 60% mortality at 1 DAT, respectively, but 93.3 and 76.7% mortality at 3 DAT. Hydramethylnon exhibited 0 and 93.3% mortality at 0.159 mg cm-2 at 1 and 3 DAT, respectively, whereas 6.7% mortality at 3 DAT was observed at 0.079 mg cm-2. In a fumigation test with females, (E)-anethole was much more effective in closed cups than in open ones, indicating that the insecticidal activity of the compound was largely attributable to fumigant action. (E)-Anethole and DDVP caused 100% mortality at 0.398 and 0.051 mg cm-2 4 and 1 h after treatment, respectively. (E)-Anethole showed 46.7% mortality at 0.199 mg cm-2 at 3 DAT, whereas deltamethrin and hydramethylnon at 0.796 mg cm-2 was ineffective for 3-day period. As naturally occurring insect-control agents, the I verum fruit-derived materials described could be useful for managing populations of B germanica.

Allylbenzene Derivatives↗

PubMed2002

Effects of formulation on the run-off of imidacloprid from turf.

Imidacloprid is a pesticide often used to control insect pests on residential lawns and golf courses. To investigate its potential to be transported into non-target aquatic systems by rainfall events, imidacloprid was applied as 5.0 g kg-1 GR and 750 g kg-1 WP to 12 plots planted with bermuda grass set on a 5% slope. The dimethylamine salt of the herbicide 2,4-D was applied simultaneously to the plots in order to normalize the results to prior trials conducted with other pesticides. At a rate of 2.5 cm h-1, 5-cm rainfall events were simulated at 24 and 48 h after application and 2.5-cm events were simulated at 96 and 192 h. After each event water was collected from each plot and analyzed by HPLC for residual insecticide or herbicide. Approximately 1.4% of the insecticide formulated as WP and 1.9% of that formulated as GR was lost from the plots after four run-off events. Of the total mass lost, 64% and 30% respectively occurred in the first and second run-off events for the WP formulation versus 75% and 20% for the GR formulation. These values compare with a total mass export of 2.6% for 2,4-D, of which 95% and 4% of the loss occurred respectively in the first and second run-off events. The maximum concentration of imidacloprid detected in run-off water was 0.49 mg litre-1 and occurred during the first run-off event.

2,4-Dichlorophenoxyacetic Acid↗