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Selective effects of insecticides on nigrostriatal dopaminergic nerve pathways.

A degeneration of the nigrostriatal pathway is a primary component of Parkinson's disease (PD), and we have investigated the actions of insecticides on this pathway. For in vivo exposures, C57BL/6 mice were treated three times over a 2-week period with heptachlor, the pyrethroids deltamethrin and permethrin, or chlorpyrifos. One day after the last treatment, we observed that heptachlor and the pyrethroids increased maximal [3H]dopamine uptake in striatal synaptosomes from treated mice, with dose-dependent changes in Vmax displaying a bell-shaped curve. Western blot analysis confirmed increased levels of dopamine transporter (DAT) protein in the striatum of mice treated with heptachlor and permethrin. In contrast, we observed a small, but statistically significant decrease in dopamine uptake by 100 mg/kg chlorpyrifos. For heptachlor, doses that upregulated DAT expression had little or no effect on serotonin transport. Permethrin did cause an upregulation of serotonin transport, but required a 30-fold greater dose than that effective on dopamine uptake. Other evidence of specificity was found in transmitter release assays, where heptachlor and deltamethrin released dopamine from striatal terminals with greater potency than other transmitter types. These findings confirm that insecticides possess specificity for effects on striatal dopaminergic neurotransmission.

Animals↗

Neonicotinoids: insecticides acting on insect nicotinic acetylcholine receptors.

Imidacloprid is increasingly used worldwide as an insecticide. It is an agonist at nicotinic acetylcholine receptors (nAChRs) and shows selective toxicity for insects over vertebrates. Recent studies using binding assays, molecular biology and electrophysiology suggest that both alpha- and non-alpha-subunits of nAChRs contribute to interactions of these receptors with imidacloprid. Electrostatic interactions of the nitroimine group and bridgehead nitrogen in imidacloprid with particular nAChR amino acid residues are likely to have key roles in determining the selective toxicity of imidacloprid. Chemical calculation of atomic charges of the insecticide molecule and a site-directed mutagenesis study support this hypothesis.

Animals↗

Community-level disruptions among zooplankton of pond mesocosms treated with a neem (azadirachtin) insecticide.

A natural, plant-derived insecticide, neem, is being evaluated as an alternative insect pest control product for forestry in Canada. As part of the process to investigate the environmental safety of neem-based insecticides, a mesocosm experiment was conducted to assess the effects of neem on natural zooplankton communities. Replicate (n=5), shallow (<1 m) forest pond enclosures were treated with Neemix 4.5, at concentrations of 0.035 (the expected environmental concentration), 0.18, 0.70, and 1.75 mg/l active ingredient, azadirachtin. Zooplankton communities were quantitatively sampled over a 4-month experimental period in treated and control enclosures, and water samples were collected to track azadirachtin concentrations. Concentrations in water declined linearly with estimated DT(50) values of 25-29 days. Trends in abundance over time among populations of cladocerans, copepods, and rotifers were found to differ significantly among treatments. At the two highest test concentrations, adverse effects were obvious with significant reductions in several cladoceran species, and near elimination of the three major copepod species present. More subtle effects at the two lowest test concentrations were determined by comparing the community structure of enclosures across treatment levels and over time through an analytical process based on the multivariate statistical software, PRIMER. Significant effects on community structure were detected at both of these lower concentrations, including the expected environmental concentration of 0.035 mg/l azadirachtin. Differential responses among species (some increases, some decreases) caused detectable disruptions in community structure among zooplankton of treated enclosures. Perturbations to zooplankton communities were sufficient to cause measurable differences in system-level metabolism (midday dissolved oxygen concentrations) at all but the lowest test concentration.

Animals↗

Effects of the pyrethroid insecticide cypermethrin on a freshwater community studied under field conditions. II. Direct and indirect effects on the species composition.

The effects of cypermethrin, a commonly used pyrethroid insecticide, were studied in small in situ enclosures situated in an eutrophic lake over an 11-day period. The experimental design used a regression principle that included three untreated controls and a gradient of six unreplicated cypermethrin concentrations, ranging from 0.01 to 6 microg/l. This paper is the second in a series of two and describes the effects on the species composition of the crustacean, rotifer, periphyton and phytoplankton communities. Multivariate ordination technique (redundancy analysis (RDA) combined with Monte Carlo permutation tests) showed that exposure to cypermethrin caused significant changes in the species composition of the communities. Changes in the structure of the communities were observed following exposure to a nominal concentration of 0.13 microg cypermethrin per litre above. The direct acute effect of exposure to cypermethrin was a rapid decrease of many species of crustacean zooplankton. The alterations in crustacean species composition were probably due to variations in susceptibility to the direct toxic effects of cypermethrin. No effects concentration (NEC) for individual zooplankton species were calculated using inverse regression and revealed that copepod nauplii were the most sensitive (NEC=0.01 microg/l) of the crustacean groups examined. The observed alterations of the species composition of the autotrophic communities as well as of the rotifers were most likely caused indirectly by cypermethrin, mediated through the direct negative effects of the insecticide on the crustacean grazers. The results of this experiment provide further knowledge about the direct and indirect effects of pesticide stress on the ecosystem level. They also show that there is a variation in sensitivity between different species of zooplankton under natural conditions and thus exemplify the necessity of multispecies approaches in the risk assessment of pesticides.

Animals↗

Analysis of the promoters for the beta-esterase genes associated with insecticide resistance in the mosquito Culex quinquefasciatus.

Resistance to organophosphorus (OP) insecticides in the mosquito Culex quinquefasciatus is primarily due to the amplification and over-expression of non-specific esterases. Co-amplification of two esterase genes, estalpha2(1) and estbeta2(1), is the most common resistance genotype. In both resistant and susceptible mosquitoes the alpha- and beta-esterase genes are oriented in a head-to-head arrangement, the intergenic sequences containing promoter elements for the divergent transcription of both esterases. Transient transfection of luciferase reporter gene constructs into a C. quinquefasciatus cell line was used to study these promoters. A control vector was constructed with the strong Drosophila actin 5c promoter driving expression of beta-galactosidase. The beta-esterase promoters from both insecticide resistant and -susceptible insects were highly active in directing luciferase expression. Transfections with panels of deletions revealed several regions where binding sites for positive and negative regulatory elements are located, and candidate transcription factor sites have been identified. Gel shift assays have identified one DNA-protein interaction that is stronger with the resistant than with the equivalent but slightly altered susceptible sequence. The arthropod initiator site TCAGT 135bp upstream of the ATG in both beta-esterase promoters is essential for transcription initiation, but a putative TATA box is not involved.

Animals↗

Bacillus thuringiensis insecticidal Cry1Aa toxin binds to a highly conserved region of aminopeptidase N in the host insect leading to its evolutionary success.

Bacillus thuringiensis insecticidal protein, Cry1Aa toxin, binds to a specific receptor in insect midguts and has insecticidal activity. Therefore, the structure of the receptor molecule is probably a key factor in determining the binding affinity of the toxin and insect susceptibility. The cDNA fragment (PX frg1) encoding the Cry1Aa toxin-binding region of an aminopeptidase N (APN) or an APN family protein from diamondback moth, Plutella xylostella midgut was cloned and sequenced. A comparison between the deduced amino acid sequence of PX frg1 and other insect APN sequences shows that Cry1Aa toxin binds to a highly conserved region of APN family protein. In this paper, we propose a model to explain the mechanism that causes B. thuringiensis evolutionary success and differing insect susceptibility to Cry1Aa toxin.

Amino Acid Sequence↗

Neutrophil function in workers exposed to organophosphate and carbamate insecticides.

Neutrophil function in 40 workers occupationally exposed to carbamate and organophophate insecticides were examined and compared to those of non-exposed individuals. Phagocytosis and intracellular killing of Candida albicans and Candida pseudotropicalis by neutrophils were studied. Two species of Candida were used since in individuals with myeloperoxidase deficiency neutrophils are unable to kill Candida albicans, while Candida pseudotropicalis can be effectively lysed. Phagocytosis of both antigens was normal in all the workers studied. On the other hand, there was a considerable reduction in the ability of neutrophils from exposed workers to kill Candida albicans whereas Candida pseudotropicalis was effectively lysed. This finding indicates some interference with the myeloperoxidase activity in the exposed population. The levels of cholinesterase activity in all workers were normal. These results demonstrate that exposure to carbamates and organophophates insecticides may lead to changes in neutrophil function even in workers presenting no impairment in the cholinesterase activity.

Adult↗

Injection of household spray insecticide.

During a three-week period, two patients who had attempted suicide by injecting themselves with commercially available household spray insecticides were seen in our emergency department. Both presented with cellulitis at and adjacent to the injection sites, and both were admitted for intravenous antibiotics, warm soaks, and elevation. In both patients abscesses subsequently developed in the areas of cellulitis. It is not clear whether the pathologic processes in these two patients were primarily due to inoculation of microorganisms or to the effects of the insecticide per se.

Abscess↗

Polychlorinated biphenyl congeners and organochlorine insecticides in the water column and sediments of Daya Bay, China.

Samples of sub-surface water, suspended particulate matter (SPM) and surface sediments collected from Daya Bay, China have been analysed for 12 polychlorinated biphenyl (PCB) congeners and 18 organochlorine insecticides, using gas chromatography electron capture detection. Total PCB levels varied from 91.1 to 1355.3 ng l-1 in water and from 0.85 to 27.37 ng g-1 dry weight in sediments. The levels of total organochlorine insecticides were in the range 143.3-5104.8 ng l-1 in water and 2.43-86.25 ng g-1 dry weight in sediment. None of the target compounds were detected in SPM. The levels of total hexachlorocyclohexanes in water varied from 35.5 to 1228.6 ng l-1, whilst in sediments they ranged from 0.32 to 4.16 ng g-1 dry weight. For the sum of dichlorodiphenyltrichloroethanes (DDTs), their levels were in the range 26.8-975.9 ng l-1 in water, and 0.14-20.27 ng g-1 dry weight in sediments. The distribution profiles of these contaminants in water and sediments suggest that there are a number of sources contributing to total contaminant burden in the bay, including soil runoffs, wastewater discharges, sewage outfalls and shipping activites. Ratios of DDT/(DDE + DDD) in the water and sediments indicate recent inputs of such chemicals into the day. The results, therefore, provide important information on the current contamination status of a key aquacultural area in China, and point to the need for urgent actions to stop the use of persistent agrochemicals such as DDT and lindane.

Aquaculture↗

Unidirectional cross-tolerance between the carbamate insecticide propoxur and the organophosphate disulfoton in mice.

Previous studies have shown that subchronic treatment of mice with the organophosphate insecticide, disulfoton, or the carbamate insecticide, propoxur, leads to the development of tolerance to their toxicity. Tolerance to disulfoton was due to a decrease in the number of muscarinic cholinergic receptors, while tolerance to propoxur appeared to be due to an induction of hepatic microsomal enzymes. In the present study we investigated if cross-tolerance between disulfoton and propoxur would occur. Cross-tolerance was evaluated by measuring acute toxicities, cholinesterase and carboxylesterase inhibition and hypothermic and antinociceptive effects. Mice tolerant to propoxur were cross-tolerant to the hypothermic and anticholinesterase effects of disulfoton. Similarly, when mice were pretreated with the microsomal enzyme inducer, phenobarbital, the toxicity of disulfoton was decreased. Mice made tolerant to disulfoton were cross-tolerant to the organophosphate chlorpyrifos, but were more sensitive than controls to the toxicity of propoxur. The acute toxicity of the organophosphate malathion was also increased in disulfoton-tolerant mice. Propoxur is metabolized by mixed function oxidases and possibly by a carboxylesterase. While hepatic microsomal enzymes appeared to be unchanged in disulfoton-tolerant mice, brain and liver carboxylesterase activities were significantly inhibited. Pretreatment of mice with the specific carboxylesterase inhibitor triorthotolylphosphate is known to greatly potentiate the toxicity of malathion and also potentiated, to a lesser extent, the toxicity of propoxur.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

In vitro genotoxic effects of the insecticide deltamethrin in human peripheral blood leukocytes: DNA damage ('comet' assay) in relation to the induction of sister-chromatid exchanges and micronuclei.

Deltamethrin, a synthetic dibromo-pyrethroid insecticide, is extensively used in agriculture, forestry and in household products because of its high activity against a broad spectrum of insect pests (both adults and larvae), its low animal toxicity and its lack of persistence in the environment. Data on the genotoxicity and carcinogenicity of deltamethrin are rather controversial, depending on the genetic system or the assay used. The aim of this study was to further evaluate the potential genotoxic activity of deltamethrin. The in vitro genotoxicity of deltamethrin has been evaluated by assessing the ability of the insecticide to damage DNA (as evaluated using the single-cell microgel-electrophoresis or 'comet' assay) or induce sister-chromatid exchanges (SCE) and micronuclei (MN) in human peripheral blood leukocytes. All treatments were conducted with and without the presence of an external bioactivation source (+/- S9mix). The results indicate that deltamethrin, in the presence of metabolic activation (+ S9mix), is able to induce DNA damage (double- and single-strand breaks, alkali-labile sites and open excision repair sites) as revealed by the increasing tail moment values observed with increasing doses. The frequency of SCE and MN were not statistically increased in deltamethrin-treated cells as compared to controls, both with and without S9mix. However, lower deltamethrin doses were tested, as compared to 'comet' assay, because of cytotoxicity.

Adult↗

In vitro insecticidal effects of fipronil and beta-cyfluthrin on larvae of the blowfly Lucilia sericata.

The insecticidal effects of the phenylpyrazole, fipronil, and a pyrethroid, beta-cyfluthrin, on larvae of the blowfly Lucilia sericata were determined in laboratory assays. When first stage larvae of L. sericata were reared on homogenized pig liver which had been treated with known amounts of test compounds, both fipronil and beta-cyfluthrin induced significant levels of mortality compared to acetone and water controls. However, fipronil was approximately 10 times more toxic than beta-cyfluthrin to L. sericata larvae following ingestion. Beta-cyfluthrin had little effect on mortality until concentrations of approximately 0.5 ppm were reached. In contrast, fipronil effected L. sericata mortality at a concentration of 0.05 ppm and 100% mortality was reached by 0.5 ppm. The lethal concentration (LC50) value for beta-cyfluthrin was 1.56 ppm as compared to 0.14 ppm for fipronil. Following contact of first and third stage larvae with cloth impregnated with known amounts of test compound, the mortality profiles of fipronil and beta-cyfluthrin were similar. At short contact times, the LC50 values for fipronil were lower than those for beta-cyfluthrin. However, at the highest contact time evaluated for the first stage larvae, 300 s, there was a reversal in this trend. The results suggest that the phenylpyrazole fipronil may represent a new potential insecticide for development against blowfly strike of sheep.

Administration, Topical↗

Insecticidal activity of essential oils to the tobacco cutworm, Spodoptera litura.

Twenty-one essential oils were tested for insecticidal activity via topical administration to third instar larvae of the tobacco cutworm, Spodoptera litura. Oils of Satureia hortensis, Thymus serpyllum and Origanum creticum produced >90% larval mortality at 24 h at a dose of 100 microg per larva. The LD(50) value for S. hortensis (48.4 microg) was comparable to that for Thymus vulgaris (46.9 microg). The monoterpenoid phenols thymol and carvacrol, major constituents of the oils of Thymus and Satureia species, likely account for the insecticidal action of the species.

Animals↗

Toxicokinetic and toxicodynamic aspects of organophosphorus (OP) insecticide poisoning.

As the majority of OP insecticides are lipophilic and not ionised they are absorbed rapidly following inhalation or ingestion; dermal absorption is slower but severe poisoning may still ensue if exposure is prolonged. Following absorption, OP compounds accumulate in fat, liver, kidneys and salivary glands. Phosphates (P=O) are biologically active as AChE inhibitors, whereas phosphorothioates (P=S) need bioactivation to their phosphate anologues (oxons) to become biologically active. Oxons inhibit AChE by phosphorylation of the serine hydroxyl group in the substrate-binding domain of the enzyme. The rate of spontaneous reactivation of alkyl phosphorylated AChE depends on the chemical structure of the OP compound. Spontaneous reactivation of dimethyl phosphorylated AChE proceeds quite rapidly. However, unless oximes are employed, there is no such expectation of rapid recovery for patients intoxicated with diethyl phosphoryl insecticides.

Absorption↗

Quantitative analysis of polychlorinated biphenyls, organochlorine insecticides, polycyclic aromatic hydrocarbons, polychlorinated hydrocarbons and polynitrohydrocarbons in spiked samples of soil, water and plasma by selected-ion monitoring gas chromatography-mass spectrometry.

A broad range of pollutants such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated hydrocarbons (PCHs), polynitrohydrocarbons (PNHs), polychlorinated biphenyls (PCBs) and organochlorine (OCs) insecticides were simultaneously analyzed in spiked soil, water or plasma samples by using gas chromatography-mass spectrometry (GC-MS). Water and plasma samples containing the pollutants were extracted by a solid-phase extraction (SPE) method using florisil columns. The soil samples, fortified with the toxicants, were extracted with water, methanol or dichloromethane (DCM). The water extract was processed by the SPE method. The methanol and DCM samples were dried, dissolved in acetonitrile and subjected to the SPE extraction. The extracted samples were analyzed by GC-MS programmed to monitor selected ions. The deuterium labelled compounds were used as the internal standards. The chromatographic profile of total ions indicated complete separation of some compounds such as isophorone, naphthalene, all PCBs, most OC insecticides and PNHs; high Mr PAHs and some PCHs were partially or incompletely separated. The chromatographic profile of individual ion indicated good separation of each ion. The minimum detection limit ranged from 1 to 4 pg injected when 1 or 2 ions were monitored or from 20 to 200 pg injected when 20 ions were monitored. The SPE method that provided 60-105% recovery of pollutants from water samples, provided only 2-60% recovery from plasma samples. This may be due to the binding of pollutants to plasma proteins. Water recovered 1-30%, while methanol or DCM recovered 65-100% of the pollutants added to the soil samples. The use of internal standards corrected for the loss of pollutants from plasma or soil.

Environmental Pollutants↗

Insecticidal activity against Aedes aegypti larvae of some medicinal South American plants.

The insecticidal activity of 11 extracts from nine South American medicinal plants has been studied using the Aedes aegypti larvicidal assay. Eight of the 11 plant extracts studied showed toxicity against the A. aegypti larvae (LC(50)<500 microg/ml). The dichloromethane extracts of Abuta grandifolia and Minthostachys setosa demonstrated high larvicidal activity, the most active being the dichloromethane extract of A. grandifolia, with an LC(50)=2.6 microg/ml (LC(100)= 8.1 microg/ml), indicating an activity 2-fold higher than beta-asarone, a natural botanical insecticide used as a positive control (LC(100)=16 microg/ml). On the other hand, the dichloromethane extract of M. setosa was quite potent against A. aegypti larvae showing an LC(50)=9.2 microg/ml (LC(100)=25.2 microg/ml). The results obtained suggest that the extracts of A. grandifolia and M. setosa are promising as larvicides against A. aegypti larvae and could be useful in the search for new larvicidal natural compounds.

Aedes↗

Argentine plants as potential source of insecticidal compounds.

CH2Cl2 and MeOH extracts of 15 Argentine plants used locally as insecticides, were evaluated for their insecticidal activity. Chenopodium multifidum L. (Chenopodiaceae); Flaveria bidentis (L.) O.K. (Compositae); Aristolochia argentina Gris. (Aristolochiaceae) and Tagetes erecta L. (Compositae) showed a significant activity against Sitophilus oryzae.

Analysis of Variance↗