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Selective effects of cyclodiene insecticides on dopamine release in mammalian synaptosomes.

Cyclodiene insecticides release labeled neurotransmitter in striatal and cortical synaptosome preparations under nondepolarizing conditions, typically showing half-maximal potencies for release in the low micromolar range. This level of potency is similar to those reported for inhibition of 36Cl- influx at the gamma-aminobutyric acid (GABA)(A) receptor, their consensus target site. A wide variety of other GABA(A) antagonists, including picrotoxinin and bicuculline, did not cause significant dopamine release, which obviated direct involvement of the GABA(A) receptor as a possible site of action. Release assays with different transmitters indicated that striatal dopaminergic terminals are severalfold more sensitive to release than other neurotransmitter types. The selective sensitivity of nigrostriatal dopaminergic nerve terminals to insecticidal organochlorines provides biochemical evidence supporting an epidemiological linkage between exposure to environmental toxicants and Parkinsonism.

Animals↗

Foetal and maternal rat brain acetylcholinesterase: isoenzymes changes following insecticidal carbamate derivatives poisoning.

Pregnant rats (18th day) were orally given insecticidal carbamates with anticholinesterasic properties: 50 mg/kg carbaryl, 0.1 mg/kg aldicarb, 2.5 mg/kg carbofuran and 20 mg/kg pirimicarb. The acetylcholinesterase (AChE) isoenzymes from the brain of mothers and their foetuses were separated by electrophoresis on polyacrylamide ge. The four carbamate derivatives caused a significant lowering of the percentage of the least mobile isoenzyme (isoenzyme 1) in mother. Aldicarb, carbaryl and pirimicarb also caused a significant decrease in the percentage of the first foetal isoenzyme. The second foetal isoenzyme underwent a decrease after treatment with aldicarb or carbofuran. The distribution of the foetal isoenzymes was seen to be different to that of the dams in both control and treated animals. The difference in sensitivity of the cerebral AChE to the insecticides under investigation is thought to depend on differences of the fixation of carbamate derivatives on the foetal and maternal isoenzymes.

Acetylcholinesterase↗

Insecticide resistance and malathion carboxylesterase in the sheep blowfly, Lucilia cuprina.

Resistance to the organophosphorus insecticide malathion in genetically related strains of the Australian sheep blowfly Lucilia cuprina was examined. Separate lines of blowflies were established by homozygosis of the fourth chromosome of the parental RM strain. Both the RM and the derived resistant (der-R) strains are approximately 100 times more resistant to malathion than the related susceptible der-S strain, resistance being correlated with a 45- to 50-fold increase in a malathion carboxylesterase (MCE) activity. MCE has a pH optimum ranging between 6.6 and 8.0 and is strongly inhibited by the carboxylesterase inhibitors triphenyl phosphate, paraoxon, and diisopropylfluorophosphate. Subcellular fractionation revealed that MCE was localized predominantly to the cytosol and mitochondria in both resistant and susceptible blowflies. A single MCE was purified to homogeneity from RM blowflies. It has a pI of 5.5, is a monomer of 60.5 kDa, and hydrolyzes malathion with a Vmax of 755 nmol/min/mg protein and a Km of 11.0 microM. L. cuprina have thus evolved a remarkable MCE which is faster and more efficient at hydrolyzing a specific insecticide than any other insect esterase vet described.

Amino Acid Sequence↗

[Residue analysis of pyrethroid insecticides in cereal grains, milled fractions and bread].

A procedure is described for the residual analysis of eight pyrethroid insecticides at levels of 2-5 micrograms/kg. Residues are extracted from cereal grains, flour or bread with a mixture of acetonitrile/water (2 + 1) and are partitioned into petroleum ether. Clean-up is performed by gel permeation chromatography on Bio-Beads S-X3 and an additional Florisil column. Electron capture gas liquid chromatography on a non-polar SE-30 capillary column separates the isomers of the compounds. Under the conditions used in the experiments with cyfluthrin, cypermethrin, deltamethrin, fenvalerate, and permethrin, the compound levels in rye and wheat did not decrease significantly. Losses during milling and baking were small. Consequently, pyrethroid insecticides remain effective in grain stored over a long period of time. However, considerable residues result in the milled fractions as well as in different types of bread.

Bread↗

Effects of the insecticides carbofuran and fenvalerate on adenylate parameters in bluegill sunfish (Lepomis macrochirus).

Bluegills (Lepomis macrochirus) were exposed to two sublethal concentrations of carbofuran, a carbamate insecticide, and fenvalerate, a synthetic pyrethroid insecticide, in continuous flow bioassay systems for 30 days. Adenylate parameters (ATP concentration, total adenylates concentration, and adenylate energy charge) were monitored in gill, liver, muscle, and stomach tissues. There were significant decreases in adenylate energy charge of gill and stomach tissue relative to initial values after 10 days of exposure to 30 micrograms/L carbofuran. There was also a significant decline in adenylate energy charge in gills of fish exposed to 0.2 micrograms/L fenvalerate relative to initial values. These values returned to normal by the end of the experiment. The biological significance of these changes is uncertain. Adenylate parameters may not be sufficiently sensitive to reflect sublethal effects of pesticides which affect the nervous system of fishes.

Adenosine Monophosphate↗

Influence of insecticidal plant materials used during storage on sensory attributes and instrumental hardness of dry edible beans (Phaseolus vulgaris L.).

Three plant products with known insecticidal properties, a dry extract of flowers of Chrysanthemum cinerariaefolium (Trevir.) Vis. produced in Rwanda, an ethanol extract of seeds of neem, Azadirachta indica A. Juss, and crushed leaves of Tetradenia riparia Hochst Codd, a traditional Rwandan medicine, were mixed with beans, Phaseolus vulgaris L., for storage protection. These plant-protected beans were compared with "off the shelf' beans that were being sold to consumers by the Rwandan National Agricultural Products Marketing Organization (OPROVIA). A trained sensory panel determined that beans treated with neem and C. cinerariaefolium were as acceptable after 8 months storage as those being sold throughout Rwanda by the marketing organization. Beans marketed by this organization were all treated with the standard insecticide application in Rwanda, 0.01% weight/weight pirimiphos methyl in a powder formulation. Instrumental hardness (% hard-to-cook/mean gram force) after 20 months of storage was acceptable for beans stored with neem or with C. cinerariaefolium or with the conventional government application of pirimiphos methyl. Use of either neem or C. cinerariaefolium for storage protection should not affect consumer acceptance of dry beans.

Fabaceae↗

Production of polyclonal antibodies towards the immunodetection of insecticide phosalone.

Hapten synthesis for the production of specific insecticide phosalone polyclonal antibodies was carried out starting from an intermediate of the phosalone synthesis, 6-chloro-2-benzoxazolone 1. Two haptens containing different spacers have been prepared: N-5-carboxypentyl-6-chloro-2-benzoxazolone 7 and N-(2-oxo-3-aza-5-carboxypentyl)-6-chloro-2-benzoxazolone 12. Each of these two haptens conjugated to bovine serum albumine (BSA) was used to immunize four rabbits. Immunoassays of phosalone were performed with ELISA using solid-phase bound hapten thyroglobulin conjugate and horseradish peroxidase labelled goat antirabbit IgG. The more sensitive response was observed when the antiserum obtained from the rabbit immunized with the hapten-BSA conjugate containing the N-2-oxo-3-aza-5-carboxypentyl spacer was in competition with the same hapten coupled to thyroglobulin. An identical response was obtained under the same conditions when using benzoxazolone instead of phosalone as competitor, showing a good recognition of the specific aromatic part of the organophosphate insecticide phosalone. Reduction of coating conjugate concentration (from 2 to 0.05 micrograms/well) and also the use of heterologous coating protein instead of homologous did improve the sensitivity, resulting in a concentration of phosalone required to inhibit binding by 50% of 2 mg/l and a detection limit of 0.02 mg/l.

Animals↗

Partition distribution of insecticides as a critical factor affecting their rates of absorption from water and relative toxicities to fish.

Loaches, Misgurnus anguillicaudatus (Cantor), a common fish in Taiwan, were treated with DDT, dieldrin, and monocrotophos by continuous exposure in aqueous solutions (or suspensions) and by injection. DDT and dieldrin were 150 and 220 times more toxic, respectively, than monocrotophos, to the fish exposed in aqueous solutions (24-hr LC50), but only 1/9 and 1/4 as toxic as monocrotophos by injection (24-hr LD50). Results of GLC analyses indicate that, at the end of 24-hr exposure, 96.5% of DDT, 92.7% of dieldrin, and 14.3% of monocrotophos were absorbed by loaches from aqueous solutions. The initial rates of absorption for DDT and dieldrin were about 10 to 20 times faster than that for monocrotophos. The large differences in relative toxicity may be due to partition distribution which in turn caused differences in absorption, as DDT and dieldrin are lipophilic and monocrotophos is hydrophilic. Statistical analysis of the relationship between fish toxicities and partition coefficients supports the present finding. The coefficient of correlation is 0.70 between parition coefficients (benzene/water) and toxicities to fish (rainbow trout) of 12 organophosphorus insecticides, 0.74 between coefficients and corrected fish toxicities, and 0.96 between partition coefficients and corrected fish toxicities for organophosphates only. Results of analyses are significant at less than 1% probability level. Similar correlation was also obtained between partition coefficients for hexane/water and toxicities of 8 organophosphorus and 5 organochlorine insecticides to rainbow trout.

Absorption↗

Insecticide and nutrient transport in water, related to agricultural land use of a stream basin in Ontario, Canada.

Transport by stream water of insecticides and nutrients in Big Creek, Norfolk County, Ontario, Canada, was examined by combining concentrations of substrates with flow data. Big Creek has its headwaters in dairy cattle country, its central basin area is mainly devoted to tobacco growing, and its lower reaches contain mixed farming, corn and vegetables, etc., before it flows into Lake Erie. Three sampling sites were chosen to represent these 3 different land uses. Generally concentrations of substrates in the water were quite uniform at the 3 sites resulting in transport of quantities in proportion to stream flow. Certain anomalies occurred and are discussed. The 2 chief insecticides found were DDT and dieldrin. Analyses for potassium, calcium and magnesium indicated that these nutrient losses into the stream were area-rather than usage-dependent. Midseason variations in loss of nitrogen and phosphorus may be the result of agricultural practices in the three areas represented in this study. The largest quantities of all nutrients lost occurred early in the season before crops were established.

Agriculture↗

Toxicological studies on the organophosphorous insecticide methyl-ISP.

Methyl-ISP, a newly developed organophosphorous insecticide, is used in China to treat and protect plants from pest infestation. Our studies demonstrated that methyl-ISP is metabolized rapidly in rat and mouse. Its toxicity was low, no obvious accumulative toxicity, chronic toxicity, teratogenicity, mutagenicity, reproductive toxicity or delayed neurotoxicity could be observed. It is therefore concluded that methyl-ISP is relatively safe to animals and human subjects. methyl-ISP is now employed to replace the other commonly used insecticide hexachlorobenzene (666) in agriculture. A preliminary study was performed to elucidate the mechanism of intoxication at subcellular levels.

Animals↗

Comparative study on the effects of three insecticides (fipronil, imidacloprid, selamectin) on developmental stages of the cat flea (Ctenocephalides felis Bouché 1835): a light and electron microscopic analysis of in vivo and in vitro experiments.

The effects of three insecticides (fipronil, imidacloprid and selamectin) on developmental stages of cat fleas (Ctenocephalides felis) were studied in vivo, in vitro and by means of light and electron microscopy. The results were documented by video. Adult fleas were attached to the skin of dogs that had been treated 7 days before with one of the three compounds. Furthermore, adult fleas were exposed exclusively to the hair and skin debris of such treated dogs or were placed on filter papers that had been impregnated with one of these three compounds or with the blood of treated dogs. Larval fleas were exposed to hair of treated dogs, to debris obtained by combing treated dogs, to dried blood samples of treated dogs or were placed onto filter papers impregnated with one of the three compounds. In these experiments with adult and larval fleas, it was noted that none of the three insecticides had a repellent effect on adult or larval fleas. Imidacloprid was the only compound that acted exclusively by body contact, and was apparently taken up by adult and larval fleas via the thin, non-sclerotized intersegmental membranes of the flea's body, shown when flea stages were exposed to hairs taken from dogs treated with one of the compounds or placed onto drug-impregnated filter papers. Imidacloprid killed larvae and adult fleas within 1 h, while it took at least 24 h until all adult fleas had died on fipronil- or selamectin-treated dogs, thus allowing longer feeding periods, increasing the risk of transmission of flea-derived diseases. Flea larvae covered with debris from dogs topically treated 7 days before with fipronil, imidacloprid or selamectin died, like the untreated control, within 16-28 h after exposure. This was, however, probably mainly due to a drying effect. Adult and larval fleas exposed to filter papers impregnated with the blood of treated dogs survived longer than 7 days, as did the untreated controls. All three drugs apparently acted on nerves and muscles and thus stopped motility.

Animals↗

[Genotoxic effects of insecticides in current use on mucosal epithelial cells from human tonsil tissue].

BACKGROUND: Vector control is of critical medical importance in disease prevention, as reflected in sections 17 and 18 of the German Protection Against Infection Act. In the past, a large number of biocides were found to be hazardous to human health and were banned from the market, subsequently being replaced by new active ingredients and galenic forms. Many of these new insecticides are available in spray or nebuliser form. Whether these preparations have genotoxic effects on mucosal epithelial cells of the upper aerodigestive tract has thus far not been investigated. MATERIALS AND METHODS: We used the comet assay, as a well-established genotoxicity test, to investigate whether malathion, diazinon, pyridostigmine bromide, piperonyl butoxide, silafluofen, and fipronil had genotoxic effects on tonsil specimens taken from 85 patients. RESULTS: All substances tested proved to have a strong genotoxic effect on mucosal epithelial cells taken from human tonsil tissue. We found clear differences between substance groups. CONCLUSIONS: Sufficient doses of a wide range of insecticides are indispensable in many areas of human life, especially for the prevention of diseases. Depending on the method of application, however, ingestion or inhalation of these substances can damage mucosal epithelial cells of the upper aerodigestive tract. Further epidemiological studies should be undertaken to investigate whether this involves potential health hazards in at-risk populations.

Adolescent↗

Development of an EnFET for the detection of organophosphorous and carbamate insecticides.

A biosensor for the detection of insecticides based on an ion-sensitive field-effect transistor (ISFET) was developed. The resulting device combines the simplicity of potentiometric sensors and the use of associated electronic systems as powerful tools for the acquisition and the processing of data. The enzyme acetylcholinesterase (AChE) was entrapped in a membrane placed on the gate of the ISFET forming an enzyme field-effect transistor (EnFET). The biosensor is applied to the determination of pesticides in spiked real samples. Organophosphorous and carbamate insecticides were measured with a detection limit of 10(-8) mol L(-1). The measurement is based on the production of hydrogen ions due to the hydrolysis of acetylthiocholine by the enzyme. The resulting local pH change is picked up by the underlying pH-sensitive ISFET and transduced as potential variations. The preparation of the membrane is simple and reproducible. The analysis in spiked real samples was performed in tap water and showed detection limits comparable to those obtained by other researchers.

Acetylcholinesterase↗

Determination of organophosphorus insecticides in natural waters using SPE-disks and SPME followed by GC/FTD and GC/MS.

Two methods for the analysis of ten organophosphorus insecticides in natural waters using solid phase extraction disks containing C18 and SDB and solid phase microextraction fibers containing polyacrylate (PA) are developed. Bromophos ethyl, bromophos methyl, dichlofenthion, ethion, fenamiphos, fenitrothion, fenthion, malathion, parathion ethyl and parathion methyl were determined by GC/MS and GC/FTD. The SPE-disks require only 1000 mL of sample and provide a method limit of detection in the range of 0.01-0.07 microgram/L and recovery rates from 60.7 to 104.1%. The solid phase microextraction (SPME) technique requires 2-5 mL of water sample and provides a method limit of detection in the range of 0.01 to 0.05 microgram/L for all detectors and the recoveries compared to distilled water ranged from 86.2 to 119.7%. The proposed methods were applied to the trace level screening determination of insecticides in river water samples originating from different Greek regions.

Calibration↗

Multiple mutations and gene duplications conferring organophosphorus insecticide resistance have been selected at the Rop-1 locus of the sheep blowfly, Lucilia cuprina.

Sequences of the esterase gene alpha E7 were compared across 41 isogenic (IV) strains of the sheep blowfly, Lucilia cuprina, and one strain of the sibling species, L. sericata. The 1.2-kb region sequenced includes sites of two insecticide resistance mutations. Gly137Asp confers resistance to organophosphorus insecticides (OPs), particularly preferring diethyl OPs such as diazinon, while Trp251Leu prefers dimethyl OPs, and particularly malathion, with the additional presence of carboxylester moieties. We found that there are just eight haplotypes among the 41 chromosomes studied: two Gly137Asp containing haplotypes, two Trp251Leu containing haplotypes, and four susceptible haplotypes, including the L. sericata sequence. While phylogenetic analysis of these haplotypes suggests that the Asp137 and Leu251 mutations each arose at least twice, evidence for recombination was detected across the region, therefore single origins for these resistance mutations cannot be ruled out. Levels of linkage disequilibrium in the data are high and significant hitchhiking is indicated by Fay and Wu' s H test but not the Tajima test. A test of haplotype diversity indicates a paucity of diversity compared with neutral expectations. Both these results are consistent with a very recent selective sweep at the Lc alphaE7 locus. Interestingly, gene duplications of three different combinations of OP resistant haplotypes were identified in seven of the isogenic (IV) strains. All three types of duplication involve an Asp137 and a Trp251 haplotype. To examine whether more haplotypes existed before the hypothesised selective sweep, fragments of alpha E7 surrounding the resistance mutations were amplified from pinned material dating back to before OPs were used. Four new sequence haplotypes, not sampled in the survey of extant haplotypes, were obtained that are all associated with susceptibility. This is suggestive of a higher historical level of susceptible allelic diversity at this locus.

Animals↗

A method to quantify and analyze the foraging activity of honey bees: relevance to the sublethal effects induced by systemic insecticides.

The assessment of agropharmaceuticals' side effects requires more realistic simulations of field conditions than those deduced from the dose-lethality relation obtained under laboratory conditions. Because the presence of sublethal doses or concentrations may also alter the behavior of foraging insects, we attempted to devise a quantifiable and accurate protocol for evidencing various alterations in free-flying bees. Such a protocol was illustrated by testing new classes of systemic insecticides. The protocol focused on video recording to quantify the foraging activity of small colonies of honey bees confined in insect-proof tunnels. The basis of the protocol was not the colony itself but the change in each colony on a specific day and between days. First, the paradigms of attendance at a safe feeding source were established by observing 8 control colonies at different times of the season during 5 days after the necessary forager training was accomplished. Second, on three different colonies we considered the paradigms on the control day before contamination and during 4 days after the feeding source was contaminated. During the same period, one more colony was exclusively fed with safe food to serve as control. Two plant-systemic insecticides were tested at contamination levels 70 times lower than the 50% of the lethal concentration. Imidacloprid, at 6 microg/kg, clearly induced a decrease in the proportion of active bees. Fipronil, at 2 microg/kg, induced an additional decrease in attendance at the feeder. Such levels are still higher than the corresponding lowest observable effect concentration (LOEC). Our protocol, which provided intermediate conditions between field and laboratory conditions, allowed the quantification, with an enhanced level of sensitivity, of sublethal effects on foraging bees.

Agriculture↗

Organochlorine insecticides in mudflats of Hong Kong, China.

Organochlorine (OC) insecticides were measured in different environmental compartments (including seawater, sediment, and biota) at six sites within the Mai Po Marshes Nature Reserve (MPMNR) and at one site in A Chau (AC) in Hong Kong in June 2002 (wet season) and January 2003 (dry season). MPMNR is a Wetland of International Importance (Ramsar site) situated in the western part of Hong Kong, whereas AC is a relatively remote island in Starling Inlet on the eastern side of the New Territories of Hong Kong. Generally, concentrations of OC insecticides in water, sediment, and biota were greater at MPMNR than at AC. Hexachlorobenzene, dichloro-diphenyl-trichloroethane, and hexachlorocyclohexanes were detected in all samples collected from MPMNR. Preliminary risk assessments were performed by comparing the measured concentrations of OCs in different environmental compartments with the corresponding guidelines promulgated by various environmental protection authorities. Concentrations of some OCs in sediments from certain locations in MPMNR were consistently greater than their corresponding sediment guideline values. These results suggest that the sediment OC concentrations in the sediments of MPMNR might pose a risk to aquatic organisms. Despite this, concentrations of OCs in biota samples were much lower than the relevant guideline values promulgated by government agencies to protect the health of human consumers. It is postulated that the management practices adopted at MPMNR have been effective in limiting the bioaccumulation window (duration) for OCs. This situation may have been aided by a frequent exchange of water between the tidal shrimp ponds at MPMNR and the open sea, again attributable to management practices. The low-cost maintenance practices adopted at MPMNR may potentially be applicable to wetlands in other southeastern Asian countries, thus allowing sustainable fishery production and wildlife conservation.

Animals↗

The impact of insecticides on beneficial arthropods in cotton agro-ecosystems in South Africa.

The unique diversity of beneficial arthropods in South Africa can be regarded as an important natural resource in agro-ecosystems as it plays an important role in the natural control of insect pests. Insecticide applications reduce the ability of these beneficials to regulate cotton pests. In the absence of insecticides, average daily predation rates of 37% and 30% of bollworm eggs and larvae respectively were found in exclusion experiments. By minimizing the number of pesticide applications the combination of the direct negative effect of pesticide application on predator populations and secondary pesticide effects such as the stimulation of red spider mite populations as a result of predator suppression, can be avoided. This would open the option to fully utilize the full pest control potential of the natural enemy complex.

Animals↗