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Characterization of [3H]ethynylbicycloorthobenzoate ([3H]EBOB) binding and the action of insecticides on the gamma-aminobutyric acid-gated chloride channel in cultured cerebellar granule neurons.

[3H]Ethynylbicycloorthobenzoate ([3H]EBOB) is a recently developed specific radioligand for the noncompetitive blocker site of the gamma-aminobutyric acid (GABA) receptor, which is the target of many polychlorocycloalkane and heterocyclic insecticides. This study characterizes [3H]EBOB binding and the action of insecticides on the GABA-gated chloride channel of cerebellar granule neurons in primary culture. Specific [3H]EBOB binding increases rapidly between days 3 and 7 and then remains constant for days 7 to 12 in culture. A single, high-affinity and saturable binding site is involved with Kd = 0.51 nM and Bmax = 239 fmol/mg protein. GABA and muscimol are 6- to 12-fold more potent as inhibitors of [3H]EBOB binding to membranes from cultured cerebellar granule neurons than from cerebellar tissue, possibly because of localization of specific subunits in the granule cells. GABA-stimulated 36Cl- influx is also inhibited by seven insecticidal compounds (including EBOB, 12-ketoendrin and alpha-endosulfan) as well as picrotoxinin and t-butylbicyclophosphorothionate with IC50 levels ranging from 290 to 15300 nM, which correlate well (r = 0.92) with their inhibitory potencies for [3H]EBOB binding (1 to 773 nM). These findings validate the use of [3H]EBOB binding in cultured cerebellar granule cells for pharmacological studies on insecticides and other toxicants acting at the noncompetitive blocker or coupled site of the GABAA receptor/chloride channel complex.

Animals↗

The insecticide-resistance problem: a review of developments in 1956 and 1957.

The author reviews the growth of the insecticide-resistance problem throughout the world during the period between July 1956 and November 1957, and the developments in research on the subject during the same period.Three new resistant species have been discovered-Anopheles subpictus, Chrysomyia putoria and Rhipicephalus sanguineus-and eight new types of resistance in already resistant species have been observed. Moreover, the geographical area covered by certain resistant insect populations has considerably increased.THE RESEARCH ACCOMPLISHMENTS DURING THE PERIOD UNDER REVIEW INCLUDE: systems of detecting resistance in the field by standard test methods; confirmation of two distinct types of resistance to chlorinated-hydrocarbon insecticides in mosquitos and bed-bugs as well as in houseflies; evidence that DDT-resistance in the housefly, Anopheles sundaicus and Aëdes aegypti is due mainly to a single genetic factor associated with the ability to dehydrochlorinate DDT, and that dieldrin-resistance of Anopheles gambiae also derives from a single factor present even in untouched populations; a fuller understanding of the physiological mechanism of BHC-resistance in the housefly; and demonstration that selection pressure from organo-phosphorus compounds induces resistance to themselves and to chlorinated-hydrocarbon insecticides.

Aedes↗

Fate and effects of the insecticide Dursban 4E in indoor Elodea-dominated and macrophyte-free freshwater model ecosystems: I. Fate and primary effects of the active ingredient chlorpyrifos.

The fate of the insecticide Dursban 4E (active ingredient chlorpyrifos) and its effect on crustaceans and insects was studied in indoor experimental freshwater ecosystems that intended to mimick drainage ditches. A single dose (simulating aerial drift) was applied to achieve nominal chlorpyrifos concentrations of 5 or 35 micrograms/L. Two experiments were performed, one in which all model ecosystems were dominated by the macrophyte Elodea nuttallii, and one using systems devoid of macrophytes. In macrophyte-dominated systems, Elodea vegetation adsorbed a large proportion of the dose applied and hampered the mixing of the insecticide in the water (at least up till day 8). Only a small proportion became incorporated in the sediment. In open water systems the insecticide was rapidly mixed in the water, and the sediment played a very significant role as sink for chlorpyrifos. In both Elodea-dominated and open water systems 50% of the dose applied had disappeared on day 8 post-treatment. The rate of disappearance of chlorpyrifos was relatively rapid in water and macrophytes, and relatively slow in the sediment. Of the arthropods in the zooplankton Cladocera were more susceptible than Copepoda. Significant effects (p less than or equal to 0.05) on Cladocera occurred relatively late in Elodea-dominated systems (in week 4 post-application) in contrast to open water systems (week 1), which is in accordance with the observed differences in the fate of chlorpyrifos. Daphnia pulex, D. longispina and Simocephalus vetulus recovered in the model ecosystems when chlorpyrifos concentrations were lower than 0.1-0.2 micrograms/L, which is in agreement with results of laboratory protocol tests performed with these cladocerans. Among the macroscopic Arthropoda the apparent order of susceptibility was amphipods greater than insects greater than isopods. The isopod Asellus aquaticus was more sensitive to the application of the insecticide than the closely related species Proasellus coxalis. In treated open water systems the latter even increased significantly in numbers. Cage experiments in the model ecosystems performed with several species of Arthropoda indicate that laboratory protocol tests may give a reasonable prediction of short-term direct effects of chlorpyrifos for the same species inhabiting more complex aquatic systems.

Animals↗

Action of polychlorocycloalkane insecticides on binding of [35S]t-butylbicyclophosphorothionate to Torpedo electric organ membranes and stereospecificity of the binding site.

Binding of [35S]t-butylbicyclophosphorothionate ([35S]TBPS) to Torpedo electric organ membranes was characterized. A dose- and pH-dependent binding (100.8 pmol/mg protein) was detected with a single affinity (Kd of 0.9 microM) in the presence of 150 mM KCl at pH 6.8. Other anions such as Br- and I- also increased binding affinity, but to a lower degree than Cl-, which increased the affinity by two- to threefold. In presence of 150 mM KCl, [35S]TBPS binding was inhibited noncompetitively by Zn2+ and by 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS) (IC50 of 9 microM). The gamma-isomer of hexachlorocyclohexane (BHC) was much more potent in inhibiting this [35S]TBPS binding and the inhibition was competitive (Ki = 40 nM). Like binding of [35S]TBPS to the gamma-aminobutyric acid (GABA) receptor, its binding to Torpedo membranes was inhibited by pentobarbital, mephobarbital, and hexobarbital (IC50 of 85, 225, and 300 microM), respectively), but not by phenobarbital. Binding was not inhibited by diazepam, GABA, bicuculline, or avermectin B1a, ligands that bind to the GABAA receptor. [35S]TBPS binding was inhibited by BHC isomers with the following decreasing order of potency alpha = gamma greater than sigma greater than beta, and by cyclodiene insecticides. Endrin was more potent than dieldrin, but endosulfan I and II had similar effects. The data suggest that the binding site for polychlorocycloalkane insecticides on this protein is much less stereoselective than that of the Cl- channel of the GABAA receptor. Also, even though this Torpedo protein has higher affinity for insecticides, such as gamma-BHC, than does the GABAA receptor, it is the latter whose specificity correlates best with polychlorocycloalkane toxicity. Nevertheless, because of its high affinity for gamma-BHC such a protein in muscles or brain may be an important target for the action of this insecticide.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Cytogenetic effects of pesticides. III. Induction of micronuclei in mouse bone marrow by the insecticides cypermethrin and rotenone.

The production of micronuclei in mouse bone marrow by the pyrethroid insecticide, cypermethrin and the botanical insecticide, rotenone was examined. Three routes of administration were used for the insecticides: intraperitoneal, oral and dermal. The different routes of treatment with cypermethrin and rotenone caused toxicity of marrow as indicated by a significant increase in the percentage of polychromatic erythrocytes (PEs) over that of the control. Cypermethrin showed mutagenic potential as evidenced by a positive response in the micronucleus assay. Oral administration of the insecticide at a dietary level of 900 ppm for 7 and 14 consecutive days as well as double and multiple (total 4) dermal treatments (360 mg/kg body wt.) induced a statistically significant increase in the frequency of PEs with micronuclei. The conducted intraperitoneal (i.p.) treatments with cypermethrin: single injection at 60 and 180 mg/kg body wt., double and multiple injections (total 3) at 60 mg/kg body wt. did not affect the percentage of PEs with micronuclei. The different treatments with rotenone: single, double and multiple (i.p.) injections (total 3) at 2 and 3 mg/kg body wt., oral administration for 14 consecutive days at dietary level of 225 ppm and multiple dermal treatments (total 4) with 135 mg/kg body wt. showed no effect on the frequency of micronuclei in PEs.

Administration, Oral↗

Construction of new insecticidal Bacillus thuringiensis recombinant strains by using the sporulation non-dependent expression system of cryIIIA and a site specific recombination vector.

Bacillus thuringiensis (Bt) delta-endotoxins are safe biological insecticidal proteins whose usefulness has long been recognized. The first commercialized Bt insecticidal formulations were composed of spore-crystal preparations derived from wild-type strains. These products generally have a limited insecticidal host range and several genetically modified strains have, therefore, been constructed using transformation procedures. However, addition of a new delta-endotoxin gene to strains already harboring other delta-endotoxin genes often resulted in broader-spectrum but less potent products because they produced significantly less of each of the crystal proteins. We report expression of the coding sequence of the sporulation specific cryIC gene from the non-sporulation-dependent cryIIIA promoter. Large amounts of CryIC accumulated in various Bt strains with different genetic backgrounds. Sporulation deficient Spo0A mutants, acrystalliferous derivatives and wild-type Bt strains expressing the engineered cryIII-cryIC gene were obtained. Introduction of the cryIII-cryIC gene whose product is highly active against Spodoptera littoralis into the Kto strain harboring the cryIA(c) gene active against Ostrinia nubilalis resulted in the construction of a new strain with increased potency and broader activity spectrum than the parent strain. Large amounts of each toxin were produced and the expression of the two genes seemed to be summed, presumably because the expression systems of the two genes are different. The plasmid shuttle vector used to introduce the cryIII-cryIC gene into the different Bt hosts utilizes the specific resolution site of transposon Tn4430 to enable construction of recombinant Bt strains that are free of foreign non-Bt DNA. This should facilitate the approval and acceptance for environmental release of the insecticidal recombinant products.

Animals↗

The effects of 2,4-D herbicide and organophosphorus insecticides on growth, photosynthesis, and chlorophyll a synthesis of Chlamydomonas reinhardtii (mt +).

The effects of the herbicide (2,4-D) and six organophosphorus insecticides (Diazinon, Dimethoate, Fenitrothion, Malathion, Phenthoate and Quinalphos) on growth, photosynthesis and chlorophyll a synthesis of the fresh water green alga Chlamydomonas reinhardtii (mt +) were studied. At low concentrations (1 and 5 ppm), the herbicide and all six orgnophosphorus insecticides stimulated photosynthesis of the alga. The stimulating effects on algal growth and chlorophyll a synthesis were only observed in the presence of low concentration (1 ppm) of 2,4-D and Fenitrothion, Growth, photosynthesis and chlorophyll a synthesis of the alga were inhibited in the presence of high concentrations (10, 20 and 40 ppm) of the herbicide and all the six organophosphorus insecticides. Results also indicated that the toxicities of these organophosphorus insecticides on the tested alga were dependent on their chemical structures.

Journal Article↗

Effect of the organophosphorous insecticide, chlorpyrifos (Dursban), on growth, fecundity and mortality of Biomphalaria alexandrina and on the production of Schistosoma mansoni cercariae in the snail.

Exposure of Biomphalaria alexandrina to sublethal concentrations (0.125, 0.25 and 0.05 ppm) of the organophosphorous insecticide, chlorpyrifos (Dursban), induced a reduction in egg production and egg hatchability. Exposure of Schistosoma mansoni miracidia to the insecticide (60 min, 0.50 ppm) prior to infection of B. alexandrina did not affect the subsequent production of cercariae. However, exposure of S. mansoni-infected snails to the insecticide until day 55, from day 20 to day 62 and from day 35 to 62 following infection resulted in blockage of cercarial shedding. Cercarial shedding commenced in some snails when the treatment stopped. Exposure to the insecticide in concentrations of 0.125 and 0.25 ppm during the first 20 days following infection did not affect the subsequent production of cercariae, but exposure to 0.5 ppm during the first 20 days affected markedly the production of cercariae due to a high snail mortality. The findings indicate that the cercaria is the target stage for the activity of chlorpyrifos on the intramolluscan larval development. It is suggested that S. mansoni cercarial production in B. alexandrina may be a useful system for monitoring the effect of low concentrations of pesticides on the aquatic environment, and that the ability by chemical means to interrupt the cercarial production might be a useful tool in further analyses of important aspects of the snail/parasite relationship.

Animals↗

Midgut bacteria required for Bacillus thuringiensis insecticidal activity.

Bacillus thuringiensis is the most widely applied biological insecticide and is used to manage insects that affect forestry and agriculture and transmit human and animal pathogens. This ubiquitous spore-forming bacterium kills insect larvae largely through the action of insecticidal crystal proteins and is commonly deployed as a direct bacterial spray. Moreover, plants engineered with the cry genes encoding the B. thuringiensis crystal proteins are the most widely cultivated transgenic crops. For decades, the mechanism of insect killing has been assumed to be toxin-mediated lysis of the gut epithelial cells, which leads to starvation, or B. thuringiensis septicemia. Here, we report that B. thuringiensis does not kill larvae of the gypsy moth in the absence of indigenous midgut bacteria. Elimination of the gut microbial community by oral administration of antibiotics abolished B. thuringiensis insecticidal activity, and reestablishment of an Enterobacter sp. that normally resides in the midgut microbial community restored B. thuringiensis-mediated killing. Escherichia coli engineered to produce the B. thuringiensis insecticidal toxin killed gypsy moth larvae irrespective of the presence of other bacteria in the midgut. However, when the engineered E. coli was heat-killed and then fed to the larvae, the larvae did not die in the absence of the indigenous midgut bacteria. E. coli and the Enterobacter sp. achieved high populations in hemolymph, in contrast to B. thuringiensis, which appeared to die in hemolymph. Our results demonstrate that B. thuringiensis-induced mortality depends on enteric bacteria.

Animals↗

Modelled impact of insecticide-contaminated dung on the abundance and distribution of dung fauna.

Deterministic models assessed the effects that contaminated dung from insecticide-treated cattle had on populations of three hypothetical species of dung fauna that dispersed randomly and could double their numbers every 1-28 weeks at low density. Insecticide was allowed to kill 2-98 % of adults and prevent 16-100% of breeding in pats produced immediately after cattle treatment, with toxicity declining to < 1% in pats produced 2-23 days later. Treatment intervals were 10-40 days. The modelled impact of insecticide was affected little by approximately four-fold variations in: length and density dependence of the attractive life span of pats, frequency of pat occupation by immature adults, distribution of pat toxicity during treatment interval, and changes in dispersal rates due to age and population density. Of greater importance were variations in: pat toxicity, treatment interval, frequency of pat occupation by breeding adults, density dependence of recruitment and death, natural adversity and mortality in dormancy, general rate of dispersal, and the size and shape of the area with treated cattle. Overall, it seemed that wide variations in the impact of contamination will occur in the field, but in many situations the risk to dung fauna can be substantial, especially for slow breeding beetles, and muscoids contacting insecticide on cattle. Risk extends outside the treated areas, for a distance equal to several daily displacements of the insects. Untreated refuges for species survival should be compact blocks at least 25 daily displacements wide.

Animals↗

Evidence for similarities between cyclodiene type insecticides and picrotoxinin in their action mechanisms.

Evidence has been obtained to indicate that cyclodiene-type insecticides, e.g., heptachlor epoxide and gamma-BHC, mimic the action of picrotoxinin. These insecticides inhibit the GABA (gamma-aminobutyric acid)-stimulated chloride uptake in the coxal muscle of the American cockroach, and directly compete against [3H]a-dihydropicrotoxinin for binding in the rat brain synaptosomes. Moreover, several cyclodiene-resistant insect strains are also resistant to picrotoxinin. This cross-resistance is specific to picrotoxinin and does not extend to other neuroexcitants. These insecticides, like picrotoxinin, cause central nerve excitation by stimulating transmitter release. Similarity in molecular structures also has been pointed out. These results indicate that some of the nerve excitation symptoms that insecticides cause are likely due to their interaction with picrotoxinin receptor.

Animals↗

Recovery of Triatoma infestans populations after insecticide application: an experimental field study.

The capacity of populations of Triatoma infestans (Klug) (Hemiptera: Reduviidae) to survive and recover was assessed after application of insecticide (gamma-HCH at a rate of 0.5 g a.i./m2) at different seasons. T. infestans populations were maintained in experimental chicken houses under natural climatic conditions in a region of Argentina endemic for Chagas disease transmitted by these bugs. Based on previous studies of T. infestans populations in these habitats, each experimental group was set up with a total of 626 T. infestans, comprising 390 eggs, 204 nymphs of particular stages, fourteen male and eighteen female adults. The chicken houses were dismantled and rebuilt at monthly intervals to study the vector population changes over a period of 33 months. When the insecticide was applied during winter, spring or summer, populations of T. infestans recovered to untreated or precontrol levels during the next reproductive season (i.e. during the hot season, October-March). In contrast, populations treated during autumn (March) remained at very low densities for 2 years and then increased rapidly to surpass the untreated populations. All populations of the bugs fell to very low numbers (sometimes less than twenty individuals) after gamma-HCH applications, but none was driven to extinction. In all cases, the density of surviving populations was independent of their density before treatment. The fact that all treated populations recovered within 1-3 years, to at least the density of untreated populations, shows the high reproductive potential of T. infestans to recover from very low population densities. Moreover, the additive effect of climatic-induced mortality and insecticide-induced mortality is only apparent when insecticides are applied just before the onset of the cold winter months during which reproductive rates are at their lowest.

Animals↗

Comparison of Disulfide Contents and Solubility at Alkaline pH of Insecticidal and Noninsecticidal Bacillus thuringiensis Protein Crystals.

We compared two insecticidal and eight noninsecticidal soil isolates of Bacillus thuringiensis with regard to the solubility of their proteinaceous crystals at alkaline pH values. The protein disulfide contents of the insecticidal and noninsecticidal crystals were equivalent. However, six of the noninsecticidal crystals were soluble only at pH values of >/=12. This lack of solubility contributed to their lack of toxicity. One crystal type which was soluble only at pH >/=12 (strain SHP 1-12) did exhibit significant toxicity to tobacco hornworm larvae when the crystals were presolubilized. In contrast, freshly prepared crystals from the highly insecticidal strain HD-1 were solubilized at pH 9.5 to 10.5, but when these crystals were denatured, by either 8 M urea or autoclave temperatures, they became nontoxic and were soluble only at pH values of >/=12. These changes in toxicity and solubility occurred even though the denatured HD-1 crystals were morphologically indistinguishable from native crystals. Our data are consistent with the view that insecticidal crystals contain distorted, destabilized disulfide bonds which allow them to be solubilized at pH values (9.5 to 10.5) characteristic of lepidopteran and dipteran larval midguts.

Journal Article↗

Insecticidal juvenile hormone analogs stimulate the production of male offspring in the crustacean Daphnia magna.

Juvenile hormone analogs (JHAs) represent a class of insecticides that were designed specifically to disrupt endocrine-regulated processes relatively unique to insects. Recently we demonstrated that the crustacean juvenoid hormone methyl farnesoate programs oocytes of the crustacean Daphnia magna to develop into males. We hypothesized that insecticidal JHAs might mimic the action of methyl farnesoate, producing altered sex ratios of offspring. Daphnids were exposed chronically (3 weeks) to sublethal concentrations of methyl farnesoate, the JHA pyriproxyfen, and several nonjuvenoid chemicals to discern whether excess male offspring production is a generic response to stress or a specific response to juvenoid hormones. Only methyl farnesoate and pyriproxyfen increased the percentage of males produced by exposed maternal organisms. As previously reported with methyl farnesoate, acute exposure (24 hr) to either pyriproxyfen or the JHA methoprene caused oocytes maturing in the ovary to develop into males. We performed experiments to determine whether combined effects of a JHA and methyl farnesoate conformed better to a model of concentration addition (indicative of same mechanism of action) or independent joint action (indicative of different mechanisms of action). Combined effects conformed better to the concentration-addition model, although some synergy, of unknown etiology, was evident between the insecticides and the hormone. These experiments demonstrate that insecticidal JHAs mimic the action of the crustacean juvenoid hormone methyl farnesoate, resulting in the inappropriate production of male offspring. The occurrence of such an effect in the environment could have dire consequences on susceptible crustacean populations.

Animals↗

[Expression and synergism of two cry insecticidal protein genes in Pseudomonas fluorescens].

Several engineered Pseudomonas fluorescens(Pf) strains were constructed mainly based on a Pseudomonas plasmid pJMS6 alpha-lac and two insecticidal crystal protein genes of Bacillus thuringiensis, cry1Ac and cry2Aa, and the host Pf strain, P303, which was with highly antifungal activity to some plant disease fungi and colonizing ability on a wide range of plants. The DNA introduction was confirmed by PCR-RFLP and Southern blot. The 132 kD insecticidal protein was detected in IPP101 and IPP202 by SDS-PAGE and rhombic insecticidal protein crystals of them were observed through electron microscope, also indicating that cry1Ac gene was highly expressed. The results of insecticidal bioassay indicated that IPP101 was more toxic than IPP201, and IPP202 was the most toxic among the 3 strains. LC50 to the neonates of cotton boll worm(Helicoverpa armigera) were 0.02604, 0.00812 and 0.00186 mL/g feed, consecutively. In IPP202, two gene products showed significant synergism, with the co-toxicity coefficient of 332.8.

Animals↗

Occupational exposure to insecticides and their effects on the auditory system.

The objective of this investigation was to study the effects of occupational exposure to organophosphates and pyrethroid insecticides on the central auditory system. The study group consisted of 98 workers exposed to insecticides and 54 non-exposed workers. Data on work history, medical history, present diseases, occupational and non-occupational exposure to noise or chemicals and lifestyle factors were obtained through an interview. Central auditory system functions were assessed through frequency patterns and duration patterns testing. Fifty-six percent of the exposed workers had hearing dysfunction at the central level and its relative risk was 7.58 for the group with exposure to insecticides (95% CI 2.9- 19.8) when compared to the non-exposed group. The group exposed to insecticides and noise had a relative risk for central disorders of 6.5 (95% CI 2.2-20.0) when compared to the non-exposed group and 9.8 (95% CI 1.4-64.5) when compared to the group exposed only to noise. The finding suggests that exposure to organophosphates and pyrethroid products can induce damage to central auditory system. Further research is needed on the ototoxic mechanisms of these chemicals, and on hearing loss prevention measurements that are applicable and adequate to such risks and populations.

Journal Article↗

[Laboratory study on the immediate and persistent insecticide activity of the pyrethroid lambda-cyhalothrin on nymphs of IV instar Triatoma infestans].

Lambda-cyhalothrin (LCT) is a new broad-spectrum synthetic pyrethroid insecticide with efficient immediate and persistent activity against a great variety of arthropods harmful both to human and animal health and to vegetal production. In many countries this insecticide has been successfully used in the control of infectious diseases vectors, such as mosquitoes, triatomine bugs (transmitters of Trypanosoma cruzi, agent of Chagas' disease) and other arthropods. In order to test the immediate and persistent activity of LCT under laboratory conditions (27 degrees C and 75-80% humidity) on laboratory reared Triatoma infestans--the most resistant instar of this insect to the commonest insecticides used--a series of six experiments was performed in 1-year period. Previous to experiments, the interior surface (400 cm2) of 36 specially manufactured cardboard boxes was uniformly sprayed with 2 ml of the following substrates: 12 with 0.6% LCT suspension, 12 with 1.0% LCT suspension and 12 with water (controls). Twenty four hours, 30 - 90 - 150 - 270 and 360 days later, in two pairs of each six boxes, lots of 10 T. infestans IV nymphs were deposited inside of them. In each of the six experiments, daily observations of the conditions of the insects (unaffected, affected in terms of various grades of progressive uncoordination, or dead) after exposure to the insecticide were performed to end with the register of these conditions 96 hours later. Moreover, after 7 and 10 days of exposure in the immediate and persistent effect of LCT respectively a 100% of mortality was verified.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cleanup with two Florisil columns for gas chromatographic determination of multiple pyrethroid insecticides in products of animal origin.

A gas chromatographic method was developed for the simultaneous determination of 9 pyrethroid insecticides in products of animal origin. The multiresidues of the pyrethroids in different samples were extracted with acetone-petroleum ether (1 + 1), and extracts were cleaned up on a Florisil partition column and a Florisil adsorption column. Four animal tissue samples were examined at 0.05-0.25 ppm fortification levels. The average recoveries of all insecticides were 76.9-88.0%, and the coefficients of variation were < 4.6% for all insecticides except permethrin. The detection limits of the method were ca 5 ppb for all insecticides but permethrin, which had a detection limit of ca 10 ppb.

Animals↗