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Stereoselective formation of a cholesterol ester conjugate from fenvalerate by mouse microsomal carboxyesterase(s).

In accordance with in vivo findings, of the four chiral isomers of fenvalerate (S-5602 Sumicidin, Pydrin, [RS]-alpha-cyano-3-phenoxybenzyl [RS]-2-(4-chlorophenyl)isovalerate), only the [2R, alpha S]-isomer (B-isomer) yielded cholesteryl [2R]-2-(4-chlorophenyl)isovalerate (CPIA-cholesterol ester) in the in vitro study using several tissue homogenates of mice, rats, dogs, and monkeys. There were species differences in the extent of CPIA-cholesterol-ester formation, with mouse tissues showing relatively higher activity than those of other animals. The kidney, brain, and spleen of mice showed relatively higher capacities to form this ester compared to other tissues, and the enzyme activity was mainly localized in microsomal fractions. The CPIA-cholesterol ester did not seem to be produced by three known biosynthetic pathways of endogenous cholesterol esters--acyl-CoA:cholesterol O-acyltransferase (ACAT), lecithin:cholesterol O-acyltransferase (LCAT), and cholesterol esterase. Carboxyesterase(s) of mouse kidney microsomes solubilized by digitonin hydrolyzed only the B alpha-isomer of fenvalerate, yielding CPIA, whereas they yielded the corresponding cholesterol ester in the presence of artificial liposomes containing cholesterol. Thus, it appears that the stereoselective formation of the CPIA-cholesterol ester results from the stereoselective formation of the CPIA-carboxyesterase complex only from the B alpha-isomer, which subsequently undergoes cleavage by cholesterol to yield the CPIA-cholesterol ester.

Animals↗

Effects of six months' feeding of cypermethrin on the blood and liver of albino rats.

A cypermethrin-mixed diet was fed uninterrupted to male albino rats for six months to evaluate toxicity in nontarget organisms. The rats consumed cypermethrin at a dose of 420 mg active ingredient (AI) per kilogram body weight per day. At the end of the stipulated period, the blood and liver were analyzed for insecticidal toxicity. The hemoglobin content and white blood cell (WBC) count remained unaltered, while the red blood cell (RBC) count and packed-cell volume (PCV) decreased significantly. The blood serum lactate dehydrogenase (LDH), isocitrate dehydrogenase (ICDH), and amylase activities were elevated 61%, 30%, and 46%, respectively, after six months of insecticide feeding, suggesting liver and possibly pancreas malfunction. The glutamate oxaloacetate transaminase (GOT) and creatine phosphokinase (CPK) activities, on the other hand, decreased 37% and 40%, respectively. The blood serum protein and free amino acids (FAA) content increased 12% and 31%, respectively, while cholesterol content decreased 49%. Consequent to cypermethrin administration the hepatic GOT, LDH, and ICDH activities increased 250%, 20%, and 30%, respectively. The soluble proteins, FAA, and glucose contents exhibited significant increases of 28%, 61%, and 71%, respectively. Histological changes were marked by hypertrophied hepatic cells and nuclei.

Amino Acids↗

Online trace enrichment to determine pyrethroids in river water by HPLC with column switching and photochemical induced fluorescence detection.

The potential of online trace enrichment on a highly apolar short column in LC was evaluated for the determination of pyrethroids in river water. Twelve millilitres of water samples, modified with 8 mL ACN (ACN/water 40:60, v/v), were passed through 50 x 4.6 mm ID first separation column packed with 5 microm Hypersil Elite C18. Pesticides were preconcentrated in this column while the matrix background was eluted to waste. Separation of pesticides was performed on a 3.5 microm symmetric C18 column (250 x 4.6 mm ID) with an ACN step gradient as mobile phase and fluorescence detection was used after postcolumn derivatization by using UV light. The use of photochemically induced fluorescence for detection improved sensitivity and selectivity. Quantification limits ranged from 0.05 to 0.1 microg/L and pesticide recoveries at two concentration levels (0.1 and 0.5 microg/L) were between 93.1 and 118.6%, with RSD between 2.5 and 7.5% (n = 3) in river water samples. No matrix effect was detected.

Automation↗

Method of analysis of a selected group of pyrethroids in soil samples using off-line flow-through extraction and on-column direct large-volume injection in reversed phase high performance liquid chromatography.

An analytical method using flow-through extraction of a soil sample filled in a short HPLC column with methanol or methanol-water mixtures and large-volume injection in RP-HPLC has been developed for the simultaneous determination of residues of three pyrethroids--kadethrin, cypermethrin, and permethrin--from soil samples. The developed RP-HPLC method enables separation of four diastereoisomers of cypermethrin into three peaks and resolution of two diastereoisomers of permethrin. The UV photometric detection limits of direct on-column large-volume injection of 1.00 mL of extract were 30 ng/mL of kadethrin, 37 ng/mL of total content of cypermethrin, and 65 ng/mL of trans-permethrin, which corresponds to a pyrethroid soil content of around 0.3 mg/kg. Effects of extractant flow rate and optimal extractant volume on the percentage recovery of pyrethroids from Slovak soil samples were studied. Recovery studies were performed at 0.5- 5.0 microg/g fortification level of kadethrin and 1.0-2.5 microg/g fortification level of cypermethrin and permethrin in a soil sample. Recoveries ranged from 83 to 90% for kadethrin, from 87 to 94% for total cypermethrin, and from 85 to 98% for trans-permethrin. This work comprises a basic study aimed at elaboration of an RP-HPLC method of direct analysis of pyrethroids in a soil matrix at low concentration levels achieved by a "solid sample injection" in HPLC--on-line interfacing of analyte extraction, extract clean-up, and analysis.

Chromatography, High Pressure Liquid↗

Involvement of a sodium channel mutation in pyrethroid resistance in Cydia pomonella L, and development of a diagnostic test.

Populations of the codling moth, Cydia pomonella L (Lepidoptera, Tortricidae) have developed resistance to several classes of insecticide such as benzoylureas, juvenile hormone analogues, ecdysone agonists and pyrethroids, but the corresponding resistance mechanisms have not been extensively studied. Knockdown resistance (kdr) to pyrethroid insecticides has been associated with point mutations in the para sodium channel gene in a great variety of insect pest species. We have studied two susceptible strains (S and Sv) and two resistant strains (Rt and Rv) of C pomonella that exhibited 4- and 80-fold resistance ratios to deltamethrin, respectively. The region of the voltage-dependent sodium channel gene which includes the position where kdr and super-kdr mutations have been found in Musca domestica L was amplified. The kdr mutation, a leucine-to-phenylalanine replacement at position 1014, was found only in the Rv strain. In contrast, the super-kdr mutation, a methionine-to-threonine replacement at position 918, was not detected in any C pomonella strain. These data allowed us to develop a PCR-based diagnostic test (PASA) to monitor the frequency of the kdr mutation in natural populations of C pomonella in order to define appropriate insecticide treatments in orchards.

Amino Acid Sequence↗

Impact of intraguild predation and lambda-cyhalothrin on predation efficacy of three acarophagous predators.

This laboratory study reports the interaction of three predators found in commercial apple orchards in Quebec, Hyaliodes vitripennis (Say) (Hemiptera: Miridae), Harmonia axyridis Pallas (Coleoptera: Coccinellidae) and Amblyseius fallacis (Garman) (Acarina: Phytoseiidae). First, intraguild predation between H vitripennis and the two other predators was characterized in the absence and presence of their extraguild prey, Tetranychus urticae Koch. The results showed an asymmetrical interaction in favour of the larger predator and the levels of intraguild predation were weak for the two predatory combinations. Presence of the phytophagous mite reduced the intensity of intraguild predation in the predatory combination of H axyridis and H vitripennis. Second, the effects of intraguild predation and the application of lambda-cyhalothrin on predation efficacy of the predators were evaluated. The application of the insecticide reduced prey consumption of H vitripennis and H axyridis but did not affect that of A fallacis. Combination of predators and an insecticide application resulted in two different situations depending on the species involved: a reduced predation efficacy for the combination of H vitripennis and H axyridis due to a knockdown effect caused by the insecticide, and no effect on T urticae consumption for H vitripennis and A fallacis. It is suggested that an integrated pest management program based on H vitripennis, A fallacis and lambda-cyhalothrin may be evaluated to repress phytophagous mites in Quebec orchards.

Acaridae↗

Inheritance of resistance and cross-resistance to deltamethrin in Plutella xylostella (Lepidoptera: Plutellidae) from Pakistan.

A field population of Plutella xylostella (L) from Pakistan was found to be highly resistant to deltamethrin (>500-fold) but had little or no resistance to spinosad, fipronil, indoxacarb, abamectin, Bacillus thuringiensis (Bt) var kurstaki, Bt var aizawai or Cry1Ac when compared with a susceptible laboratory population, Lab-UK. A sub-population was selected for six generations (laboratory G3-G8) with deltamethrin (delta-SEL), while a second sub-population was left unselected (UNSEL). Bioassays at G9 found that selection with deltamethrin gave resistance ratios of >230 compared with UNSEL (>6730 compared with Lab-UK). The delta-selected population showed no apparent cross-resistance to spinosad, fipronil or indoxacarb. Logit regression analysis of F1 reciprocal crosses between delta-SEL and Lab-UK indicated that resistance to deltamethrin in the delta-SEL population was inherited as an autosomal, incompletely dominant (D(LC) = 0.67) trait. A direct test of monogenic inheritance based on a back-cross of F1 progeny with delta-SEL suggested that resistance to deltamethrin was controlled by more than one factor.

Animals↗

Relationship between biochemical biomarkers and pre-copulatory behaviour and mortality in Gammarus pulex following pulse-exposure to lambda-cyhalothrin.

The impact of the pyrethroid lambda-cyhalothrin was studied in an in-stream mesocosm placed in a natural riffle of a Danish stream. Twice during summer 2002, the natural macroinvertebrate community was exposed in situ to a 30-min pulse of lambda-cyhalothrin. During exposure, nets caught macroinvertebrates in drift. Exposed and unexposed individuals of the amphipod Gammarus pulex (L) were transferred to aquaria in the laboratory and monitored for biochemical changes (ie biomarkers), pre-copulatory behaviour and mortality. Biochemical biomarkers were identified by screening extracts of exposed and unexposed G pulex using high-performance planar chromatography (HPPC). Biochemical biomarkers were detected 3 h after pulse-exposure, and one biomarker was persistent up to 7 days after exposure. Pre-copulatory behaviour (ie pair formation) was significantly impaired up to 5 days after exposure, and had not fully recovered at the end of the observation period. EC10(0.5 h) and EC50(0.5 h) values for pre-copulatory behaviour were 0.04 and 0.20 microg litre(-1), respectively. Mortality was significant at 0.35 microg litre(-1) with an LC50(0.5 h) of 5.69 microg litre(-1). There was a significant relationship between two biomarkers and mortality. The study demonstrated that pulse-exposure at expected environmental concentrations can reduce local populations of G pulex, resulting in severe impact on populations with limited possibilities of re-colonisation.

Amphipoda↗

Monooxygenase mediated pyrethroid detoxification in sea lice (Lepeophtheirus salmonis).

The role of monooxygenases in detoxification of the pyrethroids cypermethrin and deltamethrin was examined. Four strains of sea lice (Lepeophtheirus salmonis Krøyer) with normal or moderately reduced sensitivity towards the pyrethroids were tested in bioassays by exposure to the pyrethroid alone and in combination with an oxygenase inhibitor, piperonyl butoxide (PBO). The normal (baseline) sensitivity was considered as the sensitivity range for the two most sensitive strains. Pre-treatment with PBO elevated the sensitivity (P < 0.01) compared with groups exposed to the pyrethroid only. A positive, but not statistically significant, correlation between the activity of haem peroxidases and the pyrethroid concentration immobilizing 50% of the parasites was demonstrated (rho = 0.500 for deltamethrin and rho = 0.310 for cypermethrin). The results indicate that cytochrome P450 monooxygenases are involved in detoxification of pyrethroids in sea lice. 14C-Deltamethrin was absorbed in a lesser amount in a group of sea lice exposed to a mixture of the compound and PBO than in a group exposed to 14C-deltamethrin alone. A significant difference could be demonstrated both immediately after exposure (P < 0.01) and 24 h after exposure (P < 0.05). No significant differences were found between groups pre-treated with PBO and groups exposed to 14C-deltamethrin only. 14C-Deltamethrin was taken up mainly through the cuticle, especially the cuticle on the extremities of the ventral surface, and subsequently distributed throughout the body of the parasite.

Animals↗

Resistance of Thrips tabaci to pyrethroid and organophosphorus insecticides in Ontario, Canada.

Onion thrips, Thrips tabaci Lindeman, were collected from commercial onion fields in 2001, 2002 and 2003 to assess resistance to lambda-cyhalothrin, deltamethrin and diazinon. In 2001, six of eight adult populations were resistant to lambda-cyhalothrin, with resistance ratios (RR) ranging from 2 to 13.1 and four of these were also resistant to deltamethrin, with RR ranging from 19.3 to 120. Three of four adult populations were resistant to diazinon with RR ranging from 2.5 to 165.8. In 2002, four of seven nymphal populations and three of six adult populations were resistant to deltamethrin, with RR ranging from 4.3 to 72.5 and 9.4 to 839.2, respectively. Only one of six nymphal populations and one of five adult populations were resistant to diazinon, with RR of 5.6 and 2.3, respectively. In 2003 diagnostic dose bioassays, 15 of 16 onion thrips populations were resistant to lambda-cyhalothrin and all were resistant to deltamethrin. Eight of the 16 were resistant to diazinon. These results indicate that insecticide resistance is widespread in onion thrips in commercial onion fields in Ontario.

Animals↗

Effect of pretreatment with piperonyl butoxide on pyrethroid efficacy against insecticide-resistant Helicoverpa armigera (Lepidoptera: Noctuidae) and Bemisia tabaci (Sternorrhyncha: Aleyrodidae).

Pyrethroid resistance in B-type Bemisia tabaci Gennadius and Australian Helicoverpa armigera Hübner field populations is primarily conferred by esterase isoenzymes which metabolise and sequester pyrethroid insecticides. It has been shown previously that pyrethroid resistance-associated esterases in H. armigera are inhibited by the insecticide synergist piperonyl butoxide (PBO) over a 22-h period. It is demonstrated here that similar inhibition can be obtained against B-type B. tabaci. Small-scale field trials showed excellent levels of pyrethroid control when insects were pretreated with PBO and then dosed with pyrethroid during the time of maximum esterase inhibition. These results demonstrate that PBO can restore pyrethroid efficacy in the field against both B-type B. tabaci and resistant H. armigera.

Animals↗

Impact of cypermethrin on feeding behaviour and mortality of the spider Pardosa amentata in arenas with artificial 'vegetation'.

Pesticides can modify invertebrate movement and feeding behaviour which could reduce predation in agroecosystems. Previous assays have exposed the spider Pardosa amentata (Clerck) to the synthetic pyrethroid cypermethrin and monitored prey items consumed in small containers (requiring very little movement to capture prey). The current study used larger arenas containing artificial 'vegetation' (a plastic analogue) to encourage spiders to hunt and capture prey. The period 24 h after exposure produced greatest variability in prey item consumption between treatments and was used to examine treatment effects. At this time, cypermethrin reduced prey consumption rates but these effects did not persist. Findings did not suggest that the presence of artificial vegetation in arenas modified prey consumption rates, which was consistent for individuals treated with cypermethrin and a control group. This is despite the majority of pesticide-treated individuals exhibiting both ataxia and paralysis of the hind legs (these effects persisting for a maximum of 3 and 6 days respectively). These findings were consistent for both sexes. Spider longevity under starvation conditions was not significantly reduced by cypermethrin exposure but overall females survived longer than males. The findings are discussed in the context of the arenas used and the ecology of this common predator.

Animals↗

Dynamics of pesticide residues in the autumn Chinese cabbage (Brassica chinensis L.) grown in open fields.

Residues of the pesticides chlorpyrifos, dimethoate, cyhalothrin, cypermethrin, fenvalerate, deltamethrin and chlorothalonil in the autumn Chinese cabbage (Brassica chinensis L.) were studied using gas chromatography. The results indicated that the residues were dependent on the frequency and rate of pesticide application and on the weather conditions immediately following spraying. When the Chinese cabbages were treated once at normal rates, the half-lives of the seven pesticides in the vegetable were 4.7, 5.3, 2.9, 3.5, 5.4, 4.3 and 4.0 days respectively. Thus, based on the recommended preharvest intervals, the residual levels of all pesticides were within the national standards. When the Chinese cabbages were treated four times at maximal rates at 5 day intervals, the half-lives of chlorpyrifos, cypermethrin and chlorothalonil in the vegetable were 3.6, 2.9 and 5.9 days respectively, so that, based on the recommended preharvest intervals, the residual levels of chlorpyrifos and chlorothalonil exceeded the national standards for pollution-free vegetables, but the residual level of cypermethrin was within the national standard. Effects of rain on pesticide final residues in plants treated four times were more significant than in those treated once.

Agriculture↗

Biochemical analysis of a chlorfenapyr-selected resistant strain of Tetranychus urticae Koch.

Tetranychus urticae Koch has recently developed resistance to chlorfenapyr in Australia and Japan, but no attempt has yet been made to describe the biochemical mechanisms involved in chlorfenapyr resistance. In this study a laboratory-selected chlorfenapyr-resistant strain was investigated. Resistance to chlorfenapyr was associated with a strong increase in esterase activity and P450 mono-oxygenase (MO) activity but a decrease in 3,3',5,5'-tetramethylbenzidine (TMBZ) peroxidation activity. Differences in esterase activities between susceptible and resistant strains increased with increasing carbon number of the aliphatic side-chain of the nitrophenol substrate. A 4.4-fold increase in the O-deethylation of 7-ethoxy-4-trifluoromethyl coumarin (7-EFC) mediated by P450 MOs was detected. Remarkably, the resistant strain showed only half of the total TMBZ peroxidation activity found in the susceptible strain. The activity of these enzymes was further determined on different crosses and back-crosses of both strains. Results indicated that activities correlated with chlorfenapyr susceptibility and could be considered as biochemical markers. Esterase isozymes of both strains and their crosses were separated with isoelectric focusing (IEF) and visualised after activity staining. It was clear that two distinct zones of enhanced esterase activity were present in the chlorfenapyr-resistant strain (EST 11, pI = 4.88 and EST 16, pI = 4.71). EST 11 was identified with inhibitors as a carboxylesterase. The relative presence and intensity of these esterase zones changed in the different crosses and could be seen as a marker for chlorfenapyr resistance. Glutathione-S-transferase and glucose-6-phosphate dehydrogenase activities were not significantly different between strains. A twofold decrease in TMBZ peroxidase activity in the resistant strain could reflect decreased activation of chlorfenapyr. On the basis of these results the involvement of P450 MOs and esterases in the activation and detoxification of chlorfenapyr in T. urticae is challenged and discussed.

Animals↗

Persistence and efficacy of two diatomaceous earth formulations and a mixture of diatomaceous earth with natural pyrethrum against Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) on wheat and maize.

Laboratory tests were conducted to assess the insecticidal and residual effects of three diatomaceous earth (DE) formulations, Insecto, PyriSec and SilicoSec, against Tribolium confusum Jacquelin du Val on wheat and maize. Quantities of wheat and maize were treated with the above formulations at 500, 1000 and 1500 mg kg(-1) and stored at 25 degrees C and 55% relative humidity (RH). Samples were taken on the day of storage and every 30 days until completion of a 360 day period of storage. Adults of T. confusum were exposed to these samples at 25 degrees C and 55% RH and the mortality was measured after 24 and 48 h and 7 and 14 days of exposure. Adult mortality was higher on wheat than on maize. At the beginning of the storage period, mortalities after 14 day exposure on maize treated with the highest rate were 60, 63 and 81% for Insecto, PyriSec and SilicoSec respectively, while on wheat the mortality was 100% for all DEs. On the same commodities 360 days after treatment the respective mortality figures for wheat were 99, 98 and 100%, while in the case of maize they did not exceed 7%. Generally, mortality at exposures < or =48 h decreased with increasing storage time. Furthermore, although mortality on wheat increased with dose, the increase in DE dose from 1000 to 1500 mg kg(-1) resulted in only a small increase in mortality. Thus a DE treatment of 1000 mg kg(-1) was shown to provide long-term protection of wheat against T. confusum, although higher DE application rates and exposure intervals are needed for a satisfactory level of protection of maize against this pest.

Animals↗

Susceptibility of adult Coccinella septempunctata (Coleoptera: Coccinellidae) to insecticides with different modes of action.

Five insecticides (pyriproxifen, imidacloprid, deltamethrin + heptenophos, lambda-cyhalothrin and Bacillus thuringiensis Berliner subsp. tenebrionis) were examined in the laboratory for their acute detrimental side-effects at field rates on adult seven-spot ladybird beetle, Coccinella septempunctata L. The toxicity of the preparations was determined by measuring the acute surface contact effects (dried spray on leaves of Philadelphus coronarius L.), except for B. thuringiensis where mixed pollen was treated. Four to six concentrations were tested (pyriproxifen 12.5, 25, 50, 100, 200, 400 mg AI litre(-1); imidacloprid 62.4, 125, 250, 500 mg AI litre(-1); deltamethrin + heptenophos 26.4, 53.1, 106.3, 212.5 mg AI litre(-1); lambda-cyhalothrin 1.1, 3.4, 10, 30 mg AI litre(-1); B. thuringiensis 1.5, 3.0, 12.0, 48, 192, 768 mg AI litre(-1)), with 22 adults exposed per concentration. All tests were conducted in the laboratory of the Plant Protection Department (University of Debrecen, Hungary) at 22-25 degrees C, 40-60% RH, under a 16:8 h light:dark photoperiod in 1998-1999. Data were analyzed by probit analysis, probit transformation and analysis of variance. According to different categories of evaluation, pyriproxifen, imidacloprid and B. thuringiensis subsp. tenebrionis seem to be safe for C. septempunctata adults but the other two preparations were moderately harmful to them, which requires further semi-field or field tests to measure their real effect under field conditions.

Animals↗

Delayed cuticular penetration and enhanced metabolism of deltamethrin in pyrethroid-resistant strains of Helicoverpa armigera from China and Pakistan.

The penetration and metabolism of [14C]deltamethrin was studied in susceptible and resistant Chinese and Pakistani strains of Helicoverpa armigera (Hübner), which were resistant to deltamethrin by 330- and 670-fold, respectively. The penetration of deltamethrin into resistant individuals was significantly slower than into susceptible individuals over a 24-h period. The time taken for 50% penetration of the applied deltamethrin was 1 h for the susceptible strain and 6 h for both of the resistant strains. The internal radioactivity was reduced by the larvae of resistant strains much faster than by the susceptible larvae. After 48 h, 40% of the penetrated deltamethrin was still inside the larvae of the susceptible strain, in comparison with 1.5-5% in the Pakistani strain and 8-14% in the Chinese strain. Both of the resistant strains produced methanol-soluble and water-soluble metabolites, but the susceptible strain produced methanol-soluble metabolites only. By 12, 24 and 48 h after dosing, the amount of methanol-soluble metabolites excreted by the resistant strains was almost double that of the susceptible strain. Both of the resistant strains also excreted 5-7% of the penetrated dose as a water-soluble metabolite after 48 h. In comparison with the Chinese strain, the Pakistani strain exhibited slower penetration, lower internal content and faster excretion of deltamethrin, which correlated with the higher resistance of the Pakistani strain. These findings show that the resistant Pakistani and Chinese strains of H. armigera possess mechanisms of reduced cuticular penetration and enhanced metabolism of deltamethrin and perhaps other pyrethroids.

Animals↗

Resistance to cyfluthrin and tetrachlorvinphos in the lesser mealworm, Alphitobius diaperinus, collected from the eastern United States.

The lesser mealworm, Alphitobius diaperinus (Panzer), is an important pest in poultry facilities. The toxicity of cyfluthrin and tetrachlorvinphos to five strains of the lesser mealworm was compared with the toxicity to a susceptible laboratory strain. Bioassays were carried out with both larvae and adults. For the susceptible strain, cyfluthrin and tetrachlorvinphos had similar toxicity to adults, but cyfluthrin was 5 times more toxic to larvae when compared with tetrachlorvinphos. High levels of resistance to tetrachlorvinphos in two beetle strains were detected in both larvae and adults, although these strains were heterogeneous and still contained susceptible individuals. Resistance to cyfluthrin ranged from 1.7- to 9.5-fold for adults and from 0.5- to 29-fold for larvae at the LC(95). Overall, the patterns of resistance did not mirror the insecticide use patterns reported at these facilities. The implications of these results to management of the lesser mealworms are discussed.

Animals↗