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Purification and characterization of a carboxylesterase involved in insecticide resistance from the mosquito Culex quinquefasciatus.

A carboxylesterase (EC 3.1.1.1) involved in organophosphate insecticide resistance has been purified and characterized from the mosquito Culex quinquefasciatus. The monomeric enzyme has M(r) of 67,000 and a pI of 5.2. It hydrolysed medium-chain-length mono- and di-acylglycerols in addition to xenobiotic esters. Kinetic constants determined for four insecticides, temephos, chlorpyrifos, fenitrothion and propoxur indicate the rates of acylation and the affinities of binding of the insecticides to this carboxylesterase are important. This supports the major role of the A2 carboxylesterase is the sequestration of the insecticide with a minor role in the slow turnover of the insecticide.

Animals↗

Widespread insecticide resistance in head lice to the over-the-counter pediculocides in England, and the emergence of carbaryl resistance.

BACKGROUND: Increasing resistance to insecticides used for the control of head lice infestation has been documented over the last decade. Treatment failure and tolerance to insecticides have been validated in a number of studies undertaken in several U.K. centres. OBJECTIVES: To establish the extent of insecticide resistance in head lice and acetylcholinesterase activity in the presence of carbaryl in head lice. METHODS: Head lice were collected from school children in four centres across England (Exmouth, Loughborough, Leeds and South Shields), and tested in their response to the insecticides permethrin, phenothrin, malathion and carbaryl. Data were compared with information collected in Bristol and Bath in 1998 and with susceptible body lice. The activity of louse acetylcholinesterase was measured with and without carbaryl in head lice collected in Bristol, Leeds, Loughborough and in body lice. The efficacy of a 1% carbaryl lotion was compared in children in Bristol and Leeds. RESULTS: Compared with body lice, head lice from all six centres were significantly different in their response (P < 0.0001) to permethrin, phenothrin and malathion after 2-h exposure tests. There were significant differences in louse acetylcholinesterase activity in body lice, and head lice collected in Loughborough and Bristol in the presence or absence of carbaryl (P < 0.001), indicating enzyme inhibition. However, the difference for lice from Leeds was not significant (P = 0.363) suggesting that the enzyme was resistant to carbaryl. Eighty-nine per cent of children treated in Leeds with carbaryl were cured compared with 100% in Bristol. CONCLUSIONS: The data suggest head lice resistance is present in many parts of England to over-the-counter products containing synthetic insecticides (permethrin, phenothrin and malathion). They further suggest that resistance is starting to develop to carbaryl in head lice in Leeds and that extensive use of this product would lead to significant resistance.

Acetylcholinesterase↗

Expression and induction of three family 4 cytochrome P450 (CYP4)* genes identified from insecticide-resistant and susceptible western corn rootworms, Diabrotica virgifera virgifera.

We have previously determined that cytochrome P450-based oxidation is involved in resistance to the insecticides methyl parathion and carbaryl in geographically distinct Nebraska western corn rootworm populations. Three new family 4 cytochrome P450 (CYP4) gene fragments (CYP4AJ1, CYP4G18 and CYP4AK1) were cloned and sequenced from insecticide-resistant and -susceptible western corn rootworms. Insecticide bioassays indicated the resistant population employed in this study was significantly resistant to the insecticides methyl parathion and carbaryl. CYP4AJ1 and CYP4G18 were cloned from both genomic PCR and RT-PCR products, although only CYP4AJ1 contains an intronic region. Alignments of inferred amino acid sequences with other homologous insect CYP4 genes indicates a high degree of similarity. Northern analysis concurrently employing mixed probes representing each of the three rootworm CYP4 fragments identified increased mRNA transcript signals (i) in resistant rootworms and (ii) following induction by the P450 inducer pentamethyl benzene. These results support our previous documentation of P450-based insecticide resistance and suggest increased CYP4 transcript abundance can serve as a molecular resistance-associated marker.

Amino Acid Sequence↗

Evaluation of insecticide-treated cattle as a barrier to re-invasion of tsetse to cleared areas in northeastern Zimbabwe.

A field trial in Zimbabwe investigated the efficacy of insecticide-treated cattle as a barrier to prevent the re-invasion of tsetse, Glossina morsitans and G. pallidipes (Diptera: Glossinidae), into cleared areas. The original tsetse barrier consisted of insecticide-treated odour-baited targets, at an operational density of four to five targets per km2, supported by insecticide-treatments of cattle with either deltamethrin dip (Decatix, Coopers) at two-weekly intervals, or deltamethrin pouron (Spoton, Coopers) at monthly intervals, in a band approximately 20 km wide from the re-invasion front. Tsetse catch, and trypanosomiasis incidence in nine sentinel herds was recorded for 7-8 months, respectively, before the targets were removed, leaving only the insecticide treatment of the local cattle to stem the re-invasion of tsetse. After the removal of the target barrier, the tsetse readily invaded the trial area and the incidence of trypanosomiasis in sentinel herds increased, while their PCVs decreased. After seven months without the targets in place, trypanosomiasis prevalence in the local stock had reached alarmingly high levels; the trial was terminated prematurely and the target barrier re-deployed. Immediately after the re-deployment of the target barrier, the tsetse catch in the trial area reverted to acceptable levels along the re-invasion front, and trypanosomiasis incidence in the sentinel cattle decreased. It is concluded that, under the conditions of the field trial, the insecticidal treatment of local cattle did not in itself form an effective barrier to tsetse re-invasion. By contrast, the target barrier performed as was predicted by mathematical and experimental analysis, and readily cleared the tsetse infestation and reduced trypanosomosis incidence in the trial area.

Animals↗

Insecticide-treated plastic tarpaulins for control of malaria vectors in refugee camps.

Spraying of canvas tents with residual pyrethroid insecticide is an established method of malaria vector control in tented refugee camps. In recent years, plastic sheeting (polythene tarpaulins) has replaced canvas as the utilitarian shelter material for displaced populations in complex emergencies. Advances in technology enable polythene sheeting to be impregnated with pyrethroid during manufacture. The efficacy of such material against mosquitoes when erected as shelters under typical refugee camp conditions is unknown. Tests were undertaken with free-flying mosquitoes on entomological study platforms in an Afghan refugee camp to compare the insecticidal efficacy of plastic tarpaulin sprayed with deltamethrin on its inner surface (target dose 30 mg/m2), tarpaulin impregnated with deltamethrin (initially > or = 30 mg/m2) during manufacture, and a tent made from the factory impregnated tarpaulin material. Preliminary tests done in the laboratory with Anopheles stephensi Liston (Diptera: Culicidae) showed that 1-min exposure to factory-impregnated tarpaulins would give 100% mortality even after outdoor weathering in a temperate climate for 12 weeks. Outdoor platform tests with the erected materials (baited with human subjects) produced mosquito mortality rates between 86-100% for sprayed or factory-impregnated tarpaulins and tents (average approximately 40 anophelines and approximately 200 culicines/per platform/night), whereas control mortality (with untreated tarpaulin) was no more than 5%. Fewer than 20% of mosquitoes blood-fed on human subjects under either insecticide-treated or non-treated shelters. The tarpaulin shelter was a poor barrier to host-seeking mosquitoes and treatment with insecticide did not reduce the proportion blood-feeding. Even so, the deployment of insecticide-impregnated tarpaulins in refugee camps, if used by the majority of refugees, has the potential to control malaria by killing high proportions of mosquitoes and so reducing the average life expectancy of vectors (greatly reducing vectorial capacity), rather than by directly protecting refugees from mosquito bites. Mass coverage with deltamethrin-sprayed or impregnated tarpaulins or tents has strong potential for preventing malaria in displaced populations affected by conflict.

Afghanistan↗

Maintenance and sustained use of insecticide-treated bednets and curtains three years after a controlled trial in western Kenya.

In large experimental trials throughout Africa, insecticide-treated bednets and curtains have reduced child mortality in malaria-endemic communities by 15%-30%. While few questions remain about the efficacy of this intervention, operational issues around how to implement and sustain insecticide-treated materials (ITM) projects need attention. We revisited the site of a small-scale ITM intervention trial, 3 years after the project ended, to assess how local attitudes and practices had changed. Qualitative and quantitative methods, including 16 focus group discussions and a household survey (n = 60), were employed to assess use, maintenance, retreatment and perceptions of ITM and the insecticide in former study communities. Families that had been issued bednets were more likely to have kept and maintained them and valued bednets more highly than those who had been issued curtains. While most households retained their original bednets, none had treated them with insecticide since the intervention trial was completed 3 years earlier. Most of those who had been issued bednets repaired them, but none acquired new or replacement nets. In contrast, households that had been issued insecticide-treated curtains often removed them. Three (15%) of the households issued curtains had purchased one or more bednets since the study ended. In households where bednets had been issued, children 10 years of age and younger were a third as likely to sleep under a net as were adults (relative risk (RR) = 0. 32; 95% confidence interval (95%CI) = 0.19, 0.53). Understanding how and why optimal ITM use declined following this small-scale intervention trial can suggest measures that may improve the sustainability of current and future ITM efforts.

Bedding and Linens↗

Carbohydrate binding and resistance to proteolysis control insecticidal activity of Griffonia simplicifolia lectin II.

Griffonia simplicifolia leaf lectin II (GSII), a plant defense protein against certain insects, consists of an N-acetylglucosamine (GlcNAc)-binding large subunit with a small subunit having sequence homology to class III chitinases. Much of the insecticidal activity of GSII is attributable to the large lectin subunit, because bacterially expressed recombinant large subunit (rGSII) inhibited growth and development of the cowpea bruchid, Callosobruchus maculatus (F). Site-specific mutations were introduced into rGSII to generate proteins with altered GlcNAc binding, and the different rGSII proteins were evaluated for insecticidal activity when added to the diet of the cowpea bruchid. At pH 5.5, close to the physiological pH of the cowpea bruchid midgut lumen, rGSII recombinant proteins were categorized as having high (rGSII, rGSII-Y134F, and rGSII-N196D mutant proteins), low (rGSII-N136D), or no (rGSII-D88N, rGSII-Y134G, rGSII-Y134D, and rGSII-N136Q) GlcNAc-binding activity. Insecticidal activity of the recombinant proteins correlated with their GlcNAc-binding activity. Furthermore, insecticidal activity correlated with the resistance to proteolytic degradation by cowpea bruchid midgut extracts and with GlcNAc-specific binding to the insect digestive tract. Together, these results establish that insecticidal activity of GSII is functionally linked to carbohydrate binding, presumably to the midgut epithelium or the peritrophic matrix, and to biochemical stability of the protein to digestive proteolysis.

Binding Sites↗

Insecticide urinary metabolites in nonoccupationally exposed populations.

The wide use of insecticides in agricultural and residential settings has resulted in environmental contamination, leading to increased concern about exposure of the population and possible chronic effects on health. This review summarizes the studies that have measured urinary metabolites to assess exposure of nonoccupationally exposed population to nonpersistent insecticides, organophosphates (OPs), carbamates, and pyrethroids. Electronic search yielded 36 different studies performed in a small number of countries for the last 20 years, most of them dealing with OP urinary metabolites. Dialkylphosphates, specific metabolites of OPs, and specific metabolites of pyrethroids or carbamates, have been investigated. Results indicate that a wide range of the population, adults as well as children, is exposed to OPs and to a lesser extent to pyrethroids and carbamates. Levels are one to several orders of magnitude lower than those in occupational studies. The contribution of the different sources of insecticide exposure remains uncertain. Food contamination, as well as environmental and residential contamination, appears to influence exposure, especially in the case of children. Residential use of insecticides, having pets, and living near gardens or fields have all been inconstantly related to higher urinary metabolite levels. Occupational exposure of the parents, especially of the agricultural workers, seems to be a predictive factor of higher exposure of their children. More studies investigating every source and pathway of exposure of randomized population samples and in other countries than the United States, in particular in developing countries, could improve our knowledge of factors influencing insecticide exposure of the population.

Animals↗

Population-level effects of the neem insecticide, Neemix, on Daphnia pulex.

Although natural insecticides from the neem tree are generally perceived as less harmful to the environment than synthetic insecticides, new evidence indicates that these products may pose a risk to certain nontarget organisms. In this paper, acute and chronic effects of commercial neem insecticides on the aquatic invertebrate, Daphnia pulex were examined. The acute toxicity of two commercial neem insecticides, Neemix, Azatin and the experimental insecticide, RH-9999 to D. pulex was investigated using traditional 48 hr concentration-mortality estimates. Neemix and Azatin were equitoxic with LC50's of 0.68 and 0.57 ppm; RH-9999 was significantly less toxic with an LC50 of 13 ppm. A 10 d population growth study was conducted for Neemix and a Neemix formulation blank (Neemix devoid of the active ingredients) to determine whether the active ingredients of Neemix and/or components of the formulation were responsible for toxicity. D. pulex populations went to extinction after exposure to a Neemix concentration of 0.45 ppm azadirachtin (equivalent to the acute LC7). Neemix No Observable Effect Concentration (NOEC) and Lowest Observable Effect Concentration (LOEC) values for population growth were 0.045 and 0.15 ppm azadirachtin, respectively. The mean number of offspring per surviving female (Ro) declined in a concentration-dependent manner after exposure to Neemix with no offspring being produced after exposure to 0.45 ppm. Neemix NOEC and LOEC values for reproduction were 0.045 and 0.15 ppm, respectively. The formulation blank caused no mortality in the individuals used to start the population growth study but reduced reproduction and population growth accounting for 47% of the toxicity caused by Neemix at a concentration of 0.15 ppm. Thus, the formulation contributes substantially to the toxicity of Neemix but neem components are also toxic to D. pulex. Because the NOEC for population growth and reproduction were higher than the estimated environmental concentration of 0.035 ppm (a measure developed for forest pest mananagement), Neemix should pose little risk to populations of D. pulex.

Animals↗

Implications of gene amplification for evolution and management of insecticide resistance.

Simulations were used to compare evolution of insecticide resistance predicted by a conventional two-allele model with predictions from three- and four-allele models that assume resistance is based on gene amplification. Results were similar between models when insecticide concentration was low or moderate. In contrast, when 10% of the population was not exposed to insecticide each generation, high insecticide concentrations slowed resistance development in the two-allele model, but caused rapid development of high levels of resistance in the three- and four-allele models. The presence of a third allele at an initial frequency as low as 10-7 doubled or tripled the rate of resistance development in some cases. Attempts to slow evolution of resistance by overwhelming it with high concentrations of insecticides are not likely to succeed if gene amplification or other mechanisms generate alleles that confer high levels of resistance.

Alleles↗

Toxicity of several insecticide formulations against adult German cockroaches (Dictyoptera: Blattellidae).

Toxicity of bendiocarb, chlorpyrifos, cyfluthrin, cypermethrin, fenvalerate, hydramethylnon, malathion, propetamphos, propoxur, and pyrethrins against the adult German cockroaches, Blattella germanica (L.), was investigated. At LD50, cyfluthrin was the most toxic insecticide to adult males (0.53 microgram/g), adult females (1.2 micrograms/g), and gravid females (0.85 microgram/g). Malathion was the least toxic insecticide to adult males (464.83 micrograms/g), adult females (335.83 micrograms/g), and gravid females (275.90 micrograms/g). Males and gravid females were generally more sensitive than nongravid females to the insecticides that we tested. In tests with malathion, however, males were more tolerant. The order of toxicity of the insecticide classes varied among the stages of adult German cockroaches. The order of toxicity for males and nongravid females was pyrethroids greater than pyrethrins = organophosphates (except malathion) greater than carbamates = amidinohydrazone. The order of toxicity for gravid females was pyrethroids greater than pyrethrins = organophosphates (except malathion) greater than carbamates greater than amidinohydrazone. These differences in toxicity suggest that sex differences should be considered when determining insecticide toxicity for German cockroaches.

Animals↗

Detection of insecticide resistance in the German cockroach (Dictyoptera: Blattellidae) with glue-toxin traps.

Glue that contained an insecticide was evaluated for its ability to yield useful toxicological data for German cockroaches, Blattella germanica (L.). Toxicities of three classes of insecticides were evaluated by topical applications (LD50) and exposure to insecticide-impregnated glue (LC50). Cockroaches that were resistant to topical insecticide applications were also resistant to the glue formulation. Reliability was greatest when mortality was scored 40 to 48 h after the cockroaches were placed on the glue. This method should be adaptable for insecticide resistance monitoring of German cockroaches.

Adhesives↗

Impact of a nucleopolyhedrovirus bioinsecticide and selected synthetic insecticides on the abundance of insect natural enemies on maize in southern Mexico.

The impact of commonly used organophosphate (chlorpyrifos, methamidophos), carbamate (carbaryl), and pyrethroid (cypermethrin) insecticides on insect natural enemies was compared with that of a nucleopolyhedrovirus (Baculoviridae) of Spodoptera frugiperda (J. E. Smith) (Lepidoptera Noctuidae) in maize grown in southern Mexico. Analyses of the SELECTV and Koppert Side Effects (IOBC) databases on the impact of synthetic insecticides on arthropod natural enemies were used to predict approximately 75-90% natural enemy mortality after application, whereas the bioinsecticide was predicted to have no effect. Three field trails were performed in mid- and late-whorl stage maize planted during the growing season in Chiapas State, Mexico. Synthetic insecticides were applied at product label recommended rates using a manual knapsack sprayer fitted with a cone nozzle. The biological pesticide was applied at a rate of 3 x 10(12) occlusion bodies (OBs)/ha using identical equipment. Pesticide impacts on arthropods on maize plants were quantified at intervals between 1 and 22 d postapplication. The biological insecticide based on S. frugiperda nucleopolyhedrovirus had no adverse effect on insect natural enemies or other nontarget insect populations. Applications of the carbamate, pyrethroid, and organophosphate insecticides all resulted in reduced abundance of insect natural enemies, but for a relatively short period (8-15 d). Pesticide applications made to late-whorl stage maize resulted in lesser reductions in natural enemy populations than applications made at the mid-whorl stage, probably because of a greater abundance of physical refuges and reduced spray penetration of late-whorl maize.

Animals↗

Development and stability of insecticide resistance in the leafminer Liriomyza trifolii (Diptera: Agromyzidae) to cyromazine, abamectin, and spinosad.

Three populations of the leafminer, Liriomyza trifolii (Burgess), were collected from commercial ornamental production greenhouses in the United States and tested for susceptibility to three commercial insecticides. A leaf dip bioassay of leaves containing young (1-2-d-old) larvae was used. Based on larval mortality and compared with a susceptible laboratory reference colony, the three strains varied in spectrum and level of resistance to the insecticides. CA-1, collected from Gerbera daisy, was moderately resistant to cyromazine (18.1-fold) and abamectin (22.0-fold), but highly resistant to spinosad (> 188-fold). CA-2, collected from chrysanthemums, was not resistant to abamectin, had a low level of resistance to cyromazine (8.2-fold), but was extremely resistant to spinosad (1,192-fold). GA-1, collected from chrysanthemums, had very low levels of resistance to cyromazine (5.4-fold) and spinosad (1.9-fold) but was moderately resistant to abamectin (30.6-fold). When reared in the absence of insecticide selection pressure, all three strains reverted to approximately the level of the reference strain. The CA-1 strain reverted in nine generations to cyromazine; however, the lowest levels of abamectin and spinosad resistance reverted to was 3.1-fold at F8 and 3.2 at the F10, respectively. The CA-2 strain reverted in five generations to both cyromazine and spinosad. GA-1 reverted in five generations to abamectin. Based on the results, resistance to these three insecticides was unstable. Additionally, there was no cross-resistance among these three insecticides.

Animals↗

Effects of ground cover treatments and insecticide use on population density and damage caused by Lygus lineolaris (Heteroptera: Miridae) in apple orchards.

We conducted a 2-yr study in commercial apple orchards in Nova Scotia to assess the effects of ground cover treatments and insecticides on population density and fruit injury caused by tarnished plant bug, Lygus lineolaris (Palisot de Beauvois). The design was a split-plot with insecticides applied to whole orchard blocks and ground cover treatments applied to plots nested within orchard blocks. Ground cover treatments were 1) standard herbicide use, 2) enhanced weed control in tree rows, and 3) treatment two plus use of a selective herbicide in laneways. Treatments had few significant effects on vegetation in the tree row, but in laneways, known dicot hosts of L. lineolaris were suppressed and nonhost grasses promoted with treatment 3. Ground cover treatments did not affect cumulative captures of adult tarnished plant bugs on white sticky traps located in the plots but did affect captures in sweep nets. Split-plot ANOVA indicated no significant effect of insecticides on injury in either year, but ground cover treatments were significant in 2001. The lowest ranking rates of injury in both years were in orchards treated before bloom with a pyrethroid insecticide, either cyhalothrin-lambda or cypermethrin. The highest ranking rate of injury occurred in an orchard where insecticide was not applied until after bloom despite a high prebloom capture of L. lineolaris adults on orchard perimeter sticky traps. Fruit injury values for the ground cover treatment 3 were 63.3% (n.s.) and 50.0% (P < 0.05), respectively, of those in the standard treatment in 2000 and 2001.

Animals↗

Insecticide susceptibility in mosquitoes (Diptera: Culicidae) from French Polynesia.

Susceptibility to six organophosphate (OP), two pyrethroid (PY), and one carbamate (C) insecticides was investigated in Culex pipiens quinquefasciatus Say, Aedes aegypti (L.), and Aedes polynesiensis Marks larvae from the island of Tahiti. Cx. p. quinquefasciatus and Ae. aegypti were compared with susceptible reference strains treated simultaneously. A low, but significant, resistance to bromophos (4.6x), chlorpyrifos (5.7x), fenthion (2.4x), fenitrothion (5.0x), temephos (4.3x) and permethrin (2.1x) was found in Cx. p. quinquefasciatus, and to malathion (1.5x), temephos (2.3x), permethrin (1.8x) and propoxur (1.7x) in Ae. aegypti. Cx. p. quinquefasciatus was shown to possess over-produced esterases A2 and B2, which are known to be involved in resistance to OPs in other countries. Ae. polynesiensis was less resistant than the Ae. aegypti reference strain to all insecticides except temephos (1.8x) and permethrin (6.7x). To determine whether Ae. polynesiensis had developed resistance to these insecticides in Tahiti, a geographical survey covering 12 islands of the Society, Tuamotu, Tubuai, Marquesas, and Gambier archipelagoes was undertaken with three insecticides (temephos, deltamethrin, and permethrin). Two- to threefold variations in LC50S were observed among collections. Results are discussed in relationship to the level of insecticide exposure on the different islands.

Animals↗

Application of desiccant and insecticidal soap treatments to control Ixodes scapularis (Acari: Ixodidae) nymphs and adults in a hyperendemic woodland site.

The desiccant Drione (1% pyrethrin) and Safer's insecticidal soap (0.2% pyrethrin) formulated with and without isopropyl alcohol (ROH) were field tested against nymphal and adult populations of Ixodes scapularis Say in a hyperendemic woodlot in Westchester County, New York. Drione, insecticidal soap, and insecticidal soap with ROH provided equivalent levels of control for nymphs and adults. Compared with untreated plots, nymphal populations were significantly reduced 1 (93.3-100%) and 2 (66.4-85.7%) wk following treatment, and adult populations were significantly reduced 1 wk (53.5-62.9%) following treatment. Factors possibly contributing to the moderate and shorter period of adult control are discussed. Drione and insecticidal soap are effective alternatives to residual insecticides and could be an important component of an integrated tick management program on residential properties in areas where Lyme disease is endemic. Both products lack residual activity and would require repeated applications to maintain sufficient levels of control throughout the tick season.

1-Propanol↗

Experimental hut comparisons of nets treated with carbamate or pyrethroid insecticides, washed or unwashed, against pyrethroid-resistant mosquitoes.

The efficacy against mosquitoes (Diptera: Culicidae) of a bednet treated with carbamate insecticide [carbosulfan capsule suspension (CS) 200 mg/m(2)] was compared with four types of pyrethroid-treated nets in veranda-trap huts at Yaokoffikro near Bouaké, Côte d'Ivoire, where the malaria vector Anopheles gambiae Giles carries the kdr gene (conferring pyrethroid resistance) at high frequency and Culex quinquefasciatus Say is also pyrethroid resistant. Pyrethroids compared were lambdacyhalothrin CS 18 mg/m(2), alphacypermethrin water dispersible granules (WG) 20 mg/m(2), deltamethrin 50 mg/m(2) (Permanet) and permethrin emulsifiable concentrate (EC) 500 mg/m(2). Insecticidal power and personal protection from mosquito bites were assessed before and after the nets were used for 8 months and hand washed five times in cold soapy water. Before washing, all treatments except permethrin significantly reduced blood-feeding and all had significant insecticidal activity against An. gambiae. The carbosulfan net gave significantly higher killing of An. gambiae than all pyrethroid treatments except the Permanet. Against Culex spp., carbosulfan was more insecticidal and gave a significantly better protective effect than any of the pyrethroid treatments. After washing, treated nets retained various degrees of efficacy against both mosquito genera - but least for the carbosulfan net. Washed nets with three types of pyrethroid treatment (alphacypermethrin, lambdacyhalothrin, permethrin) gave significantly higher mortality rates of Culex than in huts with the same pyrethroid-treated nets before washing. After five washes, the Permanet, which is sold as a long-lasting insecticidal product, performed no better than the other nets in our experimental conditions.

Animals↗