Long-term use of permethrin-impregnated nets does not increase Anopheles gambiae permethrin tolerance

Abstract. Previous use of permethrin-impregnated bednets (mosquito nets) and curtains in four Kenyan villages for one year, 1990-91, raised the permethrin LT₅₀ of Anopheles gambiae to 2.4-fold above its baseline value, designated permethrin tolerance (PT), as measured by exposure to 0.25% permethrin-impregnated papers in W.H.O. test-kits. During 1992-93, with ongoing use of permethrin-impregnated nets and curtains, PT regressed slightly compared with the contemporary susceptibility level of An. gambiae from non-intervention villages, to 1.8-fold in 1992 and only 1.6-fold in 1993. Thus the selection pressure of impregnated nets for PT in Angambiae appears to be minimal in our study villages, although the impact of permethrin was demonstrated by a significantly lower parous-rate of Angambiae females in the intervention (63–66%) than in non-intervention (79%) villages, and by reduced malaria transmission (reported elsewhere).
In a selected stock of An. gambiae from the study area, PT did not affect the susceptibility to deltamethrin, fenitrothion, propoxur or DDT. Bioassays described herein provide easy procedures for field-monitoring of mosquito susceptibility/tolerance/resistance to insecticides used for net impregnation in operational programmes.     

Reduced susceptibility of Anopheles gambiae to permethrin associated with the use of permethrin-impregnated bednets and curtains in Kenya

Abstract. Susceptibility of the malaria vector Anopheles gambiae to permethrin decreased following the installation of mosquito nets impregnated with 0.5 g permethrin per square metre in four villages near Kisumu, Kenya. During the first year that permethrin-impregnated bednets and curtains were in place, the exposure time to 50% mortality (LT₅₀) increased 2.5-fold from 13 to 33min, while the LT₅₍₎ for An.gambiae was unchanged in two other villages where no intervention measures were used. Two years after permethrin-impregnated mosquito nets were distributed the LT₅₀s for An.gambiae were 28, 28 and 16min, respectively, in the villages with bednets, curtains and with no such intervention.
Using a colony of An.gambiae derived from females collected in the villages using permethrin-impregnated moquito nets, we lengthened the LT_5n from 28 to 41 min in two generations by exposing all females to permethrin-treated papers for 60 min and rearing offspring of the survivors.
Permethrin-impregnated bednets and curtains are intended to reduce vectorial capacity. Reduced susceptibility to permethrin could counter this beneficial effect.     

Village trial of bednets impregnated with wash-resistant permethrin compared with other pyrethroid formulations

Abstract. A village-scale field trial of pyrethroid-impregnated mosquito nets was undertaken in The Gambia, West Africa, in the Mandinka village of Saruja (13^o13'N, 14^o55'W) during July-November 1989. Nearly all the villagers possessed and used their own bednets. Anopheles gambiae is the main vector of human malaria in the area.
An experimental wash-resistant formulation of permethrin was compared with standard emulsifiable concentrate (EC) formulations of permethrin and lambda-cyhalothrin, versus placebo-treated bednets. Target concentrations of pyrethroids on bednets were permethrin 500mg/m² and lambda-cyhalothrin 25 mg/m². The experimental design involved random allocation of a treatment to one net per family. Whereas 68% of people questioned said they washed their nets fortnightly, observations during the 16-week trial period showed that only 4/130 (3%) of nets involved in the trial had been washed as frequently as once per month.
Early morning searches for mosquitoes under bednets (1 day/week for 16 weeks) found significantly more mosquitoes (60% An. gambiae ) in placebo-treated nets than in pyrethroid-treated nets. The numbers found with each of the three pyrethroid treatments did not differ significantly from each other. Insecticidal efficacy of the treatments was tested by bioassays using wild-caught unfed mosquitoes exposed to netting for 3min. Linear regression analysis of bioassay mortality against number of times that a net had been washed by villagers showed that nets impregnated with the wash-resistant permethrin retained their insecticidal properties better than nets impregnated with lambda-cyhalothrin or with the standard permethrin formulation.     

Efficacy of mosquito nets treated with a pyrethroid-organophosphorous mixture against Kdr- and Kdr+ malaria vectors (Anopheles gambiae)

In order to prevent the resistance of Anopheles gambiae s.l. to pyrethroids from spreading too quickly and to lengthen the effectiveness of insecticide impregnated mosquito nets, it has recently been suggested to use mixtures of insecticides that have different modes of action. This study presents the results obtained with tulle mosquito nets treated with bifenthrin (a pyrethroid) and chlorpyrifos-methyl (an organophosphorous) both separately and in mixture on two strains of An. gambiae, one sensitive to all insecticides, and the other resistant to pyrethroids. The values of KDt50 and KDt95 and the mortality induced with the mixture of bifenthrin (25 mg/m2) and chlorpyrifos-methyl (4.5 mg/m2) show a significant synergistic effect on the strain of An. gambiae susceptible to insecticides. However, the tested combination does not induce any synergistic effect on the VKPR strain selected with permethrin, but only enhances the effectiveness of the two insecticides taken separately.     

Efficacy of permethrin-impregnated curtains for malaria vector control

Preliminary results obtained by the use of permethrin-impregnated curtains against the Afrotropical malaria vectors Anopheles gambiae Giles s.l. and An.funestus Giles are reported and discussed. Field trials were carried out in villages near Ouagadougou, Burkina Faso. Houses were provided with curtains made from 100% cotton netting, impregnated with permethrin at the dose of 1 g a.i./m2, to cover the doorway, the window(s) and the space under the eaves. Entomological data collected during the period 1985-86 showed residual permethrin activity for about a year, and almost complete prevention of indoor-resting mosquitoes. Increased exit-rate and mortality-rate of house-entering malaria vectors were also obtained. Utilization of this malaria vector control method in primary health care programmes is advocated.     

Experimental hut trials of bednets impregnated with synthetic pyrethroid or organophosphate insecticide for mosquito control in The Gambia

1. Nylon bednets impregnated with different insecticides were evaluated in 1988 against wild adult mosquito populations, mostly Mansonia africana (Theobald) and Anopheles gambiae Giles sensu lato, entering experimental verandah-trap huts in The Gambia. Each bednet had six 10 x 10 cm holes made in the walls to simulate torn conditions and permit female mosquitoes to enter and feed on sleepers. 2. Individual net treatments, determined by gas chromatography of net samples from before and after 12 weeks use of the bednets, were: permethrin 670 +/- 159 and 405 +/- 190 mg/m2 (40% loss), cypermethrin 37 +/- 8 and 16 +/- 9 mg/m2 (57% loss), deltamethrin 10 +/- 7 and 10 +/- 8 mg/m2 (no loss), lambda-cyhalothrin 2.6 +/- 0.9 and 1.6 +/- 0.5 mg/m2 (38% loss), pirimiphos-methyl 4017 +/- 117 and 1160 +/- 319 mg/m2 (71% loss). 3. Washing three times in the traditional manner with local cow-fat soap reduced the initial dosages by about 85% of cypermethrin and lambda-cyhalothrin, 99.8% of pirimiphos-methyl and left no detectable residues of deltamethrin or permethrin. 4. The unwashed permethrin-treated bednet reduced the number of mosquitoes entering a hut by 60% of An.gambiae s.l. and 68% of Mansonia spp. This deterrency was less pronounced with the other insecticides and was lost by washing the bednets. 5. Each insecticide, especially lambda-cyhalothrin and pirimiphosmethyl, caused significant mortality rates of mosquitoes that entered huts with impregnated bednets, and prevented the majority of An. gambiae s.l. and Mansonia females from bloodfeeding. Washing completely removed the efficacy of deltamethrin and permethrin treated bednets, whereas nets treated with cypermethrin, lambda-cyhalothrin or pirimiphos-methyl remained significantly insecticidal after washing. 6. Aerial toxicity from the pirimiphos-methyl treated bednet killed 80% of An.gambiae s.l. confined overnight in the hut at the end of the trial, whereas the pyrethroid-treated bednets gave negligible mortality rates of mosquitoes. 7. Sleepers using the bednets had no medical symptoms significantly associated with any of the treatments. On the contrary, from 216 interviews, 4/10 complaints were associated with the use of untreated nets (P approximately 0.05), perhaps because sleepers were kept awake by mosquitoes and became more aware of any ailments. 8. It is concluded that permethrin tends mainly to deter mosquitoes from house-entry, enhancing personal protection, whereas the other insecticides kill higher proportions of the endophilic mosquitoes, which would give better community protection against malaria transmission.     

Pyrethroid resistance in Anopheles gambiae s.s. and Anopheles arabiensis in western Kenya: phenotypic,metabolic and target site characterizations of three populations

Field and laboratory investigations revealed phenotypic, target site and metabolic resistance to permethrin in an Anopheles gambiae s.s. (Diptera: Culicidae) population in Bungoma District, a region in western Kenya in which malaria is endemic and rates of ownership of insecticide‐treated bednets are high. The sensitivity of individual An. gambiae s.l. females as indicated in assays using World Health Organization (WHO) test kits demonstrated reduced mortality in response to permethrin, deltamethrin and bendiocarb. Estimated time to knock‐down of 50% (KDT₅₀) of the test population in Centers for Disease Control (CDC) bottle bioassays was significantly lengthened for the three insecticides compared with that in a susceptible control strain. Anopheles arabiensis from all three sites showed higher mortality to all three insecticides in the WHO susceptibility assays compared with the CDC bottle assays, in which they showed less sensitivity and longer KDT₅₀ than the reference strain for permethrin and deltamethrin. Microplate assays revealed elevated activity of β‐esterases and oxidases, but not glutathione‐S‐transferase, in An. gambiae s.s. survivors exposed to permethrin in bottle bioassays compared with knocked down and unexposed individuals. No An. arabiensis showed elevated enzyme activity. The 1014S kdr allele was fixed in the Bungoma An. gambiae s.s. population and absent from An. arabiensis, whereas the 1014F kdr allele was absent from all samples of both species. Insecticide resistance could compromise vector control in Bungoma and could spread to other areas as coverage with longlasting insecticide‐treated bednets increases.     

Towards a species-selective acetylcholinesterase inhibitor to control the mosquito vector of malaria, Anopheles gambiae

Anopheles gambiae is the major mosquito vector of malaria in sub-Saharan Africa. At present, insecticide-treated nets (ITNs) impregnated with pyrethroid insecticides are widely used in malaria-endemic regions to reduce infection; however the emergence of pyrethroid-resistant mosquitoes has significantly reduced the effectiveness of the pyrethroid ITNs. An acetylcholinesterase (AChE) inhibitor that is potent for An. gambiae but weakly potent for the human enzyme could potentially be safely deployed on a new class of ITNs. In this paper we provide a preliminary pharmacological characterization of An. gambiae AChE, discuss structural features of An. gambiae and human AChE that could lead to selective inhibition, and describe compounds with 130-fold selectivity for inhibition of An. gambiae AChE relative to human AChE.     

Experimental hut trials of permethrin-impregnated mosquito nets and eave curtains against malaria vectors in Tanzania

Permethrin impregnated netting was tested against Tanzanian populations of Anopheles arabiensis Patton, An.gambiae Giles and An.funestus Giles in experimental huts fitted with traps to catch samples of the mosquitoes existing during the night. Treated bednets killed some mosquitoes and increased the tendency of survivors to exit during the night. Treated cotton did not perform so well as treated nylon bednets. An impregnated bednet in which holes had been cut, to simulate a torn net, reduced the number of mosquitoes which fed and survived approximately as well as an intact untreated net. Treated curtains around the eaves of experimental huts did not perform so well as bednets but caused considerable reductions in the number of mosquitoes which fed and survived. However, there was no such effect when treated netting was placed around the eaves of a dwelling house. When one child slept under a treated net and another slept outside the net in the same hut, the number of bites on the latter child was less than if neither child had been under a net. Various aspects of the applicability of permethrin impregnated nets on a community basis are discussed.     

Effects of community-wide use of lambdacyhalothrin-impregnated bednets on malaria vectors in rural Sierra Leone

Abstract. The effect of community-wide use of bednets treated with lambdacyhalothrin 10mg/m² on the malaria vector Anopheles gambiae (forest form) was evaluated in Sierra Leone. Sixteen similar villages near the town of Bo were randomly allocated either to remain without nets or to receive treated bednets for all inhabitants, with effect from June 1992. Mosquitoes were sampled using human biting catches on verandas, light-trap catch (beside an occupied untreated bednet), window exit-trap catch and pyrethrum spray collections. During the first year of intervention (June 1992 to July 1993) the treated bednets provided personal protection for people sleeping under them, but had very little impact on densities of An. gambiae collected on human bait. The human blood index (HBI) of An.gambiae was not affected (HBI = 99% in villages with and without nets). An.gambiae parous rates were significantly reduced in all intervention villages, but malaria sporozoite rates fell in only some of the villages. These results are intermediate between those obtained from other projects in Tanzania and Burkina Faso, where treated bednets reduced man-biting, parity and sporozoite rates, versus The Gambia where treated bednets had no significant impact on any of these factors. Possible reasons for these contrasted findings are discussed.     

Olyset Net efficacy against pyrethroid-resistant Anopheles gambiae and Culex quinquefasciatus after 3 years' field use in C te d'Ivoire

Pyrethroid-impregnated bednets are advocated for personal protection against malaria vectors. To avoid the need for periodic re-treatment, it would be advantageous to have nets that retain insecticidal efficacy for years and withstand repeated washing. Such a type of commercially produced bednet with permethrin 2% incorporated in polyethylene fibres (trademark Olyset Net supplied by Sumika Life-Tech Co., Osaka, Japan) was evaluated against mosquitoes in veranda-trap huts at Yaokoffikro, near Bouaké, C te d'Ivoire, by standard WHOPES phase II procedures. Four Olyset Nets were compared with a standard untreated polyester net as control. They comprised three examples previously used in a village for over 3 years (one washed, one dirty, one very dirty) and a previously unused Olyset Net, newly unwrapped, from the same original batch. Bioassays with 3 min exposure of susceptible Anopheles gambiae Giles (Diptera: Culicidae) gave >99% mortality of female mosquitoes tested on the 'new' Olyset Net. The used Olyset Nets gave mortality rates averaging 83% for the washed net, 85% for the dirty net and 55% for the very dirty net (within 24-h following 3 min exposure). Thus, Olyset Nets were found to remain remarkably effective against susceptible An. gambiae for at least 3 years under field conditions. Wild pyrethroid-resistant populations of Culex quinquefasciatus Say and An. gambiae (savanna cytotype with 96% kdr) were assessed during June-August 1999 for their responses to sleepers protected by nets in the experimental huts. With regard to hut entry by foraging female mosquitoes, Olyset Nets showed some deterrency against An. gambiae (44% reduction by the new net, approximately 20% by the dirty nets, none by the washed net), but not against Cx. quinquefasciatus. Among mosquitoes entering the hut with untreated control net, 30-34% tried to leave (exophily) but were caught in the verandah trap. The permethrin repellency of Olyset Nets increased exophily by 19% for An. gambiae and 14% for Cx. quinquefasciatus. Blood-feeding rates were 16% An. gambiae and 35% Cx. quinquefasciatus in the hut with sleeper under the untreated net (showing considerable prevention of biting), 22-26% of both species in huts with washed or dirty used Olyset Nets (not significantly different from control), while the biting success rate of Cx. quinquefasciatus (but not kdr An. gambiae) was more than halved by the 'new' Olyset Net. Mortality rates of pyrethroid-resistant An. gambiae and Cx. quinquefasciatus from the huts were, respectively, 3% and 8% with the untreated polyester net, 27.5% and 17% with the 'new' Olyset, 15% and 17.5% with the washed Olyset, 16-25% and 17-20% with dirty old Olyset Nets. Kill differences between nets are significantly different for both An. gambiae and Cx. quinquefasciatus. Unfortunately the washed used Olyset Net showed least activity against resistant mosquitoes, despite its greatest activity against susceptible An. gambiae. In each case there was evidence that a high proportion of mosquitoes failed to feed through the net (many of them dying from starvation when they could not leave the closed hut), with indications that dirty Olyset nets enhanced this protective value.     

The influence of permethrin-impregnated bednets and mass drug administration on the incidence of Plasmodium falciparum malaria in children in Sabah, Malaysia

A small-scale trial was carried out in the Upper Kinabatangan district of Sabah, Malaysia, to determine the effect of using permethrin-impregnated bednets on malaria transmission. A total of 306 nylon bednets with cotton borders, impregnated at a dose estimated to have been 0.062 g permethrin/m2 of nylon netting, were distributed to 139 households in five villages. At the time of distributing bednets, mass drug administration with Fansidar plus primaquine was also administered to the human population to clear all parasitaemias due to Plasmodium falciparum Welch. In another village, for comparison, mass drug administration was the only intervention. After intervention measures in December 1984 and January 1985, the parasite rates in children declined in all villages during the first month, significantly more in the villages with impregnated bednets than in the control, thus proving that the nets had an impact on malaria. However, after about 2 months, parasite rates started to increase again. After 4-6 months, parasite rates in the villages with bednets approached the rate in the control village without nets. The increase in parasite rates was paralleled by a significant deterioration in the quality, physical condition and the degree of non-utilization of bednets. Entomological evaluation proved the efficacy of permethrin-impregnated nets for controlling Anopheles balabacensis Baisas and other anophelines. Bioassays (1 h exposure) of permethrin-impregnated bednets gave 100% mortality initially and 44-61% mortality after 85-106 days. Mosquito collections in treated bednets were significantly reduced for at least 217 days. The project failed to achieve prolonged suppression of malaria transmission for a combination of entomological, sociological and practical reasons which are discussed in relation to the objectives and implementation of future bednet studies.     

Insecticide resistance monitoring of field‐collected Anopheles gambiae s.l. populations from Jinja,eastern Uganda,identifies high levels of pyrethroid resistance

Insecticide resistance in the malaria vector Anopheles gambiae s.l. (Diptera: Culicidae) threatens insecticide‐based control efforts, necessitating regular monitoring. We assessed resistance in field‐collected An. gambiae s.l. from Jinja, Uganda using World Health Organization (WHO) biosassays. Only An. gambiae s.s. and An. arabiensis (?70%) were present. Female An. gambiae exhibited extremely high pyrethroid resistance (permethrin LT₅₀ > 2 h; deltamethrin LT₅₀ > 5 h). Female An. arabiensis were resistant to permethrin and exhibited reduced susceptibility to deltamethrin. However, while An. gambiae were DDT resistant, An. arabiensis were fully susceptible. Both species were fully susceptible to bendiocarb and fenitrothion. Kdr 1014S has increased rapidly in the Jinja population of An. gambiae s.s. and now approaches fixation (?95%), consistent with insecticide‐mediated selection, but is currently at a low frequency in An. arabiensis (0.07%). Kdr 1014F was also at a low frequency in An. gambiae. These frequencies preclude adequately‐powered tests for an association with phenotypic resistance. PBO synergist bioassays resulted in near complete recovery of pyrethroid susceptibility suggesting involvement of CYP450s in resistance. A small number (0.22%) of An. gambiae s.s. ×An. arabiensis hybrids were found, suggesting the possibility of introgression of resistance alleles between species. The high levels of pyrethroid resistance encountered in Jinja threaten to reduce the efficacy of vector control programmes which rely on pyrethroid‐impregnated bednets or indoor spraying of pyrethroids.     

The effect of permethrin-impregnated bednets on a population of Anopheles farauti in coastal Papua New Guinea

The effect of introducing bednets impregnated with 0.4 g/m2 permethrin on local populations of the malaria vector mosquitoes Anopheles farauti Laveran and An. koliensis Owen was monitored in a coastal village of Papua New Guinea. Whole-night landing collections were undertaken for 25 consecutive nights before and 21 nights after the introduction of the nets. Capture-recapture experiments and resting collections were also performed before and after the introduction of the nets. Following the introduction of treated nets, the biting population of An. farauti, the predominant vector, dropped from an average of 689 to 483 per man-night and the oviposition cycle became irregular, although survival rates (determined by time series analysis of the landing catches and log regression of recapture rates) were not significantly affected. The densities of An. farauti resting in and around houses and the human blood index of the engorged females also decreased significantly after introduction of the treated bednets. The population of An. koliensis dropped prior to the introduction of the nets. However, the number of nulliparous females in the landing catches remained more or less constant which implies that, in this species, survival rates were affected by the nets but that recruitment to the population was not.     

The kdr pyrethroid resistance gene in Anopheles gambiae: tests of non-pyrethroid insecticides and a new detection method for the gene

The organophosphate pirimiphos-methyl and the carbamate carbosulfan were evaluated in comparison to the pyrethroid alphacypermethrin and the 'near-pyrethroid' etofenprox against pyrethroid resistant Anopheles gambiae and Culex spp. in an experimental hut station located in central C?te d'Ivoire. Bednets were impregnated with the above mentioned compounds and randomly allocated to the huts. On 40 consecutive mornings, after sleepers had occupied the huts overnight, mosquitoes were collected from the huts, identified and scored as live or dead (including delayed mortality). An. gambiae s.l. that had been collected were tested for the presence of the kdr allele in heterozygous or homozygous form. Both non-pyrethroid treatments caused very high mortality, whereas mortality with alpha-cypermethrin and etofenprox generally did not differ from the levels observed with untreated control nets in this experiment. The nets had holes cut in them and there was considerable bloodfeeding on the sleepers, which was only significantly reduced for An. gambiae by carbosulfan and alpha-cypermethrin. PCR genotyping suggested that there was selection for the kdr resistance allele by the pyrethroid treated nets. Organophosphates and carbamates may therefore present an alternative to be used on bednets especially in areas of pyrethroid resistance, but the safety of these insecticides will have to be carefully considered.     

Pyrethroid-treated bednet effects on mosquitoes of the Anopheles gambiae complex in The Gambia

The response of Anopheles gambiae complex mosquitoes to men sleeping under insecticide-impregnated or untreated bednets in six verandah trap huts was studied during the dry season in The Gambia. With this type of hut it was possible to collect live and dead indoor-resting mosquitoes and estimate the number of wild mosquitoes which entered, bloodfed on man, and exited each night. Bednets were treated with emulsions targetted to leave deposits of 25 mg/m2 lambda-cyhalothrin, or 5, 50 or 500 mg/m2 permethrin, diluted from emulsifiable concentrates (EC), or a blank formulation similar to the EC except that the permethrin was omitted; the sixth net was left untreated. Nets and sleepers were rotated between huts on different nights, the design being based on a series of Latin squares and conducted double-blind. Permethrin-impregnated bednets deterred mosquitoes from entering the huts. The degree of deterrency was proportional to the dosage of permethrin. This effect was also caused by the blank formulation and therefore attributed to other components of the formulation, rather than to the permethrin itself. The net impregnated with 500 mg permethrin per square metre gave the best individual protection, reducing mosquito bloodfeeding by 91% compared with untreated nets. However, lambda-cyhalothrin was proportionately more insecticidal than permethrin at doses of equivalent deterrency. At this stage of research, it remains conjectural whether chemical deterrency or killing of malaria vectors is better for community protection.     

Comparison of different insecticides and fabrics for anti-mosquito bednets and curtains

Abstract. Various formulations of six insecticides (a carbamate and five pyrethroids), were impregnated into bednets and curtains made from cotton, polyester, polyethylene or polypropylene fabric. For bioassays of insecticidal efficacy, female Anopheles gambiae mosquitoes were made to walk on the fabrics for 3 min and mortality was scored after 24 h. The main concentrations tested were: bendiocarb 400 mg/m², cyfluthrin 30–50 mg/m², deltamethrin 15–25 mg/m², etofenprox 200 mg/m², lambda-cyhalothrin 5–15 mg/m² and permethrin 200–500 mg/m². Field trials in Tanzania used experimental huts (fitted with verandah traps) entered by wild free-flying Anopheles gambiae, An. funestus and Culex quinquefasciatus mosquitoes. Results of testing the impregnated fabrics in experimental huts showed better personal protection provided by bednets than by curtains. Permethrin cis:trans isomer ratios 25:75 and 40:60 were equally effective, and the permethrin rate of 200 mg/m² performed as well as 500 mg/m². Bioassay data emphasized the prolonged insecticidal efficacy of lambda-cyhalothrin deposits, except on polyethylene netting. Most of the impregnated nets (including the ‘Olyset’ net with permethrin incorporated during manufacture of the polyethylene fibre) and an untreated intact net performed well in preventing both Anopheles and Culex mosquitoes from feeding on people using them overnight in the experimental huts. Anopheles showed high mortality rates in response to pyrethroid-treated nets, but only bendiocarb treated curtains killed many Culex. Holed nets treated with either cyfluthrin (5 EW formulation applied at the rate of 50 mg a.i./m²) or lambda-cyhalothrin (2.5 CS formulation at 10 mg a.i./m²) performed well after 15 months of domestic use. Treatment with deltamethrin SC or lambda-cyhalothrin CS at the very low rate of 3 mg/m² gave good results, including after washing and re-treatment.     

Evaluation of horn flies (Diptera: Muscidae) from a pyrethroid susceptible colony for general and permethrin esterase activities

In this study we describe a nonradioactive single-fly microassay for permethrin hydrolysis. We used this assay with a microplate assay for general esterase activity to evaluate the permethrin hydrolyzing and general esterase activities of aging pyrethroid-susceptible male and female horn flies, Haematobia irritans (L.). We found substantial gender- and age-related differences regarding general esterase activity, permethrin sensitivity, and permethrin hydrolyzing activity within the colony. Extracts of female flies collected 48 h after receiving their first blood meal yielded significantly greater esterase activity than male extracts. Aging female flies were more tolerant of permethrin than were male flies. In addition, a positive correlation was found to exist between the general esterase activity of aging females and their ability to hydrolyze permethrin.     

The impact of permethrin-impregnated bednets on malaria vectors of the Kenyan coast

Abstract. The effects of introducing permethrin-impregnated bednets on local populations of the malaria vector mosquitoes Anopheles funestus and the An.gambiae complex was monitored during a randomized controlled trial at Kilifi on the Kenyan coast. Pyrethrum spray collections inside 762 households were conducted between May 1994 and April 1995 after the introduction of bednets in half of the study area. All-night human bait collections were performed in two zones (one control and one intervention) for two nights each month during the same period. PCR identifications of An.gambiae sensu lato showed that proportions of sibling species were An.gambiae sensu stricto > An.merus > An.arabiensis.
Indoor-resting densities of An.gambiae s.l. and the proportion of engorged females decreased significantly in intervention zones as compared to control zones. However, the human blood index and Plasmodium falciparum sporozoite rate remained unaffected. Also vector parous rates were unaltered by the intervention, implying that survival rates of malaria vectors were not affected. The human-biting density of An.gambiae s.l. , the predominant vector, was consistently higher in the intervention zone compared to the control zone, but showed 8% reduction compared to pre-intervention biting rates - versus 94% increase in the control zone.
Bioassay, susceptibility and high-performance liquid chromatography results all indicated that the permethrin content applied to the nets was sufficient to maintain high mortality of susceptible vectors throughout the trial. Increased rates of early outdoor-biting, as opposed to indoor-biting later during the night, were behavioural or vector composition changes associated with this intervention, which would require further monitoring during control programmes employing insecticide-treated bednets.     

Insecticide resistance in the Anopheles gambiae complex in Benin: a nationwide survey

Benin has embraced World Health Organization-recommended preventive strategies to control malaria. Its National Malaria Control Programme is implementing and/or coordinating various actions and conducting evaluation trials of mosquito control strategies. Mosquito control is based on the use of insecticide-treated nets and indoor residual spraying, but the efficacy of these strategies to control malaria vectors is endangered by insecticide resistance. Here, we present the results of a nationwide survey on the status of insecticide susceptibility and resistance in Anopheles gambiae s.l. (Diptera: Culicidae) carried out in Benin in 2006-2007 (i.e. before extensive vector control was undertaken). Overall, our study showed that the S molecular form of An. gambiae s.s. predominates and is widely distributed across the country, whereas the frequency of the M form shows a strong decline with increasing latitude. Susceptibility to DDT, permethrin, carbosulfan and chlorpyrifos-methyl was assessed; individual mosquitoes were identified for species and molecular forms, and genotyped for the kdr and ace-1 loci. Full susceptibility to chlorpyrifos-methyl was recorded and very few samples displayed resistance to carbosulfan. High resistance levels to permethrin were detected in most samples and almost all samples displayed resistance to DDT. The kdr-Leu-Phe mutation was present in all localities and in both molecular forms of An. gambiae s.s. Furthermore, the ace-1(R) mutation was predominant in the S form, but absent from the M form. By contrast, no target modification was observed in Anopheles arabiensis. Resistance in the An. gambiae S molecular form in this study seemed to be associated with agricultural practices. Our study showed important geographic variations which must be taken into account in the vector control strategies that will be applied in different regions of Benin. It also emphasizes the need to regularly monitor insecticide resistance across the country and to adapt measures to manage resistance.