Pyrethroid resistance in tropical bedbugs,Cimex hemipterus,associated with use of treated bednets

When Tanzanian villages were provided with pyrethroid-treated bednets, bedbugs (Cimicidae) disappeared; however, after about 6 years they have re-appeared in these villages. Using a newly devised test-kit, susceptibility tests of bedbugs Cimex hemipterus (Fabricius) from five of these villages showed that there is resistance to permethrin and alphacypermethrin in bedbugs from each of the villages, in contrast to those from five villages without treated nets. Circumstantial evidence indicates that bedbug resistance to pyrethroid insecticides may evolve more readily in villages with incomplete coverage rates of treated bednets, allowing bedbug infestations to become re-established. Bedbugs have not returned to a village where nearly all the beds have been provided with pyrethroid-treated bednets for 14 years.     

Anopheles gambiae gonotrophic cycle duration, biting and exiting behaviour unaffected by permethrin-impregnated bednets in The Gambia

Abstract. Permethrin-impregnated bednets protect children against malaria in The Gambia, where Anopheles gambiae complex mosquitoes are the main vectors of malaria. However, no effect has been found on mosquito density, parous rates or sporozoite rates in An.gambiae sensu lato populations; only a reduction in the numbers of mosquitoes resting indoors in rooms with treated bednets. A possible explanation for this paradox is that exposure to treated bednets leads to changed vector behaviour such as a shift in biting time, a diversion to biting outdoors instead of indoors, to biting animals instead of humans, or to increased duration of the gonotrophic cycle. To investigate these possibilities, we observed the biting and exiting behaviour of An.gambiae in ten pairs of villages, in half of which the residents used permethrin-treated bednets. The possible influence of treated bednets on the gonotrophic cycle length was evaluated by mark-release-recapture experiment. No significant difference was found between villages with treated and untreated bednets in the indoor/outdoor ratio of human biting, in mean biting times or in human blood indices of An.gambiae females found resting indoors in the mornings. The proportions of unfed, fed or gravid An.gambiae females collected in exit traps, and the number of females exiting showed no significant differences between rooms with treated and untreated bednets. Indications for a gonotrophic cycle length of 2 days were found. No evidence for any change in duration of the gonotrophic cycle in relation to exposure to treated bednets was found, although the number of recaptures was low in the villages with treated bednets. Since equal numbers of infective An.gambiae were found in villages with treated or untreated bednets, and no changes in mosquito behaviour were detected, we cannot account for how children are protected against malaria by treated bednets. One possibility is that mosquitoes divert to bite other hosts, including adults.     

Permethrin-impregnated bednets reduce nuisance arthropods in Gambian houses

The prevalence of bedbugs (Cimex hemipterus L.), chicken ticks (Argas persicus Oken) and headlice (Pediculus capitis De Geer) was surveyed in a rural area of The Gambia. At the beginning of the study 37.5% of children's beds were infested with bedbugs and 3.9% with chicken ticks, whilst the prevalence rate of pediculosis in children under 10 years old was 28.8%. Both bedbugs and headlice were clustered within compounds. Headlice prevalence increased with hair length and they were more common on girls than boys. Following this cross-sectional survey all bednets in the sixteen hamlets were either dipped in permethrin or a placebo. About 4 months later it was found that bedbugs and chicken ticks had disappeared from homes in which the bednets had been impregnated with permethrin. There was no reduction in hamlets with placebo-treated bednets. The rate of acquiring headlice between the two surveys was reduced by 91.1% in children who slept under insecticide-treated bednets compared with children with placebo-treated bednets. There were also significantly fewer day-flying and crawling insects, except earwigs, in homes of children who slept under insecticide-treated bednets compared with those with placebo-treated nets. These additional benefits of permethrin-treated bednets should contribute to their widespread acceptance and utilization by the community for personal protection.     

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.     

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.     

Impact of permethrin-treated bednets on malaria transmission by the Anopheles gambiae complex in The Gambia

Malaria vector mosquitoes belonging to the Anopheles gambiae complex were studied in four hamlets in The Gambia. All inhabitants were given bednets treated either with a placebo (milk) in two hamlets or with the pyrethroid insecticide permethrin (500 mg/m2) in two other hamlets. Malaria transmission occurred mainly during a few weeks of the rainy season, in September and October 1987. The indoor resting densities of mosquitoes in permethrin-treated hamlets were reduced, and we estimated over 90% reduction in biting on man by An. gambiae Giles sensu stricto in these hamlets. No mosquitoes were found under permethrin-treated bednets compared with eighty-one recovered from placebo-treated bednets. Mosquitoes exited more readily from rooms where permethrin-treated bednets were used than from rooms with placebo-treated nets. The annual mean probability that a child would receive an infective bite was estimated to be 0.09 in hamlets with insecticide-treated bednets, compared with 1.9 where placebo-treated bednets were used. Permethrin-treated bednets are therefore recommended as a means of effectively reducing the risk of exposure to malaria transmission, particularly in areas of low seasonal transmission.     

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.     

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.     

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.     

A field study of mating between two species of bedbug in northern KwaZulu, South Africa

The interspecific cross-mating of female Cimex lectularius Linnaeus with male Cimex hemipterus (Fabricius) results in a visible mark developing in the female ectospermalege. This mark has been used to record the frequency of mating between female C.lectularius and male C.hemipterus in sympatric wild populations in a DDT-sprayed area of northern KwaZulu, South Africa. A high percentage (11/16 = 69%) of C.lectularius females were marked, showing that these two species mate in nature.     

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.     

Dosage differential effects of permethrin impregnated into bednets on pyrethroid resistant and susceptible genotypes of the mosquito Anopheles stephensi

Abstract Effects of bednets impregnated with permethrin 200 mg and 500mg/m² on pyrethroid resistant and susceptible strains of Anopheles stephensi and their F hybrid progeny were studied, using free-flying female mosquitoes of these three genotypes, in a room with a human subject under a polyester net, having one of his arms in contact with the treated netting. Unexpectedly an apparently higher feeding rate, but lower knockdown and mortality rates, of mosquitoes were obtained for each of the three genotypes with the higher concentration of 500mg/m² compared with the lower dose of 200mg/m². At the lower dose there was 100% mortality 24 h after exposure of all three genotypes, suggesting that there would not be selection for resistance at this dose. However, at the higher dose there was significantly higher mortality of the susceptible strain than of the F hybrids, suggesting incomplete recessiveness of this resistance and that there would therefore be effective selection for resistance by this dose.
When female mosquitoes were confined in bioassay cones on treated netting, the resistant strain of An.stephensi showed significantly less irritability (scored as the time until first flight take-off) in response to each dose, as compared with the susceptible strain and F, hybrids. The higher dose provoked more irritation of each genotype; this could explain the greater knockdown and mortality rates of mosquitoes exposed to the lower dose which was less irritating and hence more effectively insecticidal. Thus a dose of 200mg/m² is preferable to 500mg/m² for malaria vector control.     

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.     

Deltamethrin-impregnated bednets reduce human landing rates of sandfly vector Lutzomyia longipalpis in Amazon households

The entomological efficacy of using 25% deltamethrin EC insecticide-treated bednets (ITNs) was evaluated against the sandfly Lutzomyia longipalpis Lutz and Neiva (Diptera: Psychodidae), the principal vector of zoonotic visceral leishmaniasis (ZVL) in Latin America. A crossover field study in Amazon Brazil (Marajó Island, Pará State) demonstrated that, compared with untreated nets, the insecticide increased the barrier effect of nets by 39% (95% confidence interval [CI] 34-44%), reduced human landing rates by 80% (95% CI 62-90%) and increased the 24-h mortality rate from 0% to 98% (95% CI 93-99%) inside ITNs. The presence of an ITN also reduced the human landing rate on unprotected persons outside the net in the same room by 56% (95% CI 52-59%), and increased 24-h mortality to 68% (95% CI 62-73%) compared to 0.4% (0.1-2.0%) in untreated houses. The reduction in human landing rates in ITN rooms was associated with a doubling in the proportion of sandflies alighting on walls compared with that in untreated rooms, which was attributed to insecticide-induced excito-repellency. There was no evidence that sandflies were diverted onto unprotected hosts. Human landing catches inside houses peaked between 19.00 hours and 23.00 hours and declined steadily to zero at 02.00 hours and thereafter. House-to-house questionnaires established that only 34% of households owned at least one net (median two, range 1-8), only 20% of the population slept under a net (33% of 0-5-year-old children), and the majority (73%) of the population slept in hammocks. Combined data pertaining to sleeping times for children and sandfly activity period indicate that > 50% of sandfly bites inside houses, and substantially more outside houses, were taken before a third of children were potentially protected by a net. This study demonstrates the clear entomological efficacy of ITNs against Lu. longipalpis in this endemic region. The effectiveness of ITNs at preventing ZVL infection and disease has still to be evaluated.     

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.     

Permethrin-impregnated bednets are more effective than DDT house-spraying to control malaria in Solomon Islands

Abstract. A field trial compared DDT house-spraying with permethrin-impregnated bednets for malaria control in Solomon Islands from 1987 to 1991. Mortality-rates of malaria vector Anopheles farauti in exit window traps were 11.6% from an untreated hut, 10.1% from a hut sprayed with DDT 2 g/m², and 98% of those from a hut in which the occupants used bednets treated with permethrin 0.5 g/m². Since bioassays of the DDT-sprayed walls (15 min exposure in W.H.O. standard test cones) gave 77% mortality of Anfarauti , it was concluded that the insignificant impact of DDT could be explained by the exophilic behaviour of endophagic vectors, whereas the greater impact of permethrin was attributed to the more effective exposure of Anfarauti females to the impregnated bednets - attracted by the occupants. The parous rate was higher indoors, except in the area with permethrin-impregnated bednets. It was therefore concluded that permethrin-impregnated bednets reduced the mean longevity of Anfarauti and hence its vectorial capacity. The circumsporozoite (CS) antigen positivity rate of Anfarauti in the DDT area was 0.18% outdoors, significantly less than 1.42% indoors. In the comparison area CS rates were 0.65% outdoors and 0.75% indoors. CS antigen was not detected in Anfarauti from the bednet area, indicating the apparent prevention of malaria transmission. As DDT spraying was so much less effective, it was discontinued in 1993 and permethrin-impregnated bednets are now the principal malaria control method in Solomon Islands.     

Shift in species composition in the Anopheles gambiae complex after implementation of long‐lasting insecticidal nets in Dielmo,Senegal

Long‐lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) are the cornerstones of malaria vector control. However, the effectiveness of these control tools depends on vector ecology and behaviour, which also largely determine the efficacy of certain Anopheles mosquitoes (Diptera: Culicidae) as vectors. Malaria vectors in sub‐Saharan Africa are primarily species of the Anopheles gambiae complex, which present intraspecific differences in behaviour that affect how they respond to vector control tools. The focus of this study is the change in species composition in the An. gambiae complex after the implementation of LLINs in Dielmo, Senegal. The main findings referred to dramatic decreases in the proportions of Anopheles coluzzii and An. gambiae after the introduction of LLINs, and an increase in the proportion of Anopheles arabiensis. Two years after LLINs were first introduced, An. arabiensis remained the most prevalent species and An. gambiae had begun to rebound. This indicated a need to develop additional vector control tools that can target the full range of malaria vectors.     

Behaviour of Anopheles albimanus in relation to pyrethroid-treated bednets

Abstract. Responses of the malaria vector Anopheles albimanus to pyrethroid impregnated bednets made of cotton or nylon, compared with untreated nets, were investigated in houses occupied by two people inside and/or outside two bednets, in coastal Chiapas, México. The pyrethroid used was lambdacyhalothrin 30mga.i./m². Bioassay mortality rates of An.albimanus exposed to treated nets for 3 or 15min, rose from 40–55% to 90–100% for nylon nets 3–19 weeks post-treatment, but were consistently lower for treated cotton nets. An.albimanus females (collected unfed on human bait) were released in houses surrounded by curtains for trapping mosquitoes that exited from the house. Floor sheets were used in and around each experimental house for retrieving any mosquitoes knocked-down and/or killed. During post-treatment assessment for 17 weeks, An.albimanus blood-feeding success rates were 23–24% with untreated nets, 14–18% with treated cotton nets and 8–15% with treated nylon nets, significantly reduced when both human baits were inside the treated bednets, but not when one or more baits were outside the treated bednet(s) within the house. Proportions of mosquitoes leaving houses <3 h post-release were 53–59% from houses with untreated bednets versus 65–78% with treated bednets. Except in one case (when both humans were outside treated cotton nets), these increased early exit rates were significant, whether or not the human baits were inside the treated bednets indoors. Mortality rates of An.albimanus females exiting overnight (22.00–06.00 hours) averaged 15–39% from houses with treated cotton and 16–46% with treated nylon nets, very significantly greater than the control mean rates of 6–8% mortality with untreated nets. Observations on wild-caught An. albimanus females marked with fluorescent powder and released indoors revealed that few mosquitoes (3–11 %) actually contacted the bednets unless both human baits remained under them - when contact rates were 22% on treated nylon, 23% on treated cotton and 42% on untreated nets (P= 0.05). The mean resting time was significantly longer on untreated (14.4min) than on treated nylon (5.8min) or cotton (9.5 min) bednets, whereas mean resting times on other surfaces indoors were 16.5–19.8min. Proportions exiting within 2h of release were significantly more from houses with treated houses (33–35%) than with untreated nets (8%). However, mortality rates of mosquitoes that landed on treated nets were very significantly greater (90–100%) than after landing on untreated nets (10%). Thus, despite some excito-repellency, lambdacyhalothrin-impregnated bednets (especially made of nylon) proved to be effective as an alternative to house-spraying against An.albimanus.     

Elevated oxidase and esterase levels associated with permethrin tolerance in Anopheles gambiae from Kenyan villages using permethrin-impregnated nets

The permethrin tolerance (PT) of a population of the mosquito Anopheles gambiae (Diptera: Culicidae) increased following the introduction of permethrin-impregnated nets for malaria control in certain villages near Kisumu, western Kenya. Using a biochemical test that indirectly measures oxidases associated with permethrin resistance, we found that this population had higher oxidase levels than a comparison population from villages without impregnated nets. Mosquitoes from a colony of An. gambiae selected for PT, the RSP (reduced susceptibility to permethrin) strain, were exposed to permethrin with or without the oxidase inhibitor piperonyl butoxide (PB). Significantly higher mortality rates occurred when permethrin was synergized by PB, presumably by suppression of oxidases responsible for PT. An unselected (UNS) colony of An. gambiae that was more susceptible than RSP in a permethrin-susceptibility bioassay (i.e. LT50 22 min for UNS, vs. 42min for RSP) was compared with the RSP colony for levels of oxidases and esterases. The levels of both enzymes were very significantly higher in the RSP strain (P<0.0001). We speculate that use of impregnated nets selected for higher oxidase and esterase levels in An. gambiae to metabolize permethrin acquired from the nets. Both oxidase and esterase mechanisms could confer cross-resistance to other pyrethroids.