Evolutionary plasticity of monooxygenase-mediated resistance

The cytochrome P450 monooxygenases are an important metabolic system involved in the detoxification of xenobiotics, and are thus one of the major mechanisms by which insects evolve insecticide resistance. However, comparatively little is known about the evolutionary constraints of this insecticide resistance mechanism. We investigated the genetic basis of resistance in a strain of house fly (NG98) from Georgia, USA that had evolved 3700-fold resistance to the pyrethroid insecticide permethrin, and compared this to other permethrin resistant strains of house flies from the US and Japan. Resistance in NG98 was due to kdr on autosome 3 and monooxygenase-mediated resistance on autosomes 1, 2, and 5. These results indicate that the genes which evolve to produce monooxygenase-mediated resistance to permethrin are different between different populations, and that the P450 monooxygenases have some degree of plasticity in response to selection. Monooxygenase-mediated resistance appears to evolve using different P450s, and possibly different regulatory signals controlling P450 expression, even in strains selected with the same insecticide.     

Genetic and biochemical studies of resistance to permethrin in a pyrethroid-resistant strain of the housefly (Musca domestica L.)

One or more weak factors of resistance on autosome 2, and barely detectable resistance on autosome 3, confer moderate resistance to several pyrethroids (5–13-fold) in the field-collected Ipswich strain of houseflies. In these flies, which unlike other pyrethroid-resistant strains lack kdr or super-kdr, pyrethroid resistance probably developed in response to prolonged treatment of buildings for animals with pyrethrins synergised with piperonyl butoxide. Substrains, isolated genetically from Ipswich flies and with resistance only on autosome 2, degraded permethrin more rapidly than susceptible flies and produced larger amounts of very polar metabolites. In this, they differed from flies with kdr or super-kdr which resembled susceptible flies in their metabolism of permethrin. NIA 16388 (propyl prop-2-ynyl phenylphosphonate) was a better synergist and reduced the metabolism of permethrin more than piperonyl butoxide in both the susceptible and resistant insects. The slight increase in synergism and minimal decrease in metabolism when piperonyl butoxide was applied with NIA 16388 indicated that the latter also inhibited detoxication that was sensitive to piperonyl butoxide.     

Cross-Resistance to Imidacloprid in Strains of German Cockroach (Blattella germanica) and House Fly (Musca domestica)

The toxicity of a promising new insecticide, imidacloprid, was evaluated against several susceptible and resistant strains of German cockroach and house fly. Imidacloprid rapidly immobilized German cockroaches followed by a period of about 72 h during which some cockroaches recovered. After 72 h there was no further recovery. Imidacloprid-treated houseflies were immobilized more slowly than treated cockroaches, with the maximum effect observed after 72 h, and there was no recovery. Based upon 72-h LD₅₀ values imidacloprid was moderately toxic to German cockroaches (LD₅₀ values were 6–8 ng mg^-1) and had only low toxicity to house flies (LD₅₀ 140 ng mg^-1). Piperonyl butoxide (PBO) blocked the observed recovery in German cockroaches. PBO also greatly enhanced the 72-h LD₅₀ of imidacloprid from 43- to 59-fold in cockroaches and 86-fold in house flies. Two strains of German cockroach (Baygon-R and Pyr-R) showed >4-fold cross-resistance to imidacloprid. This cross-resistance could not be suppressed by PBO, suggesting that P450 monooxygenase-mediated detoxication is not responsible for this cross-resistance. Variation in the level of synergism observed with PBO (between strains) suggests the ‘basal’ level of monooxygenase-mediated detoxication of imidacloprid is quite variable between strains of German cockroach. The AVER and LPR strains of house fly showed significant cross-resistance to imidacloprid. PBO reduced the level of cross-resistance in AVER from >4·2-fold to 0·5-fold (i.e. the AVER strain LD₅₀ was half that of the susceptible strain when both were treated with PBO), but PBO did not suppress the cross-resistance in LPR. These data suggest monooxygenases are the mechanism responsible for cross-resistance to imidacloprid in AVER, but not in the LPR strain. © of SCI.     

Indoxacarb resistance in the house fly, Musca domestica

Indoxacarb (DPX-MP062) is a recently introduced oxadiazine insecticide with activity against a wide range of pests, including house flies. It is metabolically decarbomethoxylated to DCJW. Selection of field collected house flies with indoxacarb produced a New York indoxacarb-resistant (NYINDR) strain with >118-fold resistance after three generations. Resistance in NYINDR could be partially overcome with the P450 inhibitor piperonyl butoxide (PBO), but the synergists diethyl maleate and S,S,S-tributyl phosphorothioate did not alter expression of the resistance, suggesting P450 monooxygenases, but not esterases or glutathione S-transferases are involved in the indoxacarb resistance. Conversely, the NYINDR strain showed only 3.2-fold resistance to DCJW, and this resistance could be suppressed with PBO. Only limited levels of cross-resistance were detected to pyrethroid, organophosphate, carbamate or chlorinated hydrocarbon insecticides in NYINDR. Indoxacarb resistance in the NYINDR strain was inherited primarily as a completely recessive trait. Analysis of the phenotypes vs. mortality data revealed that the major factor for indoxacarb resistance is located on autosome 4 with a minor factor on autosome 3. It appears these genes have not previously been associated with insecticide resistance.     

Various mechanisms responsible for permethrin metabolic resistance in seven field-collected strains of the German cockroach from Iran, Blattella germanica (L.) (Dictyoptera: Blattellidae)

Insecticide resistance in the German cockroach can be mediated by a number of mechanisms, the most common being enhanced enzymatic metabolism. Seven field-collected strains of German cockroach, Blattella germanica (L.) with various levels of resistance to pyrethroids, five out of which were also cross-resistant to DDT were used in this study. The investigation of possible mechanisms responsible for permethrin resistance was carried out using the synergists PBO, DEF and DMC and biochemical assays, including general esterases, glutathione S-transeferases and monooxygenases assays, using an automated microtitre plate reader. PBO and DEF, the inhibitors of cytochrome p450 monooxygenases and general esterases, respectively, affected permethrin resistance to varying degrees depending on the strain. DDT resistance in five strains were not completely eliminated by the synergist DMC, an inhibitor of glutathione S-transferase enzymes, suggesting that a further non-metabolic resistance mechanism such as kdr-type may be present. This suggestion was further supported by GST assay data, where a little elevation in GST activity was detected in only two strains. The synergist data supported by biochemical assays implicated that cytochrome p450 monooxygenases or hydrolases are involved in permethrin resistance in some strains. However, these results implicated both enhanced oxidative and hydrolytic metabolism of permethrin as resistance mechanism in the other strains. The results of synergist and biochemical studies implicated that all the field-collected permethrin resistant strains have developed diverse mechanisms of resistance, although these strains have been collected from the same geographic area. The change in resistance ratios of some strains by using PBO or DEF is discussed. It is of interest to note that because resistance to permethrin was not completely eliminated by DEF and PBO, it is likely that one or more additional mechanisms are involved in permethrin resistance in every strain studied.     

Pyrethroid resistance and cross‐resistance in the German cockroach,Blattella germanica (L)

A German cockroach (Blatella germanica (L)) strain, Apyr‐R, was collected from Opelika, Alabama after control failures with pyrethroid insecticides. Levels of resistance to permethrin and deltamethrin in Apyr‐R (97‐ and 480‐fold, respectively, compared with a susceptible strain, ACY) were partially or mostly suppressed by piperonyl butoxide (PBO) and S,S,S,‐tributylphosphorotrithioate (DEF), suggesting that P450 monooxygenases and hydrolases are involved in resistance to these two pyrethroids in Apyr‐R. However, incomplete suppression of pyrethroid resistance with PBO and DEF implies that one or more additional mechanisms are involved in resistance. Injection, compared with topical application, resulted in 43‐ and 48‐fold increases in toxicity of permethrin in ACY and Apyr‐R, respectively. Similarly, injection increased the toxicity of deltamethrin 27‐fold in ACY and 28‐fold in Apyr‐R. These data indicate that cuticular penetration is one of the obstacles for the effectiveness of pyrethroids against German cockroaches. However, injection did not change the levels of resistance to either permethrin or deltamethrin, suggesting that a decrease in the rate of cuticular penetration may not play an important role in pyrethroid resistance in Apyr‐R. Apyr‐R showed cross‐resistance to imidacloprid, with a resistance ratio of 10. PBO treatment resulted in no significant change in the toxicity of imidacloprid, implying that P450 monooxygenase‐mediated detoxication is not the mechanism responsible for cross‐resistance. Apyr‐R showed no cross‐resistance to spinosad, although spinosad had relatively low toxicity to German cockroaches compared with other insecticides tested in this study. This result further confirmed that the mode of action of spinosad to insects is unique. Fipronil, a relatively new insecticide, was highly toxic to German cockroaches, and the multi‐resistance mechanisms in Apyr‐R did not confer significant cross‐resistance to this compound. Thus, we propose that fipronil could be a valuable tool in integrated resistance management of German cockroaches. © 2001 Society of Chemical Industry     

Involvement of esterases in diazinon resistance and biphasic effects of piperonyl butoxide on diazinon toxicity to Haematobia irritans irritans (Diptera: Muscidae)

Resistance to insecticides remains a major problem for the successful control of the horn fly, Haematobia irritans irritans (L.), one of the most important pests of cattle in many countries including the United States. The organophosphate (OP) insecticide diazinon has been used to control pyrethroid-resistant populations of the horn fly. There are only a few reported cases of horn fly resistance to diazinon in the United States and Mexico. Piperonyl butoxide (PBO) has been used successfully as a synergist of pyrethroid insecticides to control horn flies. PBO-synergized diazinon products are also available for horn fly control in the United States, although PBO is known to inhibit the bio-activation of certain OP insecticides including diazinon. A study was conducted to evaluate the effect of PBO on diazinon toxicity to horn flies using a filter paper bioassay technique. These bioassays in both the susceptible and diazinon-resistant horn fly strains revealed a biphasic effect of PBO on diazinon toxicity to horn flies. PBO inhibited diazinon toxicity when the PBO concentration used was high (5%), and no effect was observed when PBO concentration was intermediate (2%). However, at low concentrations (1% and lower), PBO significantly synergized diazinon toxicity. We demonstrated that enhanced esterase activity was associated with survivability of horn flies exposed to diazinon alone. PBO has been shown to inhibit esterase activity in other insect species. However, results of biochemical assays with esterases from this study suggest that PBO did not have significant effect on the overall esterase activity in the horn fly. The observed synergistic effect of PBO at lower concentrations on diazinon toxicity to horn flies could not be explained by reduced esterase activity due to PBO inhibition. It is likely that PBO synergized diazinon toxicity at lower concentrations by facilitating penetration of diazinon through the cuticle and/or inhibiting the oxidative detoxification of diazinon, and reduced diazinon toxicity at high PBO concentration by inhibiting the bio-activation of diazinon.     

Resistance in the mosquito, Culex quinquefasciatus, and possible mechanisms for resistance

Two mosquito strains of Culex quinquefasciatus (Say), MAmCq(G0) and HAmCq(G0), were collected from Mobile and Huntsville, Alabama, respectively. MAmCq(G0) and HAmCq(G0) were further selected in the laboratory with permethrin for one and three generations, respectively. The levels of resistance to permethrin in MAmCq(G1) (after one-generation selection) and HAmCq(G3) (after three-generation selection) increased rapidly. Resistance to permethrin in MAmCq(G1) and HAmCq(G3) was partially suppressed by piperonyl butoxide (PBO), S,S,S-tributylphosphorotrithioate (DEF) and diethyl maleate (DEM), inhibitors of cytochrome P450 monooxygenases, hydrolases and glutathione S-transferases (GST), respectively, suggesting these three enzyme families are important in conferring permethrin resistance in both strains. A substitution of leucine to phenylalanine (L to F) resulting from a single nucleotide polymorphism (SNP), termed the kdr mutation, in the para-homologous sodium channel gene has been reported as a very common mutation associated with pyrethroid resistance of insects. A 341-bp sodium channel gene fragment, where the kdr mutation resides, was generated by PCR from genomic DNAs of Cx. quinquefasciatus strains. We found that the kdr mutation was present in both permethrin-selected and unselected HAmCq and MAmCq mosquito populations, suggesting that the kdr mutation plays the role in permethrin resistance. There was no significant change in the frequency and heterozygosity of the A to T SNP for the kdr allele between permethrin-selected and unselected MAmCq and HAmCq mosquitoes, indicating that other mechanisms are involved in the evolution of resistance in mosquitoes selected by permethrin in the laboratory.     

Selection of Anopheles stephensi with DDT and dieldrin and cross-resistance spectrum to pyrethroids and fipronil

The field strain of Anopheles stephensi, the main malaria vector in south of Iran, was colonized in laboratory and selected with DDT and dieldrin in two separate lines for 3 generations to a level of 19.5- and 14-fold for DDT and dieldrin resistance, respectively. Synergist tests with chlorofenethol (DMC) and piperonyl butoxide (PBO) on the selected strains indicated that dehydrochlorination and oxidative detoxification might be the underlying mechanisms involved in the resistance to dieldrin and DDT in selected strains. DDT selection decreased susceptibility to DDT and pyrethroids including lambdacyhalothrin, permethrin deltamethrin and cyfluthrin. The result also showed that selection with dieldrin caused negative and positive cross-resistance to pyrethroid and fipronil, respectively. Based on these results, it can be concluded that besides metabolic resistance mechanisms, other factors such as mutation in γ aminobutyric acid (GABA) and voltage-gated sodium channels (Kdr) might be involved.     

Molecular analysis of resistance in a deltamethrin-resistant strain of Musca domestica from China

We investigated the molecular basis of resistance in a strain of house fly (BJD) from Beijing, China. This strain showed 567-fold resistance to commonly used deltamethrin. Flies were 64-fold resistant to deltamethrin synergized by piperonyl butoxide (PBO). The 5′-flanking sequence of the cytochrome P450 gene CYP6D1 in BJD strain had a 15-bp insert as in the LPR strain. Two mutations (kdr, super-kdr) in the voltage sensitive sodium channel (VSSC) were also detected in the BJD strain. Our results showed that a combination of resistance alleles for CYP6D1 and VSSC existed in deltamethrin resistant house flies in China.     

Assessing insecticide resistance and aversion to methomyl-treated toxic baits in Musca domestica L (Diptera: Muscidae) populations in southern California

Progeny of house flies (Musca domestica L) from ten California poultry operations, three dairies and one horse-riding facility were tested for methomyl- and muscalure-treated bait resistance using up to three different assays: a topical assay, a no-choice feeding assay and a choice feeding assay. LD50 resistance factors from the topical assay, compared with a locally-derived susceptible colony, ranged between 1 and 4. LC50 resistance factors from the no-choice feeding assay ranged mostly between 2 and 5, with one value of 18. Measurable LT50 resistance ratios for female flies in the choice feeding assay ranged from 43 to 159; two populations had <10% mortality at 48h and could not be measured. LT50 resistance ratios for male flies in the choice feeding assay ranged from 26 to 96, and one population was too resistant to measure. A behavioral assay tested the feeding preference of male and female flies provided a dish of sugar and a dish of methomyl- and muscalure-treated bait. Of eight strains tested, females from seven strains and males from six strains showed significant preference for sugar over bait. Behavioral factors appear to be important in the severe resistance of house flies to baits in California.     

Pyrethroid tolerance in Culex pipiens pipiens var molestus from Marin County, California

In May 2001 a sample of Culex pipiens pipiens variety molestus Forskål from Marin County, California, collected as larvae and reared to adults, was found to show reduced resmethrin and permethrin knock‐down responses in bottle bioassays relative to a standard susceptible Cx pipiens quinquefasciatus Say colony (CQ1). Larval susceptibility tests, using CQ1 as standard susceptible, indicated that the Marin mosquitoes had LC₅₀ resistance ratios of 18.3 for permethrin, 12 for deltamethrin and 3.3 for pyrethrum. A colony of Marin was established and rapidly developed higher levels of resistance in a few generations after exposure to permethrin as larvae. These selected larvae were shown to cross‐resist to lambda‐cyhalothrin as well as to DDT. However, adult knock‐down time in the presence of permethrin, resmethrin and pyrethrum was not increased after increase in tolerance to pyrethroids as larvae. Partial and almost complete reversion to susceptibility as larvae was achieved with S, S, S‐tributylphosphorotrithioate and piperonyl butoxide (PBO), respectively, suggesting the presence of carboxylesterase and P450 monooxygenase mediated resistance. Insensitive target site resistance (kdr) was also detected in some Marin mosquitoes by use of an existing PCR‐based diagnostic assay designed for Cx p pipiens L mosquitoes. Carboxylesterase mediated resistance was supported by use of newly synthesized novel pyrethroid‐selective substrates in activity assays. Bottle bioassays gave underestimates of the levels of tolerance to pyrethroids of Marin mosquitoes when compared with mortality rates in field trials using registered pyrethroid adulticides with and without PBO. This study represents the first report of resistance to pyrethroids in a feral population of a mosquito species in the USA. Copyright © 2003 Society of Chemical Industry     

Linkage of genes for sodium channel and cytochrome P450 (CYP6B10) in Heliothis virescens

Genetic linkage of hscp (heliothis sodium channel protein) and CYP6B10 was discovered in Heliothis virescens. The hscp gene encodes the sodium channel target of pyrethroid insecticides and cytochrome P450 genes encode important enzymes involved in detoxication of various pesticides. Previously, two mechanisms, nerve insensitivity due to sodium channel and synergism by propynyl aryl ethers, were observed in pyrethroid-resistant H virescens and were not separated by repeated back-crossing. We hypothesized genetic linkage of target site insensitivity and monooxygenase-mediated detoxication. Single nucleotide polymorphisms were discovered in IIS6 of hscp; Hpy of hscp and CYP6B10. Segregation of these and other markers was tested in backcrosses. We observed cosegregation of hscp to CYP6B10, but both genes assorted independently of y, ye and sex. Genes y and ye assorted independently of each other. This was the first observation of linkage between genes controlling detoxication and sodium ion channel insensitivity in a species known to express high levels of pyrethroid resistance. Linkage was not likely because this species has 31 chromosomes; therefore, we will investigate the possibility of a resistance cassette. We expect similar linkage in other noctuid pests.     

The effect of a piperonyl butoxide/tau-fluvalinate mixture on pollen beetle (Meligethes aeneus) and honey bees (Apis mellifera)

BACKGROUND: Previous work has characterised pyrethroid resistance in pollen beetle (Meligethes aeneus F.) as principally an oxidative mechanism. Piperonyl butoxide (PBO) can synergise this resistance in the field, but its effects on the honey bee are thought to be unacceptable. RESULTS: A field trial in Poland was conducted to show that a mixture of PBO and tau‐fluvalinate at the registered rate gave increased and longer‐lasting control of resistant pollen beetle. Four days after spraying with tau‐fluvalinate, only 20% of pollen beetles were controlled, compared with 70% if the tau‐fluvalinate/PBO mixture was used. No detriment to honey bee health was observed using the same mixture. CONCLUSIONS: PBO, if used in conjunction with a pyrethroid of relatively low bee toxicity, can successfully overcome pyrethroid resistance in pollen beetle without incurring an increased loss of honey bees, even if they are present at the time of spraying. Copyright © 2012 Society of Chemical Industry     

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

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

Monitoring insecticide resistance in house flies (Diptera: Muscidae) from New York dairies

House flies were collected from dairies across New York state and the levels of resistance to seven insecticides were measured using standard laboratory assays with three to five diagnostic concentrations. The highest levels of resistance were found for tetrachlorvinphos, permethrin and cyfluthrin. Although levels of resistance to methomyl were somewhat lower, they were among the highest ever reported for field‐collected house flies. Resistance to pyrethrins was limited primarily to the lowest diagnostic concentration. House flies were susceptible to fipronil at all dairies, suggesting that this material would be highly effective for fly control. The levels of resistance were similar at all the dairies, irrespective of their insecticide use, suggesting substantial movement of flies between facilities. Relative to resistance levels found at New York dairies in 1987, resistance levels had increased for permethrin, were unchanged for tetrachlorvinphos and had decreased for dimethoate. To identify a single diagnostic concentration that could be used in the laboratory assays to assess accurately resistance levels in future studies, we carried out a ‘simulated’ field bioassay using formulated materials. A diagnostic concentration for each insecticide is proposed on the basis of a comparison of our bioassays. © 2001 Society of Chemical Industry     

Evaluation of field resistance in field-collected mosquito Culex quinquefasciatus Say through quantification of ULV permethrin/PBO formulation in field bioassays

BACKGROUND

Pyrethroids are among the most applied adulticides worldwide to control mosquito vectors for prevention of arboviral diseases transmission. However, pesticide resistance development in a mosquito population could lead to decreased control efficacy. While most studies investigate the resistant genotype (i.e. kdr, CYP450, etc.) as explanatory variables, few field efficacy studies have measured pesticide quantities deposited at different distances from the sprayer in association with observed mosquito mortality. The current study determined field delivered amounts of an applied ULV permethrin/PBO formulation (31% permethrin + 66% piperonyl butoxide) by GC/MS and estimated practical resistance ratios using caged mosquito females.

RESULTS

For field samples, the extraction method recovered 78 ± 3.92–108 ± 8.97% of the permethrin/PBO formulation when utilizing the peaks of PBO from GC/MS to estimate the concentrations of adulticide deposited near the mosquito cages. The field bioassay showed that the spatial distribution of permethrin/PBO formulation was heterogeneous among three pseudo-replicates within the same distance. Within the quantifiable permethrin/PBO range of 15.7–51.4 ng/cm², field-collected mosquito mortalities started at 64% and linearly increased reaching 100% only in two areas, while all Sebring susceptible mosquitoes died. The field LC₉₅ resistance ratio (RR) of F₀ Cx. quinquefasciatus ranged from 2.65–3.51, falling within the 95% CI of RR₉₅ estimated by laboratory vial assays. Tests with and without PBO indicated P450's enzymes contributed to field resistance.

CONCLUSION

Results showed the suitability of the collection and quantification method to estimate the field resistance ratio at the applied pesticide rate. Pesticide quantification would also allow the association of the known frequencies of resistance mechanisms (e.g. kdr, CYP450) with field mortalities to estimate the resistance level conferred by such mechanisms. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Susceptibility of house flies (Diptera: Muscidae) exposed to commercial insecticides on painted and unpainted plywood panels

House flies were collected from dairies in New York state and the levels of resistance to commercially available insecticide formulations were measured on painted and unpainted plywood panels. Dimethoate was ineffective on all surfaces. The wettable powder permethrin formulation was more toxic than the emulsifiable concentrate formulation. The wettable powder cyfluthrin formulation was also more toxic than the recently developed liquid formulation. In general, the best house fly control was obtained on flat latex painted plywood panels and the poorest control on gloss latex painted panels. It is highly unlikely that producers obtain adequate control with dimethoate and permethrin.     

Characterization of the structure-activity relationship of kdr and two variants of super-kdr to pyrethroids in the housefly (Musca domestica L.)

Structure-activity relationships (SARs) for 10 pyrethroids against susceptible, kdr and super-kdr strains of houseflies (Musca domestica L.) were investigated by Principal Components Analysis. In the three strains with kdr_Latina' all only slightly to moderately (2.6 to 26-fold) resistant to pyrethroids, no correlation between the structure and Levels of resistance could be discerned. In flies with super-kdr, SARs were influenced by the nature of the alcoholic portion of the ester. Resistance was strongest to esters of a-cyano-3-phenoxybenzyl alcohol (74 to 430-fold) and to permethrin (48 to 55-fold). It was weak (6.2 to 11-fold) to cyclopentenone derivatives, being barely stronger than for flies with kdr (2-6 to 6.3-fold). Two variants of super-kdr (3D and A2) were distinguished on the basis of their differential response to esters of 5-benzyl-3-furylmethanol. It is presumed that kdr_Latina, super-kdr_A2 and super-kdr_3D form an allelic series in which kdr_Latina represents ground level insensitivity, and the two super-kdrs the progressive extension of strong resistance to more types of ester. The strong differences in resistance to different pyrethroid esters by super-kdr flies provides scope for improving management of resistance to pyrethroid insecticides and for modifying the SAR of pyrethroids to favour weak resistance.     

Laboratory studies on the mechanisms of resistance to permethrin in a field-selected strain of house flies

Enhanced oxidative metabolism appeared to be a major factor involved in resistance to permethrin in a field strain of house flies, selected with permethrin over 4 years. This was shown in the 7.8-fold synergism by piperonyl butoxide which reduced the resistance ratio from 97 to 15. The rate of permethrin detoxication was significantly higher (P=0.05) in the resistant flies compared with a susceptible strain or resistant flies pretreated with piperonyl butoxide. The esterase inhibitor S,S,S-tributyl phosphorotrithioate did not reduce the level of resistance to permethrin in the resistant strain, although some hydrolytic metabolism was apparent. Rates of penetration were similar in susceptible and resistant flies and in resistant flies pre-treated with piperonyl butoxide. A minor unidentified resistance factor, possibly reduced sensitivity of the nervous system, may also have been present in the resistant strain.