Esterase inhibition by synergists in the western flower thrips Frankliniella occidentalis

BACKGROUND: Western flower thrips (WFT), Frankliniella occidentalis (Pergande), is among the most important crop pests in the south‐eastern region of Spain. Its increasing resistance to insecticides constitutes a serious problem, and understanding the mechanisms involved is therefore of great interest. Use of synergists to inhibit the enzymes involved in insecticide detoxification is widely used to determine their responsibility for insecticide resistance. However, they do not always act as intended or expected, and caution must be exercised when interpreting synergist results. RESULTS: Laboratory‐selected strains of WFT were used to analyse the effects of the synergists piperonyl butoxide (PBO), S,S,S‐tributyl phosphorotrithioate (DEF) and methiocarb on total esterase activity. Significant differences were found, indicating esterase activity inhibition by DEF, a lower effect for methiocarb and a small inhibition of the activity by PBO. Esterase isoenzyme inhibition by these compounds showed a similar result; this assay revealed an extreme sensitivity of Triplet A (resistance‐associated esterases) to DEF. In an in vivo assay carried out with these compounds at different incubation times, only DEF caused posterior in vitro esterase activity inhibition, with a maximum effect 1 h after treatment. CONCLUSION: In this work, only DEF shows true synergistic inhibition of WFT esterases. Copyright © 2011 Society of Chemical Industry     

Synergism studies with binary mixtures of pyrethroid, carbamate and organophosphate insecticides on Frankliniella occidentalis (Pergande)

The major mechanism of resistance to most insecticides in Frankliniella occidentalis (Pergande) is metabolic, piperonyl butoxide (PBO) suppressible, mediated by cytochrome-P450 monooxygenases and conferring cross-resistance among insecticide classes. The efficacy of insecticide mixtures of acrinathrin, methiocarb, formetanate and chlorpyrifos was studied by topical exposure in strains of F. occidentalis selected for resistance to each insecticide. The method consisted in combining increasing concentrations of one insecticide with a constant low rate of the second one as synergist. Acrinathrin activity against F. occidentalis was enhanced by carbamate insecticides, methiocarb being a much better synergist than formetanate. Monooxygenase action on the carbamates would prevent degradation of the pyrethroid, hence providing a level of synergism by competitive substrate inhibition. However, the number of insecticides registered for control of F. occidentalis is very limited, and they are needed for antiresistance strategies such as mosaics and rotations. Therefore, a study was made of the synergist effect of other carbamates not used against thrips, such as carbofuran and carbosulfan, against a susceptible strain and a field strain. Neither carbamate showed synergism to acrinathrin in the susceptible strain, but both did in the field strain, carbosulfan being a better synergist than carbofuran. The data obtained indicate that low rates of carbamates could be used as synergists to restore some pyrethroid susceptibility in F. occidentalis.     

High level methoprene resistance in the mosquito Ochlerotatus nigromaculis (Ludlow) in central California

In the summer of 1998, failures of methoprene field applications to control the mosquito Ochlerotatus nigromaculis (Ludlow) were noticed in several pastures in the outskirts of Fresno, California, USA. Effective control with methoprene had been achieved for over 20 years prior to this discovery. Susceptibility tests indicated that the Fresno Oc nigromaculis populations had developed several thousand‐fold higher LC₅₀ and LC₉₀ tolerance levels to methoprene compared with methoprene‐naïve populations. The synergists piperonyl butoxide (PBO), S,S,S‐tributyl phosphorotrithioate and 3‐octylthio‐1,1,1‐trifluoro‐2‐propanone had little synergistic effect, suggesting that the mechanism of methoprene tolerance was not mediated by P450 monooxygenase or carboxylesterase enzyme degradation. As part of initiating a resistance management strategy, partial reversion back to methoprene susceptibility was achieved in a resistant population after six consecutive applications of Bacillus thuringiensis israelensis Goldberg & Marga coupled with two oil and two pyrethrum + PBO applications. © 2002 Society of Chemical Industry     

西花蓟马抗甲氨基阿维菌素苯甲酸盐种群的交互抗性与生化抗性机制

为了解西花蓟马Frankliniella occidentalis(Pergande)对甲氨基阿维菌素苯甲酸盐的抗性风险,采用生物和生化测定方法研究了西花蓟马甲维盐抗性种群与其它杀虫剂的交互抗性和生化抗性机制。西花蓟马甲维盐抗性种群对阿维菌素有高水平交互抗性,抗性倍数为31.656,对啶虫脒有中等水平交互抗性,为12.182,对吡虫啉、溴虫腈、氯氟氰菊酯、毒死蜱和灭多威有低水平交互抗性,为5.517~8.568,而对多杀菌素无明显交互抗性。增效剂胡椒基丁醚(PBO)、马来酸二乙酯(DEM)、三丁基三硫磷酸酯(DEF)和磷酸三苯酯(TPP)对甲维盐抗性种群和田间种群均有显著增效作用。甲维盐抗性种群多功能氧化酶细胞色素P₄₅₀和b₅含量、O-脱甲基酶、谷胱甘肽S-转移酶和羧酸酯酶活性均显著提高,分别为敏感种群的3.89、3.61、5.32、4.42和1.30倍,表明多功能氧化酶、谷胱甘肽S-转移酶和羧酸酯酶等解毒代谢酶活性的提高是西花蓟马对甲维盐产生抗性的重要机制。     

Pyrethroid synergists suppress esterase-mediated resistance in Indian strains of the cotton bollworm, Helicoverpa armigera (Hübner)

The role of esterase in pyrethroid resistance was studied in the final larval instar of different strains of the cotton bollworm, Helicoverpa armigera. The resistant strains viz., Nagpur strain and the Delhi strain were found to have elevated midgut esterase activity in comparison to the susceptible strain. Nagpur strain and Delhi strain have 2.24 and 1.73-fold higher esterase activity, respectively, than that of the susceptible strain. The Native PAGE displayed important differences in the midgut esterase isozyme pattern between the susceptible and the pyrethroid-resistant strains. Out of the 10 esterase isozyme observed, susceptible strain lacked three bands, E2, E6 and E10 that were found in the resistant strains. The potency of the synergists piperonyl butoxide (PBO) and dihydrodillapiole (DDA) as esterase inhibitor were also studied both in vitro and in vivo. The in vitro results clearly show that both PBO and DDA inhibited esterase activity in the two resistant strains, while there was almost no esterase inhibition in the homogenate of the susceptible strain. The in vivo inhibition studies (topical application of PBO and DDA followed by biochemical analysis) illustrated that PBO- and DDA-esterase binding is rather slow and non permanent process. Esterase inhibition did not occur immediately after the synergist treatment but at 4 and 8 h post treatment in case of PBO and DDA, respectively. Native PAGE revealed that the in vivo esterase inhibition caused by both PBO and DDA was due to the binding of the synergist with the E6 isozyme which was not present in the susceptible strain.     

Esterase inhibitors as synergists for (+)-trans-chrysanthemate insecticide chemicals

Four synergists are used to evaluate the relative contribution of esterases and oxidases in the metabolism of four pyrethroids, the (+)-trans- and (+)-cis-isomers of resmethrin and tetramethrin, by five insect species and by mice. Three of these compounds are known pyrethroid synergists, S,S,S-tributyl phosphorotrithioate acting as an esterase inhibitor and piperonyl butoxide and O-(2-methylpropyl) O-(2-propynyl) phenylphosphonate acting as oxidase inhibitors. The fourth synergist, 1-naphthyl N-propylcarbamate, is an esterase inhibitor selected by screening 65 candidate esterase and oxidase inhibitors for maximal potency in synergizing the toxicity of trans-resmethrin to milkweed bugs. Naphthyl propylcarbamate synergizes the toxicity of trans-resmethrin and -tetramethrin to milkweed bugs, cockroaches, houseflies, cabbage loopers, and mealworms but not to mice. The persistence of trans-resmethrin in milkweed bugs treated by injection is increased by the esterase inhibitors while that of cis-resmethrin is increased by the oxidase inhibitors. The optimal synergist varies with the species and the pyrethoid, being related to both the nature of the pyrethroid alcohol moiety and the trans- or cis-configuration of the acid moiety. This probably results from species variations in the relative significance of esterases and oxidases in pyrethroid detoxification.     

三种增效剂对甜菜夜蛾防治的增效作用

甜菜夜蛾 Spodoptera exigua( Hübner)近年来在我国很多地区爆发成灾 ,发生区域南至广东、台湾、福建 ,北到东北地区 ,而且其与棉铃虫 ( H elicoverpa armigera Hübner)一样 ,都是杂食性害虫 ,寄主植物十分广泛。甜菜夜蛾爆发的原因很多 [1 ] ,其中对常规化学农药的高抗性是重要原因之一 [2 ]。利用增效剂增强农药对抗性害虫的防治效果是害虫抗性治理的主要措施之一 ,如增效醚 ( PBO)、MGK2 6 4被推荐与拟除虫菊酯类杀虫剂混合使用防治对有机磷、拟除虫菊酯类农药有抗性的田间小菜蛾 [3] 。本试验利用采自田间的甜菜夜蛾种群 ,测定了 …     

Temporal synergism can enhance carbamate and neonicotinoid insecticidal activity against resistant crop pests

BACKGROUND: Piperonyl butoxide (PBO) effectively synergises synthetic pyrethroids, rendering even very resistant insect pests susceptible, provided a temporal element is included between exposure to synergist and insecticide. This concept is now applied to carbamates and neonicotinoids. RESULTS: A microencapsulated formulation of PBO and pirimicarb reduced the resistance factor in a clone of Myzus persicae (Sulzer) from >19 000- to 100-fold and in Aphis gossypii (Glover) from >48 000- to 30-fold. Similar results were obtained for a strain of Bemisia tabaci Gennadius resistant to imidacloprid and acetamiprid, although a second resistant strain did not exhibit such a dramatic reduction, presumably owing to the presence of target-site insensitivity and the absence of metabolic resistance. Synergism was also observed in laboratory susceptible insects, suggesting that, even when detoxification is not enhanced, there is degradation of insecticides by the background enzymes. Use of an analogue of PBO, which inhibits esterases but has reduced potency against microsomal oxidases, suggests that acetamiprid resistance in whiteflies is largely oxidase based. CONCLUSION: Temporal synergism can effectively enhance the activity of carbamates and neonicotinoids against resistant insect pests. Although the extent of this enhancement is dependent upon the resistance mechanisms present, inhibition of background enzymes can confer increased sensitivity against target-site resistance as well as increased metabolism. .     

Synergism of insecticides provides evidence of metabolic mechanisms of resistance in the obliquebanded leafroller Choristoneura rosaceana (Lepidoptera: Tortricidae)

The interactions between six insecticides (indoxacarb, cypermethrin, chlorpyrifos, azinphosmethyl, tebufenozide and chlorfenapyr) and three potential synergists, (piperonyl butoxide (PBO), S,S,S-tributyl phosphorotrithioate (DEF) and diethyl maleate (DEM)) were studied by dietary exposure in a multi-resistant and a susceptible strain of the obliquebanded leafroller, Choristoneura rosaceana (Harris). The synergists did not produce appreciable synergism with most of the insecticides in the susceptible strain. Except for tebufenozide, PBO synergized all the insecticides to varying degrees in the resistant strain. A very high level of synergism by PBO was found with indoxacarb, which reduced the resistance level from 705- to 20-fold when PBO was administered alone and to around 10-fold when used in combination with DEF. DEF also synergized indoxacarb, cypermethrin, chlorpyrifos, azinphosmethyl and tebufenozide in the resistant strain. DEM produced synergism of indoxacarb, chlorpyrifos, azinphos-methyl and chlorfenapyr in the resistant strain. DEM was highly synergistic to cypermethrin, and to some extent to tebufenozide in both the susceptible and resistant strains equally, implying that detoxification by glutathione S-transferases was not a mechanism of resistance for these insecticides. The high level of synergism seen with DEM in the case of cypermethrin may be due to an increase in oxidative stress resulting from the removal of the antioxidant, glutathione. These studies indicate that enhanced detoxification, often mediated by cytochrome P-450 monooxygenases, but with probable esterase and glutathione S-transferase contributions in some cases, is the major mechanism imparting resistance to different insecticides in C. rosaceana.     

Toxicological and biochemical characterization of coumaphos resistance in the San Roman strain of Boophilus microplus (Acari: Ixodidae)

The San Roman strain of the southern cattle tick, Boophilus microplus, collected from Mexico was previously reported to have a high level of resistance to the organophosphate acaricide coumaphos. An oxidative detoxification mechanism was suspected to contribute to coumaphos resistance in this tick strain, as coumaphos bioassay with piperonyl butoxide (PBO) on larvae of this resistant strain resulted in enhanced coumaphos toxicity, while coumaphos assays with PBO resulted in reduced toxicity of coumaphos in a susceptible reference strain. In this study, we further analyzed the mechanism of oxidative metabolic detoxification with synergist bioassays of coroxon, the toxic metabolite of coumaphos, and the mechanism of target-site insensitivity with acetylcholinesterase (AChE) inhibition kinetics assays. Bioassays of coroxon with PBO resulted in synergism of coroxon toxicity in both the San Roman and the susceptible reference strains. The synergism ratio of PBO on coroxon in the resistant strain was 4.5 times that of the susceptible strain. The results suggested that the cytP450-based metabolic detoxification existed in both resistant and susceptible strains, but its activity was significantly enhanced in the resistant strain. Comparisons of AChE activity and inhibition kinetics by coroxon in both susceptible and resistant strains revealed that the resistant San Roman strain had an insensitive AChE, with a reduced phosphorylation rate, resulting in a reduced bimolecular reaction constant. These data indicate a mechanism of coumaphos resistance in the San Roman strain that involves both insensitive AChE and enhanced cytP450-based metabolic detoxification.     

Metabolic mechanisms of insecticide resistance in the western flower thrips, Frankliniella occidentalis (Pergande)

The interactions between six insecticides (methiocarb, formetanate, acrinathrin, deltamethrin, methamidophos and endosulfan) and three potential synergists (piperonyl butoxide (PBO), S,S,S-tributyl phosphorotrithioate (DEF) and diethyl maleate (DEM)) were studied by topical exposure in strains selected for resistance to each insecticide, and in a susceptible strain of Frankliniella occidentalis (Pergande). In the susceptible strain PBO produced appreciable synergism only of formetanate, methiocarb and methamidophos. Except for endosulfan, PBO synergized all the insecticides to varying degrees in the resistant strains. A very high level of synergism by PBO was found with acrinathrin, which reduced the resistance level from 3344- to 36-fold. PBO slightly synergized the carbamates formetanate (4.6-fold) and methiocarb (3.3-fold). PBO also produced a high synergism of deltamethrin (12.5-fold) and methamidophos (14.3-fold) and completely restored susceptibility to both insecticides. DEF did not produce synergism with any insecticide in the resistant strains and DEM was slightly synergistic to endosulfan (3-fold). These studies indicate that an enhanced detoxification, mediated by cytochrome P-450 monooxygenases, is the major mechanism imparting resistance to different insecticides in F occidentalis. Implications of different mechanisms in insecticide resistance in F occidentalis are discussed.     

The effect of piperonyl butoxide on pyrethroid-resistance-associated esterases in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Pyrethroid resistance in field populations of Australian Helicoverpa armigera (Hübner) is primarily a consequence of the overproduction of esterase isoenzymes which metabolise and sequester pyrethroid insecticides. Biochemical studies have shown that pyrethroid-resistance-associated esterases in H armigera are inhibited by the insecticide synergist piperonyl butoxide (PBO). Esterase inhibition by PBO did not occur immediately after dosing, but exhibited maximum inhibition 3-4 h after dosage. Esterase activity subsequently recovered until full activity was restored by 24 h. Topical bioassays using a pre-treatment of PBO showed that maximum H armigera mortality was achieved with pre-treatment times corresponding to maximum esterase inhibition. These results demonstrated that, with correct temporal application, PBO can restore pyrethroid efficacy against H armigera. It would also be expected that restoration of efficacy with other conventional insecticides, currently compromised by esterase-based resistance mechanisms, would occur.     

三种增效剂对棉铃虫增效作用

用３个不同地区的棉铃虫田间种群分别测定了３种增效剂对３种杀虫剂的增效作用。结果表明ＰＢ对氯氟氰菊酯,ＤＥＦ对辛硫磷,ＰＢ和ＴＰＰ对灭多威增效较好,而且棉铃虫的抗性水平越高,增效剂的增效作用越突出。     

Selection for resistance, reversion towards susceptibility and synergism of chlorantraniliprole and spinetoram in obliquebanded leafroller, Choristoneura rosaceana (Lepidoptera: Tortricidae)

BACKGROUND: Obliquebanded leafroller, Choristoneura rosaceana (Harris), is an economic pest of pome fruits throughout North America. Repeated applications of broad‐spectrum insecticides have resulted in the development of resistance, cross‐resistance and multiple resistance in this pest. Studies were conducted to determine the possibility of resistance evolution, stability of resistance and the effect of metabolic synergists on toxicity of new reduced‐risk insecticides chlorantraniliprole and spinetoram in C. rosaceana. RESULTS: Larvae of C. rosaceana were selected for resistance to chlorantraniliprole and spinetoram in the laboratory. Significant levels of resistance to each insecticide were observed after 12 generations of selection. In the absence of selection pressure, susceptibility of a subset of larvae from both chlorantraniliprole‐ and spinetoram‐selected populations reverted to preselection levels after five and six generations respectively, indicating that resistance to both chlorantraniliprole and spinetoram was unstable in C. rosaceana. In synergist bioassays performed after 12 generations of selection, S,S,S‐tributylphosphoro trithioate (DEF) and piperonyl butoxide (PBO) synergized the toxicity of chlorantraniliprole and spinetoram respectively, suggesting the involvement of esterases in chlorantraniliprole resistance and the involvement of mixed‐function oxidases in spinetoram resistance. CONCLUSION: These findings suggest that chlorantraniliprole and spinetoram could be incorporated into C. rosaceana resistance management programs by using rotational strategies. Copyright © 2011 Society of Chemical Industry     

The pattern of cross-resistance to insecticides and juvenile hormone analogues in a diflubenzuron-resistant strain of the cotton leaf worm Spodoptera littoralis boisd

The rate of development of resistance to diflubenzuron in a laboratory susceptible strain of the cotton leafworm Spodoptera littoralis Boisd., the pattern of cross-resistance exhibited by the resistant strain to several insecticides and juvenile hormone analogues, as well as the synergistic action of piperonyl butoxide (PB) and S, S, S-tributyl phosphorotrithioate (TBP) with insecticides or diflubenzuron on both strains, were investigated. Resistance to diflubenzuron increased slightly in the first eight selected generations and was enhanced by further selection until in generation 30, the selected strain attained the high level of resistance of 290.7-fold, compared with the parent strain. The resistant strain when challenged with either insecticides or juvenile hormone analogues at selected generations (5, 10, 15, 20, 25 and 30) exhibited different levels of resistance to several insecticides representing organochlorine and organo-phosphorus compounds, carbamates and pyrethroids, but a clear case of negative correlation was indicated between resistance to diflubenzuron and juvenile hormone analogues. With regard to the synergistic action of PB and TBP on the toxicity of either diflubenzuron or insecticides against the fourth-instar larvae of the susceptible strain, methomyl showed slight levels of synergism when it was combined with them. With the exception of cypermethrin, which was not affected by the two synergists, lower levels of synergism were observed with the compounds endrin, diflubenzuron and fenvalerate when they were combined with the same synergists. These two synergists however, antagonised the toxic action of the organophosphorus compounds phosfolan and chlorpyrifos. Against the resistant strain, endrin was moderately synergised by TBP but only slightly by PB. Slight levels of synergism were observed when methomyl, phosfolan and diflubenzuron were combined with either synergist, but both antagonised chlorpyrifos and fenvalerate. Resistance to diflubenzuron and to the other tested chemicals in the resistant strain was scarcely affected by the two synergists.     

An analogue of piperonyl butoxide facilitates the characterisation of metabolic resistance

BACKGROUND: Previous work has demonstrated that piperonyl butoxide (PBO) not only inhibits microsomal oxidases but also resistance‐associated esterases. The ability to inhibit both major metabolic resistance enzymes makes it an ideal synergist to enhance xenobiotics but negates the ability to differentiate which enzyme group is responsible for conferring resistance. RESULTS: This study examines an analogue that retains the ability to inhibit esterases but is restricted in its ability to act on microsomal oxidases, thus allowing an informed decision on resistance enzymes to be made when used in conjunction with the parent molecule. CONCLUSION: Using examples of resistant insects with well‐characterised resistance mechanisms, a combination of PBO and analogue allows identification of the metabolic mechanism responsible for conferring resistance. The relative potency of PBO as both an esterase inhibitor and an oxidase inhibitor is also discussed. Copyright © 2008 Society of Chemical Industry     

Synergist induced susceptibility of tobacco caterpillar, <Emphasis Type="Italic">Spodoptera litura</Emphasis> (Fabricius) from Kerala,India exposed to conventional insecticides

The present study was undertaken to assess the insecticide resistance developed in various field collected population of S. litura and to induce susceptibility by using the synergists. Third-instar larvae collected from three different locations of Kerala viz., Thiruvananthapuram (TVM), Pathanamthitta (PTA) and Alappuzha (ALP) were exposed to conventional insecticides like chlorpyriphos, quinalphos, lambda-cyhalothrin and cypermethrin by leaf dip bioassay and resistance ratios were calculated by using the baseline data generated for respective insecticides using susceptible strain. Resistance ratios recorded were 1965, 840 and 320 against chlorpyriphos, 605, 255 and 59 against quinalphos, 926, 250 and 108 against lambda-cyahlothrin and 2566, 534 and 396 against cypermethrin respectively for TVM, PTA and ALP populations. The effect of selected synergists viz., piperonyl butoxide (PBO), diethyl maleate (DEM) and triphenyl phosphate (TPP) was studied in combination with respective test insecticides against the highly resistant population of S.litura collected from TVM of Kerala. The population was tested with insecticide in combination of the above synergists at different ratios. When PBO, TPP and DEM at ratio of 1:4 were used the synergistic ratio was 8.47, 7.26 and 3.98 for chlorpyriphos, 6.09, 5.26 and 3.05 for quinalphos, 13.37, 4.53 and 7.39 for lambda cyhalothrin and 4.77, 3.36 and 3.40 for cypermethrin respectively. PBO showed highest synergistic activity against both the organophosphates tested followed by DEM and TPP. Highest synergistic activity against synthetic pyrethroids also was shown by PBO, followed by TPP and DEM. The results obtained from the present study revealed that PBO at 1:4 ratio showed higher synergism with the test insecticides against the resistant populations of S.litura and proved to be an effective molecule alternate for breaking the resistance against conventional organophosphates and synthetic pyrethroids.     

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.     

Assessing the performance against houseflies of indoor aerosol space sprays. Part I. Factors examined during tests with houseflies in free flight

The development of a mathematical model which, by quantifying the interaction of a population of houseflies with an ageing spray cloud, is capable of predicting the knockdown and kill performance of insecticidal aerosol formulations shown against houseflies, Musca domestica L., will be presented in a series of four separate papers. In this first paper, measurements were made of insecticide collected by houseflies during free flight using a standard room-spray technique. With pressure-packed aerosol formulations where the solvent is the principal change, it was shown that both knockdown and mortality responses of the insects were directly related to dose collected. The process of droplet collection was most significantly influenced by their size and availability, dependent upon the time after spraying, the position of insect release and the activity of individual adult M. domestica. The knockdown or mortality response of the insect to given amounts of insecticide was largely unaffected by the remaining solvent, although, with mineral oil, some inhibition was noted at low mortality levels. The amount of insecticide accumulated by houseflies in their initial flight was of prime importance in determining the overall effect. Flies remaining not knocked down at the end of the exposure period were shown to be so because they had accumulated less toxicant and not necessarily because they were more tolerant of the insecticide. The establishment of a relationship between the number of flies in flight and time after release has provided a key function for incorporation into the model.     

Low expression of nicotinic acetylcholine receptor subunit Mdα2 in neonicotinoid‐resistant strains of Musca domestica L.

BACKGROUND: Neonicotinoid action as well as resistance involves interaction with nicotinic acetylcholine receptors (nAChRs). In the housefly, neonicotinoid resistance also involves cytochrome P450, as indicated by bioassay with synergist as well as altered expression. In bioassay, synergism was only partial and indicated possible target‐site resistance. The nAChR α2 subunit is important in neonicotinoid toxicity to insects, and gene expression of the Mdα2 subunit was investigated in field populations and laboratory strains of neonicotinoid‐resistant and insecticide‐susceptible houseflies, Musca domestica L. The genomic sequence covering exon III–VII of Mdα2 was analysed for mutations. RESULTS: Gene expression profiling of Mdα2 revealed notable differences between neonicotinoid‐resistant and insecticide‐susceptible houseflies. On average, the neonicotinoid‐resistant field population 766b and the imidacloprid selected strain 791imi had 60% lower copy numbers of Mdα2 compared with the susceptible reference strain. Sequencing of exon III–VII of the Mdα2, encoding acetylcholine binding‐site regions and three out of four transmembrane domains, did not reveal any mutations explaining the increased neonicotinoid tolerance in the strains examined. CONCLUSION: Previous discoveries and the results of this study suggest that the neonicotinoid resistance mechanism in Danish houseflies involves both cytochrome P450 monooxygenase‐mediated detoxification and reduced expression of the nAChR subunit α2. Copyright © 2010 Society of Chemical Industry