Synergist Piperonyl Butoxide Research Articles

Do bednets including piperonyl butoxide offer additional protection against populations of Anopheles gambiae s.l. that are highly resistant to pyrethroids? An experimental hut evaluation in Burkina Fasov.

Malaria control is dependent on the use of longlasting insecticidal nets (LLINs) containing pyrethroids. A new generation of LLINs containing both pyrethroids and the synergist piperonyl butoxide (PBO) has been developed in response to increasing pyrethroid resistance in African malaria vectors, but questions remain about the performance of these nets in areas where levels of pyrethroid resistance are very high. This study was conducted in two settings in southwest Burkina Faso, Vallée du Kou 5 and Tengrela, where Anopheles gambiae s.l. (Diptera: Culicidae) mortality rates in World Health Organization (WHO) discriminating dose assays were < 14% for permethrin and < 33% for deltamethrin. When mosquitoes were pre-exposed to PBO in WHO tube assays, mortality rates increased substantially but full susceptibility was not restored. Molecular characterization revealed high levels of kdr alleles and elevated levels of P450s previously implicated in pyrethroid resistance. In cone bioassays and experimental huts, PBO LLINs outperformed the pyrethroid-only equivalents from the same manufacturers. Blood feeding rates were 1.6-2.2-fold lower and mortality rates were 1.69-1.78-fold greater in huts with PBO LLINs vs. non-PBO LLINs. This study indicates that PBO LLINs provide greater personal and community-level protection than standard LLINs against highly pyrethroid-resistant mosquito populations.

Prevalence of resistance to deltamethrin in Rhyzopertha dominica (F.) in eastern Australia

The pyrethroid, deltamethrin, is currently used as a grain protectant to protect stored grain from insect pests in several countries including Australia. We examined the variation in response to wheat treated with deltamethrin among 15 field populations of the lesser grain borer, Rhyzopertha dominica (F.) collected from Queensland and New South Wales in eastern Australia during 2008 and 2009, to establish whether resistance has developed in this species. Based on the results of treated wheat bioassays, using a discriminating dose of 0.5 mg kg−1, we found that 14 of the 15 field populations were resistant to deltamethrin. The distance between the two most geographically separated field populations was 450 km, and both of these populations contained resistant insects. Deltamethrin applied at 1 mg kg−1 alone or with the synergist piperonyl butoxide at 8 mg kg−1 failed to control resistant R. dominica. On the basis of our findings, we conclude that populations with deltamethrin resistant individuals are common in the eastern part of Australia, and control failures are expected when such populations are present in treated grain.

The impact of temperature on insecticide toxicity against the malaria vectors Anopheles arabiensis and Anopheles funestus

BackgroundIt is anticipated that malaria elimination efforts in Africa will be hampered by increasing resistance to the limited arsenal of insecticides approved for use in public health. However, insecticide susceptibility status of vector populations evaluated under standard insectary test conditions can give a false picture of the threat, as the thermal environment in which the insect and insecticide interact plays a significant role in insecticide toxicity.MethodsThe effect of temperature on the expression of the standard WHO insecticide resistance phenotype was examined using Anopheles arabiensis and Anopheles funestus strains: a susceptible strain and the derived resistant strain, selected in the laboratory for resistance to DDT or pyrethroids. The susceptibility of mosquitoes to the pyrethroid deltamethrin or the carbamate bendiocarb was assessed at 18, 25 or 30 °C. The ability of the pyrethroid synergist piperonyl-butoxide (PBO) to restore pyrethroid susceptibility was also assessed at these temperatures.ResultsTemperature impacted the toxicity of deltamethrin and bendiocarb. Although the resistant An. funestus strain was uniformly resistant to deltamethrin across temperatures, increasing temperature increased the resistance of the susceptible An. arabiensis strain. Against susceptible An. funestus and resistant An. arabiensis females, deltamethrin exposure at temperatures both lower and higher than standard insectary conditions increased mortality. PBO exposure completely restored deltamethrin susceptibility at all temperatures. Bendiocarb displayed a consistently positive temperature coefficient against both susceptible and resistant An. funestus strains, with survival increasing as temperature increased.ConclusionsEnvironmental temperature has a marked effect on the efficacy of insecticides used in public health against important African malaria vectors. Caution must be exercised when drawing conclusions about a chemical’s efficacy from laboratory assays performed at only one temperature, as phenotypic resistance can vary significantly even over a temperature range that could be experienced by mosquitoes in the field during a single day. Similarly, it might be inappropriate to assume equal efficacy of a control tool over a geographic area where local conditions vary drastically. Additional studies into the effects of temperature on the efficacy of insecticide-based interventions under field conditions are warranted.

Effectiveness of a long-lasting piperonyl butoxide-treated insecticidal net and indoor residual spray interventions, separately and together, against malaria transmitted by pyrethroid-resistant mosquitoes: a cluster, randomised controlled, two-by-two factorial design trial

SummaryBackgroundProgress in malaria control is under threat by wide-scale insecticide resistance in malaria vectors. Two recent vector control products have been developed: a long-lasting insecticidal net that incorporates a synergist piperonyl butoxide (PBO) and a long-lasting indoor residual spraying formulation of the insecticide pirimiphos-methyl. We evaluated the effectiveness of PBO long-lasting insecticidal nets versus standard long-lasting insecticidal nets as single interventions and in combination with the indoor residual spraying of pirimiphos-methyl.MethodsWe did a four-group cluster randomised controlled trial using a two-by-two factorial design of 48 clusters derived from 40 villages in Muleba (Kagera, Tanzania). We randomly assigned these clusters using restricted randomisation to four groups: standard long-lasting insecticidal nets, PBO long-lasting insecticidal nets, standard long-lasting insecticidal nets plus indoor residual spraying, or PBO long-lasting insecticidal nets plus indoor residual spraying. Both standard and PBO nets were distributed in 2015. Indoor residual spraying was applied only once in 2015. We masked the inhabitants of each cluster to the type of nets received, as well as field staff who took blood samples. Neither the investigators nor the participants were masked to indoor residual spraying. The primary outcome was the prevalence of malaria infection in children aged 6 months to 14 years assessed by cross-sectional surveys at 4, 9, 16, and 21 months after intervention. The endpoint for assessment of indoor residual spraying was 9 months and PBO long-lasting insecticidal nets was 21 months. This trial is registered with ClinicalTrials.gov, number NCT02288637.Findings7184 (68·0%) of 10 560 households were selected for post-intervention survey, and 15 469 (89·0%) of 17 377 eligible children from the four surveys were included in the intention-to-treat analysis. Of the 878 households visited in the two indoor residual spraying groups, 827 (94%) had been sprayed. Reported use of long-lasting insecticidal nets, across all groups, was 15 341 (77·3%) of 19 852 residents after 1 year, decreasing to 12 503 (59·2%) of 21 105 in the second year. Malaria infection prevalence after 9 months was lower in the two groups that received PBO long-lasting insecticidal nets than in the two groups that received standard long-lasting insecticidal nets (531 [29%] of 1852 children vs 767 [42%] of 1809; odds ratio [OR] 0·37, 95% CI 0·21–0·65; p=0·0011). At the same timepoint, malaria prevalence in the two groups that received indoor residual spraying was lower than in groups that did not receive indoor residual spraying (508 [28%] of 1846 children vs 790 [44%] of 1815; OR 0·33, 95% CI 0·19–0·55; p<0·0001) and there was evidence of an interaction between PBO long-lasting insecticidal nets and indoor residual spraying (OR 2·43, 95% CI 1·19–4·97; p=0·0158), indicating redundancy when combined. The PBO long-lasting insecticidal net effect was sustained after 21 months with a lower malaria prevalence than the standard long-lasting insecticidal net (865 [45%] of 1930 children vs 1255 [62%] of 2034; OR 0·40, 0·20–0·81; p=0·0122).InterpretationThe PBO long-lasting insecticidal net and non-pyrethroid indoor residual spraying interventions showed improved control of malaria transmission compared with standard long-lasting insecticidal nets where pyrethroid resistance is prevalent and either intervention could be deployed to good effect. As a result, WHO has since recommended to increase coverage of PBO long-lasting insecticidal nets. Combining indoor residual spraying with pirimiphos-methyl and PBO long-lasting insecticidal nets provided no additional benefit compared with PBO long-lasting insecticidal nets alone or standard long-lasting insecticidal nets plus indoor residual spraying.FundingUK Department for International Development, Medical Research Council, and Wellcome Trust.

Residual efficacy of Deltamethrin against Sitophilus oryzae (L.), Rhyzopertha dominica (F.), Tribolium castaneum (Herbst) and Sitotroga cerealella (Oliv.) in wheat grain

Residual efficacy of the insecticide deltamethrin, an EC formulation containing 25 g/L AI + 225 g/L PBO (piperonyl butoxide synergist), against lab populations of Sitophilus oryzae, Rhyzopertha dominica, Tribolium castaneum and Sitotroga cerealella was investigated in the laboratory by applying product water solutions (0.25 mg AI/kg) to wheat grain (at 25?1?C and 60?5% r.h.). Adult mortality on 0, 7, 14 and 30 days old deposits was estimated after 2, 7 and 14 days of exposure to treated wheat grain and additional 7 days of recovery. Progeny reduction (PR, %) was also assessed. After 2 days of exposure to deposits of all ages, deltamethrin caused only 0-10% mortality of coleopterans (up to 37% after the recovery period) and 23-30% of S. cerealella, while mortality before and after recovery from 14 days of exposure was 95-50% for S. oryzae, 97-100% for R. dominica, 99-100% for T. castaneum and 100% for S. cerealella. Progeny production of S. oryzae was highest after parents contacted with 14 days old deposit of deltamethrin (PR, 76%), and lowest after contact with fresh deposit (PR, 95%), while R. dominica and T. castaneum had no progeny on any deltamethrin deposit age in wheat (PR, 100%), and S. cerealella had only a very low progeny (PR, 99%). The results showed that the synergised deltamethrin, applied at 0.25 mg/kg in wheat grain, is a highly effective insecticide for storedproduct insect control, while a higher dose is required for successful residual control of S. oryzae.

A village level cluster-randomized entomological evaluation of combination long-lasting insecticidal nets containing pyrethroid plus PBO synergist in Southern Mali

BackgroundThere is growing concern that malaria vector resistance to pyrethroid insecticides may reduce the effectiveness of long-lasting insecticidal nets (LLINs). Combination LLINs are designed to control susceptible and pyrethroid-resistant mosquito populations through a mixture of pyrethroid with piperonyl butoxide (PBO) synergist. A cluster randomized trial with entomology outcome measures was conducted in Mali to determine the added benefit over mono-treated pyrethroid predecessors. Four LLIN treatments; permethrin + PBO, permethrin, deltamethrin + PBO, and deltamethrin, were randomly allocated to four villages each (16 villages total) and distributed to cover every sleeping place. Entomological monitoring of indoor Anopheles resting densities, host preference, vector longevity, and sporozoite rates were monitored every 2 months over 2 years in 2014 and 2015.ResultsBottle bioassays confirmed permethrin and deltamethrin resistance in Anopheles gambiae sensu lato (s.l.), (the predominant species throughout the study) with pre-exposure to PBO indicating partial involvement of oxidases. Between 2014 and 2015 the mean indoor resting density was greater in the deltamethrin + PBO LLIN arm than the deltamethrin LLIN arm at 3.05 (95% CI 3.00–3.10) An. gambiae s.l. per room per day compared with 1.9 (95% CI 1.87–1.97). There was no significant difference in sporozoite rate at 3.97% (95% CI 2.91–5.02) for the deltamethrin LLIN arm and 3.04% (95% CI 2.21–3.87) for deltamethrin + PBO LLIN arm (P = 0.17). However, when analysed by season there was some evidence that the sporozoite rate was lower in the deltamethrin + PBO LLIN arm than deltamethrin LLIN arm during the rainy/high malaria transmission seasons at 1.95% (95% CI 1.18–2.72) and 3.70% (95% CI 2.56–4.84) respectively (P = 0.01).ConclusionsWhile there was some evidence that An. gambiae s.l. sporozoite rates were lower in villages with deltamethrin + PBO LLINs during the high malaria transmission seasons of 2014–2015, there was no reduction in parity rates or indoor resting densities. There was also no evidence that permethrin + PBO LLINs provided any improved control when compared with permethrin LLINs. Combination nets may have a greater impact in areas where mixed function oxidases play a more important role in pyrethroid resistance.

Temporal distribution and insecticide resistance profile of two major arbovirus vectors Aedes aegypti and Aedes albopictus in Yaound\xe9, the capital city of Cameroon

BackgroundAedes aegypti and Ae. albopictus are the major epidemic vectors of several arbovirus diseases such as yellow fever, dengue, Zika and chikungunya worldwide. Both Aedes vectors are presents in Cameroon; however, knowledge on the dynamic of the distribution of these species across cities and their resistance profile to insecticide are limited. Here, we assessed the current distribution of Ae. aegypti and Ae. albopictus in Yaoundé, the Capital City, established the resistance profile to insecticides and explored the resistance mechanisms involved.MethodsImmature stages of Aedes were sampled in several breeding sites in December 2015 (dry season) and June 2016 (rainy season) in three central neighborhoods and four peripheral neighborhoods and reared to adult stage. The G0 adults were used for molecular identification and genotyping of F1534C mutation in Ae. aegypti. Bioassays and piperonyl butoxide (PBO) assays were carried out according to WHO guidelines.ResultsAnalysis revealed that both species Ae. aegypti and Ae. albopictus are present in all prospected sites in Yaounde. However, in the dry season Ae. aegypti is most abundant in neighborhoods located in downtown. In contrast, Ae. albopictus was found most prevalent in suburbs whatever the season and in downtown during the rainy season. Bioassay analysis showed that both Ae. aegypti and Ae. albopictus, are resistant to 0.05% deltamethrin, 0.1% bendiocarb and 4% dichlorodiphenyltrichloroethane (DDT). A decreased of susceptibility to 0.75% permethrin and a full susceptibility to malathion 5% was observed. The mortality rate was increased after pre-exposure to synergist PBO. None of Ae. aegypti assayed revealed the presence of F1534C mutation.ConclusionThese findings are useful to planning vector control programme against arbovirus vectors in Cameroon and can be used as baseline in Africa where data on Aedes resistance is very scarce to plan further works.

Insecticide resistance status of three malaria vectors, Anopheles gambiae (s.l.), An. funestus and An. mascarensis, from the south, central and east coasts of Madagascar

BackgroundInsecticide-based vector control, which comprises use of insecticide-treated bed nets (ITNs) and indoor residual spraying (IRS), is the key method to malaria control in Madagascar. However, its effectiveness is threatened as vectors become resistant to insecticides. This study investigated the resistance status of malaria vectors in Madagascar to various insecticides recommended for use in ITNs and/or IRS.MethodsWHO tube and CDC bottle bioassays were performed on populations of Anopheles gambiae (s.l.), An. funestus and An. mascarensis. Adult female An. gambiae (s.l.) mosquitoes reared from field-collected larvae and pupae were tested for their resistance to DDT, permethrin, deltamethrin, alpha-cypermethrin, lambda-cyhalothrin, bendiocarb and pirimiphos-methyl. Resting An. funestus and An. mascarensis female mosquitoes collected from unsprayed surfaces were tested against permethrin, deltamethrin and pirimiphos-methyl. The effect on insecticide resistance of pre-exposure to the synergists piperonyl-butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) also was assessed. Molecular analyses were done to identify species and determine the presence of knock-down resistance (kdr) and acetylcholinesterase resistance (ace-1R) gene mutations.ResultsAnopheles funestus and An. mascarensis were fully susceptible to permethrin, deltamethrin and pirimiphos-methyl. Anopheles gambiae (s.l.) was fully susceptible to bendiocarb and pirimiphos-methyl. Among the 17 An. gambiae (s.l.) populations tested for deltamethrin, no confirmed resistance was recorded, but suspected resistance was observed in two sites. Anopheles gambiae (s.l.) was resistant to permethrin in four out of 18 sites (mortality 68–89%) and to alpha-cypermethrin (89% mortality) and lambda-cyhalothrin (80% and 85%) in one of 17 sites, using one or both assay methods. Pre-exposure to PBO restored full susceptibility to all pyrethroids tested except in one site where only partial restoration to permethrin was observed. DEF fully suppressed resistance to deltamethrin and alpha-cypermethrin, while it partially restored susceptibility to permethrin in two of the three sites. Molecular analysis data suggest absence of kdr and ace-1R gene mutations.ConclusionThis study suggests involvement of detoxifying enzymes in the phenotypic resistance of An. gambiae (s.l.) to pyrethroids. The absence of resistance in An. funestus and An. mascarensis to pirimiphos-methyl and pyrethroids and in An. gambiae (s.l.) to carbamates and organophosphates presents greater opportunity for managing resistance in Madagascar.

Combined target site (kdr) mutations play a primary role in highly pyrethroid resistant phenotypes of Aedes aegypti from Saudi Arabia

BackgroundPyrethroid resistance is a threat to effective vector control of Aedes aegypti, the vector of dengue, Zika and other arboviruses, but there are many major knowledge gaps on the mechanisms of resistance. In Jeddah and Makkah, the principal dengue-endemic areas of Saudi Arabia, pyrethroids are used widely for Ae. aegypti control but information about resistance remains sparse, and the underlying genetic basis is unknown. Findings from an ongoing study in this internationally significant area are reported here.MethodsAedes aegypti collected from each city were raised to adults and assayed for resistance to permethrin, deltamethrin (with and without the synergist piperonyl butoxide, PBO), fenitrothion, and bendiocarb. Two fragments of the voltage-gated sodium channel (Vgsc), encompassing four previously identified mutation sites, were sequenced and subsequently genotyped to determine associations with resistance. Expression of five candidate genes (CYP9J10, CYP9J28, CYP9J32, CYP9M6, ABCB4) previously associated with pyrethroid resistance was compared between assay survivors and controls.ResultsJeddah and Makkah populations exhibited resistance to multiple insecticides and a similarly high prevalence of resistance to deltamethrin compared to a resistant Cayman strain, with a significant influence of age and exposure duration on survival. PBO pre-exposure increased pyrethroid mortality significantly in the Jeddah, but not the Makkah strain. Three potentially interacting Vgsc mutations were detected: V1016G and S989P were in perfect linkage disequilibrium in each strain and strongly predicted survival, especially in the Makkah strain, but were in negative linkage disequilibrium with 1534C, though some females with the Vgsc triple mutation were detected. The candidate gene CYP9J28 was significantly over-expressed in Jeddah compared to two susceptible reference strains, but none of the candidate genes was consistently up-regulated to a significant level in the Makkah strain.ConclusionsDespite their proximity, Makkah and Jeddah exhibit significant differences in pyrethroid resistance phenotypes, with some evidence to suggest a different balance of mechanisms, for example with more impact associated with CYP450s in the Jeddah strain, and the dual kdr mutations 989P and 1016G in the more resistant Makkah strain. The results overall demonstrate a major role for paired target site mutations in pyrethroid resistance and highlight their utility for diagnostic monitoring.

Toxicity and Metabolism of Zeta-Cypermethrin in Field-Collected and Laboratory Strains of the Neotropical Predator Chrysoperla externa Hagen (Neuroptera: Chrysopidae).

Resistance to pesticides has been studied in several insect pests, but information on the natural enemies of pests-including the Neotropical predator Chrysoperla externa Hagen (Neuroptera: Chrysopidae), a major biological control agent in South America-is lacking. We report here a comparative study between a field-collected strain of C. externa subjected to monthly sprayings of pyrethroids and neonicotinoids and a laboratory strain without exposure to pesticides. The tolerance of both strains against zeta-cypermethrin was similar, and addition of the synergist piperonyl butoxide increased the toxicity by 30% in both strains. Gas-chromatography analyses and mixed-function-oxidase measurements indicated similar values in both strains and also confirmed the key role of oxidative metabolism in this species. Because C. externa has maintained a tolerance to zeta-cypermethrin without previous pesticide exposure, this species could potentially be mass-reared and released in fields in the presence of pesticide pressure.

On nonlinear beta regression residuals

We proposed a new residual to be used in linear and nonlinear beta regressions. Unlike the residuals that had already been proposed, the derivation of the new residual takes into account not only information relative to the estimation of the mean submodel but also takes into account information obtained from the precision submodel. This is an advantage of the residual we introduced. Additionally, the new residual is computationally less intensive than the weighted residual. Recall that the computation of the latter involves an n×n matrix, where n is the sample size. Obviously, that can be a problem when the sample size is very large. In contrast, our residual does not suffer from that. It can be easily computed even in large samples. Finally, our residual proved to be able to identify atypical observations as well as the weighted residual. We also propose new thresholds for residual plots and a scheme for the choice of starting values to be used in maximum likelihood point estimation in the class of nonlinear beta regression models. We report Monte Carlo simulation results on the behavior of different residuals. We also present and discuss two empirical applications; one uses the proportion of killed grasshoppers in an assay on the grasshopper Melanopus sanguinipes with the insecticide carbofuran and the synergist piperonyl butoxide, which enhances the toxicity of the insecticide, and the other uses simulated data. The results favor the new methodology we introduce.

Residual efficacy of cypermethrin and pirimiphos-methyl against Sitotroga cerealella (Olivier) in wheat grain

Residual efficacy of EC formulations of two insecticides: cypermethrin supplemented with the synergist piperonyl butoxide, and pirimiphos-methyl, against adults of the Angoumois grain moth, Sitotroga cerealella (Olivier), was investigated in the laboratory (at 25?1?C and 55-60% r.h.) by applying water solutions of products based either on cypermethrin (1.6 mg a.i./kg of wheat grain) or pirimiphos-methyl (4 mg a.i./kg of wheat grain) to wheat grain. Insect mortality on deposits of different age: 0, 7, 14, 30, 60, 90, 120, 150 and 180 days was estimated after 2, 7 and 14 days of insect exposure to treated wheat. After 2 day exposure, cypermethrin caused mortality of up to 46% on all deposits (age 0-180 days). After 7 days of exposure, high efficacy (94-100%) was found only on deposits that were up to 90 days old, while the mortality of S. cerealella adults on all deposits following 14 days of exposure was 98-100%, and it was probably additionally enhanced by natural mortality. Pirimiphos-methyl was 98-100% effective after 2 days of exposure to deposits that were up to 30 days old, while it achieved maximum efficacy (100%) after 7 days of insect exposure to deposits aging for up to 150 days, and 94% efficacy on 180 days old deposits. After 14 days of adult exposure to all deposit ages, there were no surviving insects (efficacy 100%). The results show that pirimiphos-methyl was more effective against S. cerealella adults and had a longer residual activity than cypermethrin.

Toxicity and Physiological Actions of Carbonic Anhydrase Inhibitors to Aedes aegypti and Drosophila melanogaster.

The physiological role of carbonic anhydrases in pH and ion regulation is crucial to insect survival. We examined the toxic and neurophysiological effects of five carbonic anhydrase inhibitors (CAIs) against Aedes aegypti. The 24 h larvicidal toxicities followed this rank order of potency: dichlorphenamide > methazolamide > acetazolamide = brinzolamide = dorzolamide. Larvicidal activity increased modestly in longer exposures, and affected larvae showed attenuated responses to probing without overt tremors, hyperexcitation, or convulsions. Acetazolamide and dichlorphenamide were toxic to adults when applied topically, but were of low potency and had an incomplete effect (<50% at 300 ng/mosquito) even after injection. Dichlorphenamide was also the most toxic compound when fed to adult mosquitoes, and they displayed loss of posture and occasionally prolonged fluttering of the wings. Co-exposure with 500 ng of the synergist piperonyl butoxide (PBO) increased the toxicity of dichlorphenamide ca. two-fold in feeding assays, indicating that low toxicity was not related to oxidative metabolism. Dichlorphenamide showed mild depolarizing and nerve discharge actions on insect neuromuscular and central nervous systems, respectively. These effects were increased in low buffer salines, indicating they were apparently related to loss of pH control in these tissues. Overall, sulfonamides displayed weak insecticidal properties on Aedes aegypti and are weak lead compounds.

Toxicity and metabolic mechanisms underlying the insecticidal activity of parsley essential oil on bean weevil, Callosobruchus maculatus

Control of the bean weevil, Callosobruchus maculatus (Coleoptera: Chrysomelidae: Bruchinae), in stored cowpea beans, Vigna unguiculata (L) Walp., when accomplished, is mainly achieved by applications of phosphine. However, the long-term use of this insect control practice has contributed to the selection of resistant populations, enhancing the need to develop alternative control tools. Due to their diverse biologically active compounds, the essential oils of aromatic plants such as parsley, Petroselinum sativum L., have been suggested as suitable alternatives for controlling insect pests. Here, we evaluated the toxicity (including the metabolic mechanisms underlying the insecticidal activity) of parsley essential oil against C. maculatus. Parsley essential oil controlled C. maculatus in a concentration-dependent manner (concentration increases resulted in lower weight losses and better germination levels of beans) with a fumigant toxicity (LC50: 489.5 μL L−1 air) significantly lower than that observed for phosphine (LC50: 35.7 μL L−1 air). However, when applied at their LC10 values (Parsley essential oil: 399.3 μL L−1 air. Phosphine: 18.6 μL L−1 air), the parsley essential oil caused significantly higher reductions in the emergence of C. maculatus compared with phosphine. Additionally, application of the synergists piperonyl butoxide (PBO), triphenyl phosphate (TPP) and diethyl maleate (DEM) demonstrated that glutathione S-transferases enhanced the parsley essential oil toxicity, while cytochrome P450-dependent monooxygenases formed part of the resistance mechanisms used by C. maculatus to mitigate the toxicity of the essential oil. Our findings suggest that parsley essential oil may be integrated into the control of C. maculatus only when synergized with PBO.

Increasing insecticide resistance in Anopheles funestus and Anopheles arabiensis in Malawi, 2011-2015.

BackgroundSusceptibility of principal Anopheles malaria vectors to common insecticides was monitored over a 5-year period across Malawi to inform and guide the national malaria control programme.MethodsAdult blood-fed Anopheles spp. and larvae were collected from multiple sites in sixteen districts across the country between 2011 and 2015. First generation (F1) progeny aged 2–5 days old were tested for susceptibility, using standard WHO procedures, against pyrethroids (permethrin and deltamethrin), carbamates (bendiocarb and propoxur), organophosphates (malathion and pirimiphos-methyl) and an organochlorine (DDT).ResultsMortality of Anopheles funestus to deltamethrin, permethrin, bendiocarb and propoxur declined significantly over the 5-year (2011–2015) monitoring period. There was wide variation in susceptibility to DDT but it was not associated with time. In contrast, An. funestus exhibited 100% mortality to the organophosphates (malathion and pirimiphos-methyl) at all sites tested. There was reduced mortality of Anopheles arabiensis to deltamethrin over time though this was not statistically significant. However, mortality of An. arabiensis exposed to permethrin declined significantly over time. Anopheles arabiensis exposed to DDT were more likely to be killed if there was high ITN coverage in the mosquito collection area the previous year. There were no other associations between mosquito mortality in a bioassay and ITN coverage or IRS implementation. Mortality of An. funestus from four sites exposed to deltamethrin alone ranged from 2 to 31% and from 41 to 94% when pre-exposed to the synergist piperonyl butoxide followed by deltamethrin. For permethrin alone, mortality ranged from 2 to 13% while mortality ranged from 63 to 100% when pre-exposed to PBO.ConclusionPyrethroid resistance was detected in An. funestus and An. arabiensis populations across Malawi and has worsened over the last 5 years. New insecticides and control strategies are urgently needed to reduce the burden of malaria in Malawi.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-016-1610-1) contains supplementary material, which is available to authorized users.

The impact of pyrethroid resistance on the efficacy and effectiveness of bednets for malaria control in Africa.

Long lasting pyrethroid treated bednets are the most important tool for preventing malaria. Pyrethroid resistant Anopheline mosquitoes are now ubiquitous in Africa, though the public health impact remains unclear, impeding the deployment of more expensive nets. Meta-analyses of bioassay studies and experimental hut trials are used to characterise how pyrethroid resistance changes the efficacy of standard bednets, and those containing the synergist piperonyl butoxide (PBO), and assess its impact on malaria control. New bednets provide substantial personal protection until high levels of resistance, though protection may wane faster against more resistant mosquito populations as nets age. Transmission dynamics models indicate that even low levels of resistance would increase the incidence of malaria due to reduced mosquito mortality and lower overall community protection over the life-time of the net. Switching to PBO bednets could avert up to 0.5 clinical cases per person per year in some resistance scenarios.

Mechanisms of Insecticide Resistance in Field Populations of the Varroa Mite (Acari: Mesostigmata: Varroidae) in Florida

Summary The varroa mite (Acari: Mesostigmata: Varroidae) has developed resistance to the 2 major miticides (fluvalinate and coumaphos) registered for control of this invasive pest of bees (Hymenoptera: Apidae) in the United States. Comparative studies on miticide toxicity with and without the synergists piperonyl butoxide and formamidine indicated that enhanced metabolism by mixed-function oxidases and altered target site were the major mechanisms of resistance to organophosphorus and pyrethroid insecticides in varroa mite populations of northern Florida.

Toxicology of potassium channel-directed compounds in mosquitoes

Potential targets for new vector control insecticides are nerve and muscle potassium channels. In this study, the activities of known potassium channel blockers (4-aminopyridine, quinidine, and tetraethylammonium) and the insecticide propoxur were compared to three experimental catechols and several other compounds against Anopheles gambiae and Aedes aegypti mosquitoes. Experimental catechol 1 was the most toxic experimental compound in all of the mortality assays conducted, but was at least 100-fold and 39-fold less toxic than propoxur against Ae. aegypti and An. gambiae, respectively. Injection treatment and synergist (piperonyl butoxide) bioassays found that catechol toxicity was not unduly impacted by cuticular transport or oxidative metabolism. Electrophysiological studies showed a decrease in amplitude of evoked muscle contractions, along with an increase in twitch duration at concentrations that increased basal muscle tension (mM). High concentration effects on basal muscle tension were matched by complete depolarization of the muscle membrane potential. Effects on muscle physiology and blockage of Kv2.1 potassium channels in patch clamp experiments were generally consistent with in vivo toxicity, except for 4-aminopyridine, which suggest the involvement of other potassium channel subtypes. Extensive melanization of Anopheles larvae, but not Aedes larvae, occurred from exposure to catechol compounds. Interaction with the phenol oxidase system within insects may be the cause of this melanization, but any contribution to toxicity requires further investigation.

Insecticide resistance status of the Anopheles funestus population in Central African Republic: a challenge in the war.

BackgroundIn the Central African Republic, malaria is a major public health problem and the leading cause of death among children. This disease appears to be hyperendemic but no substantial entomological data, including data on Anopheles spp. susceptibility to insecticides, is available. This study evaluates, for the first time in the CAR, the status of insecticide resistance in the Anopheles funestus population, the second major vector of malaria in Africa.MethodsWHO standard bioassay susceptibility tests were performed on the An. funestus population using F1 generation from gravid females mosquitoes (F0) collected by manual aspirator sampling of households in Gbanikola, Bangui in October 2014 to assess: (i) An. funestus susceptibility to bendiocarb, malathion, permethrin, lamda-cyhalothrin, deltamethrin and DDT, and (ii) the effect of pre-exposure to the piperonyl butoxide (PBO) synergist on insecticide susceptibility. Additional tests were conducted to investigate metabolic resistance status (cytochrome P450 monooxygenases, glutathione S-transferases, and esterases).ResultsA high phenotypic resistance of An. funestus population to malathion, DDT and pyrethroids was observed with a mortality rate ranging from 23 to 74 %. For the pyrethroid groups, the mortality rate was 35, 31 and 23 % for lambda-cyhalothrin, deltamethrin, and permethrin, respectively. In contrast a 100 % mortality rate to bendiocarb was recorded. Knockdown time (KDT) was long for all pyrethroids, DDT and malathion with KDT50 higher than 50 min. Pre-exposure of An. funestus to PBO synergist significantly restored susceptibility to all pyrethroids (Fisher's exact test P <0.0001) but not in DDT (Fisher's exact test P = 0.724). Data from biochemical tests suggest the involvement of cytochrome P450 monooxygenases, esterases and glutatione S-transferases in the resistance of An. funestus population from Gbanikola (Wilcoxon test P <0.05).ConclusionEvidence of biochemical resistance to insecticide was detected in An. funestus population from the district of Gbanikola, Bangui. This study suggests that detoxifying enzymes are involved in insecticide resistance of An. funestus. However, despite disruptive violence, further research is urgently needed to assess the insecticide susceptibility status of An. funestus population in all CAR regions; insecticide resistance could rapidly compromise the success of malaria control programs.

CHARACTERIZATION AND EXPRESSION PROFILES OF FIVE POSSIBLE CYTOCHROME P450 GENES FROM Liposcelis entomophila (ENDERLEIN) (PSOCOPTERA: LIPOSCELIDIDAE).

In this study, the cDNAs of five cytochromes P450 genes (named CYP345P1, CYP358B1, CYP4FD2, CYP4CD2, and CYP6JN1) contained open reading frames from 1,500 to 1,554 nucleotides that encoded 499 to 517 amino acids were cloned from the psocid Liposcelis entomophila. They are characterized by predicted molecular weights from 57.67 to 59.64 kDa and theoretical isoelectric points of 5.57-9.07. Quantitative real-time PCR analysis showed these five genes were expressed at all tested developmental stages and higher expressions were observed in adults. CYP358B1 was expressed at higher levels in egg and adult compared to the larval stages. mRNA abundances of five genes were detected in both sexes and were relatively more abundant in adult females than in adult males. Synergism bioassay showed that the synergic ratio was 2.20 and 2.45 when insects were treated with the mixture of deltamethrin or malathion with the synergist piperonyl butoxide (PBO). Because PBO induces cytochrome P450s in some insects, this suggested to us that cytochromes P450 might participate in detoxification of these insecticides. The transcripts of the five cytochromes P450 genes in adult psocids could be induced to the highest level at 12h after the exposure to malathion. After exposure to deltamethrin, CYP358B1 reached maximum expression at 24h. The maximum expression of the other four genes occurred at 36 h. Treatments with the carbamate propoxur did not influence transcription of the cytochromes P450 gene. The induction profiles suggested that these five cytochrome P450 genes may be associated with deltamethrin and malathion metabolism in psocids.