Insecticide Resistance Management Research Articles

Monitoring of Target-Site Mutations Conferring Insecticide Resistance in Spodoptera frugiperda.

Simple SummaryFall armyworm, Spodoptera frugiperda, is an invasive moth species and one of the most destructive pests of maize. It is native to the Americas but recently invaded (sub)tropical regions in Africa, Asia and Oceania. Fall armyworm larvae feeding on maize plants cause substantial economic damage and are usually controlled by the application of insecticides and genetically modified (GM) maize expressing Bacillus thuringiensis (Bt) proteins, selectively targeting fall armyworm. It has developed resistance to many different classes of insecticides and Bt proteins as well; therefore, it is important to check field populations for the presence of mutations in target proteins conferring resistance. Here, we developed molecular diagnostic tools allowing us to test the frequency of resistance alleles in field-collected populations, either alive or preserved in alcohol. We tested 34 different populations collected on four different continents for the presence of mutations conferring resistance to common classes of insecticides and Bt proteins. We detected resistance mutations which are quite widespread, whereas others are restricted to certain geographies or even completely absent. The established molecular methods show robust results in samples collected across a broad geographical range and can be used to support decisions for sustainable fall armyworm control and applied resistance management.Fall armyworm (FAW), Spodoptera frugiperda, a major pest of corn and native to the Americas, recently invaded (sub)tropical regions worldwide. The intensive use of insecticides and the high adoption of crops expressing Bacillus thuringiensis (Bt) proteins has led to many cases of resistance. Target-site mutations are among the main mechanisms of resistance and monitoring their frequency is of great value for insecticide resistance management. Pyrosequencing and PCR-based allelic discrimination assays were developed and used to genotype target-site resistance alleles in 34 FAW populations from different continents. The diagnostic methods revealed a high frequency of mutations in acetylcholinesterase, conferring resistance to organophosphates and carbamates. In voltage-gated sodium channels targeted by pyrethroids, only one population from Indonesia showed a mutation. No mutations were detected in the ryanodine receptor, suggesting susceptibility to diamides. Indels in the ATP-binding cassette transporter C2 associated with Bt-resistance were observed in samples collected in Puerto Rico and Brazil. Additionally, we analyzed all samples for the presence of markers associated with two sympatric FAW host plant strains. The molecular methods established show robust results in FAW samples collected across a broad geographical range and can be used to support decisions for sustainable FAW control and applied resistance management.

Regional differences in gene regulation may underlie patterns of sensitivity to novel insecticides in Leptinotarsa decemlineata.

Agricultural insect pests frequently exhibit geographic variation in levels of insecticide resistance, which are often presumed to be due to the intensity of insecticide use for pest management. However, regional differences in the evolution of resistance to novel insecticides suggests that other factors are influencing rates of adaptation. We examined median lethal concentration (LC50 ) bioassay data spanning 15 years and six insecticides (abamectin, imidacloprid, spinosad, cyantraniliprole, chlorantraniliprole, and metaflumizone) for evidence of regional differences in Leptinotarsa decemlineata baseline sensitivity to insecticides as they became commercially available. We consistently found that larvae from Colorado potato beetle populations from the northwestern USA had the highest baseline sensitivity to novel insecticides, while populations from the eastern USA had the lowest. Comparisons of gene expression between populations from these regions revealed constitutively elevated expression of an array of detoxification genes in the East, but no evidence of additional induction when exposed to imidacloprid. Our results suggest a mechanism for geographic variation in rates of adaptation to insecticides, whereby baseline levels of gene expression determine a population's response to novel insecticides. These findings have implications for the regional development of insecticide resistance management strategies and for the fundamental question of what determines the rate of adaptation to insecticides. © 2020 Society of Chemical Industry.

Characteristics of biochemical resistance mechanism of novel insecticides in Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)

Phenacoccus solenopsis Tinsley is a major sap-sucking insect pest of many ornamental and agricultural crops in Pakistan. Insecticides are recommended to manage P. solenopsis but due to inappropriate use of insecticides, the resistance become the major concern. The study was conducted to investigate resistance against insecticides with different action modes (carbosulfan, clothianidin and chlorfenapyr) by selecting P. solenopsis populations successively for twelve generations with mentioned insecticides. Along with, possible biochemical mechanisms (acetylcholinesterase (AChE), acid and alkaline phosphatases (ACP and ALP), esterase (EST) and glutathione S-transferases (GST)) were assessed. The results described very high resistance (235-fold) to carbosulfan selected pop. (Carbo-SEL G12) and clothianidin selected pop. (Cloth-SEL G12) (112-fold) and high resistance to chlorfenapyr selected pop. (Chlor-SEL G12) (86-fold), relative to the unselected population. The activities of AChE, ACP, ALP, EST and GST enzymes gradually increased upon selection from G1 to G10. The enzymes contribute in resistance of tested insecticides were following: EST (15.61 μmol/min/mg of protein) in Carbo-SEL G12. Whereas, AChE activity (21.5 μmol/min/mg of protein) in Cloth-SEL G12 and the highest ALP enzymes activity 16.57 μmol/min/mg of protein in Chlor-SEL G12, as compared to the unselected population. Current research suggests that the detoxifying enzymes may contribute to resistance evolvement in P. solenopsis and this information may be used for the efficient management of insecticide resistance.

Fitness cost of insecticide resistance on the life-traits of a Anopheles coluzzii population from the city of Yaoundé, Cameroon

Background: Pyrethroid resistance is rapidly expanding in An. gambiae s.l. populations across Sub-Saharan Africa. Yet there is still not enough information on the fitness cost of insecticide resistance . In the present study, the fitness cost of insecticide resistance on Anopheles coluzzii population from the city of Yaoundé was investigated. Methods: A resistant An. coluzzii colony was established from field collected mosquitoes resistant to both DDT and pyrethroid and selected for 12 generations with deltamethrin 0.05%. The Ngousso laboratory susceptible strain was used as control. A total of 100 females of each strain were blood fed and allowed for individual eggs laying, and then different life traits parameters such as fecundity, fertility, larval development time, emergence rate and longevity were measured. The TaqMan assay was used to screen for the presence of the L1014F and L1014S kdr mutations. Results: Field collected mosquitoes from the F0 generation had a mortality rate of 2.05% for DDT, 34.16% for permethrin and 50.23% for deltamethrin. The mortality rate of the F12 generation was 30.48% for deltamethrin, 1.25% for permethrin and 0% for DDT. The number of eggs laid per female was lower in the resistant colony compared to the susceptible (p <0.0001). Insecticide resistant larvae were found with a significantly long larval development time (10.61±0.33 days) compare to susceptible (7.57±0.35 days). The number of emerging females was significantly high in the susceptible group compared to the resistant . The adults lifespan was also significantly high for susceptible (21.73±1.19 days) compared to resistant (14.63±0.68 days). Only the L1014F-kdr allele was detected in resistant population.. Conclusion: The study suggests that pyrethroid resistance is likely associated with a high fitness cost on An.coluzzii populations. The addition of new tools targeting specifically larval stages could improve malaria vectors control and insecticide resistance management.

Patterns of pesticide usage in agriculture in rural Tanzania call for integrating agricultural and public health practices in managing insecticide-resistance in malaria vectors

BackgroundUnrestricted use of pesticides in agriculture is likely to increase insecticide resistance in mosquito vectors. Unfortunately, strategies for managing insecticide resistance in agriculture and public health sectors lack integration. This study explored the types and usage of agricultural pesticides, and awareness and management practices among retailers and farmers in Ulanga and Kilombero districts in south-eastern Tanzania, where Anopheles mosquitoes are resistant to pyrethroids.MethodsAn exploratory sequential mixed-methods approach was employed. First, a survey to characterize pesticide stocks was conducted in agricultural and veterinary (agrovet) retail stores. Interviews to assess general knowledge and practices regarding agricultural pesticides were performed with 17 retailers and 30 farmers, followed by a survey involving 427 farmers. Concurrently, field observations were done to validate the results.ResultsLambda-cyhalothrin, cypermethrin (both pyrethroids) and imidacloprids (neonicotinoids) were the most common agricultural insecticides sold to farmers. The herbicide glyphosate (amino-phosphonates) (59.0%), and the fungicides dithiocarbamate and acylalanine (54.5%), and organochlorine (27.3%) were also readily available in the agrovet shops and widely used by farmers. Although both retailers and farmers had at least primary-level education and recognized pesticides by their trade names, they lacked knowledge on pest control or proper usage of these pesticides. Most of the farmers (54.4%, n = 316) relied on instructions from pesticides dealers. Overall, 93.7% (400) farmers practised pesticides mixing in their farms, often in close proximity to water sources. One-third of the farmers disposed of their pesticide leftovers (30.0%, n = 128) and most farmers discarded empty pesticide containers into rivers or nearby bushes (55.7%, n = 238).ConclusionSimilarities of active ingredients used in agriculture and malaria vector control, poor pesticide management practices and low-levels of awareness among farmers and pesticides retailers might enhance the selection of insecticide resistance in malaria vectors. This study emphasizes the need for improving awareness among retailers and farmers on proper usage and management of pesticides. The study also highlights the need for an integrated approach, including coordinated education on pesticide use, to improve the overall management of insecticide resistance in both agricultural and public health sectors.

Pyrethroid and Carbamate Resistance in Anopheles funestus Giles along Lake Kariba in Southern Zambia.

.Whereas data on insecticide resistance and its underlying mechanisms exist for parts of Zambia, data remain limited in the southern part of the country. This study investigated the status of insecticide resistance, metabolic mechanisms, and parasite infection in Anopheles funestus along Lake Kariba in southern Zambia. Indoor-resting mosquitoes were collected from 20 randomly selected houses within clusters where a mass drug administration trial was conducted and raised to F1 progeny. Non–blood-fed 2- to 5-day-old female An. funestus were exposed to WHO insecticide-impregnated papers with 0.05% deltamethrin, 0.1% bendiocarb, 0.25% pirimiphos-methyl, or 4% dichloro-diphenyl-trichloroethane (DDT). In separate assays, An. funestus were pre-exposed to piperonyl butoxide (PBO) to determine the presence of monooxygenases. Wild-caught An. funestus that had laid eggs for susceptibility assays were screened for circumsporozoite protein of Plasmodium falciparum by ELISA, and sibling species were identified by polymerase chain reaction. Anopheles funestus showed resistance to deltamethrin and bendiocarb but remained susceptible to pirimiphos-methyl and DDT. The pre-exposure of An. funestus to PBO restored full susceptibility to deltamethrin but not to bendiocarb. The overall sporozoite infection rate in An. funestus populations was 5.8%. Detection of pyrethroid and carbamate resistance in An. funestus calls for increased insecticide resistance monitoring to guide planning and selection of effective insecticide resistance management strategies. To prevent the development of resistance and reduce the underlying vectorial capacity of mosquitoes in areas targeted for malaria elimination, an effective integrated vector management strategy is needed.

A new knockdown resistance (kdr) mutation, F1534L, in the voltage-gated sodium channel of Aedes aegypti, co-occurring with F1534C, S989P and V1016G

BackgroundAedes aegypti is a primary vector of dengue, chikungunya and Zika infections in India. In the absence of specific drugs or safe and effective vaccines for these infections, their control relies mainly on vector control measures. The emergence of insecticide resistance in vectors, especially against pyrethroids, is a serious threat to the insecticide-based vector control programme. This study reports the presence of multiple knockdown resistance (kdr) mutations present in an Ae. aegypti population from Bengaluru (India), including a new mutation F1534L.MethodsAedes aegypti collected from Bengaluru were subjected to insecticide susceptibility tests with DDT, deltamethrin and permethrin. The DNA sequencing of partial domain II, III and IV of the voltage-gated sodium channel (VGSC) was performed to screen kdr mutations present in the population and PCR-based assays were developed for their detection. Genotyping of kdr mutations was done using PCR-based assays, allelic frequencies were determined, and tests of genetic association of kdr mutations with the insecticide resistance phenotype were performed.ResultsThe Ae. aegypti population was resistant to DDT, deltamethrin and permethrin. The DNA sequencing of the VGSC revealed the presence of four kdr mutations, i.e. S989P and V1016G in domain II and two alternative kdr mutations F1534C and F1534L in domain III. Allele-specific PCR assays (ASPCR) were developed for the detection of kdr mutations S989P and V1016G and an existing PCR-RFLP based strategy was modified for the genotyping of all three known kdr mutations in domain III (F1534L, F1534C and T1520I). Genotyping of Ae. aegypti samples revealed a moderate frequency of S989P/V1016G (18.27%) and F1534L (17.48%), a relatively high frequency of F1534C (50.61%) and absence of T1520I in the population. Mutations S989P and V1016G were in complete linkage disequilibrium in this population while they were in linkage equilibrium with kdr mutations F1534C and F1534L. The alleles F1534C and F1534L are genetically associated with permethrin resistance.ConclusionsA new kdr mutation, F1534L, was found in an Ae. aegypti population from Bengaluru (India), co-occurring with the other three mutations S989P, V1016G and F1534C. The findings of a new mutation have implications for insecticide resistance management.

Resistance monitoring to four insecticides and mechanisms of resistance in Lygus lineolaris Palisot de Beauvois (Hemiptera: Miridae) populations of southeastern USA cotton.

Economically damaging infestations of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), have become more frequent in Virginia and North Carolina cotton since 2013 and 2010, respectively. Foliar insecticide use has increased dramatically in response. Efficacy data (LC50 ) are needed to evaluate L. lineolaris susceptibility and resistance levels (RR50 ) to commonly used and recommended insecticides for managing this pest in the southeastern USA. Elevated resistance levels to acephate and bifenthrin were measured in L. lineolaris populations collected from wild and cultivated hosts in Virginia, North Carolina and northern Alabama when compared to a susceptible laboratory population. High levels of bifenthrin resistance were observed in 2018 and 2019. Mixed-function oxidase and esterase (EST) inhibitors, piperonyl butoxide and S,S,S-Tributyl phosphotrithioate, respectively, had a synergistic effect on bifenthrin with resistant populations of L. lineolaris. Bifenthrin-resistant L. lineolaris populations expressed elevated levels of cytochrome P450 (CYP450 ) monooxygenase and general EST activity. Results suggest that insecticide resistance is present in some locations and that CYP450 and EST activity in L. lineolaris contribute to pyrethroid resistance in the southeastern USA. Results can serve as a baseline for continued monitoring of L. lineolaris insecticide resistance and inform insecticide resistance management strategies that help southeastern USA cotton producers to minimize inputs and slow resistance development. © 2020 Society of Chemical Industry.

Aerobic Metabolism Alterations as an Evidence of Underlying Deltamethrin Resistance Mechanisms in Triatoma infestans (Hemiptera: Reduviidae)

Triatoma infestans (Klug, 1834), the main vector of Chagas disease in Latin America, is regularly controlled by spraying the pyrethroid deltamethrin, to which some populations have developed resistance. The three main mechanisms of resistance are 1) metabolic resistance by overexpression or increased activity of detoxifying enzymes, 2) target site mutations, and 3) cuticle thickening/modification. We use open-flow respirometry to measure real-time H2O loss rate (V˙H2O) and CO2 production rate (V˙CO2), on nymphs from susceptible and resistant populations before and after exposure to the insecticide to understand the underlying mechanisms of resistance in live insects. Lack of differences in V˙H2O between populations suggested that cuticular thickness/composition is not acting as a relevant resistance mechanism. Similarly, there was no difference in resting V˙CO2, suggesting a trade-off between resistance mechanisms and other physiological processes. The increment in V˙CO2 after application of deltamethrin was similar in both populations, which suggested that while enhanced enzymatic detoxification may play a role in resistance expression in this population, the main mechanism involved should be a passive one such as target site mutations. Open-flow respirometry provided useful evidence for evaluating the mechanisms involved in deltamethrin resistance. Using this technique could improve efficiency of scientific research in the area of insecticide resistance management, leading to a faster decision making and hence improved control results.

Indoxacarb-loaded fluorescent mesoporous silica nanoparticles for effective control of Plutella xylostella L. with decreased detoxification enzymes activities.

Plutella xylostella L. is a cosmopolitan lepidopteron insect pest for numerous vegetables and crops. The extensive use of insecticides has resulted in the emergence of resistance in P. xylostella. Thus, development of innovative strategies to overcome the insecticide resistance and control P. xylostella effectively is highly desirable. Inspired by the concept and breakthrough of nanomedical strategies to treat multidrug resistance, nanotechnology may find potential application in overcoming or delaying insecticide resistance. Carbon dots-embedded fluorescent mesoporous silica nanoparticles (FL-SiO2 NPs) were successfully developed. Indoxacarb-loaded nanoparticles (IN@FL-SiO2 NPs) were facilely prepared with loading content of 24%. The release of indoxacarb from IN@FL-SiO2 NPs was pH sensitive. IN@FL-SiO2 NPs exhibited better insecticidal activity against P. xylostella than indoxacarb technical under the same doses of active ingredient applied. Moreover, the activities of detoxification enzymes including GST, CarE, and P450 of P. xylostella were suppressed by treatment with IN@FL-SiO2 NPs. Furthermore, the entry of FL-SiO2 NPs into the midgut of P. xylostella was confirmed by CLSM observation. Although there is no absolute correlation between the enzyme activity and resistance, the change in corresponding enzyme activity can afford valuable information on the resistance situation. IN@FL-SiO2 NPs treated P. xylostella displayed higher mortality, along with decreased enzymes activities, which indicates that nano-based delivery system of insecticide could be potentially applied in insecticide resistance management. © 2020 Society of Chemical Industry.

Target Site Insensitivity Detection in Deltamethrin Resistant Culex pipiens Complex in Iran

Some mosquito species which belong to the Culex. pipiens complex are primary vectors for West Nile virus, Sindbis, Dirofilaria immitis, and many arboviruses. Knockdown resistance (kdr) mutations in the voltage-gated sodium channel (VGSC) gene of Cx. pipiens that is inherited, is one of the important threats for the efficacy of pyrethroids insecticides. Knockdown resistance (kdr) mutation, L1014F, is a well-defined mechanism of resistance to pyrethroids and DDT in many insect species. The aim of study was to determine the mechanisms of Insecticide resistance in this species. Specimens of Cx. pipiens, the major vector of West Nile virus, were obtained in Tehran, Iran by collecting larvae from polluted wastewater in Qarchak of Tehran. In 2016 Insecticide susceptibility tests were performed according to WHO methods with deltamethrin 0.05%. We focused on determination of this point mutation in the VGSC gene of Cx. pipiens by Real-time PCR. Our results revealed high levels of resistance to deltamethrin 0.05%. The lethal times i.e. LT50 and LT90 for deltamethrin were 2.1530 and 8.5117 h respectively. The result of Real-time PCR confirmed the presence of resistant genotype in all the members of tested population. This study is the first report on kdr genotyping of Cx. pipiens from Tehran and our results on the VGSC gene in position L1014F confirmed the TTA to TTT nucleotide change. This finding will provide a clue for management of insecticide resistance in mosquito which are vectors of arboviruses and decision for replacement of novel approach for vector control.

Susceptibility of fall armyworm, Spodoptera frugiperda (J.E. Smith), to eight insecticides in China, with special reference to lambda-cyhalothrin

Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is the main destructive insect pest of grain crops that occurs in all maize growing regions of the Americas. It has rapidly invaded the Southern China since January 2019. However, the current status of insecticide resistance in S. frugiperda has not been reported in China. In this study, we determined the susceptibility of eight populations of FAW to eight insecticides by an artificial diet incorporation method. The results showed that among eight insecticides, emamectin benzoate, spinetoram, chlorantraniliprole, chlorfenapyr, and lufenuron showed higher toxicity to this pest, while lambda-cyhalothrin and azadirachtin exhibited lower toxicity. Susceptibility of S. frugiperda to indoxacarb was significantly different (10.0-fold for LC50) across the various geographic populations. To investigate the biochemical mechanism of FAW to lambda-cyhalothrin, we performed the synergism tests and the results showed that piperonyl butoxide (PBO) and triphenyl phosphate (TPP) produced a high synergism of lambda-cyhalothrin effects in the two field populations. Sequencing of the gene encoding the acetylcholinesterase (AChE) gene in the two field populations identified two amino acid mutations, all of which have been shown previously to confer resistance to organophosphates (OPs) in several arthropod species. The results of this study provided valuable information for choosing alternative insecticides and for insecticide resistance management of S. frugiperda.

CHANGES IN SUSCEPTIBILITIES TO INSECTICIDES IN MUSCA DOMESTICA UNDER SELECTION WITH FIPRONIL

New knowledge about resistance and cross-resistance to insecticides of different chemical classes in insects is crucial for correct and effective pest management. This paper reports changes in the susceptibility to insecticides of different chemical classes in the first generations of houseflies Musca domestica L. when they were selected with fipronil. The selection of M. domestica was carried out by feeding adult insects with sugar containing fipronil in a dose that causes the 40–60% mortality of insects. The susceptibility of adults and larvae to insecticides was evaluated by the resistance ratio (RR). In the fipronil-selected strain of M. domestica, the susceptibility to fipronil was unchanged in adults in 3–5 generations and increased in larvae in the fifth generation (RR = 0.5). The susceptibility to deltamethrin decreased in adults of the fourth generation and in larvae of the fifth generation, and the RR was 4.5 and 4.3, respectively, that may suggest the development of cross-resistance in the future. The susceptibility to chlorfenapyr was reduced in adults of fipronil-selected flies of the fourth generation (RR = 1.9). The susceptibility to ivermectin increased in adults of fipronil-selected flies (RR = 0.67) and in larvae of the fifth generation (RR = 0.5). The susceptibility to acetamiprid in the fipronil-selected strain and the laboratory (unselected) strain of M. domestica did not differ. The results are useful for the development of insecticide resistance management programs.

'What I cannot create, I do not understand': functionally validated synergism of metabolic and target site insecticide resistance.

The putative synergistic action of target-site mutations and enhanced detoxification in pyrethroid resistance in insects has been hypothesized as a major evolutionary mechanism responsible for dramatic consequences in malaria incidence and crop production. Combining genetic transformation and CRISPR/Cas9 genome modification, we generated transgenic Drosophila lines expressing pyrethroid metabolizing P450 enzymes in a genetic background along with engineered mutations in the voltage-gated sodium channel (para) known to confer target-site resistance. Genotypes expressing the yellow fever mosquito Aedes aegypti Cyp9J28 while also bearing the paraV1016G mutation displayed substantially greater resistance ratio (RR) against deltamethrin than the product of each individual mechanism (RRcombined: 19.85 > RRCyp9J28: 1.77 × RRV1016G: 3.00). Genotypes expressing Brassicogethes aeneus pollen beetle Cyp6BQ23 and also bearing the paraL1014F (kdr) mutation, displayed an almost multiplicative RR (RRcombined: 75.19 ≥ RRCyp6BQ23: 5.74 × RRL1014F: 12.74). Reduced pyrethroid affinity at the target site, delaying saturation while simultaneously extending the duration of P450-driven detoxification, is proposed as a possible underlying mechanism. Combinations of target site and P450 resistance loci might be unfavourable in field populations in the absence of insecticide selection, as they exert some fitness disadvantage in development time and fecundity. These are major considerations from the insecticide resistance management viewpoint in both public health and agriculture.

Field resistance of Plutella xylostella (Lepidoptera: Plutellidae) to lufenuron: Inheritance and lack of cross-resistance to methoxyfenozide

Plutella xylostella (L.), commonly known as diamondback moth, is one of the most important pests of Brassicaceae in Brazil. This study aimed to assess the susceptibility of P. xylostella and pattern of resistance inheritance to lufenuron, an insect growth regulator (IGR) that impairs the insect molting, as well as the cross-resistance to methoxyfenozide (Nonsteroidal ecdysone agonist). Concentration-mortality bioassays were performed using the leaf dipping method towards field collected populations and genotypes used in the genetic crossings. Populations of this pest, from the mesoregion of Pernambuco (Brazil), were highly resistant to lufenuron. The recommended concentration of lufenuron caused more than 80% mortality of larvae of two populations [Piedade (PDD) and Recife (REC-S)]. Methoxyfenozide caused more than 80% mortality of larvae of all assessed populations, suggesting no cross-resistance between insect growth regulators. The population of Bezerros (BZR-R) was the most resistant (11,283-fold) to lufenuron compared with REC-S population (standard reference). Resistance to lufenuron was inherited as autosomal, incompletely recessive, and monofactorial trait, suggesting that a single gene or a set of genes tightly linked is involved in resistance. Methoxyfenozide is an alternative molecule suggested for the control of P. xylostella in areas where resistance to lufenuron has evolved. Furthermore, information provided in this work should fine-tune insecticide resistance management (IRM) programs to delay the evolution of resistance to chitin synthesis inhibitors like lufenuron, where resistance frequency is incipient.

Biochemical Mechanisms, Cross-resistance and Stability of Resistance to Metaflumizone in Plutella xylostella.

The diamondback moth, Plutella xylostella (L.) is an important pest of cruciferous crops worldwide. It has developed resistance to many conventional and novel insecticide classes. Metaflumizone belongs to the new chemical class of semicarbazone insecticides. To delay the development of metaflumizone resistance in P. xylostella and to guide insecticide use in the field, the biochemical mechanisms, cross-resistance spectrum, and stability of resistance to metaflumizone were studied in a laboratory-selected resistant strain (metaflu-SEL). Synergism tests with the carboxylesterase inhibitor triphenyl phosphate (TPP), the glutathione S-transferase depletor diethyl maleate (DEM), and the P450 inhibitor piperonyl butoxide(PBO) had no obvious effect on metaflumizone in the metaflu-SEL strain and the susceptible strain (SS) of P. xylostella, with synergism ratios that ranged from 1.02 to 1.86. Biochemical studies revealed that the cytochrome P450-dependent monooxygenase increased only 1.13-fold in the metaflu-SEL strain compared with the UNSEL stain; meanwhile, carboxylesterase and glutathione S-transferase activity showed no difference. These results suggest that these detoxification enzymes may be not actively involved in metaflumizone resistance. Furthermore, the metaflu-SEL population showed a moderate level of cross-resistance to indoxacarb (11.63-fold), but only very low cross-resistance to spinosad (1.75-fold), spinetoram (3.52-fold), abamectin (2.81-fold), beta-cypermethrin (0.71-fold), diafenthiuron (0.79-fold), chlorantraniliprole (2.16-fold), BT (WG-001) (3.34-fold), chlorfenapyr (0.49-fold), and chlorfluazuron (0.97-fold). Moreover, metaflumizone resistance decreased from 1087.85- to 1.23-fold in the metaflu-SEL strain after 12 generations without exposure to metaflumizone. These results are useful for formulating insecticide resistance management strategies to control P. xylostella and to delay the development of metaflumizone resistance in the field.

Insecticide resistance and malaria control: A genetics-epidemiology modeling approach

Malaria, a deadly infectious disease caused by the protozoan Plasmodium, remains a major public health menace affecting at least half the human race. Although the large-scale usage of insecticides-based control measures, notably long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS), have led to a dramatic reduction of the burden of this global scourge between the period 2000 to 2015, the fact that the malaria vector (adult female Anopheles mosquito) has become resistant to all currently-available insecticides potentially makes the current laudable global effort to eradicate malaria by 2040 more challenging. This study presents a novel mathematical model, which couples malaria epidemiology with mosquito population genetics, for assessing the impact of insecticides resistance on malaria epidemiology. Numerical simulations of the model, using data relevant to malaria transmission dynamics in the Jimma Zone of Southwestern Ethiopia, show that the implementation of a control strategy based on using LLINs alone can lead to the effective control of malaria, while also effectively managing insecticide resistance, if the LLINs coverage in the community is high enough (over 90%). It is further shown that combining LLINs with IRS (both at reduced and realistically-attainable coverage levels) can lead to the aforementioned effective control of malaria and effective management of insecticide resistance if their coverage levels lie within a certain effective control window in the LLINs-IRS coverage parameter space (this result generally holds regardless of whether or not larviciding is implemented in the community). The study identifies three key parameters of the model that negatively affect the size of the effective control window, namely parameters related with the coverage level of larviciding, the number of new adult mosquitoes that are females and the initial size of the frequency of resistant allele in the community. For the coverage of LLINs and IRS within the effective control window, an additional increase in the values of the aforementioned three parameters may lead to a shrinkage in the size of the effective control window (thereby causing the failure of the insecticides-based control).

Impact of deltamethrin selection on kdr mutations and insecticide detoxifying enzymes in Aedes aegypti from Mexico

BackgroundInsecticide resistance is a serious problem for vector control programmes worldwide. Resistance is commonly attributed to mutations at the insecticide’s target site or increased activity of detoxification enzymes.MethodsWe determined the knockdown concentration (KC50) and lethal concentration (LC50) of deltamethrin in six natural populations of adult Aedes aegypti from southeastern Mexico. These populations were then selected over five generations using the LC50 from the preceding generation that underwent selection, and the heritability of deltamethrin resistance was quantified. For each generation, we also determined the frequency of the kdr alleles L410, I1016 and C1534, and the levels of activity of three enzyme families (α- and β-esterases, mixed-function oxidases and glutathione S-transferases (GST)) associated with insecticide detoxification.ResultsThere was an increase in KC50 and LC50 values in the subsequent generations of selection with deltamethrin (FS5vs FS0). According to the resistance ratios (RRs), we detected increases in LC50 ranging from 1.5 to 5.6 times the values of the parental generation and in KC50 ranging from 1.3–3.8 times the values of the parental generation. Triple homozygous mutant individuals (tri-locus, LL/II/CC) were present in the parental generations and increased in frequency after selection. The frequency of L410 increased from 1.18-fold to 2.63-fold after selection with deltamethrin (FS5vs FS0) in the populations analyzed; for I1016 an increase between 1.19-fold to 2.79-fold was observed, and C1534 was fixed in all populations after deltamethrin selection. Enzymatic activity varied significantly over the generations of selection. However, only α- esterase activity remained elevated in multiple populations after five generations of deltamethrin selection. We observed an increase in the mean activity levels of GSTs in two of the six populations analyzed.ConclusionsThe high levels of resistance and their association with high frequencies of kdr mutations (V410L, V1016I and F1534C) obtained through artificial selection, suggest an important role of these mutations in conferring resistance to deltamethrin. We highlight the need to implement strategies that involve the monitoring of kdr frequencies in insecticide resistance monitoring and management programmes.

Insecticides, biologics and nematicides: Updates to IRAC’s mode of action classification - a tool for resistance management

Insecticide resistance has been and continues to be a significant problem for invertebrate pest control. As such, effective insecticide resistance management (IRM) is critical to maintain the efficacy of current and future insecticides. A technical group within CropLife International, the Insecticide Resistance Action Committee (IRAC) was established 35 years ago (1984) as an international association of crop protection companies that today spans the globe. IRAC's focus is on preserving the long-term utility of insect, mite, and most recently nematode control products through effective resistance management to promote sustainable agriculture and improved public health. A central task of IRAC has been the continual development and documentation of the Mode of Action (MoA) Classification scheme, which serves as an important tool for implementing IRM strategies focused on compound rotation / alternations. Updates to the IRAC MoA Classification scheme provide the latest information on the MoA of current and new insecticides and acaricides, and now includes information on biologics and nematicides. Details for these new changes and additions are reviewed herein.

Identification and detection of a novel point mutation in the Chitin Synthase gene of Culex pipiens associated with diflubenzuron resistance.

BackgroundDiflubenzuron (DFB) is one of the most used insecticides in mosquito larval control including that of Culex pipiens, the proven vector of the recent West Nile Virus epidemics in Europe. Two mutations (I1043L and I1043M) in the chitin synthase (CHS) putative binding site of DFB have been previously reported in Cx. pipiens from Italy and associated with high levels of resistance against this larvicide.Methodology/Principal findingsHere we report the identification of a third mutation at the same I1043 position of the CHS gene resulting in the substitution of Isoleucine to Phenylalanine (I1043F). This mutation has also been found in agricultural pests and has been functionally validated with genome editing in Drosophila, showing to confer striking levels (>15,000 fold) of DFB resistance. The frequency of the I1043F mutation was found to be substantially higher in Cx. pipiens mosquitoes surviving DFB doses largely exceeding the recommended field dose, raising concerns about the future efficient use of this insecticide. We monitored the presence and frequency of DFB mutations in Cx. pipiens mosquitoes from several Mediterranean countries, including Italy, France, Greece, Portugal and Israel. Among the Cx. pipiens populations collected in Northern Italy all but one had at least one of the three DFB mutations at allele frequencies reaching 93.3% for the I1043M, 64.8% for the I1043L and 10% for the I1043F. The newly reported I1043F mutation was also identified in two heterozygote individuals from France (4.2% allelic frequency). In contrast to Italy and France, no DFB resistant mutations were identified in the Cx. pipiens mosquitoes sampled from Greece, Portugal and Israel.Conclusions/SignificanceThe findings of our study are of major concern for mosquito control programs in Europe, that rely on the use of a limited number of available larvicides, and highlight the necessity for the development of appropriate Insecticide Resistance Management (IRM) programs, to ensure the sustainable use of DFB.