Insecticide Resistance Monitoring Research Articles

Genome-wide analysis of detoxification genes conferring diamide insecticide resistance in Spodoptera exigua identifies CYP9A40

For over a decade, diamide insecticides have been effective against lepidopteran pests like beet armyworm, Spodoptera exigua (Hübner, 1808). However, the evolution of resistance poses a challenge to their sustainable use. We identified an I4790M mutation in the S. exigua ryanodine receptor (RyR) gene, but its correlation with resistance varied across the field-collected Korean populations of S. exigua. RNA sequencing and differential gene expression analysis were performed to investigate other resistance mechanisms. Diamide-resistant and susceptible strains and F1 hybrids were compared by mapping RNA-seq reads to the S. exigua reference genome. CYP9A40 was identified as a critical gene in diamide resistance due to its high expression in the resistant strains. Synergist bioassays with piperonyl butoxide supported the role of P450s in diamide metabolic resistance in S. exigua. A strong positive correlation between CYP9A40 over-expression levels (up to 80-fold) and diamide LC50 values was obtained for field-collected populations uniformly showing a 100% frequency of the RyR I4790M target-site resistance allele. To validate the function of CYP9A40 in diamide detoxification, we recombinantly expressed the gene and tested its ability to bind and degrade chlorantraniliprole as a substrate. The results confirmed its catalytic role in diamide metabolism. CYP9A40 has been identified and validated to confer metabolic resistance in Korean S. exigua populations. It works alongside the RyR target-site I4790M mutation to enhance diamide resistance. These mechanisms offer insights for resistance monitoring and support insecticide resistance management programs to improve control strategies for S. exigua.

Molecular Diagnostics for Monitoring Insecticide Resistance in Lepidopteran Pests

Chemical control methods to prevent crop damage have long been directly implicated in the selection of lepidoptera insect populations resistant to insecticides. More recently, new products featuring different modes of action (MoA), developed to mitigate the negative effects of control management on both producers and the environment, are rapidly losing efficacy due to the emergence of insect resistance. Among these, certain resistances are associated with molecular changes in the genomes of pest insects that are valuable for developing molecular markers for diagnostic tools, particularly the point mutations. Molecular diagnosis represents an innovative solution for insecticide resistance management (IRM) practices, allowing for the effective monitoring of insecticide resistance. This approach facilitates decision making by enabling the timely alternation between different modes of action (MoAs). In this context, this review focuses on the major lepidopteran pests that affect globally significant crops, discussing the impacts of insecticide resistance. It gathers literature on diagnostic methods; provides a comparative overview of the advantages of different techniques in terms of efficiency, cost, precision, sensitivity, and applicability; and highlights several novel diagnostic tools. Additionally, this review explores the coffee leaf miner, Leucoptera coffeella, as an applied model to illustrate potential approaches for more effective and sustainable control strategies.

Area-wide survey and monitoring of insecticide resistance in the brown planthopper, Nilaparvata lugens (Stål), from 2020 to 2023 in China

The brown planthopper (BPH), Nilaparvata lugens (Stål), is a notorious pest affecting Asian rice crops. The evolution of insecticide resistance in BPH has emerged as a significant challenge in effectively managing this pest. This study revealed the resistance status of BPH to nine insecticides in ten provinces and Shanghai City in China from 2020 to 2023. Monitoring results showed that the resistance of BPH to triflumezopyrim, nitenpyram, and dinotefuran increased rapidly. The average resistance ratio of BPH to triflumezopyrim increased from 2.5 to 7.1 fold, nitenpyram from 18.3 to 37.7 fold, and dinotefuran from 119.5 to 268.1 fold. All populations remained extremely high resistance to imidacloprid, thiamethoxam, and buprofezin. Most field populations of BPH maintained moderate resistance to chlorpyrifos and sulfoxaflor, and high resistance to pymetrozine by rice stem dipping method. However, considering the reproduction-inhibiting character of pymetrozine, susceptible to low resistance levels to pymetrozine were monitored by Insecticide Resistance Action Committee (IRAC) NO.005 method. This result indicated that pymetrozine might lose efficacy in the control of application generation, but it could significantly inhibit the reproduction of field populations of BPH. Additionally, we compared the expression levels of 11 nicotinic acetylcholine receptor (nAChR) genes, the targets of nAChR competitive modulators, in four field populations (FY23, YH23, LJ23, LP23) and susceptible strain. The expression level of nAChR α4 was significantly reduced in all field populations, while α1, α2, α6, and α7 were significantly reduced in some field populations. Our findings provide valuable information for resistance management strategies in N. lugens and offer new insights into the resistance mechanisms of nAChR competitive modulators.

A regional One Health approach to the risk of invasion by Anopheles stephensi in Mauritius.

Anopheles stephensi is an invasive malaria vector in Africa that threatens to put an additional 126 million people at risk of malaria if it continues to spread. The island nation of Mauritius is highly connected to Asia and Africa and is at risk of introduction due to this connectivity. For early detection of An. stephensi, the Vector Biology and Control Division under the Ministry of Health in Mauritius, leveraged a well-established Aedes program, as An. stephensi is known to share Aedes habitats. These efforts triggered multisectoral coordination and cascading benefits of integrated vector and One Health approaches. Beginning June 2021, entomological surveys were conducted at points of entry (seaport, airport) and on ships transporting livestock in collaboration with the Civil Aviation Department, the Mauritian Port Authority and National Veterinary Services. A total of 18, 39, 723 mosquito larval surveys were respectively conducted in the airport, seaport, and other localities in Mauritius while two, 20, and 26 adult mosquito surveys were respectively conducted in the airport, seaport, and twenty-six animal assembly points. Alongside adult mosquito surveys, surveillance of vectors of veterinary importance (e.g.- Culicoides spp.) was also carried out in collaboration with National Parks and Conservation Service and land owners. A total of 8,428 adult mosquitoes were collected and 1,844 larval habitats were positive for mosquitoes. All collected mosquitoes were morphologically identified and 151 Anopheles and 339 Aedes mosquitoes were also molecularly characterized. Mosquito species detected were Aedes albopictus, Anopheles arabiensis, An. coustani, An. merus, Culex quinquefasciatus, Cx. thalassius and Lutzia tigripes. Anopheles stephensi was not detected. The One Health approach was shared with the French Agricultural Research Centre for International Development (CIRAD), strengthening collaboration between Mauritius and Réunion Island on vector surveillance at entry points and insecticide resistance monitoring. The Indian Ocean Commission (IOC) was also alerted to the risk of An. stephensi, leading to regional efforts supporting trainings and development of a response strategy to An. stephensi bringing together stakeholders from Comoros, Madagascar, Mauritius, Réunion Island and Seychelles. Mauritius is a model system showing how existing public health entomology capabilities can be used to enhance vector surveillance and control and create multisectoral networks to respond to any emerging public and veterinary health vector-borne disease threat.

A tiny sample rapid visual detection technology for imidacloprid resistance in Aphis gossypii by CRISPR/Cas12a

Insecticide resistance monitoring is essential for guiding chemical pest control and resistance management policies. Currently, rapid and effective technology for monitoring the resistance of tiny insects in the field is absent. Aphis gossypii Glover is a typical tiny insect, and one of the most frequently reported insecticide-resistant pests. In this study, we established a novel CRISPR/Cas12a-based rapid visual detection approach for detecting the V62I and R81T mutations in the β1 subunit of the nAChR in A. gossypii, to reflect target-site resistance to imidacloprid. Based on the nAChR β1 subunit gene in A. gossypii, the V62I/R81T-specific RPA primers and crRNAs were designed, and the ratio of 10 μM/2 μM/10 μM for ssDNA/Cas12a/crRNA was selected as the optimal dosage for the CRISPR reaction, ensuring that Cas12a only accurately recognizes imidacloprid-resistance templates. Our data show that the field populations of resistant insects possessing V62I and R81T mutations to imidacloprid can be accurately identified within one hour using the RPA-CRISPR/Cas12a detection approach under visible blue light at 440–460 nm. The protocol for RPA-CRISPR detection necessitates a single less than 2 mm specimen of A. gossypii tissues to perform RPA-CRISPR detection, and the process only requires a container at 37 °C and a portable blue light at 440–460 nm. Our research represents the first application of RPA-CRISPR technology in insecticide resistance detection, offers a new method for the resistance monitoring of A. gossypii or other tiny insects, helps delay the development of resistance to imidacloprid, improves the sustainability of chemical control, and provides theoretical guidance for managing pest resistance.

For Malaria Elimination, India Needs to Integrate Its Research and Control Arms.

India has targeted malaria elimination by 2030. The national malaria control program has positioned its strategies in this direction. Substantial support in the form of dynamic research inputs leading to policy formulation and change is needed to steer the country towards malaria elimination. Indian Council of Medical Research (ICMR), India's nodal research body, has been generating evidence and helping to frame several policies ranging from malaria management to vector control operations. Since the country is preparing for malaria elimination, the connection between the programmatic needs and the research agenda needs further strengthening. Typically, the national malaria control program handles the implementation of programmatic activities, while the national research body, ICMR, conducts research studies to generate evidence. We propose a virtual integration of the activities conducted by these two entities to maximize the potential for translating research findings into programmatic policies. Joint monitoring of drug and insecticide resistance, codevelopment and utilization of more innovative surveillance systems, data-backed mitigation responses, and overcoming last-mile challenges are reasons for the virtual amalgamation of the two bodies. Timely translation of research outputs into policy, co-opting of workforce and material resources, joint capacity building, and synergistic advocacy are benefits of the proposed new alliance for more efficient operations. The close functioning will provide impetus to narrow down current gaps and disrupt traditional barriers, galvanizing the country toward malaria elimination.

Preliminary monitoring of knockdown resistance (kdr) mutation in Anopheles stephensi: insights from a malarious area in Southeastern Iran

BackgroundAnopheles stephensi is recognized as the main malaria vector in Iran. In recent years, resistance to several insecticide classes, including organochlorine, pyrethroids, and carbamate compounds, has been reported for this medically important malaria vector. The main objective of the present study was to evaluate the insecticide susceptibility status of An. stephensi collected from the southern part of Iran, and to clarify the mechanism of resistance, using bioassay tests and molecular methods comparing the sequence of susceptible and resistant mosquitoes.MethodsMosquito larvae were collected from various larval habitats across six different districts (Gabrik, Sardasht, Tidar, Dehbarez, Kishi and Bandar Abbas) in Hormozgan Provine, located in the southern part of Iran. From each district standing water areas with the highest densities of Anopheles larvae were selected for sampling, and adult mosquitoes were reared from them. Finally, the collected mosquito species were identified using valid keys. Insecticide susceptibility of An. stephensi was tested using permethrin 0.75%, lambdacyhalothrin 0.05%, deltamethrin 0.05%, and DDT 4%, following the World Health Organization (WHO) test procedures for insecticide resistance monitoring. Additionally, knockdown resistance (kdr) mutation in the voltage-gated sodium channel (vgsc) gene was sequenced and analysed among resistant populations to detect possible molecular mechanisms of observed resistance phenotypes.ResultsThe susceptibility status of An. stephensi revealed that resistance to DDT and permethrin was found in all districts. Furthermore, resistance to all tested insecticides in An. stephensi was detected in Gabrik, Sardasht, Tidar, and Dehbarez. Analysis of knockdown resistance (kdr) mutations at the vgsc did not show evidence for the presence of this mutation in An. stephensi.ConclusionBased on the results of the current study, it appears that in An. stephensi from Hormozgan Province (Iran), other resistance mechanisms such as biochemical resistance due to detoxification enzymes may be involved due to the absence of the kdr mutation or non-target site resistance. Further investigation is warranted in the future to identify the exact resistance mechanisms in this main malaria vector across the country.

Insecticide resistance status and bar-codingof dengue vectors in three districts of Tamil Nadu, India.

Occurrence and distribution of vector population are crucial for entomological study in context of prevention, control and elimination of vector-borne diseases. To update some entomological aspects in three districts of Tamil Nadu state namely Kumbakonam, Nagapattinam and Thriuvarur districts, the study was undertaken. The objective of the study was to understand the prevalence of mosquitoes; to assess insecticide resistance and phylogenetic analysis of dengue vectors [Aedes (Ae.) aegypti and Ae.albopictus]. The immature stages of mosquitoes were collected from different localities by standard WHO methods marking with GPS and mapping was done using ArcGIS 10.4 software for all three districts. Insecticide resistance test was conducted using WHO susceptibility test kits. The F1 generation of female adult mosquitoes of Ae. aegypti and Ae. albopictus were exposed to DDT 4% and Malathion 5% with the control paper of Risella oil and olive oil respectively. Further, genomic DNA of individual mosquito was isolated, and the sequencingwas done through Eurofins, Bangalore, India. The FASTA sequence was analyzed and the phylogenic tree was constructed using the Maximum likelihood method in Molecular Evolutionary Genetics Analysis (MEGA) software (version 10.0). A totally 5307 specimens were collected through expanded survey in all three study areas. The collection yielded 16 species from six genera of mosquitoes. In total collection, the Ae. albopictus was the dominant species in Kumbakonam and Thiruvarur districts and Ae. aegypti was dominant in Nagapattinam district. The predominant breeding sources were discarded tyre with rainwater, plastic cups, coconut shells, aluminum vessels, sliver containers, bottles, grinding stones and earthen pots etc. The study revealed high pupal indices in all three study areas. Insecticide resistance monitoring revealed possible resistance in Ae. aegypti against DDT in all three districts whereas against Malathion, possible resistance was recorded in Kumbakonam and Nagapattinam and in Thiruvarur district, the species was found to be susceptible. Ae. albopictus showed resistance against DDT in all three districts but susceptible to Malathion. The sequences obtained for Dengue vectors showed 99% similar with Genbank. The phylogenetic tree was constructed using COI region sequences. Certainly, observed the different genetic relationship among Ae. aegypti and Ae. albopictus between the study areas. The study confirmed the presence of Ae. aegypti and Ae. albopictus in all three districts. The study further revealed that these vectors are susceptible to Malathion but resistance to DDT. The continue surveillance of dengue vector and monitoring of insecticide resistance will strengthen the control programme for appropriate vector control measurements.

Monitoring insecticide resistance in Bemisia tabaci mitotypes in the South Punjab region of Pakistan

Monitoring insecticide resistance in Bemisia tabaci mitotypes in the South Punjab region of Pakistan

Potential of MALDI-TOF MS biotyping to detect deltamethrin resistance in the dengue vector Aedes aegypti.

Insecticide resistance in mosquitoes is spreading worldwide and represents a growing threat to vector control. Insecticide resistance is caused by different mechanisms including higher metabolic detoxication, target-site modification, reduced penetration and behavioral changes that are not easily detectable with simple diagnostic methods. Indeed, most molecular resistance diagnostic tools are costly and labor intensive and then difficult to use for routine monitoring of insecticide resistance. The present study aims to determine whether mosquito susceptibility status against the pyrethroid insecticides (mostly used for mosquito control) could be established by the protein signatures of legs and/or thoraxes submitted to MALDI-TOF Mass Spectrometry (MS). The quality of MS spectra for both body parts was controlled to avoid any bias due to unconformity protein profiling. The comparison of MS profiles from three inbreeds Ae. aegypti lines from French Guiana (IRF, IR03, IR13), with distinct deltamethrin resistance genotype / phenotype and the susceptible reference laboratory line BORA (French Polynesia), showed different protein signatures. On both body parts, the analysis of whole protein profiles revealed a singularity of BORA line compared to the three inbreeding lines from French Guiana origin, suggesting that the first criteria of differentiation is the geographical origin and/or the breeding history rather than the insecticide susceptibility profile. However, a deeper analysis of the protein profiles allowed to identify 10 and 11 discriminating peaks from leg and thorax spectra, respectively. Among them, a specific peak around 4870 Da was detected in legs and thoraxes of pyrethroid resistant lines compared to the susceptible counterparts hence suggesting that MS profiling may be promising to rapidly distinguish resistant and susceptible phenotypes. Further work is needed to confirm the nature of this peak as a deltamethrin resistant marker and to validate the routine use of MS profiling to track insecticide resistance in Ae. aegypti field populations.

Insecticide Susceptibilities and Enzyme Activities of Four Stink Bug Populations in Mississippi, USA.

In Mississippi, the Pentatomidae complex infesting soybean is primarily composed of Euschistus servus, Nezara viridula, Chinavia hilaris, and Piezodorus guildinii. This study employed spray bioassays to evaluate the susceptibilities of these stink bugs to seven commonly used formulated insecticides: oxamyl, acephate, bifenthrin, λ-cyhalothrin, imidacloprid, thiamethoxam, and sulfoxaflor. Stinks bugs were collected from soybeans in Leland, MS, USA during 2022 and 2023, as well as from wild host plants in Clarksdale, MS. There was no significant difference in the susceptibility of C. hilaris to seven insecticides between two years, whereas P. guildinii showed slightly increased susceptibility to neonicotinoids in 2023. Among all four stink bug species, susceptibility in 2022 was ranked as P. guildinii ≤ C. hilaris ≈ N. viridula, while in 2023, it was ranked as P. guildinii ≤ C. hilaris ≤ E. Servus. Additionally, populations of E. servus and P. guildinii collected from Clarksdale exhibited high tolerance to pyrethroids and neonicotinoids. Moreover, populations of E. servus and P. guildinii from SIMRU-2022 and Clarksdale-2023 showed elevated esterase and cytochrome P450 activity, respectively. These findings from spray bioassays and enzyme activity analyses provide a baseline for monitoring insecticide resistance in Pentatomidae and can guide insecticide resistance management strategies for Mississippi soybean.

A Framework for Unsupervised Profiling of Malaria Vectors' Insecticide Resistance Using Machine Learning Technique.

Background: There is a need to identify different insecticide resistance profiles that represent circumscription-encapsulation of knowledge about malaria vectors' insecticide resistance to increase our understanding of malaria vectors' insecticide resistance dynamics. Methods: Data used in this study are part of the aggregation of over 20,000 mosquito collections done between 1957 and 2018. We applied two data preprocessing steps. We developed three clustering machine learning models based on the K-means algorithm with three selected datasets. The elbow method was used to fine-tune the hyperparameters. We used the silhouette score to assess the clustering results produced by each of the three models. The proposed framework incorporates continuous learning, allowing the machine learning models to learn continuously. Results: For the first model, the optimal number of clusters (profiles) k was 17. For the second model, we found four profiles. For the third model, the optimal number of profiles was 7. Discussion: We found that the insecticide resistance profiles have dynamic resistance levels with respect to the insecticide component, species component, location component, and time component. This profiling task provided knowledge about the evolution of malaria vectors' insecticide resistance in the African continent by encapsulating the information on the complex interaction between the different dimensions of malaria vectors' insecticide resistance into different profiles. Policy makers can use the knowledge about the different profiles found from the analysis of available insecticide resistance monitoring data (through profiling) by using our proposed approach to set up malaria vector control strategies that consider the locations, species present in those locations, and potentially efficient insecticides.

Effects of mosquito age and batch size on knockdown and mortality of laboratory-reared Anopheles dirus, Anopheles minimus, and wild-caught Anopheles harrisoni (Diptera: Culicidae) exposed to transfluthrin using WHO tube and CDC bottle bioassays.

WHO tube and CDC bottle bioassays are currently available for insecticide resistance monitoring and malaria transmission research. Multiple parameters including mosquito density, age, and nutritional status may affect the readout in these bioassays' tests. This study aims to assess the effects of experimental factors on knockdown and mortality measurements in dominant malaria vectors in Thailand following exposure to sublethal and lethal doses of transfluthrin. The effects of (i) 3 different mosquito batch sizes (5, 10, and 20 individuals) and (ii) 2 age groups (3-5 and 20-23 days old) on outcomes measured using the WHO tube (14.7 µg/cm2) and CDC bottle bioassay discriminating concentration (0.006 µg/cm2) against 2 laboratory strains: Anopheles dirus Peyton & Harrison and Anopheles minimus Theobald (species A) and wild-caught Anopheles harrisoni Harbach & Manguin (species C). Our results showed higher knockdown at 1-h exposure using WHO tube and CDC bottle bioassays containing 20 individuals compared to batches containing 10 and 5 individuals. Older mosquitoes showed greater susceptibility than younger test population, especially for An. mininus. Our study supports WHO recommendations for using 3- to 5-day-old mosquitoes. It also validates Praulin et al. (2022) proposal to divide the cohort into smaller batches with more test replicates when it is not practicable to test 25 mosquitoes per replicate.

Malaria transmission potential of Anopheles gambiae s.l. in indoor residual spraying areas with clothianidin 50 WG in northern Benin

The study objective was to assess the frequency of the kdr-L995F and ace-1 G280S genetic mutations in Anopheles gambiae s.l. mosquitoes and examine their ability to transmit Plasmodium falciparum in areas where indoor residual spraying (IRS) was implemented with Clothianidin 50 WG. The study was conducted in six communes in the Alibori and Donga departments of which four were IRS-treated and two were untreated and served as control. Post-IRS monthly samples of adult mosquitoes were collected in study communes using human landing catches (HLC). An. gambiae s.l. specimens were processed to detect kdr-L995F and ace-1 G280S mutations via PCR as well as Plasmodium falciparum infectivity through CSP ELISA. Our data revealed a high and similar allelic frequency for the kdr-L995F mutation in both treated and control communes (79% vs. 77%, p = 0.14) whilst allelic frequency of the ace-1 G280S mutation was lower across the study area (2–3%, p = 0.58). The sporozoite rate was 2.6% and 2.4% respectively in treated and untreated communes (p = 0.751). No association was found between Plasmodium falciparum infection in Anopheles gambiae s.l. vectors and carriage of kdr-L995F and ace-1 G280S mutations regardless of genotypes. The study findings underline the need for an integrated approach to malaria control, combining different control methods to effectively target transmission. Regular monitoring of insecticide resistance and genetic mutations is essential to guide control strategies.

Molecular monitoring of insecticide resistance in major disease vectors in Armenia

BackgroundArmenia is considered particularly vulnerable to life-threatening vector-borne diseases (VBDs) including malaria, West Nile virus disease and leishmaniasis. However, information relevant for the control of the vectors of these diseases, such as their insecticide resistance profile, is scarce. The present study was conducted to provide the first evidence on insecticide resistance mechanisms circulating in major mosquito and sand fly populations in Armenia.MethodsSampling sites were targeted based mainly on previous historical records of VBD occurrences in humans and vertebrate hosts. Initially, molecular species identification on the collected vector samples was performed. Subsequently, molecular diagnostic assays [polymerase chain reaction (PCR), Sanger sequencing, PCR-restriction fragment length polymorphism (RFLP), quantitative PCR (qPCR)] were performed to profile for major insecticide resistance mechanisms, i.e. target site insensitivity in voltage-gated sodium channel (vgsc) associated with pyrethroid resistance, acetylcholinesterase (ace-1) target site mutations linked to organophosphate (OP) and carbamate (CRB) resistance, chitin synthase (chs-1) target site mutations associated with diflubenzuron (DFB) resistance and gene amplification of carboxylesterases (CCEs) associated with resistance to the OP temephos.ResultsAnopheles mosquitoes were principally represented by Anopheles sacharovi, a well-known malaria vector in Armenia, which showed no signs of resistance mechanisms. Contrarily, the knockdown resistance (kdr) mutations V1016G and L1014F/C in the vgsc gene were detected in the arboviral mosquito vectors Aedes albopictus and Culex pipiens, respectively. The kdr mutation L1014S was also detected in the sand fly, vectors of leishmaniasis, Phlebotomus papatasi and P. tobbi, whereas no mutations were found in the remaining collected sand fly species, P. sergenti, P. perfiliewi and P. caucasicus.ConclusionsThis is the first study to report on molecular mechanisms of insecticide resistance circulating in major mosquito and sand fly disease vectors in Armenia and highlights the need for the establishment of systematic resistance monitoring practices for the implementation of evidence-based control applications.Graphical

Resistance of mosquitoes to Lambda-Cyhalothrin and DDT in a Niger Delta Region of Nigeria.

The escalation of insecticide resistance across the World Health Organization (WHO) African region calls for the routine monitoring of insecticide resistance in mosquitoes. While pyrethroids are recommended for use and dichlorodiphenyltrichloroethane (DDT) has been prescribed, it is imperative to reevaluate their efficacy across diverse geographical settings. The extent of resistance among mosquitoes to DDT and lambda-cyhalothrin remains understudied in Ika North East, Delta State, Nigeria, where the yellow fever virus was reported in 2019. We investigated the susceptibility of female mosquitoes to DDT (4%) and lambda-cyhalothrin (0.05%) in this specific study area. Adult female mosquitoes of Anopheles gambiae, Culex quinquefasciatus, and Aedes albopictus, aged between 2 and 4 days, were subjected to these insecticides utilizing the WHO bioassay method. The assessment of knockdown was done between 10 and 60 min, then mortality after 24 h. C. quinquefasciatus and A. albopictus mosquitoes exposed to DDT and lambda-cyhalothrin exhibited high susceptibility, resulting in complete mortality (100%); however, A. gambiae displayed resistance, with mortality rates of 19% and 76%, respectively . Notably, A. gambiae mosquitoes exposed to piperonyl butoxide (PBO)-DDT and PBO-lambda-cyhalothrin exhibited enhanced mortality, reaching 95% (indicating suspected resistance) and 100% (indicating susceptibility), respectively. Knockdown time (KDT) for 50% in mosquitoes exposed to lambda-cyhalothrin ranged from 3.94 to 33.51 min. Similarly, KDT model for 95% ranged from 19.04 to 84.15 min. Among the tested mosquito species, Culex mosquitoes exhibited the shortest knockdown resistance time for lambda-cyhalothrin, recorded at 3.94 min. Similarly, the KDT for DDT ranged from 24.97 to 187.06 min for 50% mortality and from 61.04 to 431.03 min for 95% mortality, respectively. Anopheles mosquitoes exposed to PBO + DDT recorded the lowest KDT. Our study underscores the potential of exercising caution in the use of DDT and lambda-cyhalothrin insecticides for the control of mosquitoes due to emerging resistance.

Fine-scale monitoring of insecticide resistance in Aedes aegypti (Diptera: Culicidae) from Sri Lanka and modeling the phenotypic resistance using rational approximation

BackgroundThe unplanned and intensified use of insecticides to control mosquito-borne diseases has led to an upsurge of resistance to commonly used insecticides. Aedes aegypti, the main vector of dengue, chikungunya, and Zika virus, is primarily controlled through the application of adulticides (pyrethroid insecticides) and larvicides (temephos). Fine spatial-scale analysis of resistance may reveal important resistance-related patterns, and the application of mathematical models to determine the phenotypic resistance status lessens the cost and usage of resources, thus resulting in an enhanced and successful control program.MethodsThe phenotypic resistance for permethrin, deltamethrin, and malathion was monitored in the Ae. aegypti populations using the World Health Organization (WHO) adult bioassay method. Mosquitoes' resistance to permethrin and deltamethrin was evaluated for the commonly occurring base substitutions in the voltage-gated sodium channel (vgsc) gene. Rational functions were used to determine the relationship between the kdr alleles and the phenotypic resistant percentage of Ae. aegypti in Sri Lanka.ResultsThe results of the bioassays revealed highly resistant Ae. aegypti populations for the two pyrethroid insecticides (permethrin and deltamethrin) tested. All populations were susceptible to 5% malathion insecticide. The study also revealed high frequencies of C1534 and G1016 in all the populations studied. The highest haplotype frequency was detected for the haplotype CC/VV, followed by FC/VV and CC/VG. Of the seven models obtained, this study suggests the prediction models using rational approximation considering the C allele frequencies and the total of C, G, and P allele frequencies and phenotypic resistance as the best fits for the area concerned.ConclusionsThis is the first study to our knowledge to provide a model to predict phenotypic resistance using rational functions considering kdr alleles. The flexible nature of the rational functions has revealed the most suitable association among them. Thus, a general evaluation of kdr alleles prior to insecticide applications would unveil the phenotypic resistance percentage of the wild mosquito population. A site-specific strategy is recommended for monitoring resistance with a mathematical approach and management of insecticide applications for the vector population.Graphical

Baseline susceptibility of Anopheles gambiae to clothianidin in northern Ghana

BackgroundClothianidin, an insecticide with a novel mode of action, has been deployed in the annual indoor residual spraying programme in northern Ghana since March 2021. To inform pragmatic management strategies and guide future studies, baseline data on local Anopheles gambiae sensu lato (s.l.) susceptibility to the clothianidin insecticide were collected in Kpalsogu, a village in the Northern region, Ghana.MethodsPhenotypic susceptibility of An. gambiae mosquitoes to clothianidin was assessed using the World Health Organization (WHO) insecticide resistance monitoring bioassay. The WHO cone bioassays were conducted on mud and cement walls sprayed with Sumishield 50 wettable granules (WG) (with clothianidin active ingredient). Daily mortalities were recorded for up to 7 days to observe for delayed mortalities. Polymerase chain reaction (PCR) technique was used to differentiate the sibling species of the An. gambiae complex and also for the detection of knock down resistance genes (kdr) and the insensitive acetylcholinesterase mutation (ace-1).ResultsThe WHO susceptibility bioassay revealed a delayed killing effect of clothianidin. Mosquitoes exposed to the cone bioassays for 5 min died 120 h after exposure. Slightly higher mortalities were observed in mosquitoes exposed to clothianidin-treated cement wall surfaces than mosquitoes exposed to mud wall surfaces. The kdr target-site mutation L1014F occurred at very high frequencies (0.89–0.94) across all vector species identified whereas the ace-1 mutation occurred at moderate levels (0.32–0.44). Anopheles gambiae sensu stricto was the most abundant species observed at 63%, whereas Anopheles arabiensis was the least observed at 9%.ConclusionsAnopheles gambiae s.l. mosquitoes in northern Ghana were susceptible to clothianidin. They harboured kdr mutations at high frequencies. The ace-1 mutation occurred in moderation. The results of this study confirm that clothianidin is an effective active ingredient and should be utilized in malaria vector control interventions.

Susceptibility status and synergistic activity of DDT and Lambda-cyhalothrin on Anopheles gambiae and Aedes aegypti in Delta State, Nigeria.

The detection of insecticide resistance in male mosquitoes has been treated with less importance in monitoring insecticide resistance spread in mosquitoes. There are no studies on the susceptibility and synergistic activity of DDT and lambda-cyhalothrin on male Anopheles gambiae and Aedes aegypti in Delta State, Nigeria. Even though studies have extensively reported resistance in female mosquitoes, the susceptibility of male mosquitoes to insecticide classes should be ascertained. In this study, we tested the susceptibility status and synergistic activity of DDT and Lambda-cyhalothrin on An. gambiae and Ae. aegypti in Delta State, Nigeria, in order to ascertain the level of resistance and knockdown. In addition, we modelled the knockdown time using Probit analysis model. WHO bioassay method was used to expose two days old adult mosquitoes to 4% DDT and 0.05% lambda-cyhalothrin. The results showed that An. gambiae mosquitoes exposed to DDT and lambda-cyhalothrin were confirmed resistant (61% and 53% respectively). However, pre-exposing the resistant mosquito population to piperonyl butoxide (4%) showed an increase in mortality to 90% (possible resistance) in DDT and 98% (susceptible) in lambda-cyhalothrin. Ae. aegypti mosquitoes exposed to DDT were susceptible (98%) while those exposed to lambda-cyhalothrin were confirmed resistant (87%) and this increased to complete mortality (100%) in PBO+lambda-cyhalothrin population. Furthermore, the results showed that the knockdown time (KDT50 and KDT95) in An. gambiae exposed to DDT was 39.5-71.2 minutes and 124.5-146.4 minutes respectively, while that of lambda-cyhalothrin was 33.0-81.8 minutes and 64.0-124.4 minutes respectively. In Ae. aegypti, KDT50 and KDT95 was 23.9 and 61.7minutes for DDT exposure whereas it was 5.6-15.3 minutes and 36.1-72.3 minutes for lambda-cyhalothrin exposure. It can be concluded that male An. gambiae mosquitoes exposed to the insecticides were resistant and the causes may be linked to certain resistant genes in the mosquitoes. The chances of transferring resistance are possible in wild species and molecular-based studies on the resistant gene in male mosquitoes as well as the tendencies of transfer are required to establish this focus.

Insecticide resistance monitoring in Korean local populations of diamondback moth (Plutella xylostella) (I)

Insecticide resistance monitoring in Korean local populations of diamondback moth (Plutella xylostella) (I)