Western Kenyan Anopheles gambiae s.s. showing intense permethrin resistance harbor distinct microbiota

AbstractBackgroundInsecticide resistance poses a growing challenge to malaria vector control in Kenya and around the world. Following evidence of associations between the mosquito microbiota and insecticide resistance, we comparatively characterized the microbiota of An. gambiae s.s. from Tulukuyi village, Bungoma, Kenya, with differing permethrin resistance profiles.MethodsUsing the CDC bottle bioassay, 133 2-3 day-old, virgin, non-blood fed female F1 progeny of field-caught An. gambiae s.s. were exposed to five times (107.5μg/ml) the discriminating dose of permethrin. Post bioassay, 50 resistant and 50 susceptible mosquitoes were subsequently screened for kdr East and West mutations, and individually processed for microbial analysis using high throughput sequencing targeting the universal bacterial and archaeal 16S rRNA gene.Results47% of the samples tested (n=133) were resistant, and of the 100 selected for further processing, 99% were positive for kdr East and 1% for kdr West. Overall, 84 bacterial taxa were detected across all mosquito samples, with 36 of these shared between resistant and susceptible mosquitoes. A total of 20 were unique to the resistant mosquitoes and 28 were unique to the susceptible mosquitoes. There were significant differences in bacterial composition between resistant and susceptible individuals (F=2.33, P=0.001), with presence of Sphingobacterium, Lysinibacillus and Streptococcus (all known pyrethroid-degrading taxa), and the radiotolerant Rubrobacter, being significantly associated with resistant mosquitoes. On the other hand, the presence of Myxococcus, was significantly associated with susceptible mosquitoes.ConclusionThis is the first report of distinct microbiota in An. gambiae s.s. associated with intense pyrethroid resistance. The findings highlight differentially abundant bacterial taxa between resistant and susceptible mosquitoes, and further suggest a microbe-mediated mechanism of insecticide resistance in mosquitoes. Our results also indicate fixation of the kdr East mutation in this mosquito population, precluding further analysis of its associations with the mosquito microbiota, but presenting the hypothesis that any microbe-mediated mechanism of insecticide resistance would be likely of a metabolic nature. Overall, this study lays initial groundwork for understanding microbe-mediated mechanisms of insecticide resistance in African malaria vectors, and potentially identifying novel microbial markers of insecticide resistance that could supplement existing vector surveillance tools.

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Western Kenyan Anopheles gambiae showing intense permethrin resistance harbour distinct microbiota

Malaria Journal ◽

10.1186/s12936-021-03606-4 ◽

2021 ◽

Vol 20 (1) ◽

Cited By ~ 1

Author(s):

Diana Omoke ◽

Mathew Kipsum ◽

Samson Otieno ◽

Edward Esalimba ◽

Mili Sheth ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

High Throughput Sequencing ◽

Pyrethroid Resistance ◽

Sensu Stricto ◽

Malaria Vector Control ◽

Rrna Gene ◽

Vector Surveillance ◽

Bacterial Composition ◽

Microbial Analysis

Abstract Background Insecticide resistance poses a growing challenge to malaria vector control in Kenya and around the world. Following evidence of associations between the mosquito microbiota and insecticide resistance, the microbiota of Anopheles gambiae sensu stricto (s.s.) from Tulukuyi village, Bungoma, Kenya, with differing permethrin resistance profiles were comparatively characterized. Methods Using the CDC bottle bioassay, 133 2–3 day-old, virgin, non-blood fed female F1 progeny of field-caught An. gambiae s.s. were exposed to five times (107.5 µg/ml) the discriminating dose of permethrin. Post bioassay, 50 resistant and 50 susceptible mosquitoes were subsequently screened for kdr East and West mutations, and individually processed for microbial analysis using high throughput sequencing targeting the universal bacterial and archaeal 16S rRNA gene. Results 47 % of the samples tested (n = 133) were resistant, and of the 100 selected for further processing, 99 % were positive for kdr East and 1 % for kdr West. Overall, 84 bacterial taxa were detected across all mosquito samples, with 36 of these shared between resistant and susceptible mosquitoes. A total of 20 bacterial taxa were unique to the resistant mosquitoes and 28 were unique to the susceptible mosquitoes. There were significant differences in bacterial composition between resistant and susceptible individuals (PERMANOVA, pseudo-F = 2.33, P = 0.001), with presence of Sphingobacterium, Lysinibacillus and Streptococcus (all known pyrethroid-degrading taxa), and the radiotolerant Rubrobacter, being significantly associated with resistant mosquitoes. On the other hand, the presence of Myxococcus, was significantly associated with susceptible mosquitoes. Conclusions This is the first report of distinct microbiota in An. gambiae s.s. associated with intense pyrethroid resistance. The findings highlight differentially abundant bacterial taxa between resistant and susceptible mosquitoes, and further suggest a microbe-mediated mechanism of insecticide resistance in mosquitoes. These results also indicate fixation of the kdr East mutation in this mosquito population, precluding further analysis of its associations with the mosquito microbiota, but presenting the hypothesis that any microbe-mediated mechanism of insecticide resistance would be likely of a metabolic nature. Overall, this study lays initial groundwork for understanding microbe-mediated mechanisms of insecticide resistance in African mosquito vectors of malaria, and potentially identifying novel microbial markers of insecticide resistance that could supplement existing vector surveillance tools.

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Current status of insecticide resistance in malaria vectors in the Asian countries: a systematic review

F1000Research ◽

10.12688/f1000research.46883.2 ◽

2022 ◽

Vol 10 ◽

pp. 200

Author(s):

Dewi Susanna ◽

Dian Pratiwi

Keyword(s):

Systematic Review ◽

Insecticide Resistance ◽

Vector Control ◽

Malaria Vector ◽

Pyrethroid Resistance ◽

Current Trend ◽

Current Status ◽

Malaria Vectors ◽

Malaria Vector Control ◽

Publication Date

Background: The application of insecticides for malaria vector control has led to a global problem, which is the current trend of increased resistance against these chemicals. This study aimed to review the insecticide resistance status was previously determined in Asia and how to implement the necessary interventions. Moreover, the implications of resistance in malaria vector control in this region were studied. Methods: This systematic review was conducted using a predefined protocol based on PRISMA-retrieved articles from four science databases, namely ProQuest, Science Direct, EBSCO, and PubMed in the last ten years (2009 to 2019). The searching process utilized four main combinations of the following keywords: malaria, vector control, insecticide, and Asia. In ProQuest, malaria control, as well as an insecticide, were used as keywords. The following criteria were included in the filter, namely full text, the source of each article, scholarly journal, Asia, and publication date as in the last ten years. Results: There were 1408 articles retrieved during the initial search (ProQuest=722, Science Direct=267, EBSCO=50, PubMed=285, and Scopus=84). During the screening, 27 articles were excluded because of duplication, 1361 based on title and abstract incompatibility with the inclusion criteria, and 20 due to content differences. In the final screening process, 15 articles were chosen to be analyzed. From the 15 articles, it is known that there was organochlorine (DDT), organophosphate (malathion), and pyrethroids resistance in several Anopheles species with a less than 80% mortality rate. Conclusion: This review found multiple resistance in several Anopheles includes resistance to pyrethroid. The reports of pyrethroid resistance were quite challenging because it is considered effective in the malaria vector control. Several countries in Asia are implementing an insecticide resistance management (IRM) strategy against malaria vectors following the Global Plan for IRM.

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A pyrethroïd-treated bed net increases host attractiveness for Anopheles gambiae s.s. carrying the kdr allele in a dual-choice olfactometer

10.1101/077552 ◽

2016 ◽

Cited By ~ 1

Author(s):

Angélique Porciani ◽

Malal Diop ◽

Nicolas Moiroux ◽

Tatiana Kadoke-Lambi ◽

Anna Cohuet ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Sub Saharan Africa ◽

Malaria Vectors ◽

Malaria Vector Control ◽

Anopheles Gambiae S.S ◽

Host Seeking ◽

Seeking Behavior ◽

Sub Saharan ◽

The Impact

AbstractThe use of long lasting insecticide nets (LLINs) treated with pyrethroïd is known for its major contribution in malaria control. However, LLINs are suspected to induce behavioral changes in malaria vectors, which may in turn drastically affect their efficacy against Plasmodium sp. transmission. In sub Saharan Africa, where malaria imposes the heaviest burden, the main malaria vectors are widely resistant to pyrethroïds, the insecticide family used on LLINs, which also threatens LLIN efficiency. There is therefore a crucial need for deciphering how insecticide-impregnated materials might affect the host-seeking behavior of malaria vectors in regards to insecticide resistance. In this study, we explored the impact of permethrin-impregnated net on the host attractiveness for Anopheles gambiae mosquitoes, either susceptible to insecticides, or carrying the insecticide resistance conferring allele kdr. Groups of female mosquitoes were released in a dual-choice olfactometer and their movements towards an attractive odor source (a rabbit) protected by insecticide-treated (ITN) or untreated nets (UTN) were monitored. Kdr homozygous mosquitoes, resistant to insecticides, were more attracted by a host behind an ITN than an UTN, while the presence of insecticide on the net did not affect the choice of susceptible mosquitoes. These results suggest that permethrin-impregnated net is detectable by malaria vectors and that the kdr mutation impacts their response to a LLIN protected host. We discuss the implication of these results for malaria vector control.

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Impact of insecticide resistance in Anopheles arabiensis on malaria incidence and prevalence in Sudan and the costs of mitigation

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1713814114 ◽

2017 ◽

Vol 114 (52) ◽

pp. E11267-E11275 ◽

Cited By ~ 12

Author(s):

Hmooda Toto Kafy ◽

Bashir Adam Ismail ◽

Abraham Peter Mnzava ◽

Jonathan Lines ◽

Mogahid Shiekh Eldin Abdin ◽

...

Keyword(s):

Insecticide Resistance ◽

Pyrethroid Resistance ◽

Malaria Incidence ◽

Disease Incidence ◽

Indoor Residual Spraying ◽

Sub Saharan Africa ◽

Malaria Vectors ◽

Incidence And Mortality ◽

Additional Protection ◽

The Impact

Insecticide-based interventions have contributed to ∼78% of the reduction in the malaria burden in sub-Saharan Africa since 2000. Insecticide resistance in malaria vectors could presage a catastrophic rebound in disease incidence and mortality. A major impediment to the implementation of insecticide resistance management strategies is that evidence of the impact of resistance on malaria disease burden is limited. A cluster randomized trial was conducted in Sudan with pyrethroid-resistant and carbamate-susceptible malaria vectors. Clusters were randomly allocated to receive either long-lasting insecticidal nets (LLINs) alone or LLINs in combination with indoor residual spraying (IRS) with a pyrethroid (deltamethrin) insecticide in the first year and a carbamate (bendiocarb) insecticide in the two subsequent years. Malaria incidence was monitored for 3 y through active case detection in cohorts of children aged 1 to <10 y. When deltamethrin was used for IRS, incidence rates in the LLIN + IRS arm and the LLIN-only arm were similar, with the IRS providing no additional protection [incidence rate ratio (IRR) = 1.0 (95% confidence interval [CI]: 0.36–3.0; P = 0.96)]. When bendiocarb was used for IRS, there was some evidence of additional protection [interaction IRR = 0.55 (95% CI: 0.40–0.76; P < 0.001)]. In conclusion, pyrethroid resistance may have had an impact on pyrethroid-based IRS. The study was not designed to assess whether resistance had an impact on LLINs. These data alone should not be used as the basis for any policy change in vector control interventions.

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Phenotypic Insecticide Resistance in Anopheles gambiae (Diptera: Culicidae): Specific Characterization of Underlying Resistance Mechanisms Still Matters

Journal of Medical Entomology ◽

10.1093/jme/tjaa195 ◽

2020 ◽

Author(s):

Adandé A Medjigbodo ◽

Luc S Djogbenou ◽

Aubin A Koumba ◽

Laurette Djossou ◽

Athanase Badolo ◽

...

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Resistance Mechanisms ◽

Malaria Vectors ◽

Effective Control ◽

Anopheles Coluzzii ◽

Anopheles Gambiae S.S ◽

Diagnostic Dose ◽

Pirimiphos Methyl ◽

Old Adult

Abstract An effective control of malaria vectors requires an extensive knowledge of mechanisms underlying the resistance-phenotypes developed by these vectors against insecticides. We investigated Anopheles gambiae mosquitoes from Benin and Togo for their intensity of insecticide resistance and we discussed the involvement of genotyped mechanisms in the resistance-phenotypes observed. Three- to five-day-old adult mosquitoes emerged from field and laboratory An. gambiae larvae were assayed using WHO tube intensity tests against various doses of deltamethrin: 1× (0.05%); 2× (0.1%); 5× (0.25%); 7.5× (0.375%) and those of pirimiphos-methyl: 0.5× (0.125%); 1× (0.25%). Members of An. gambiae complex were screened in field populations using polymerase chain reaction (PCR) assays. The presence of kdrR(1014F/1014S) and ace-1R(119S) mutations was also investigated using TaqMan and PCR-RFLP techniques, respectively. Anopheles gambiae from field were very resistant to deltamethrin, whereas KisKdr and AcerKdrKis strains displayed 100% mortality rates at 2× the diagnostic dose. In contrast, the field mosquitoes displayed a low resistance-intensity against 1× the diagnostic dose of pirimiphos-methyl, whereas AcerKis and AcerKdrKis strains showed susceptibility at 0.5× the diagnostic dose. Anopheles gambiae s.s., Anopheles coluzzii, and Anopheles arabiensis were identified. Allelic frequencies of kdrR (1014F) and ace-1R (119S) mutations in the field populations varied from 0.65 to 1 and 0 to 0.84, respectively. The field An. gambiae displayed high-resistance levels against deltamethrin and pirimiphos-methyl when compared with those of the laboratory An. gambiae-resistant strains. These results exhibit the complexity of underlying insecticide resistance mechanisms in these field malaria vectors.

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PCR-RFLP Detection and Genogroup Identification of Piscirickettsia salmonis in Field Samples

Pathogens ◽

10.3390/pathogens9050358 ◽

2020 ◽

Vol 9 (5) ◽

pp. 358

Author(s):

Pamela Aravena ◽

Rodrigo Pulgar ◽

Javiera Ortiz-Severín ◽

Felipe Maza ◽

Alexis Gaete ◽

...

Keyword(s):

High Throughput Sequencing ◽

In Silico Analysis ◽

Cost Effective ◽

Rrna Gene ◽

Bacterial Composition ◽

Bacterial Dna ◽

Phenotypic Differences ◽

Field Samples ◽

Pcr Rflp ◽

Digestion Pattern

Piscirickettsia salmons, the causative agent of piscirickettsiosis, is genetically divided into two genomic groups, named after the reference strains as LF-89-like or EM-90-like. Phenotypic differences have been detected between the P. salmonis genogroups, including antibiotic susceptibilities, host specificities and pathogenicity. In this study, we aimed to develop a rapid, sensitive and cost-effective assay for the differentiation of the P. salmonis genogroups. Using an in silico analysis of the P. salmonis 16S rDNA digestion patterns, we have designed a genogroup-specific assay based on PCR-restriction fragment length polymorphism (RFLP). An experimental validation was carried out by comparing the restriction patterns of 13 P. salmonis strains and 57 field samples obtained from the tissues of dead or moribund fish. When the bacterial composition of a set of field samples, for which we detected mixtures of bacterial DNA, was analyzed by a high-throughput sequencing of the 16S rRNA gene amplicons, a diversity of taxa could be identified, including pathogenic and commensal bacteria. Despite the presence of mixtures of bacterial DNA, the characteristic digestion pattern of the P. salmonis genogroups could be detected in the field samples without the need of a microbiological culture and bacterial isolation.

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Addressing Key Gaps in Implementation of Mosquito Larviciding to Accelerate Malaria Vector Control in Southern Tanzania: Results of a Stakeholder Engagement Process in Local District Councils

10.21203/rs.3.rs-98596/v1 ◽

2020 ◽

Author(s):

Salum Abdallah Mapua ◽

Marceline F. Finda ◽

Ismail H. Nambunga ◽

Betwel J. Msugupakulya ◽

Kusirye Ukio ◽

...

Keyword(s):

Malaria Control ◽

Public Engagement ◽

Program Effectiveness ◽

Indoor Residual Spraying ◽

Malaria Vectors ◽

Malaria Vector Control ◽

Production Plant ◽

Vector Surveillance ◽

Community Members ◽

Key Informants

Abstract Background: Larval source management was historically one of the most effective malaria control methods but is now widely deprioritized in Africa, where insecticide treated nets (ITNs) and indoor residual spraying (IRS) are preferred. However, in Tanzania, following initial successes in urban Dar-es-Salaam starting early-2000s, the government now encourages larviciding in both rural and urban councils nationwide to complement other efforts; and a biolarvicide production-plant has been established outside the commercial capital. This study investigated key obstacles and opportunities relevant to effective rollout of larviciding for malaria control, with a focus on the meso-endemic region of Morogoro, southern Tanzania. Methods: Key-informants were interviewed to assess awareness and perceptions regarding larviciding among designated health officials (malaria focal persons, vector surveillance officers and ward health officers) in nine administrative councils (N = 27). Interviewer-administered questionnaires were used to assess awareness and perceptions of community members in selected areas regarding larviciding (N = 490). Thematic content analysis was done and descriptive statistics used to summarize the findings.Results: A majority of malaria control officials had participated in larviciding at least once over the previous three years. A majority of community members had neutral perceptions towards positive aspects of larviciding, but overall support for larviciding was high, although several challenges were expressed, notably: i) insufficient knowledge for identifying relevant aquatic habitats of malaria vectors and applying larvicides, ii) inadequate monitoring of program effectiveness, iii) limited financing, and iv) lack of personal protective equipment. Although the key-informants reported sensitizing local communities, most community members were still unaware of larviciding and its potential. Conclusion: The larviciding program was widely supported by both communities and malaria control officials, but there were gaps in technical knowledge, implementation and public engagement. To improve overall impact, it is important to: i) intensify training efforts, particularly for identifying habitats of important vectors, ii) adopt standard technical principles for applying larvicides, iii) improve financing for local implementation and iv) improve public engagement to boost community awareness and participation. These lessons could also be valuable for other malaria endemic areas wishing to deploy larviciding for malaria control or elimination.

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Insecticide resistance characteristic of Anopheles vector species successfully controlled by deployment of pyrethroid and PBO long lasting insecticidal treated nets (LLINs) in Tanzania

10.1101/2021.03.19.436139 ◽

2021 ◽

Author(s):

Johnson Matowo ◽

David Weetman ◽

Patricia Pignatell ◽

Alexandra Wright ◽

Jacques Charlwood ◽

...

Keyword(s):

Insecticide Resistance ◽

Pyrethroid Resistance ◽

Cluster Randomized Trial ◽

World Health ◽

Malaria Vectors ◽

Metabolizing Enzymes ◽

Sub Saharan ◽

Underlying Mechanisms ◽

Cluster Randomized ◽

Health Organization

Long lasting insecticidal nets (LLINs) are a proven tool to reduce malaria transmission, but in Africa efficacy is being reduced by pyrethroid resistance in the major vectors. A cluster randomized trial in Muleba district, Tanzania demonstrated that permethrin LLINs co-treated with piperonyl butoxide (PBO), a synergist that can block pyrethroid-metabolizing enzymes in the mosquito, had much greater efficacy than pyrethroid-only nets. Insecticide resistance profiles and underlying mechanisms were investigated in Anopheles gambiae and An. funestus from Muleba during the trial. Diagnostic dose bioassays using permethrin, together with intensity assays, suggest pyrethroid resistance that is both strong and very common, but not extreme. Transcriptomic analysis found multiple P450 genes over expressed including CYP6M2, CYP6Z3, CYP6P3, CYP6P4, CYP6AA1 and CYP9K1 in An. gambiae and CYP6N1, CYP6M7, CYP6M1 and CYP6Z3 in An. funestus. Indeed, very similar suites of P450 enzymes commonly associated with resistant populations elsewhere in Africa were detected as over expressed suggesting a convergence of mechanisms across Sub-Saharan African malaria vectors. The findings give insight into factors that may correlate with pyrethroid PBO LLIN success, broadly supporting model predictions, but revision to guidelines previously issued by the World Health Organization is warranted.

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Relative Contributions of Genetic and Non-Genetic Factors on Mosquito Microbiota

10.21203/rs.3.rs-38080/v1 ◽

2020 ◽

Cited By ~ 1

Author(s):

Haikel Nasser Bogale ◽

Matthew V. Cannon ◽

Kalil Keita ◽

Denka Camara ◽

Yaya Barry ◽

...

Keyword(s):

Insecticide Resistance ◽

Bacterial Diversity ◽

Blood Meal ◽

High Throughput Sequencing ◽

Genetic Factors ◽

Resistance Locus ◽

Microbial Composition ◽

Bacterial Composition ◽

Relative Contribution ◽

Main Driver

Abstract Background The commensal microbiota of mosquitoes impacts their development, immunity, and competency, and could provide a target for alternative entomological control approaches. However, despite the importance of the mosquito/microbiota interactions, little is known about the relative contribution of genetic and non-genetic factors in shaping the bacterial communities of mosquitoes. Methods We used a high-throughput sequencing based assay to characterize the bacterial composition and diversity of 665 individual field-caught mosquitoes, as well as their species, genotype at an insecticide resistance locus, blood meal composition, and the eukaryotic parasites and viruses they carry. We then used these data to rigorously estimate the individual effect of each parameter on the bacterial diversity as well as their relative contribution to the microbial composition. Results Overall, multivariate analyses did not reveal any significant contribution of the mosquito species, insecticide resistance or blood meal to the bacterial composition of the mosquitoes surveyed. Infection with parasites and viruses only contributed very marginally and the main driver of the bacterial diversity was the location where each mosquito was collected which explained roughly 20% of the variance observed. Conclusions This analysis shows that, when confounding factors are taken into account, the sites where the mosquitoes are collected is the main driver of the bacterial diversity of wild-caught mosquitoes, although further studies will be needed to determine how the specific components of the local environment affecting the bacterial composition.

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Monitoring of Insecticide Resistance in Anopheles culicifacies in Twelve Districts of Madhya Pradesh, Central India (2017–2019)

Journal of Tropical Medicine ◽

10.1155/2022/4404027 ◽

2022 ◽

Vol 2022 ◽

pp. 1-8

Author(s):

Ashok K. Mishra ◽

Praveen K Bharti ◽

Gyan Chand ◽

Aparup Das ◽

Himanshu Jayswar ◽

...

Keyword(s):

Insecticide Resistance ◽

Vector Control ◽

Indoor Residual Spraying ◽

Central India ◽

Control Programme ◽

Madhya Pradesh ◽

Control Measures ◽

Malaria Vectors ◽

Malaria Vector Control ◽

The Impact

Background. Indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs) are malaria vector control measures used in India, but the development of insecticide resistance poses major impediments for effective vector control strategies. As per the guidelines of the National Vector Borne Disease Control Programme (NVBDCP), the study was conducted in 12 districts of Madhya Pradesh to generate data on insecticide resistance in malaria vectors. Methods. The susceptibility tests were conducted on adult An. culicifacies as per the WHO standard technique with wild-caught mosquitoes. The blood-fed female mosquitoes were exposed in 3 to 4 replicates on each occasion to the impregnated papers with specified discriminating dosages of the insecticides (DDT: 4%, malathion: 5%, deltamethrin: 0.05%, and alphacypermethrin: 0.05%), for one hour, and mortality was recorded after 24-hour holding. Results. An. culicifacies was found resistant to DDT 4% in all the 12 districts and malathion in 11 districts. The resistance to alphacypermethrin was also observed in two districts, and possible resistance was found to alphacypermethrin in seven districts and to deltamethrin in eight districts, while the vector was found susceptible to both deltamethrin and alphacypermethrin in only 3 districts. Conclusion. An. culicifacies is resistant to DDT and malathion and has emerging resistance to pyrethroids, alphacypermethrin, and deltamethrin. Therefore, regular monitoring of insecticide susceptibility in malaria vectors is needed for implementing effective vector management strategies. However, studies to verify the impact of IRS with good coverage on the transmission of disease are required before deciding on the change of insecticide in conjunction with epidemiological data.

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