Anopheles Gambiae Research Articles

The effect of larval exposure to acids and detergents on the life history of the major malaria vector Anopheles arabiensis Patton (Diptera: Culicidae).

Anopheles arabiensis, a highly adaptable member of the Anopheles gambiae complex, poses a challenge for control efforts due to its outdoor biting and resting behaviour. Consequently, indoor insecticide-based control methodsare ineffective against An. arabiensis. Furthermore, An. arabiensis are adapting to breeding in polluted waters, and may be contributing to residual malaria and malaria in urban areas. There have been some advances in understanding the effect of rural pollutants on Anopheles mosquitoes, but the effect of urban pollutants is poorly understood. Thus, in this study, the effect of acidic pollutants [nitric acid (HNO3) and hydrochloric acid (HCl)] and alkaline pollutants (phosphate-free and phosphate-containing detergent) on two laboratory-reared An. arabiensis strains - an insecticide susceptible strain (SENN) and an insecticide-resistant strain selected from SENN (SENN-DDT) - were determined. The median lethal concentration (LC50) and larval exposure on larval development, adult longevity and insecticide tolerance were evaluated. Nitric acid and phosphate-containing detergent were found to be more toxic than HCl and phosphate-free detergent in terms of LC50 values. Detergent exposure (both phosphate-containing and phosphate-free) increased adult longevity of both strains. Nitric acid reduced larval development time in both SENN and SENN-DDT, whereas HCl reduced larval development time in SENN only. By contrast, both phosphate-containing and phosphate-free detergents increased larval development time of both strains. Furthermore, HNO3 and phosphate-containing detergent increased insecticide tolerance the most. The two An. arabiensis strains responded to urban pollutants differently. Thus, this study provides insight into the adaptation of An. arabiensis to acidic and alkaline urban pollutants. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The bioefficacy of a novel VECTRON™ T500 indoor residual spray formulation in an experimental huts trial against Anopheles gambiae s.l. populations

Background: The emergence of insecticide resistance and its spread through populations of malaria vectors has decreased the number of insecticides available for control. Insecticide resistance has been observed in vector populations across sub-Saharan Africa in malaria endemic areas. Therefore, new compounds with different modes of action are needed that can be used in the management of resistance. The current study assessed the bioefficacy of the new indoor residual spray formulation, VECTRON™ T500 against Klypson 500 WG and water against laboratory reared and wild populations of Anopheles gambiae s.l.. The comparative experimental hut trial was implemented between June 2022 and December 2022 to determine the efficacy of VECTRON™ T500, containing the active ingredient (ai) broflanilide as a 50 % wettable powder (WP). The efficacy of VECTRON™ T500 was compared with a positive control, Klypson 500 WG, a wettable granule (WG) formulation that contains 50 % clothianidin. Cement and mud walls were sprayed with VECTRON™ T500, Klypson 500 WG with water sprayed as a negative control. The two insecticides and negative control were evaluated monthly for six months against laboratory and the field-derived An. gambiae s.l. using the standard WHO cone bioassays. Each wall had two cones on each day of testing. VECTRON™ T500 was sprayed on both surface types at a rate of 100mg ai/m2 whilst Klypson 500 WG was applied at a rate of 300 mg ai/m2. For both wall surface types, the vector from the laboratory-reared and the wild populations exhibited a low knockdown effect to both VECTRON™ T500 and Klypson 500 WG. A total of 3,840 mosquitoes were used of which 2,880 (75 %) were susceptible colony of An. gambiae s.s. and 960 (25 %) were wild An. gambiae s.l.. The VECTRON™ T500 induced a mortality of 97.8 %-98.1 % in the laboratory population and 83.2-95.0 % wild population mosquitoes on cement and mud-walled huts respectively while Klypson 500 WG ranged from 89.6-99.0 % for wild and 99.0-99.3 % for the laboratory population mosquitoes on cement and mud walls respectively. The knockdown due to VECTRON™ T500 was 7.08 % in the sixth month, while for Klypson 500 WG was 16.04-17.50 %. The monthly wall cone bioassay mortality with VECTRON™ T500 remained over 80 % for 6 months post-spraying for both laboratory and wild populations. The findings of this study have shown VECTRON™ T500 to have extended efficacy against malaria vector mosquitoes when applied to cement and mud walls. The evaluated new IRS formulation, VECTRON™ T500, performed equally with the positive control, Klypson 500 WG, regarding its impact on vector mortality.

Mosquitoes integrate visual and acoustic cues to mediate conspecific interactions in swarms

Male mosquitoes form aerial aggregations, known as swarms, to attract females and maximize their chances of finding a mate. Within these swarms, individuals must be able to recognize potential mates and navigate the social environment to successfully intercept a mating partner. Prior research has almost exclusively focused on the role of acoustic cues in mediating the male mosquito's ability to recognize and pursue females. However, the role of other sensory modalities in this behavior has not been explored. Moreover, how males avoid collisions with one another in the swarm while pursuing females remains poorly understood. In this study, we combined free-flight and tethered-flight simulator experiments to demonstrate that swarming Anopheles coluzzii mosquitoes integrate visual and acoustic information to track conspecifics and avoid collisions. Our tethered experiments revealed that acoustic stimuli gated mosquito steering responses to visual objects simulating nearby mosquitoes, especially in males that exhibited a strong response toward visual objects in the presence of female flight tones. Additionally, we observed that visual cues alone could trigger changes in mosquitoes' wingbeat amplitude and frequency. These findings were corroborated by our free-flight experiments, which revealed that Anopheles coluzzii modulate their thrust-based flight responses to nearby conspecifics in a similar manner to tethered animals, potentially allowing for collision avoidance within swarms. Together, these results demonstrate that both males and females integrate multiple sensory inputs to mediate swarming behavior, and for males, the change in flight kinematics in response to multimodal cues might allow them to simultaneously track females while avoiding collisions.

Susceptibility status of female anopheles’ mosquito to organophosphate and carbamate insecticides in market environs in Itu LGA, Akwa Ibom State, Nigeria

Malaria remains a serious public health problem and is still responsible for death of individuals globally with the female Anopheles gambiae established to be one of the major vectors which aid the transmission of this disease. This study examined the susceptibility and resistance of An. gambiae to carbamate and organophosphate which has not been frequently used in vector control in Nigeria. The CDC (Centres for Disease Control and Prevention) bottle bioassay was employed to measure the susceptibility of An. gambiae to insecticides. The procedure involved 2–5-day old An. gambiae adult mosquito populations starved from blood. In this study, we employed a diagnostic time of 30 minutes to administer doses of 12.5µg/bottle of Bendiocarb and Propoxur (Carbamates) and 20µg Pirimiphos-methyl (Organophosphate) to Anopheles spp. populations. The knockdown effect of the three insecticides indicated that bendiocarb and propoxur (carbamates) had a mortality of 98% while pirimiphos-methyl (organophosphate) had 81% at 30 minutes, regarded as the HUH for measuring resistance/susceptibility of insecticides. Further exposure at 60 minutes yielded 100% mortality for both bendiocarb and propoxur while 98% mortality was applicable in pirimiphos-methyl insecticides. An. gambiae was susceptible to bendiocarb and propoxur insecticides and resistant to pirimiphos-methyl. Invariably, vector control decision-making in the area will be influenced by the result of the study being that it is a baseline data/information. It is paramount that before any vector control strategies are implemented, sufficient susceptibility/resistance data be taken into thought

Genetic characterisation of candidate ecdysteroid kinases in Drosophila melanogaster.

Ecdysteroids are major hormones in insects and control moulting, growth, reproduction, physiology, and behaviour. The biosynthesis of ecdysteroids such as 20-hydroxyecdysone (20E) from dietary sterols is well characterised, but ecdysteroid catabolism is poorly understood. Ecdysteroid kinases (EcKs) mediate the reversible phosphorylation of ecdysteroids, which has been implicated in ecdysteroid recycling during embryogenesis and reproduction in various insects. However, to date only two EcK-encoding genes have been identified, in the silkworm Bombyx mori and the mosquito Anopheles gambiae. Previously, we identified two ecdysteroid kinase-like (EcKL) genes-Wallflower (Wall) and Pinkman (pkm)-in the model fruit fly Drosophila melanogaster that are orthologs of the ecdysteroid 22-kinase gene BmEc22K. Here, using gene knockdown, knockout and misexpression, we explore Wall and pkm's possible functions and genetically test the hypothesis that they encode EcKs. Wall and pkm null mutants are viable and fertile, suggesting they are not essential for development or reproduction, whereas phenotypes arising from RNAi and somatic CRISPR appear to derive from off-target effects or other artefacts. However, misexpression of Wall results in dramatic phenotypes, including developmental arrest, and defects in trachea, cuticle and pigmentation. Wall misexpression fails to phenocopy irreversible ecdysteroid catabolism through misexpression of Cyp18a1, suggesting Wall does not directly inactivate 20E. Additionally, Wall misexpression phenotypes are not attenuated in Cyp18a1 mutants, strongly suggesting Wall is not an ecdysteroid 26-kinase. We hypothesise that the substrate of Wall in this misexpression experiment and possibly generally is an unknown, atypical ecdysteroid that plays essential roles in Drosophila development, and may highlight aspects of insect endocrinology that are as-yet uncharacterised. We also provide preliminary evidence that CG5644 encodes an ecdysteroid 22-kinase conserved across Diptera.

Community participatory mapping of malaria mosquito breeding sites in Mozambique

BackgroundThe community involvement and the people’s knowledge allow detailed information about the distribution, location, and identification of mosquito breeding-sites. Information which is fundamental for their efficient management and elimination. Since participatory mapping has proven to be an effective tool to identify health determinants, the study aimed to apply the methodology to identify and map potential mosquito breeding-sites in Tambai, Nhamatanda, Mozambique.MethodsA study was conducted using an open-question guide. Discussions were held with 94 participants within ten focus groups, selected in collaboration with local community leaders. A thematic content analysis was performed. Descriptive statistics were used to characterize sociodemographic data. Geographic Positioning System (GPS) was used to compare and map potential breeding-sites. Children under 5 years of age who tested positive for malaria, were georeferenced to the maps.ResultsParticipants were aware of causes and transmission of malaria, no major differences between groups were observed regarding knowledge and identification of principal potential breeding sites. Gender and age determined specific information, number, and diversity of identified potential breeding sites. A total of 125 potential breeding-sites (36 permanent and 89 temporary) were mapped.ConclusionsSeveral potential mosquito breeding-sites were identified, located throughout the community, often near house conglomerates and malaria cases. Community participatory mapping could be used to identify potential mosquito breeding-sites by the national malaria control programmes to establish an efficient larval surveillance system, while improving community engagement and control strategies.Trial registration: ClinicalTrials.gov ID: NCT04419766.

First detection of kdr L1014F allele in Anopheles ziemanni and Anopheles pharoensis in Cameroon and distribution of the allele in members of the Anopheles gambiae complex

BackgroundKnockdown resistance (kdr) is one of the primary resistance mechanisms present in anopheline species. Although this mutation is largely spread across the Anopheles gambiae s.l. members, its prevalence in other species is still not well documented.MethodsThe present study investigated the distribution and allelic frequencies of kdr in An. gambiae s.l., An. pharoensis, and An. ziemanni samples collected in 2022 and 2023 in nine sites spread across five ecogeographical settings in Cameroon. Members of the An. gambiae complex were identified molecularly by polymerase chain reaction (PCR). kdr L1014F and L1014S alleles were screened by PCR and confirmed by sequencing.ResultsAn. gambiae (49.9%), An. coluzzii (36.5%), and An. arabiensis (13%) were identified, and the frequency of the kdr L1014F was high in both An. gambiae and An. coluzzii in all sites. The kdr L1014F allele was detected for the first time in 8 out of 14 An. ziemanni samples examined and in 5 out of 22 An. pharoensis samples examined. The kdr L1014S allele was scarce and found only in the heterozygote “RS” state in An. arabiensis and An. gambiae in Yangah and Santchou.ConclusionsThe present study sheds light on the rapid expansion of the kdr L1014F allele in malaria vectors in Cameroon and stresses the need for surveillance activities also targeting secondary malaria vectors to improve the control of malaria transmission.Graphical

Uridine diphosphate (UDP)-glycosyltransferases (UGTs) are associated with insecticide resistance in the major malaria vectors Anopheles gambiae s.l. and Anopheles funestus

Malaria remains one of the highest causes of morbidity and mortality, with 249 million cases and over 608,000 deaths in 2022. Insecticides, which target the Anopheles mosquito vector, are the primary method to control malaria. The widespread nature of resistance to the most important insecticide class, the pyrethroids, threatens the control of this disease. To reverse the stall in malaria control there is urgent need for new vector control tools, which necessitates understanding the molecular basis of pyrethroid resistance. In this study we utilised multi-omics data to identify uridine-diphosphate (UDP)-glycosyltransferases (UGTs) potentially involved in resistance across multiple Anopheles species. Phylogenetic analysis identifies sequence similarities between Anopheline UGTs and those involved in agricultural pesticide resistance to pyrethroids, pyrroles and spinosyns. Expression of five UGTs was characterised in An. gambiae and An. coluzzii to determine constitutive over-expression, induction, and tissue specificity. Furthermore, a UGT inhibitor, sulfinpyrazone, restored susceptibility to pyrethroids and DDT in An. gambiae, An. coluzzii, An. arabiensis and An. funestus, the major African malaria vectors. Taken together, this study provides clear association of UGTs with pyrethroid resistance as well as highlighting the potential use of sulfinpyrazone as a novel synergist for vector control.

Larvicidal and Synergistic Properties of Fixed Oils Derived from Plants and their Capacity to inhibit Pupation and Emergence of Malaria Vector (Anopheles gambiae)

Malaria vectors have evolved resistance to almost all WHO-recommended insecticides, which compromises vector control. This study aimed to evaluate the insecticidal activity of four doses of fixed oils from Jatropha curcas and Ricinus communis on Anopheles gambiae in western Burkina Faso. Biological tests of susceptibility to oil doses were carried out on two populations of the An. gambiae complex from August to October 2022 using the WHO standard protocol. The synergistic or antagonistic effects of the extracts’ combinations were evaluated by comparing “sums of effects” to “effects of sums” according to the WHO protocol. R software enabled probit and ANOVA analyses. All oils showed larvicidal activity on susceptible strains of An. gambiae, Kisumu. The LC50 of the combination of the two oils was lower (54.09±1.03 ppm), followed by the oil of J. curcas (58.8±1.03 ppm) and that of R. communis (139.0±1.04 ppm) on the field strain of An. gambiae. J. curcas oil was more toxic on both strains, leading to 100% mortality at 48h and 72h of exposure. Synergistic insecticidal effects after 24h and additive effects after 48h of the combined oil at 50 and 150 ppm resulted in 41.75% to 91.66% mortality of larvae and reduced pupation from 2.66% to 0.00% and reduced the emergence of An. gambiae from 1.16 to 0.00%. J. curcas and R. communis’ oils contain linalool, tannins, alkaloids, saponins, and terpenes. Applying J. curcas or R. communis oil or a combined oil at 50 and 150 ppm as a spray could constitute an effective strategy for integrated control of An. gambiae mosquitoes.

Longitudinal survey of insecticide resistance in a village of central region of Burkina Faso reveals co-occurrence of 1014F, 1014S and 402L mutations in Anopheles coluzzii and Anopheles arabiensis

BackgroundPyrethroid resistance is one of the major threats for effectiveness of insecticide-treated bed nets (ITNs) in malaria vector control. Genotyping of mutations in the voltage gated sodium channel (VGSC) gene is widely used to easily assess the evolution and spread of pyrethroid target-site resistance among malaria vectors. L1014F and L1014S substitutions are the most common and best characterized VGSC mutations in major African malaria vector species of the Anopheles gambiae complex. Recently, an additional substitution involved in pyrethroid resistance, i.e. V402L, has been detected in Anopheles coluzzii from West Africa lacking any other resistance alleles at locus 1014. The evolution of target-site resistance mutations L1014F/S and V402L was monitored in An. coluzzii and Anopheles arabiensis specimens from a Burkina Faso village over a 10-year range after the massive ITN scale-up started in 2010.MethodsAnopheles coluzzii (N = 300) and An. arabiensis (N = 362) specimens collected both indoors and outdoors by different methods (pyrethrum spray catch, sticky resting box and human landing collections) in 2011, 2015 and 2020 at Goden village were genotyped by TaqMan assays and sequencing for the three target site resistance mutations; allele frequencies were statistically investigated over the years.ResultsA divergent trend in resistant allele frequencies was observed in the two species: 1014F decreased in An. coluzzii (from 0.76 to 0.52) but increased in An. arabiensis (from 0.18 to 0.70); 1014S occurred only in An. arabiensis and slightly decreased over time (from 0.33 to 0.23); 402L increased in An. coluzzii (from 0.15 to 0.48) and was found for the first time in one An. arabiensis specimen. In 2020 the co-occurrence of different resistance alleles reached 43% in An. coluzzii (alleles 410L and 1014F) and 32% in An. arabiensis (alleles 1014F and 1014S).ConclusionsOverall, an increasing level of target-site resistance was observed among the populations with only 1% of the two malaria vector species being wild type at both loci, 1014 and 402, in 2020. This, together with the co-occurrence of different mutations in the same specimens, calls for future investigations on the possible synergism between resistance alleles and their phenotype to implement local tailored intervention strategies.

Semi-field evaluation of electrocuting eave tubes for the control of endophagic mosquitoes in south-east Tanzania

BackgroundEave spaces are major entry points through which malaria vectors enter houses. Interventions that target mosquitoes at the eaves have recently been developed. However, most of these interventions are based on insecticides for which resistance has been reported. Here we evaluated the efficacy of mosquito electrocuting eave tubes (MEETs) against Anopheles gambiae sensu stricto (An. gambiae s.s.) and Anopheles funestus s.s. under semi-field conditions.MethodsExperiments were conducted in two semi-field chambers, each containing one experimental hut. Six electrocuting eave tubes were installed in each hut to assess their impact on laboratory-reared An. gambiae s.s. and An. funestus s.s.. Each species was assessed separately over 10 nights by releasing 200 unfed females per night into each chamber. One volunteer slept in each hut from 7 p.m. to 5 a.m. Mosquitoes were collected indoors and outdoors using mouth and Prokopack aspirators.ResultsThe placement of MEETs significantly reduced the nightly An. gambiae s.s. indoor and outdoor biting, by 21.1% and 37.4%, respectively. Indoor-biting An. funestus s.s. were reduced by 87.5% while outdoor-biting numbers of An. funestus s.s. declined by 10.4%.ConclusionsMEETs represent a promising tool for controlling mosquitoes at the point of house entry. Further validation of their potential under natural field conditions is necessary. Several advantages over insecticide-based eave tubes are indicated and discussed in this article.Graphical

Field-evolved resistance to neonicotinoids in the mosquito, Anopheles gambiae, is associated with downregulation and mutations of nicotinic acetylcholine receptor subunits combined with cytochrome P450-mediated detoxification.

Neonicotinoid insecticides act selectively on their nicotinic receptor targets leading to variable sensitivity among arthropods. This study aimed to investigate the molecular mechanisms underlying contrasting susceptibility to neonicotinoids observed in wild populations of two mosquito sibling species. Bioassays and a synergism test revealed that the sister taxa, Anopheles gambiae and An. coluzzii, from Yaounde, Cameroon, rely on cytochrome P450s to detoxify neonicotinoids and develop resistance. However, contrary to An. coluzzii, An. gambiae populations are evolving stronger resistance to several active ingredients facilitated by mutations and reduced expression of nicotinic acetylcholine receptors. Six mutations were detected in coding sequences of the β1 and α6 subunits, including two substitutions in one of the loops that modulate ligand binding and sensitivity. Allele frequencies were strongly correlated with a susceptibility gradient between An. coluzzii and An. gambiae suggesting that the mutations may play a key role in sensitivity. Messenger RNA expression levels of the β1, α3, and α7 subunits decreased dramatically, on average by 23.27, 17.50, 15.80-fold, respectively, in wild An. gambiae populations compared to a susceptible insectary colony. By contrast, only the β2 and α9-1 subunits were moderately downregulated (5.28 and 2.67-fold change, respectively) in field-collected An. coluzzii adults relative to susceptible colonized mosquitoes. Our findings provide critical information for the application and resistance management of neonicotinoids in malaria prevention.

Ontogenetic transfer of microplastics in natural populations of malaria mosquitoes in Western Siberia

AbstractThe uptake, accumulation, and ontogenetic transfer of microplastics (MPs) in bloodsucking mosquitoes of the genus Anopheles, vectors of helminth and protozoa parasites in humans and animals, were studied under laboratory conditions. In the experiment, 2‐μm‐diameter fluorescent polystyrene spheres were counted in mosquitoes of all stages, from larvae to adults. Larvae from a natural population readily ingested MPs with food, accumulating on average 3.9 × 106 particles per larva within 3 days. The concentration of MPs decreased significantly in mosquitoes during their transition from stage to stage. The average number of detected MPs attained 110 particles per pupa and 3.0 particles per adult. MP uptake by mosquitoes did not affect their survival, whereas the rate of metamorphosis in MP‐consuming mosquitoes was slightly higher. Our data confirmed that MPs can be transferred from feeding larvae to non‐feeding pupae and adult flying Anopheles mosquitoes from natural populations in aquatic ecosystems.

A study on Anopheles gambiae s.l. biting activities in wonji sugar estate, Eastern Oromia Ethiopia: a challenge to the current intervention tools

A study on Anopheles gambiae s.l. biting activities in wonji sugar estate, Eastern Oromia Ethiopia: a challenge to the current intervention tools

Homing gene drives can transfer rapidly between Anopheles gambiae strains with minimal carryover of flanking sequences

CRISPR-Cas9 homing gene drives are designed to induce a targeted double-stranded DNA break at a wild type allele (‘recipient’), which, when repaired by the host cell, is converted to the drive allele from the homologous (‘donor’) chromosome. Germline localisation of this process leads to super-Mendelian inheritance of the drive and the rapid spread of linked traits, offering a novel strategy for population control through the deliberate release of drive individuals. During the homology-based DNA repair, additional segments of the recipient chromosome may convert to match the donor, potentially impacting carrier fitness and strategy success. Using Anopheles gambiae strains with variations around the drive target site, here we assess the extent and nature of chromosomal conversion. We show both homing and meiotic drive contribute as mechanisms of inheritance bias. Additionally, over 80% of homing events resolve within 50 bp of the chromosomal break, enabling rapid gene drive transfer into locally-adapted genetic backgrounds.

Multi-omics analysis of antiviral interactions of Elizabethkingia anophelis and Zika virus

The microbial communities residing in the mosquito midgut play a key role in determining the outcome of mosquito pathogen infection. Elizabethkingia anophelis, originally isolated from the midgut of Anopheles gambiae possess a broad-spectrum antiviral phenotype, yet a gap in knowledge regarding the mechanistic basis of its interaction with viruses exists. The current study aims to identify pathways and genetic factors linked to E. anophelis antiviral activity. The understanding of E. anophelis antiviral mechanism could lead to novel transmission barrier tools to prevent arboviral outbreaks. We utilized a non-targeted multi-omics approach, analyzing extracellular lipids, proteins, metabolites of culture supernatants coinfected with ZIKV and E. anophelis. We observed a significant decrease in arginine and phenylalanine levels, metabolites that are essential for viral replication and progression of viral infection. This study provides insights into the molecular basis of E. anophelis antiviral phenotype. The findings lay a foundation for in-depth mechanistic studies.

Larvicidal Potency of Ashes of Two Insecticidal Plants against the Activities of Anopheles coluzzii and Culex quinquefasciatus Mosquitoes

Larvicidal Potency of Ashes of Two Insecticidal Plants against the Activities of Anopheles coluzzii and Culex quinquefasciatus Mosquitoes

Host-specific effects of a generalist parasite of mosquitoes

Microsporidians are obligate parasites of many animals, including mosquitoes. Some microsporidians have been proposed as potential agents for the biological control of mosquitoes and the diseases they transmit due to their detrimental impact on larval survival and adult lifespan. To get a more complete picture of their potential use as agents of biological control, we measured the impact of Vavraia culicis on several life-history traits of Aedes aegypti and Anopheles gambiae. We measured the infection dynamics and clearance rate for the two species, and we assessed sexual dimorphism in infection dynamics within each species. Our results show differences in infection dynamics, with Ae. aegypti life-history traits being more affected during its aquatic stage and exhibiting higher clearance of the infection as adults. In contrast, An. gambiae was unable to clear the infection. Additionally, we found evidence of sexual dimorphism in parasite infection in An. gambiae, with males having a higher average parasite load. These findings shed light and improve our knowledge of the infection dynamics of V. culicis, a microsporidian parasite previously recognized as a potential control agent of malaria.

Organophosphate and carbamate susceptibility profiling of Anopheles gambiae sl. across different ecosystems in southern Benin

Background To overcome the spread of high pyrethroid resistance in the main malaria vectors and malaria disease persistence, it is crucial to look for effective and better resistance management strategies. Understanding the phenotypic profile of Anopheles gambiae sl. against alternatives insecticides like organophosphates and carbamates is crucial. Methods Anopheles larvae and pupae were collected from the breeding sites in rice fields, pineapple crop areas, and peri-urban areas. WHO susceptibility tests were conducted on unfed female mosquitoes aged 3–5 days old. Mosquitoes were exposed to malathion 5%, pirimiphos-methyl 0.25%, and bendiocarb 0.1% using the standard WHO protocol. Polymerase chain reaction (PCR) techniques were used to detect species, kdr and Ace-1 mutations. Results Anopheles gambiae sl. from Sèdjè-Dénou rice field population was resistant to bendiocarb (0.1%) with a mortality rate of 72.2% whereas Anopheles gambiae sl. populations from Zinvié-Dokomey (rice field), Zè-Tozounmè (pineapple field), and Adjagbo (peri-urban area) were suspected to be resistant with mortality rates of 90%, 93.5%, 95.4% respectively. However, all of them were susceptible to organophosphates (malathion and pirimiphos-methyl) with a mortality rate of 100%. PCR assay revealed that 100% of the mosquitoes tested were Anopheles coluzzii. The frequencies of Ace-1R mutation in all Anopheles coluzzii populations tested were low (3–27%). Conclusions Organophosphates (malathion and pirimiphos-methyl) have maintained their efficacy against Anopheles coluzzii populations from Sèdjè-Dénou (rice field), Zè Tozounmè (pineapple field), Zinvié Dokomey (rice field), or Adjagbo (peri-urban area). The good efficacy of these organophosphates against Anopheles coluzzii populations from the southern part of Benin are observed in the current study. The use of pirimiphos-methyl for IRS in this part of the country would be a successful alternative for malaria control in this area.

The expanded immunoregulatory protease network in mosquitoes is governed by gene co-expression.

Serine protease cascades regulate key innate immune responses. In mosquitoes, these cascades involve clip-domain serine proteases and their non-catalytic homologs (CLIPs), forming a complex network whose make-up and structural organization is not fully understood. This study assessed the impact of 85 CLIPs on humoral immunity in Anopheles gambiae. By coupling RNAi with assays measuring antimicrobial activity and melanization, we identified 27 CLIPs as immunoregulators that together form two distinct subnetworks. CLIPs regulating antimicrobial activity were found to control infection resistance, as knockdowns reduced bacterial load and improved survival. Furthermore, our analysis of CLIP gene expression unveiled a novel immunoregulatory mechanism reliant on protease baseline co-expression rather than infection-induced upregulation. These findings underscore that despite its complexity mosquito immune regulation may be targeted for malaria interventions.