Search

Increased levels of insecticide resistance in major malaria vectors such as Anopheles funestus threaten the effectiveness of insecticide-based control programmes. Understanding the landscape features impacting the spread of resistance makers is necessary to design suitable resistance management strategies. Here, we examined the influence of the highest mountain in West Africa (Mount Cameroon; 4095 m elevation) on the spread of metabolic and target-site resistance alleles in An . funestus populations. Vector composition varied across the four localities surveyed along the altitudinal cline with major vectors exhibiting high parity rate (80.5%). Plasmodium infection rates ranged from 0.79% ( An. melas ) to 4.67% ( An. funestus ). High frequencies of GSTe2 R (67-81%) and Rdl R (49-90%) resistance alleles were observed in An. funestus throughout the study area, with GSTe2 R frequency increasing with altitude, whereas the opposite is observed for Rdl R . Patterns of genetic diversity and population structure analyses revealed high levels of polymorphisms with 12 and 16 haplotypes respectively for GSTe2 and Rdl . However, the reduced diversity patterns of resistance allele carriers revealed signatures of positive selection on the two genes across the study area irrespective of the altitude. Despite slight variations associated with the altitude, the spread of resistance alleles suggest that control strategies could be implemented against malaria vectors across mountainous landscapes.

Growing insecticide resistance in malaria vectors is threatening the effectiveness of insecticide-based interventions, including Long Lasting Insecticidal Nets (LLINs). However, the impact of metabolic resistance on the effectiveness of these tools remains poorly characterized. Using experimental hut trials and genotyping of a glutathione S-transferase resistance marker (L119F- GSTe2 ), we established that GST-mediated resistance is reducing the efficacy of LLINs against Anopheles funestus . Hut trials performed in Cameroon revealed that Piperonyl butoxide (PBO)-based nets induced a significantly higher mortality against pyrethroid resistant An. funestus than pyrethroid-only nets. Blood feeding rate and deterrence were significantly higher in all LLINs than control. Genotyping the L119F- GSTe2 mutation revealed that, for permethrin-based nets, 119F- GSTe2 resistant mosquitoes have a greater ability to blood feed than susceptible while the opposite effect is observed for deltamethrin-based nets. For Olyset Plus, a significant association with exophily was observed in resistant mosquitoes (OR = 11.7; p < 0.01). Furthermore, GSTe2 -resistant mosquitoes (cone assays) significantly survived with PermaNet 2.0 (OR = 2.1; p < 0.01) and PermaNet 3.0 (side) (OR = 30.1; p < 0.001) but not for Olyset Plus. This study shows that the efficacy of PBO-based nets (e.g., blood feeding inhibition) against pyrethroid resistant malaria vectors could be impacted by other mechanisms including GST-mediated metabolic resistance not affected by the synergistic action of PBO. Mosaic LLINs incorporating a GST inhibitor (diethyl maleate) could help improve their efficacy in areas of GST-mediated resistance.

Background: Despite the increased report of insecticide resistance in malaria vectors, its impact on mosquito's life-traits after exposure to insecticide-treated nets remains under investigated. Here, we assessed the effects of exposure to PermaNet 2.0 on several life traits of An. gambiae s.l. and An. funestus s.l. field mosquitoes in Cameroon., Methodology: Female Anopheles mosquitoes were collected indoor using electric aspirators in southern Cameroon (Obout) in 2016. After assessing the resistance status of F1 from the field collected-mosquitoes, progeny of the first generation (An. funestus s.l.) and seventh generation (An. gambiae s.l.) were used to assess the long-term effect of exposure to PermaNet 2.0 on several life-traits of these vectors (longevity, blood feeding ability, fecundity and fertility) in comparison to untreated net. In addition, the L119F-GSTe2 mutation associated with DDT/pyrethroids resistance in An. funestus was genotyped to assess its association with increased life-span post-exposure., Principal Findings: Both An. funestus and An. gambiae were resistant to pyrethroids and DDT with a greater level in the latter. Pyrethroid-only nets PermaNet 2.0 (17.5% mortality) and Olyset (0% mortality) exhibited a significantly reduced efficacy against An. funestus in contrast to a greater efficacy for PBO-based Nets Olyset Plus (65% mortality), PermaNet 3.0 top (100% mortality). In both species, mosquitoes that survived exposure to PermaNet 2.0 exhibited a significantly reduced longevity than those non-exposed (6.95 days vs 12.46 for An. funestus P<0.001; 8.87 vs 11.25 days for An. gambiae; P<0.001). However, no significant difference was observed for blood feeding and fecundity in both species. In addition, molecular analysis of the L119F-GSTe2 mutation revealed that this mutation is associated with an increase in the chance of surviving after exposure to this net in An. funestus., Conclusions: These results show that although the PermaNet 2.0 presents a reduced efficacy against resistant populations, it remains efficient after exposure by reducing the life expectancy of the vectors which could contribute in the reduction of malaria incidence., Competing Interests: The authors have declared that no competing interests exist.

Background: The epidemiological profiles of vector-borne diseases, such as malaria, are strongly associated with landscape components. The reduction of malaria burden in endemic and epidemic regions mainly depends on knowledge of the malaria-transmitting mosquito species, populations and behavioural characteristics, as well as malaria exposure risks. This work aimed at carrying out a holistic study in order to characterise Anopheles species in relation to human malaria in seven wetlands along the lower section of the volcanic chain of Cameroon., Results: Eight malaria vectors: Anopheles arabiensis, An. coluzzii, An. funestus (s.s.), An. gambiae, An. hancocki, An. melas, An. nili and An. ziemanni, were found biting humans. Anopheles gambiae was widespread; however, it played a secondary role in the Ndop plain where An. ziemmani was the primary vector species (79.2%). Anophelines were more exophagic (73.6%) than endophagic (26.4%), showing a marked nocturnal activity (22:00-4:00 h) for An. coluzzii and An. gambiae while An. funestus (s.s.) was mostly caught between 1:00 and 6:00 h and An. ziemanni having an early evening biting behaviour (18:00-00:00 h). Female Anopheles were mostly observed to have relative high parity rates (≥ 70%), with the exception of the Meanja site where species parity varies from 46 to 55%. Overall, the transmission level was low with entomological inoculation rates estimated to 0.7 infected bites per person per month (ib/p/mth) in Tiko and Ndop, 1.4 ib/p/mth in Mamfe and 2.24 ib/p/mth in Santchou., Conclusions: The present study represents detailed Anopheles vector characterisation from an understudied area along the volcanic chain of Cameroon with endemic malaria transmission. The significant differences in bionomics and Anopheles species distribution within the studied wetlands, highlights the importance of providing baseline data and an opportunity to assess the outcome of ongoing malaria control interventions in the country.