Growing problems of pyrethroid resistance in Anopheles funestus have intensified efforts to identify alternative insecticides. Many agrochemicals target the GABA receptors, but cross-resistance from dieldrin resistance may preclude their introduction.Dieldrin resistance was detected in An. funestus populations from West (Burkina Faso) and central (Cameroon) Africa, but populations from East (Uganda) and Southern Africa (Mozambique and Malawi) were fully susceptible to this insecticide. Partial sequencing of the dieldrin target site, the γ-aminobutyric acid (GABA) receptor, identified two amino acid substitutions, A296S and V327I. The A296S mutation has been associated with dieldrin resistance in other species. The V327I mutations was detected in the resistant sample from Burkina Faso and Cameroon and consistently associated with the A296S substitution. The full-length of the An. funestus GABA-receptor gene, amplified by RT-PCR, generated a sequence of 1674 bp encoding 557 amino acid of the protein in An. funestus with 98% similarity to that of Anopheles gambiae. Two diagnostic assays were developed to genotype the A296S mutation (pyrosequencing and PCR-RFLP), and use of these assays revealed high frequency of the resistant allele in Burkina Faso (60%) and Cameroon (82%), moderate level in Benin (16%) while low frequency or absence of the mutation was observed respectively in Uganda (7.5%) or 0% in Malawi and Mozambique.The distribution of the RdlR mutation in An. funestus populations in Africa suggests extensive barriers to gene flow between populations from different regions.

BackgroundThe insecticide resistance status of the malaria vector Anopheles funestus and the underlying resistance mechanisms remain uncharacterised in many parts of Africa, notably in Benin, West Africa. To fill this gap in our knowledge, we assessed the susceptibility status of a population of this species in Pahou, Southern Benin and investigated the potential resistance mechanisms.Methodology/Principal FindingsWHO bioassays revealed a multiple resistance profile for An. funestus in Pahou. This population is highly resistant to DDT with no mortality in females after 1h exposure to 4%DDT. Resistance was observed against the Type I pyrethroid permethrin and the carbamate bendiocarb. A moderate resistance was detected against deltamethrin (type II pyrethroids). A total susceptibility was observed against malathion, an organophosphate. Pre-exposure to PBO did not change the mortality rates for DDT indicating that cytochrome P450s play no role in DDT resistance in Pahou. No L1014F kdr mutation was detected but a correlation between haplotypes of two fragments of the Voltage-Gated Sodium Channel gene and resistance was observed suggesting that mutations in other exons may confer the knockdown resistance in this species. Biochemical assays revealed elevated levels of GSTs and cytochrome mono-oxygenases in Pahou. No G119S mutation and no altered acetylcholinesterase gene were detected in the Pahou population. qPCR analysis of five detoxification genes revealed that the GSTe2 is associated to the DDT resistance in this population with a significantly higher expression in DDT resistant samples. A significant over-expression of CYP6P9a and CYP6P9b previously associated with pyrethroid resistance was also seen but at a lower fold change than in southern Africa.ConclusionThe multiple insecticide resistance profile of this An. funestus population in Benin shows that more attention should be paid to this important malaria vector for the implementation and management of current and future malaria vector control programs in this country.

BackgroundMalaria burden is high in Nigeria, yet information on the major mosquito vectors is lacking especially in the Sudan savannah region of the country. In order to facilitate the design of future insecticide-based control interventions in the region, this study has established the resistance profile of An. gambiae s.l. populations in two northern Nigeria locations and assessed the contribution of target site resistance mutations.MethodsLarval collection was conducted in two localities in Sudan savannah (Bunkure and Auyo) of northern Nigeria between 2009 and 2011, from which resulting adult, female mosquitoes were used for insecticides bioassays with deltamethrin, lambda-cyhalothrin, DDT and malathion. The mosquitoes were identified to species level and molecular forms and then genotyped for the presence of L1014F-kdr, L1014S-kdr and ace-1R mutations.ResultsWHO bioassays revealed that An. gambiae s.l. from both localities were highly resistant to lambda-cyhalothrin and DDT, but only moderately resistant to deltamethrin. Full susceptibility was observed to malathion. An. gambiae, M form (now An. coluzzii), was predominant over An. arabiensis in Auyo and was more resistant to lambda-cyhalothrin than An. arabiensis. No ‘S’ form (An. gambiae s.s.) was detected. A high frequency of 1014 F mutation (80.1%) was found in An. coluzzii in contrast to An. arabiensis (13.5%). The presence of the 1014 F kdr allele was significantly associated with resistance to lambda-cyhalothrin in An. coluzzii (OR = 9.85; P < 0.001) but not in An. arabiensis. The L1014S-kdr mutation was detected in a single An. arabiensis mosquito while no ace-1R mutation was found in any of the mosquitoes analysed.ConclusionsThe predominance of An. coluzzii and its resistance profile to main insecticides described in this study can guide the implementation of appropriate vector control interventions in this region of Nigeria where such information was previously lacking.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2334-14-441) contains supplementary material, which is available to authorized users.

The susceptibility of Aedes aegypti Linnaeus 1762, Culex quinquefasciatus Say 1823 and Anopheles gambiae Giles 1902 was tested on 0.75 % permethrin pyrethroid in the laboratory. All Larval and pupal stages of the mosquitoes were collected from different localities within Zaria, Nigeria and reared to adulthood in the laboratory. The adults that emerged were tested on 0.75 % permethrin strips using WHO Standard Test Kit. Mortalities were recorded at five minutes interval for 1 hr and maintained for 24hrs post-exposure on 7 % sucrose solution after which a final mortality was recorded. The Knockdown times (KDT50 and KDT95) were determined by Probit Analysis. 100 % mortality was obtained for Ae. aegypti and Cx. quinquefasciatus after 1hr and 24hr exposures, respectively, whereas An. gambiae recorded 92.11 % and 90.91 % mortalities after 1hr and 24hr exposure, respectively. The KDT50 and KDT95 did not differ significantly (P>0.05) between Ae. aegypti and Cx. quinquefasciatus but differed significantly (P<0.05) with An. gambiae. The study shows that Ae. aegypti and C. quinquefasciatus are susceptible to permethrin whereas An. gambiae is resistant to the insecticide in the area.

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