The spread of insecticide resistance genes in Anopheles gambiae Giles sensu stricto threatens to compromise vector-based malaria control programs. Two mutations at the same locus in the voltage-gated sodium channel gene are known to confer knockdown resistance (kdr) to pyrethroids and DDT. Kdr-e involves a leucine-serine substitution, and it was until recently thought to be restricted to East Africa, whereas kdr-w, which involves a leucine-phenylalanine substitution, is associated with resistance in West Africa. In this study, we analyze the frequency and relationship between the kdr genotypes and resistance to type I and type II pyrethroids and DDT by using WHO test kits in both the Forest-M and S molecular forms of An. gambiae in Cameroon. Both kdr-w and kdr-e polymorphisms were found in sympatric An. gambiae, and in many cases in the same mosquito. Kdr-e and kdr-w were detected in both forms, but they were predominant in the S form. Both kdr-e and kdr-w were closely associated with resistance to DDT and weakly associated with resistance to type II pyrethroids. Kdr-w conferred greater resistance to permethrin than kdr-e. We also describe a modified diagnostic designed to detect both resistant alleles simultaneously.

BackgroundAs part of a study to determine the impact of insecticide resistance on the effectiveness of long-lasting insecticide treated nets (LLINs) in the north of Cameroon, the unexpectedly high density and anthropophilic behaviour of Anopheles rufipes lead us to investigate this species bionomics and role in human malaria parasite transmission.MethodsFor four consecutive years (2011–2014), annual cross-sectional sampling of adult mosquitoes was conducted during the peak malaria season (September-October) in three health districts in northern Cameroon. Mosquitoes sampled by human landing catch and pyrethrum spray catch methods were morphologically identified, their ovaries dissected for parity determination and Anopheles gambiae siblings were identified by molecular assay. Infection with P. falciparum and blood meal source in residual fauna of indoor resting anopheline mosquitoes were determined by enzyme-linked-immunosorbent assays.Results Anopheles gambiae (sensu lato) (s.l.) comprised 18.4% of mosquitoes collected with An. arabiensis representing 66.27% of the sibling species. The proportion of An. rufipes (2.7%) collected was high with a human-biting rate ranging between 0.441 and 11.083 bites/person/night (b/p/n) and an anthropophagic rate of 15.36%. Although overall the members of An. gambiae complex were responsible for most of the transmission with entomological inoculation rates (EIR) reaching 1.221 infective bites/person/night (ib/p/n), An. arabiensis and An. coluzzii were the most implicated. The roles of An. funestus, An. pharoensis and An. paludis were minor. Plasmodium falciparum circumsporozoite protein rate in Anopheles rufipes varied from 0.6 to 5.7% with EIR values between 0.010 and 0.481 ib/p/n.ConclusionsThe study highlights the epidemiological role of An. rufipes alongside the members of the An. gambiae complex, and several other sympatric species in human malaria transmission during the wet season in northern Cameroon. For the first time in Cameroon, An. rufipes has been shown to be an important local malaria vector, emphasising the need to review the malaria entomological profile across the country as pre-requisite to effective vector management strategies.

Insecticides are a key component of vector-based malaria control programmes in Cameroon. As part of ongoing resistance surveillance efforts, Anopheles gambiae s.l. female mosquitoes were exposed to organochlorine (DDT), a carbamate (bendiocarb), an organophosphate (malathion), and three pyrethroids (deltamethrin, lambda-cyhalothrin and permethrin) in WHO bioassay test kits. Results indicated a higher level of resistance (reduced mortality and knockdown effect) to DDT and pyrethroids in populations of A. gambiae s.s. than in A. arabiensis. The West and East African knockdown resistance (kdr) mutations were found in both species but at much higher frequencies in A. gambiae s.s. The West Africa kdr mutant was also more frequent in the A. gambiae S form than in the M form. No resistance to bendiocarb and malathion was found. Carbamate and organophosphorous compounds could thus be used as alternatives in locations in Cameroon where pyrethroid-resistant populations are found.

BackgroundMalaria transmission in Cameroon is mediated by a plethora of vectors that are heterogeneously distributed across the country depending on the biotope. To effectively guide malaria control operations, regular update on the role of local Anopheles species is essential. Therefore, an entomological survey was conducted between August 2010 and May 2011 to evaluate the role of the local anopheline population in malaria transmission in three villages of the Ndop health district in the northwest region of Cameroon where malaria is holoendemic, as a means to acquiring evidence based data for improved vector intervention.MethodsMosquitoes were sampled both indoor and outdoor for four consecutive nights in each locality during each month of survey. Sampling was done by the human landing catch method on volunteers. Anopheles species were identified morphologically and their ovaries randomly dissected for parity determination. Infection with Plasmodium falciparum was detected by Circumsporozoite protein ELISA. Members of An. gambiae complex were further identified to molecular level by PCR and RFLP PCR.ResultsAn. ziemanni was the main malaria vector and whether outdoor or indoor. The man biting rate for the vectors ranged from 6.75 to 8.29 bites per person per night (b/p/n). The entomological inoculation rate for this vector species was 0.0278 infectious bites per person per night (ib/p/n) in Mbapishi, 0.034 ib/p/n in Mbafuh, and 0.063 ib/p/n in Backyit. These were by far greater than that for An. gambiae. No difference was observed in the parity rate of these two vectors. PCR analysis revealed the presence of only An. colluzzi (M- form).ConclusionsAn. ziemanni is an important local malaria vector in Ndop health district. The findings provide useful baseline information on the anopheles species composition, their distribution and role in malaria transmission that would guide the implementation of integrated vector management strategies in the locality.

Cross-sectional entomological surveys were carried out during the 2006 dry and rainy seasons in Lagdo, Cameroon to measure the impact of rice cultivation on malaria transmission and to monitor vector susceptibility to insecticides. Adult anopheline mosquitoes were captured on human volunteers and by pyrethrum spray collections. A total of 4740 mosquitoes was collected during the study. Anopheles arabiensis was the major species and the main malaria vector in all study sites, followed by A. funestus. Malaria transmission was high in the non-irrigated zone of Mayo Mbocki, whereas in the irrigated area of Gounougou it was below detection level during the dry season and high during the rainy season. Insecticide susceptibility tests performed on A. gambiae s.l. populations detected resistance to lambdacyhalothrin and to a lower extent to deltamethrin. All survivors were A. arabiensis. None of the surviving mosquitoes carried the kdr mutation, suggesting an alternative resistance mechanism.

BackgroundDevelopment of large scale agro-industries are subject to serious environmental modifications. In malaria endemic areas this would greatly impact on the transmission paradigm. Two cross-sectional entomological surveys to characterize the Anopheles fauna and their entomological inoculation rates were conducted during May 2010 (peak rainy season) and December 2010 (peak dry season) in the intense rubber cultivated area of Niete in southern forested Cameroon.MethodsMosquitoes were sampled by night collections on human volunteers, identified morphologically and members of the Anopheles gambiae complex further identified to species and molecular form. Parity status was determined following the dissection of the ovaries. Plasmodium falciparum circumsporozoite antigen indices were estimated after the identification of CS antigen by ELISA and the average entomological inoculation rates determined.ResultsA total of 1187 Anopheles was collected, 419 (35.3%) in the rainy season and 768 (64.7%) in the dry season. Species found were the M molecular form of An. gambiae s.s (66.8%), An. ziemanni (28.3%), An. paludis (4.7%), An. smithii (0.2%). An. gambiae M-form was the principal species in the dry (56.2%) and wet (86.2%) seasons. Average overall entomological inoculation rate for the malaria vectors varied between the dry season (1.09 ib/p/n) and the rainy season (2.30 ib/p/n).ConclusionsMalaria transmission in Niete occurs both in the dry and rainy season with the intensities peaking in the dry season. This is unlike previous studies in other areas of southern forested Cameroon where transmission generally peaks in the rainy season. Environmental modifications due to agro-industrial activities might have influenced vector distribution and the dynamics of malaria transmission in this area. This necessitates the possible implementation of control strategies that are related to the eco-geography of the area.

The spread of insecticide resistance genes in Anopheles gambiae Giles sensu stricto threatens to compromise vector-based malaria control programs. Two mutations at the same locus in the voltage-gated sodium channel gene are known to confer knockdown resistance (kdr) to pyrethroids and DDT. Kdr-e involves a leucine-serine substitution, and it was until recently thought to be restricted to East Africa, whereas kdr-w, which involves a leucine-phenylalanine substitution, is associated with resistance in West Africa. In this study, we analyze the frequency and relationship between the kdr genotypes and resistance to type I and type II pyrethroids and DDT by using WHO test kits in both the Forest-M and S molecular forms of An. gambiae in Cameroon. Both kdr-w and kdr-e polymorphisms were found in sympatric An. gambiae, and in many cases in the same mosquito. Kdr-e and kdr-w were detected in both forms, but they were predominant in the S form. Both kdr-e and kdr-w were closely associated with resistance to DDT and weakly associated with resistance to type II pyrethroids. Kdr-w conferred greater resistance to permethrin than kdr-e. We also describe a modified diagnostic designed to detect both resistant alleles simultaneously.

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