Bendiocarb and Malathion Resistance in Two Major Malaria Vector Populations in Cameroon Is Associated with High Frequency of the G119S Mutation (Ace-1) and Overexpression of Detoxification Genes

The spread of pyrethroid resistance in malaria vectors is a major threat affecting the performance of current control measures. However, there is still not enough information on the resistance profile of mosquitoes to carbamates and organophosphates which could be used as alternatives. The present study assessed the resistance profile of Anopheles gambiae s.l. to bendiocarb and malathion, at the phenotypic and molecular levels, in different eco-epidemiological settings in Cameroon. Anopheles gambiae s.l. mosquitoes were collected from four eco-epidemiological settings across the country and their susceptibility level to bendiocarb and malathion was determined using WHO tubes bioassays. The ace-1 target site G119S mutation was screened by PCR. Reverse Transcription quantitative PCR 3-plex TaqMan assays were used to quantify the level of expression of eight genes associated with metabolic resistance. Resistance to malathion and/or bendiocarb was recorded in all study sites except in mosquitoes collected in Kaélé and Njombé. The Ace-1 (G119S) mutation was detected in high frequencies (>40%) in Kékem and Santchou. Both An. gambiae and An. coluzzii were detected carrying this mutation. The cytochrome P450s gene Cyp6p3 associated with carbamate resistance and the glutathione S-transferase gene Gste2 associated with organophosphate resistance were found to be overexpressed. Genes associated with pyrethroid (Cyp6m2, Cyp9k1, Cyp6p3) and organochlorine (Gste2, Cyp6z1, Cyp6m2) and cuticle resistance (Cyp4g16) were also overexpressed. The rapid spread of resistance to organophosphates and carbamates could seriously compromise future control strategies based on IRS. It is therefore becoming important to assess the magnitude of bendiocarb and malathion resistance countrywide

Patterns of anopheline feeding/resting behaviour and Plasmodium infections in North Cameroon, 2011-2014: implications for malaria control.

BACKGROUND: Effective malaria control relies on evidence-based interventions. Anopheline behaviour and Plasmodium infections were investigated in North Cameroon, following long-lasting insecticidal net (LLIN) distribution in 2010. METHODS: During four consecutive years from 2011 to 2014, adult mosquitoes were collected indoors, outdoors and in exit traps across 38 locations in the Garoua, Pitoa and Mayo-Oulo health districts. Anophelines were morphologically and molecularly identified, then analysed for blood meal origins and Plasmodium falciparum circumsporozoite protein (Pf-CSP). Blood from children under 5 years-old using LLINs was examined for Plasmodium infections. RESULTS: Overall, 9376 anophelines belonging to 14 species/sibling species were recorded. Anopheles gambiae (s.l.) [An. arabiensis (73.3%), An. coluzzii (17.6%) and An. gambiae (s.s.) (9.1%)] was predominant (72%), followed by An. funestus (s.l.) (20.5%) and An. rufipes (6.5%). The recorded blood meals were mainly from humans (28%), cattle (15.6%) and sheep (11.6%) or mixed (45%). Pf-CSP rates were higher indoors (3.2-5.4%) versus outdoors (0.8-2.0%), and increased yearly (χ2 < 18, df = 10, P < 0.03). Malaria prevalence in children under 5 years-old, in households using LLINs was 30% (924/3088). CONCLUSIONS: The present study revealed the variability of malaria vector resting and feeding behaviour, and the persistence of Plasmodium infections regardless the use of LLINs. Supplementary interventions to LLINs are therefore needed to sustain malaria prevention in North Cameroon

Impact of vegetable crop agriculture on anopheline agressivity and malaria transmission in urban and less urbanized settings of the South region of Cameroon

Abstract Background The use of inland valley swamps for vegetable crop agriculture contributes to food security in urban and less urbanized settings in Africa. The impact of this agriculture on aggressive mosquitoes’ diversity and malaria transmission in central Africa is poorly documented. This study is aimed at assessing the impact of vegetable crop agriculture on these entomological parameters in urban and less urbanized settings of the forest area, south of Cameroon. Methods The human bait technique was used for the capture of aggressive mosquitoes from January to December 2012. For three consecutive days each month, captures were performed on volunteers in hydro-agricultural and river bank sites of Akonolinga and Yaoundé. Physico-chemical characteristics of mosquito breeding sites were recorded. Molecular alongside morpho-taxonomic techniques were used for the identification of mosquito species; ELISA test was used to reveal Plasmodium falciparum infected mosquitoes through the detection of CSP. Mosquito diversity, aggressivity and malaria transmission in sites and settings were determined and compared. Results Biting rates were higher in hydro-agricultural sites of less urbanized and urban settings (31.8 b/p/n and 28.6 b/p/n respectively) than in river banks sites (6.83 b/p/n and 3.64 b/p/n respectively; p < 0.0001). Physico-chemical parameters of breeding sites were not fundamentally different. Five anopheline species were identified; An. gambiae, An. funestus s.s., An. moucheti s.s., An. hancocki and An. nili s.s. In hydro-agricultural sites 2 species were captured in the urban setting versus 4 in the less urbanized setting, meanwhile in river bank sites, 3 species were captured in the urban setting versus 4 species in the less urbanized setting. An. nili s.s. was found in river banks only. An. hancocki was not found to insure Plasmodium falciparum Welch transmission. EIR in hydro-agricultural sites varied from 1.86 ib/p/n (urban area) to 2.13 ib/p/n (less urbanized area) with higher rates in April/May and August. Overall, EIR was higher in less urbanized areas (p < 0.0001) but the difference was nullified with the practice of vegetable crop agriculture (p = 0.2). Conclusion These results highlight the need for specific preventive measures that take into account the ecological peculiarities related to vegetable crop agriculture on hydro-agricultural lands, in order to protect inhabitants from malaria

Pyrethroid Resistance Situation across Different Eco-Epidemiological Settings in Cameroon

Rapid emergence and spread of pyrethroid resistance in Anopheles gambiae populations is among the main factors affecting malaria vector control in Cameroon, but there is still not enough data on the exact pyrethroid resistance status across Cameroon. The present study assessed pyrethroid resistance profile in different eco-epidemiological settings in Cameroon. Susceptibility bioassay tests were performed with F0 An. gambiae females aged three to five days. Mosquito susceptibility to both permethrin and deltamethrin was assessed. Species of the An. gambiae s.l. complex were identified using molecular diagnostic tools. Target site mutations conferring resistance were detected using Taqman assays. Quantitative reverse transcription-real-time PCR (qRT-PCR) 3-plex TaqMan&reg; assays were used for the quantification of detoxification genes implicated in pyrethroid resistance. An. gambiae, An. coluzzii and An. arabiensis were identified in the different settings. An. gambiae was dominant in Santchou, K&eacute;kem, B&eacute;labo, Bertoua and Njomb&eacute;, while An. coluzzii was abundant in Tibati and Ka&eacute;l&eacute;. High frequencies of the kdr L1014F allele ranging from 43% to 100% were recorded in almost all sites. The L1014S kdr allele was detected at low frequency (4.10&ndash;10%) only in mosquito populations from Njomb&eacute; and Tibati. The N1575Y mutation was recorded in Ka&eacute;l&eacute;, Santchou, Tibati and Bertoua with a frequency varying from 2.10% to 11.70%. Six Cytochrome P450 genes (Cyp6p3, Cyp6m2, Cyp9k1, Cyp6p4, Cyp6z1, and Cyp4g16) were found to be overexpressed in at least one population. Analysis of cuticular hydrocarbon lipids indicated a significant increase in CHC content in mosquito populations from Ka&eacute;l&eacute; and Njomb&eacute; compared to K&eacute;kem, B&eacute;labo and Bertoua populations. The study indicated high pyrethroid resistance across different ecological settings in Cameroon with different profile of resistance across the country. The present situation calls for further actions in order to mitigate the impact of insecticide resistance on vector control measures