A High Proportion of Malaria Vector Biting and Resting Indoors despite Extensive LLIN Coverage in Côte d'Ivoire.

Malaria is still a leading cause mortality in Côte d'Ivoire despite extensive LLINs coverage. We present the results of an entomological survey conducted in a coastal and in an inland village with the aim to estimate Anopheles gambiae sensu lato (s.l.) female's abundance indoor/outdoor and Plasmodium falciparum infection rate and analyze the occurrence of blood-feeding in relation to LLINs use. Pyrethrum spray (PSC) and window exit traps (WT) collections were carried out to target endophagic/endophilic and endophagic/exophilic females, respectively. Data on LLINs use in sampled houses were collected. (1) high levels of malaria transmission despite LLINs coverage >70% (~1 An. gambiae s.l. predicted mean/person/night and ~5% Plasmodium falciparum infection rate); (2) 46% of females in the PSC sample were blood-fed, suggesting that they fed on an unprotected host inside the house; (3) 81% of females in WT were unfed, suggesting that they were leaving the house to find an available host. Model estimates that if everyone sleeps under LLINs the probability for a mosquito to bite decreases of 48% and 95% in the coastal and inland village, respectively. The results show a high proportion of mosquito biting and resting indoors despite extensive LLINs. The biological/epidemiological determinants of accounting for these results merit deeper investigations.

Entomological monitoring data driving decision-making for appropriate and sustainable malaria vector control in Côte d'Ivoire.

BACKGROUND: Entomological surveillance provides critical information on vectors for appropriate malaria vector control and strategic decision-making. The widely documented insecticide resistance of malaria vectors in Côte d'Ivoire requires that any vector control intervention deployment be driven by entomological data to optimize its effectiveness and appropriate resource allocations. To achieve this goal, this study documents the results of monthly vector surveillance and insecticide susceptibility tests conducted in 2019 and a review of all previous entomological monitoring data used to guide vector control decision making. Furthermore, susceptibility to pirimiphos-methyl and clothianidin was assessed in addition to chlorfenapyr and pyrethroids (intensity and piperonyl butoxide (PBO) synergism) tests previously reported. Vector bionomic data were conducted monthly in four sites (Sakassou, Béoumi, Dabakala and Nassian) that were selected based on their reported high malaria incidence. Adult mosquitoes were collected using human landing catches (HLCs), pyrethrum spray catches (PSCs), and human-baited CDC light traps to assess vector density, behaviour, species composition and sporozoite infectivity. RESULTS: Pirimiphos-methyl and clothianidin susceptibility was observed in 8 and 10 sites, respectively, while previous data reported chlorfenapyr (200 µg/bottle) susceptibility in 13 of the sites, high pyrethroid resistance intensity and increased mortality with PBO pre-exposure at all 17 tested sites. Anopheles gambiae sensu lato was the predominant malaria vector collected in all four bionomic sites. Vector density was relatively higher in Sakassou throughout the year with mean biting rates of 278.2 bites per person per night (b/p/n) compared to Béoumi, Dabakala and Nassian (mean of 48.5, 81.4 and 26.6 b/p/n, respectively). The mean entomological inoculation rate (EIR) was 4.44 infective bites per person per night (ib/p/n) in Sakassou, 0.34 ib/p/n in Beoumi, 1.17 ib/p/n in Dabakala and 1.02 ib/p/n in Nassian. The highest EIRs were recorded in October in Béoumi (1.71 ib/p/n) and Nassian (3.22 ib/p/n), in July in Dabakala (4.46 ib/p/n) and in May in Sakassou (15.6 ib/p/n). CONCLUSION: Based on all results and data review, the National Malaria Control Programme developed and implemented a stratified insecticide-treated net (ITN) mass distribution in 2021 considering new generation ITNs. These results also supported the selection of clothianidin-based products and an optimal spraying time for the first indoor residual spraying (IRS) campaign in Sakassou and Nassian in 2020.

Seasonal variation and intra urban heterogeneity of the entomological risk of transmission of dengue and yellow fever in Abidjan, Côte d'Ivoire.

Dengue and yellow fever are prevalent in Côte d'Ivoire and Aedes (Stegomyia) aegypti (Linnaeus), (Diptera: Culicidae), is known as the main vector. We aimed to assess seasonal variation and spatial heterogeneity in the transmission of both arbovirus diseases in Abidjan. Entomological surveys targeting larvae of A. aegypti, were carried out between November 2015 and August 2016 covering the four climatic seasons including a cohort of 100 houses randomly selected in three neighbourhoods. A. aegypti was the predominant species (96.6%) of mosquitoes resulting from the rearing of harvested larvae, and the only vector of dengue and yellow fever recorded during the study period. The highest proportion of water storage containers (45.5%) which represented the major breeding sites infested by the larvae of A. aegypti, was observed in Anoumabo. The house indices >5% and/or Breteau indices >20 recorded in each neighbourhood, during the different climatic seasons, indicated that there was, a high and permanent, heterogeneity in the transmission risk of dengue and yellow fever between the three neighbourhoods. In terms of transmission risk, Anoumabo was the neighbourhood with the highest risk compared to the two others, then, particular attention should be paid to this site in terms of surveillance by vector control programme in Abidjan.

Is Côte D'Ivoire a new high hybridization zone for the two major malaria vectors, Anopheles coluzzii and An. gambiae (Diptera, Culicidae)?

Anopheles gambiae and An. coluzzii are very closely related and recently differentiated species representing the main malaria vectors in the Afrotropical region and responsible of up to >3 infective bites/person/night in Côte D'Ivoire, where prevention and control has stagnated in recent years. The aim of the present study was to genetically and ecologically characterize An. gambiae and An. coluzzii populations from two villages of Côte D'Ivoire, lying in the coastal forest belt and 250 km inland in the Guinean savannah mosaic belt, respectively. Results reveal high frequencies of both species in both study sites and high frequencies of hybrids (4-33%) along the whole year of sampling. Consistently with observations for the well-known high hybridization zone at the far-west of the species range, hybrid frequencies were higher in the coastal village and highest when the two species occurred at more balanced frequencies, supporting the "frequency-dependent hybridization" ecological speciation theory. Pilot genotyping revealed signatures of genomic admixture in both chromosome-X and -3. Coupled with previous reports of hybrids in the region, the results point to the coastal region of Côte D'Ivoire as a possible regions of high hybridization. Preliminary characterization of parameters relevant for malaria transmission and control (e.g. possibly higher sporozoite rates and indoor biting preferences in hybrids than in the parental species) highlight the possible relevance of the breakdown of reproductive barriers between An. gambiae and An. coluzzii not only in the field of ecological evolution, but also in malaria epidemiology and control.

Spatial Heterogeneity and Seasonal Distribution of Aedes (Stegomyia) aegypti (L) in Abidjan, Côte d'Ivoire.

Although the urban areas of Abidjan, Côte d'Ivoire have faced recurrent outbreaks of Aedes-borne arboviruses, the seasonal dynamics of local populations of the key vector Aedes aegypti remained still underexplored for an effective vector control. The current study thus assessed the seasonal dynamics and the spatial distribution of Ae. aegypti in three neighborhoods of Abidjan city. Aedes eggs were collected using ovitraps in three different neighborhoods (Anoumambo, Bromakoté, and Petit-Bassam) during the four climatic seasons of Abidjan. Aedes egg samples were immersed into distilled water, and emerged larvae were reared until the adult stage for species morphological identification. Spatial autocorrelation was measured with the Moran's Index, and areas with high egg abundance were identified. In total, 3837 eggs were collected providing 1882 adult mosquitoes in the 3 neighborhoods. All the specimens belonged to only one Aedes species, Ae. aegypti. The average of 15.89 eggs per ovitrap, 13.67 eggs per ovitrap, and 19.87 eggs per ovitrap were obtained in Anoumambo, Bromakoté, and Petit-Bassam, respectively, with no statistical difference between the three sites. A higher abundance of Ae. aegypti was observed during the long rainy season and the short dry season. The Moran analysis showed a clustered distribution of Ae. aegypti eggs during the long rainy season in the three sites and a random spatial distribution during the short dry season. Ovitraps with high number of eggs were aggregated in the peripheral part (near to the lagoon) of Anoumambo and Petit-Bassam in central Bromakoté and extending along the railway during the long rainy season. This study revealed a heterogeneous potential risk of transmission of arbovirus according to neighborhood. It provided data to better understand Ae. aegypti ecology to select appropriate periods and places for Aedes vector control actions and surveillance of arboviruses in Abidjan.

Susceptibility of Anopheles gambiae from Côte d'Ivoire to insecticides used on insecticide-treated nets: evaluating the additional entomological impact of piperonyl butoxide and chlorfenapyr.

BACKGROUND: Pyrethroid-treated mosquito nets are currently the mainstay of vector control in Côte d'Ivoire. However, resistance to pyrethroids has been reported across the country, limiting options for insecticide resistance management due to the paucity of alternative insecticides. Two types of insecticide-treated nets (ITNs), ITNs with pyrethroids and the synergist piperonyl butoxide (PBO), and Interceptor®G2 nets, a net treated with a combination of chlorfenapyr and alpha-cypermethrin, are believed to help in the control of pyrethroid-resistant mosquitoes. METHODS: The susceptibility of Anopheles gambiae sensu lato (s.l.) to pyrethroid insecticides with and without pre-exposure to PBO as well as to chlorfenapyr was investigated in fifteen sites across the country. Susceptibility tests were conducted on 2- to 4-day old adult female An. gambiae s.l. reared from larval collections. The resistance status, intensity, and effects of PBO on mortality after exposure to different concentrations of deltamethrin, permethrin and alpha-cypermethrin were determined using WHO susceptibility test kits. In the absence of a WHO-recommended standard protocol for chlorfenapyr, two interim doses (100 and 200 µg/bottle) were used to test the susceptibility of mosquitoes using the CDC bottle assay method. RESULTS: Pre-exposure to PBO did not result in full restoration of susceptibility to any of the three pyrethroids for the An. gambiae s.l. populations from any of the sites surveyed. However, PBO pre-exposure did increase mortality for all three pyrethroids, particularly deltamethrin (from 4.4 to 48.9%). Anopheles gambiae s.l. from only one site (Bettie) were susceptible to chlorfenapyr at the dose of 100 µg active ingredient (a.i.)/bottle. At the dose of 200 µg (a.i.)/bottle, susceptibility was only recorded in 10 of the 15 sites. CONCLUSION: Low mosquito mortality was found for pyrethroids alone, and while PBO increased mortality, it did not restore full susceptibility. The vector was not fully susceptible to chlorfenapyr in one third of the sites tested. However, vector susceptibility to chlorfenapyr seems to be considerably higher than for pyrethroids alone or with PBO. These data should be used cautiously when making ITN procurement decisions, noting that bioassays are conducted in controlled conditions and may not fully represent field efficacy where the host-seeking behaviours, which include free-flying activity are known to enhance pro-insecticide chlorfenapyr intoxication to mosquitoes.

Spatial Assessment of Contact Between Humans and Anopheles and Aedes Mosquitoes in a Medium-Sized African Urban Setting, Using Salivary Antibody-Based Biomarkers.

BACKGROUND: Anarchic and poorly controlled urbanization led to an increased risk of mosquito-borne diseases (MBD) in many African cities. Here, we evaluate the spatial heterogeneity of human exposure to malaria and arboviral disease vectors in an urban area of northern Senegal, using antibody-based biomarkers of exposure to Anopheles and Aedes mosquito bites. METHODS: A cross-sectional study was undertaken during the rainy season of 2014 in 4 neighborhoods of Saint-Louis, a city in northern Senegal. Among children aged 6-59 months in each neighborhood, the dried blood spot technique was used to evaluate immunoglobulin G (IgG) responses to both gSG6-P1 (Anopheles) and Nterm-34-kDa (Aedes) salivary peptides as validated biomarkers of respective mosquito bite exposure. RESULTS: IgG response levels to gSG6-P1 and Nterm-34-kDa salivary peptides varied significantly between the 4 neighborhoods (P < .0001). The level of exposure to Aedes bites also varied according to household access to sanitation services (P = .027), whereas that of exposure to Anopheles bites varied according to insecticide-treated bed net use (P = .006). In addition, spatial clusters of high contact between humans and mosquitoes were identified inside 3 neighborhoods. CONCLUSIONS: Antibody-based biomarkers of exposure to Anopheles and Aedes mosquito bites could be helpful tools for evaluating the heterogeneity of exposure to malaria and arboviral disease vectors by national control programs.

Surveys of Arboviruses Vectors in Four Cities Stretching Along a Railway Transect of Burkina Faso: Risk Transmission and Insecticide Susceptibility Status of Potential Vectors.

Background: A severe outbreak of dengue occurred in Burkina Faso in 2016, with the most cases reported in Ouagadougou, that highlights the necessity to implement vector surveillance system. This study aims to estimate the risk of arboviruses transmission and the insecticide susceptibility status of potential vectors in four sites in Burkina Faso. Methods: From June to September 2016, house-to-house cross sectional entomological surveys were performed in four cities stretching along a southwest-to-northeast railway transect. The household surveys analyzed the presence of Aedes spp. larvae in containers holding water and the World Health Organization (WHO) larval abundance indices were estimated. WHO tube assays was used to evaluate the insecticide susceptibility within Aedes populations from these localities. Results: A total of 31,378 mosquitoes' larvae were collected from 1,330 containers holding water. Aedes spp. was the most abundant (95.19%) followed by Culex spp. (4.75%). Aedes aegypti a key vector of arboviruses (ARBOV) in West Africa was the major Aedes species found (98.60%). The relative larval indices, house index, container and Breteau indexes were high, up to 70, 35, and 10, respectively. Aedes aegypti tended to breed mainly in discarded tires and terracotta jars. Except in Banfora the western city, Ae. aegypti populations were resistant to deltamethrin 0.05% in the other localities with low mortality rate under 20% in Ouagadougou whereas they were fully susceptible to malathion 5% whatever the site. Intermediate resistance was observed in the four sites with mortality rates varying between 78 and 94% with bendiocarb 0.1%. Conclusions: This study provided basic information on entomological indices that can help to monitor the risks of ARBOV epidemics in the main cities along the railway in Burkina Faso. In these cities, all larval indices exceeded the risk level of ARBOV outbreak. Aedes aegypti the main species collected was resistant to deltamethrin 0.05% and bendiocarb 0.1% whereas they were fully susceptible to malathion 5%. The monitoring of insecticide resistance is also important to be integrated to the vector surveillance system in Burkina Faso.

The current insecticide resistance status of Anopheles gambiae (s.l.) (Culicidae) in rural and urban areas of Bouaké, Côte d'Ivoire.

BACKGROUND: Several studies were carried out in experimental hut station in areas surrounding the city of Bouaké, after the crisis in Côte d'Ivoire. They reported increasing resistance levels to insecticide for malaria transmiting mosquitoes. The present work aims to evaluate the current resistance level of An. gambiae (s.l.) in rural and urban areas in the city of Bouaké. METHODS: Larvae of Anopheles gambiae (s.l.) were collected from five different study sites and reared to adult stages. The resistance status was assessed using the WHO bioassay test kits for adult mosquitoes, with eight insecticides belonging to pyrethroids, organochlorines, carbamates and organophosphates classes. Molecular assays were performed to identify the molecular forms of An. gambiae (s.l.), the L1014F kdr and the ace-1R alleles in individual mosquitoes. The synergist PBO was used to investigate the role of enzymes in resistance. Biochemical assays were performed to detect potential increased activities in mixed function oxidase (MFO) levels, non-specific esterases (NSE) and glutathione S-transferases (GST). RESULTS: High resistance levels to pyrethroids, organochlorines, and carbamates were observed in Anopheles gambiae (s.l.) from Bouaké. Mortalities ranged between 0 and 73% for the eight tested insecticides. The pre-exposure to PBO restored full or partial susceptibility to pyrethroids in the different sites. The same trend was observed with the carbamates in five sites, but to a lesser extent. With DDT, pre-exposure to PBO did not increase the mortality rate of An. gambiae (s.l.) from the same sites. Tolerance to organophosphates was observed. An increased activity of NSE and higher level of MFO were found compared to the Kisumu susceptible reference strain. Two molecular forms, S form [(An. gambiae (s.s)] and M form (An. coluzzi) were identified. The kdr allele frequencies vary from 85.9 to 99.8% for An. gambiae (s.s.) and from 81.7 to 99.6% for An. coluzzii. The ace-1R frequencies vary between 25.6 and 38.8% for An. gambiae (s.s.) and from 28.6 to 36.7% for An. coluzzii. CONCLUSION: Resistance to insecticides is widespread within both An. gambiae (s.s.) and An. coluzzii. Two mechanisms of resistance, i.e. metabolic and target-site mutation seemed to largely explain the high resistance level of mosquitoes in Bouaké. Pyrethroid resistance was found exclusively due to the metabolic mechanism.