Insecticide Resistance and Metabolic Mechanisms in Aedes aegypti from Two Agrosystems (Vegetable and Cotton Crops) in Côte d'Ivoire.

Background: The emergence of insecticide resistance in Aedes vectors mosquitoes poses a real challenge for arboviral-borne disease control. In Côte d'Ivoire, data are available on phenotypic resistance and the presence of kdr mutations in Aedes aegypti. Therefore, information on metabolic resistance in Aedes populations is very scarce. Here, we assessed the insecticide resistance status of Ae. aegypti in periurban and rural areas of Côte d'Ivoire, and we investigated the role of detoxification enzymes as possible resistance mechanisms. Materials and Methods: Aedes mosquito eggs were collected between June 2019 to April 2021 in two agricultural sites. Adults of Ae. aegypti were tested using World Health Organization tube assays, with seven insecticides belonging to pyrethroids, organochlorines, carbamates, and organophosphates classes. We determined the knockdown times (KdT50, KdT95) and resistance ratios of pyrethroids in natural populations. The synergist piperonyl butoxide (PBO) was used to investigate the role of enzymes in resistance. Biochemical assays were performed to detect potential increased activities in mixed-function oxidase levels, nonspecific esterases (NSEs), and glutathione S-transferases. Results: The results showed that Ae. aegypti populations were resistant to five insecticides with mortality of 46% and 89% for 0.75% permethrin, 68% and 92% for 0.05% deltamethrin, 57% and 89% for lambda-cyhalothrin, 41% and 47% for dichlorodiphenyltrichloroethane (DDT), 82% and 91% for chlorpyrifos-methyl in Songon-Agban and Kaforo, respectively. Susceptibility to carbamates was observed in our study sites. After exposure to PBO, the susceptibility of Ae. aegypti to pyrethroids and DDT was partially restored in Songon-Agban. Whereas in Kaforo, none increase of the mortality rates of Ae. aegypti for these four insecticides was observed after exposure to PBO. Increased activity of NSE (α-esterases) was found in Songon-Agban compared with the reference susceptible strain. Conclusion: These findings provide valuable information to support decisions for vector control strategies in Cote d'Ivoire. Also, we highlight the need for the monitoring of insecticide resistance management in Aedes vectors.

Entomological drivers of uneven malaria transmission in urban lowland areas in Bouaké, Côte d'Ivoire.

BACKGROUND: The use of urban lowlands for agriculture contributes to the food security of city- dwellers, but promotes malaria transmission. The objective of the study was to characterize the entomological drivers of malaria transmission in two lowlands (N'Gattakro and Odiennekourani) in the city of Bouaké, Côte d'Ivoire. METHODS: The human landing catch technique was used to capture mosquitoes in houses located at the edge of two lowlands in Bouaké from February to December 2019. Cultivated surfaces were calculated monthly in both lowlands for each crop type (rice and market gardening) using images acquired by a drone. The different mosquito species were identified morphologically and by PCR analysis for the Anopheles gambiae complex. Anopheles infection by Plasmodium parasites was assessed by quantitative PCR. Mosquito diversity, biting behaviour and rhythmicity, and malaria transmission were determined in each lowland and compared. RESULTS: Anopheles gambiae sensu lato (s.l.) was predominant in N'Gattakro and Culex quinquefasciatus in Odiennekourani. Four Anopheles species were identified: An. gambiae s.l. and Anopheles funestus s.l. in both lowlands, Anopheles pharoensis in N'Gattakro, and Anopheles ziemanni in Odiennekourani. Within the An. gambiae complex, three species were caught: An. gambiae sensu stricto (s.s.), Anopheles coluzzii, and Anopheles arabiensis for the first time in Côte d'Ivoire (30.1%, 69.9% and 0% in N'Gattakro, and 45.1%, 52.6% and 2.4% in Odiennekourani, respectively). Anopheles gambiae s.l. species exhibited a significant exophagic behaviour in N'Gattakro (77.1% of outdoor bites versus 52.2% in Odiennekourani). In N'Gattakro, 12.6% of captures occurred before bedtime (09.00 pm) and after waking up (05.00 am), 15.1% in Odiennekourani. The mean human biting rate was higher in N'Gattakro than in Odiennekourani (61.6 versus 15.5 bites per person per night). Overall, Anopheles infection rate was 0.68%, with 0.539 and 0.029 infected bites per person per night in N'Gattakro and Odiennekourani, respectively. CONCLUSION: The risk of malaria in urban agricultural lowland areas is uneven. The role of agricultural developments and irrigation patterns in the production of larval habitat should be explored. The exophagic behaviour of Anopheles vectors raises the question of the residual transmission that needs to be assessed to implement appropriate control strategies.

Spatiotemporal analysis of malaria incidence in Côte d'Ivoire from 2015 to 2019.

BACKGROUND: The collection of malaria cases over time allows the identification of areas with the highest incidence. Our objective was to characterize the spatial distribution of malaria in Côte d'Ivoire from 2015 to 2019 at the health district level. METHODS: Data on the number of reported malaria cases confirmed by rapid diagnostic test (RDT) in the general population, the number of patients attending medical consultations and the total population by health district and year were collected from the National Malaria Control Program in Côte d'Ivoire. Crude and adjusted incidence rates were estimated for each health district and year. Adjusted incidence rates were used to perform global (Moran's index) and local indicators of spatial autocorrelation (LISA) analyses. RESULTS: Between 2015 and 2019, mean crude incidence rates increased from 155.5‰ to 229.8‰. We observed significant heterogeneity in malaria incidence rates across the study period and within a given year. The overall Moran index showed spatial autocorrelation for every year analysed except 2017. The LISA analysis showed that the health districts with high incidence rates were concentrated in the western zone of Côte d'Ivoire. CONCLUSIONS: The use of spatial analyses to identify the areas with the highest malaria incidence rates is a relevant approach to optimize control measures in targeted areas.

High vectorial transmission of malaria in urban and rural settings in the northern, western and eastern regions of Côte d'Ivoire.

BACKGROUND & OBJECTIVES: Malaria remains a public health problem in Côte d'Ivoire. To cope with this issue, the Ministry of Health established strategies through Long-Lasting Insecticidal Nets (LLINs) and artemisinin-based medicines. To better understand the influence of periodic mass distribution of LLINs on malaria transmission, this entomological survey was conducted in three regions of Côte d'Ivoire. METHODS: Mosquitoes were sampled by Human Landing Catches (HLC) in urban and rural settings of Korhogo, Man and Abengourou. Mosquitoes were identified morphologically and by molecular methods. Plasmodium falciparum infection was assessed by ELISA, and the Entomological Inoculation Rates (EIR) were calculated for each species and setting. RESULTS: Only An. gambiae s.l. was identified in Korhogo and in Abengourou while An. gambiae s.l. and An. funestus s.l. was reported in Man. An. coluzzii was the most abundant species of the An. gambiae siblings collected in Abengourou, and in Man while An. gambiae was most abundant in Korhogo. In urban settings, malaria vectors showed high aggressiveness (>11 bites per person per night) and the annual EIR was high (83.22-438.44 infectious bites per person per year). In rural settings, malaria vectors showed also high aggressiveness (19-52 b/p/n). The annual EIR is very high (>94 ib/p/yr). However, the weakest EIR was recorded in the northern region with 94.90 ib/p/yr. INTERPRETATION & CONCLUSION: This work indicates that malaria transmission remains high and heterogeneous across Côte d'Ivoire, despite repeated mass distribution of LLINs. Also, in Man, malaria transmission is more intense with the involvement of two main vectors. Furthermore, in the village of Korhogo, the EIR remained relatively low.

First detection of the malaria vector Anopheles arabiensis in Côte d'Ivoire: urbanization in question.

BACKGROUND: Previous studies have revealed high malaria transmission in Bouaké, Côte d'Ivoire. The sociopolitical crisis in the country and the resulting environmental changes have raised the need to update existing knowledge on mosquito vector species and malaria transmission. METHODS: Adult mosquitoes were caught using the human landing catch (HLC) sampling method in Bouaké. They were collected in six districts representative of the diversity in urban landscapes. Sampling points were selected along the water network crossing the city and monitored from 2020 to 2021 to detect the presence of anopheline larvae. PCR techniques were used to ascertain the species of the Anopheles gambiae complex, Plasmodium falciparum sporozoite infection in a subset of Anopheles vectors, and insecticide resistance mechanisms in Anopheles arabiensis only. RESULTS: A total of 4599 Anopheles mosquitoes were collected and then identified. Anopheles gambiae sensu lato (s.l.) made up the majority of the whole collection (99%) while Anopheles funestus (0.7%), Anopheles ziemanni (0.2%), Anopheles pharoensis (0.2%) represented the remaining proportion of collection. Among the Anopheles gambiae complex, three species were identified namely An. gambiae sensu stricto (45.9%), Anopheles coluzzii (52.2%), and Anopheles arabiensis (1.9%). The first two species had already been collected in Bouaké, however this is the first time that An. arabiensis is reported in Côte d'Ivoire. Anopheles arabiensis was also collected during the larval surveys in a similar proportion (1.1%) in the same areas as the adults. CONCLUSIONS: This study detected the presence of An. arabiensis for the first time in Côte d'Ivoire. This species was found quite far from its usual geographic area and its expansion could be linked to the urbanization process. Although no An. arabiensis was found to be infected by Plasmodium sp., its role in malaria transmission in Bouaké has to be explored, particularly since its exophagic behaviour raises the issue of control measures and the use of insecticide-impregnated nets. The spread of this species in Côte d'Ivoire should be assessed through further research in additional towns in the country.

First evaluation of antibody responses to Culex quinquefasciatus salivary antigens as a serological biomarker of human exposure to Culex bites: A pilot study in Côte d'Ivoire.

BACKGROUND: Culex mosquitoes are vectors for a variety of pathogens of public health concern. New indicators of exposure to Culex bites are needed to evaluate the risk of transmission of associated pathogens and to assess the efficacy of vector control strategies. An alternative to entomological indices is the serological measure of antibodies specific to mosquito salivary antigens. This study investigated whether the human IgG response to both the salivary gland extract and the 30 kDa salivary protein of Culex quinquefasciatus may represent a proxy of human exposure to Culex bites. METHODOLOGY/PRINCIPAL FINDINGS: A multidisciplinary survey was conducted with children aged 1 to 14 years living in neighborhoods with varying exposure to Culex quinquefasciatus in the city of Bouaké, Côte d'Ivoire. Children living in sites with high exposure to Cx quinquefasciatus had a significantly higher IgG response to both salivary antigens compared with children living in the control site where only very few Culex were recorded. Moreover, children from any Culex-high exposed sites had significantly higher IgG responses only to the salivary gland extract compared with children from the control village, whereas no difference was noted in the anti-30 kDa IgG response. No significant differences were noted in the specific IgG responses between age and gender. Sites and the use of a bed net were associated with the level of IgG response to the salivary gland extract and to the 30 kDa antigen, respectively. CONCLUSIONS/SIGNIFICANCE: These findings suggest that the IgG response to Culex salivary gland extracts is suitable as proxy of exposure; however, the specificity to the Culex genus needs further investigation. The lower antigenicity of the 30 kDa recombinant protein represents a limitation to its use. The high specificity of this protein to the Culex genus makes it an attractive candidate and other specific antibody responses might be more relevant as a biomarker of exposure. These epidemiological observations may form a starting point for additional work on developing serological biomarkers of Culex exposure.

Influence of Host-Related Factors and Exposure to Mosquito Bites on the Dynamics of Antibody Response to Plasmodium falciparum Antigens.

Humoral immunity to Plasmodium falciparum is acquired after repeated infections, and can lead to clinical protection. This study aimed to evaluate how human-, parasite-, and environment-related determinants can modulate the dynamics of IgG responses to Plasmodium falciparum after an infection. Individuals (n = 68, average age = 8.2 years) with uncomplicated malaria were treated with ACT and followed up for 42 days. IgG responses to P. falciparum merozoite antigens (PfMSP1, PfMSP3, PfAMA1, PfGLURP-R0), to whole schizont extract (PfSchz), and to Anopheles gSG6-P1 and Aedes Nterm-34 kDa salivary peptides were measured. Regression analyses were used to identify factors that influence the dynamics of IgG response to P. falciparum antigen between D0 and D42, including demographic and biological factors and the level of exposure to mosquito bites. The dynamics of IgG response to P. falciparum differed according to the antigen. According to multivariate analysis, IgG responses to PfSchz and to PfGLURP-R0 appear to be affected by exposure to Aedes saliva and are associated with age, parasite density, and anti-Plasmodium pre-existing immune response at study inclusion. The present work shows that human exposure to Aedes saliva may contribute, in addition to other factors, to the regulation of anti-Plasmodium immune responses during a natural infection.

Diversity of Anopheles Gambiae s.l., Giles (Diptera: Culicidae) Larval Habitats in Urban Areas and Malaria Transmission in Bouaké, Côte d'Ivoire.

Malaria vectors are supposedly uncommon in urban areas owing to the lack of suitable breeding sites for their development. However, the maintenance in urban areas of traditional rural practices along with humanitarian crisis can create favorable conditions for malaria transmission. This study aimed to provide relevant entomological data on the risk of malaria transmission in the city of Bouaké, after the military-political crisis from 2002 to 2011 in Côte d'Ivoire. Adults mosquitoes were collected by human landing catches in Dar Es Salam, Kennedy and N'gattakro neighborhoods. Potential breeding sites were georeferenced and mapped using a GPS. Mosquito species were identified morphologically and by molecular methods. Plasmodium infections were detected by quantitative PCR. Anopheline larvae were found in rice and vegetable crops, puddles, and footprints. A total of 939 Anopheles gambiae s.l. were caught during the surveys. The average human biting rate was 8.8 bites/person/night. The A. gambiae s.l. species were A. gambiae s.s (89.6%) and Anopheles coluzzii (10.4%). The average infectivity rate was 0.74% and the average annual entomological inoculation rate was estimated at 19 infected bites/person/year ranging from 0 in Dar Es Salam and N'gattakro to 58 in Kennedy. The risk of malaria transmission exists in Bouaké city, although Plasmodium infections are low.

Operational Evaluation of the Effectiveness of Long-lasting Insecticidal Nets on Human-Vector Contact in an African Urban Malaria Context.

BACKGROUND: Malaria is still a major public health concern in Côte d'Ivoire despite mass distribution of long-lasting insecticidal nets (LLINs) as a key preventive strategy. This study intended to evaluate the operational effectiveness of LLINs on the level of human-vector contact using 1 antibody-based biomarker of exposure to Anopheles in urban areas. METHODS: This cross-sectional study collected socio-demographic data and use of LLINs from 9 neighborhoods in the city of Bouaké (Côte d'Ivoire). Dry blood spots performed in children aged >6 months and adults were used to evaluate immunoglobulin G (IgG) responses to the Anopheles gSG6-P1 salivary peptide. RESULTS: IgG response levels to the salivary peptide were significantly lower in individuals who declared having "always" (n = 270) slept under an LLIN compared with those who had "often" (n = 2087) and "never" (n = 88) slept under an LLIN (P < .0001). IgG response levels to gSG6-P1 between those who declared having "always" and "not always" slept under an LLIN varied according to neighborhood, socio-professional category, and age group. CONCLUSIONS: The human IgG level to this gSG6-P1 salivary peptide could be a useful tool to evaluate the actual effectiveness of LLINs and help design behavioral change interventions that are crucial for sustaining universal coverage.

Pattern of antibody responses to Plasmodium falciparum antigens in individuals differentially exposed to Anopheles bites.

BACKGROUND: In malaria-endemic areas, human populations are frequently exposed to immunomodulatory salivary components injected during mosquito blood feeding. The consequences on pathogen-specific immune responses are not well known. This study evaluated and compared the humoral responses specific to merozoite stage vaccine candidates of Plasmodium falciparum, in children differentially exposed to Anopheles bites in a natural setting. METHODS: The cross-sectional study was carried out in Bouaké (Côte d'Ivoire) where entomological data and blood samples from children (0-14 years) were collected in two sites with similar malaria prevalence. Antibody (IgG, IgG1, IgG3) responses to PfAMA1 and PfMSP1 were evaluated by ELISA. Univariate and multivariate analysis were performed to assess the relationship between the immune responses to P. falciparum antigens and exposure to Anopheles bites in the total cohort and in each site, separately. The individual level of exposure to Anopheles bites was evaluated by quantifying specific IgG response to the Anopheles gSG6-P1 salivary peptide, which represents a proxy of Anopheles exposure. RESULTS: The anti-Plasmodium humoral responses were different according to the level of exposure of children, with those highly exposed to Anopheles presenting significantly lower antibody responses to PfMSP1 in total population (IgG and IgG3) and in Petessou village (IgG, IgG1, IgG3). No significant difference was seen for PfAMA1 antigen between children differently exposed to Anopheles. In Dar-es-Salam, a neighbourhood where a high Culex density was reported, children presented very low antibody levels specific to both antigens, and no difference according to the exposure to Anopheles bites was found. CONCLUSION: These findings may suggest that immunomodulatory components of Anopheles saliva, in addition to other factors, may participate to the modulation of the humoral response specific to Plasmodium merozoite stage antigens. This epidemiological observation may form a starting point for additional work to decipher the role of mosquito saliva on the modulation of the anti-Plasmodium acquired immunity and clinical protection in combining both field and ex vivo immunological studies.

Vectorial Transmission of Malaria in Major Districts of Côte d'Ivoire.

To better understand the influence of periodic mass distribution of Long-Lasting Insecticidal Nets (LLINs) on malaria transmission, a 1-yr entomological survey was conducted in three major districts of Côte d'Ivoire. Mosquitoes were sampled by Human Landing Catches (HLC) in urban and rural areas of San Pedro and Abidjan (coastal), and in Yamoussoukro (central). Mosquitoes were identified morphologically and by molecular methods. The Plasmodium falciparum circumsporozoite (CSP) indices were measured by ELISA, and the Entomological Inoculation Rates (EIR) were calculated for each species and area. Anopheles gambiae s.l. Giles (Diptera: Culicidae) and Anopheles nili Theobald (Diptera: Culicidae) were identified in coastal districts, while An. gambiae s.l. and Anopheles funestus Giles (Diptera: Culicidae) were reported in the central district. In urban areas, malaria vectors showed a low aggressiveness (<10 bites per person per night), except in Yamoussoukro, where up to 18.9 b/p/n were recorded. The annual EIR was higher in the central urban area (138.7 infected bites per person per year) than in coastal ones (10-62 ib/p/n). In rural areas, malaria vectors were highly aggressive with an average 13 b/p/n for An. gambiae s.l, 21.2 b/p/n for An. nili and 12 b/p/n for An. funestus. The annual EIR ranged between 94.9 and 193.4 infected bites per person per year. This work indicates that, despite repeated mass distribution of LLINs, malaria transmission remains high and heterogeneous across Côte d'Ivoire. Malaria transmission was lower in coastal urban areas than in the central one, and remains high rural areas where two additional Anopheles vectors are involved in malaria transmission.

Exploring the heterogeneity of human exposure to malaria vectors in an urban setting, Bouaké, Côte d'Ivoire, using an immuno-epidemiological biomarker.

BACKGROUND: In some African cities, urban malaria is a threat to the health and welfare of city dwellers. To improve the control of the disease, it is critical to identify neighbourhoods where the risk of malaria transmission is the highest. This study aims to evaluate the heterogeneity of malaria transmission risk in one city (Bouaké) in a West African country (Côte d'Ivoire) that presents several levels of urbanization. METHODS: Two cross-sectional studies were conducted in three neighbourhoods (Dar-es-Salam, Kennedy and N'gattakro) in Bouaké during both the rainy and dry seasons. Data on insecticide-treated net (ITN) use and blood samples were collected from children aged between 6 months and 15 years to determine the parasite density and the prevalence of Plasmodium falciparum and the level of IgG against the Anopheles gSG6-P1 salivary peptide, used as the biomarker of Anopheles bite exposure. RESULTS: The specific IgG levels to the gSG6-P1 salivary peptide in the rainy season were significantly higher compared to the dry season in all neighbourhoods studied (all p < 0.001). Interestingly, these specific IgG levels did not differ between neighbourhoods during the rainy season, whereas significant differences in IgG level were observed in the dry season (p = 0.034). ITN use could be a major factor of variation in the specific IgG level. Nevertheless, no difference in specific IgG levels to the gSG6-P1 salivary peptide was observed between children who declared "always" versus "never" sleeping under an ITN in each neighbourhood. In addition, the prevalence of P. falciparum in the whole population and immune responders was significantly different between neighbourhoods in each season (p < 0.0001). CONCLUSION: This study highlights the high risk of malaria exposure in African urban settings and the high heterogeneity of child exposure to the Anopheles vector between neighbourhoods in the same city. The Anopheles gSG6-P1 salivary peptide could be a suitable biomarker to accurately and quantitatively assess the risk of malaria transmission in urban areas.

Evaluation of Malaria Urban Risk Using an Immuno-Epidemiological Biomarker of Human Exposure to Anopheles Bites.

Urban malaria is an underestimated serious health concern in African countries. This study aimed to evaluate the risk of malaria transmission in an urban area by evaluating the level of human exposure to Anopheles bites using an Anopheles salivary biomarker (gambiae Salivary Gland Protein-6 peptide 1 [gSG6-P1] peptide). Two multidisciplinary cross-sectional studies were undertaken in five sites of Bouaké city (three urban districts and two surrounding villages, used as control; Côte d'Ivoire) during the rainy season and the dry season. Blood samples were obtained from children 6 months to 14 years of age for immunological tests. The level of anti-gSG6-P1 immunoglobulin G (IgG) antibodies was significantly higher in the rainy season than the dry season in both urban and rural sites (P < 0.0001). Interestingly, children with the highest anti-gSG6-P1 IgG responses in the rainy season were infected by Plasmodium falciparum. Surprisingly, no difference of anti-gSG6-P1 IgG level was observed between urban and rural areas, for either season. The current data suggest that children in the urban city of Bouaké could be as highly exposed to Anopheles bites as children living in surrounding villages. The immunological biomarker of human exposure to Anopheles bites may be used to accurately assess the potential risk of malaria transmission in African urban settings.

Spatial and temporal variation of malaria entomological parameters at the onset of a hydro-agricultural development in central Côte d'Ivoire.

BACKGROUND: A deeper understanding of the ecology and small-scale heterogeneity of malaria transmission is essential for the design of effective prevention, control and elimination interventions. The spatial and temporal distribution of malaria vectors was investigated in five villages in close proximity to a hydro-agricultural system in Côte d'Ivoire over the course of construction and the early phase of irrigated rice farming. METHODS: The study was carried out in five villages (Raffierkro, N'Douakro, Ahougui, Kpokahankro, Koffikro) near Bouaké, central Côte d'Ivoire, between early 2007 and late 2009. In each village, mosquitoes were collected by human landing catches and identified morphologically at genus and species level, and entomological parameters were determined. Plasmodium infection was assessed by dissection and an enzyme-linked immunosorbent assay. RESULTS: A total of 19,404 mosquitoes belonging to the genus Anopheles were sampled during 328 human-night catches. Before the construction of the hydro-agricultural system, comparable densities of Anopheles gambiae were observed in all villages. In subsequent years, densities in Raffierkro and Ahougui were significantly higher than the other villages [Kruskal-Wallis (KW) test = 31.13, p < 0.001]. The density of Anopheles funestus in the five villages was comparable in the early stage of the project, while a high density was reported in Koffikro at the end (KW test = 11.91, p = 0.018). Transmission of Plasmodium falciparum is perennial in the study area. Over the course of the study, high entomological inoculation rates (EIRs) were found: 219-328 infectious bites per person per year with An. gambiae. For An. funestus considerably lower EIRs were observed (5.7-39.4). Changing patterns of An. gambiae were not correlated with malaria transmission. CONCLUSION: In this study setting, located in the bioclimatic transition zone of Côte d'Ivoire, rice cultivation was not observed to increase malaria transmission. The entomological parameters recorded until the onset of rice-growing activities in a hydro-agricultural system presented considerable heterogeneity both in space and time; a strong increase of Anopheles mosquitoes was observed in two of the five villages located in close proximity to the dam and irrigated rice fields. Malaria still is a main public health problem in all villages that require adequate control measures.

Long-term trends in Anopheles gambiae insecticide resistance in Côte d'Ivoire.

BACKGROUND: Malaria control is heavily dependent on the use of insecticides that target adult mosquito vectors via insecticide treated nets (ITNs) or indoor residual spraying (IRS). Four classes of insecticide are approved for IRS but only pyrethroids are available for ITNs. The rapid rise in insecticide resistance in African malaria vectors has raised alarms about the sustainability of existing malaria control activities. This problem might be particularly acute in Côte d'Ivoire where resistance to all four insecticide classes has recently been recorded. Here we investigate temporal trends in insecticide resistance across the ecological zones of Côte d'Ivoire to determine whether apparent pan-African patterns of increasing resistance are detectable and consistent across insecticides and areas. METHODS: We combined data on insecticide resistance from a literature review, and bioassays conducted on field-caught Anopheles gambiae mosquitoes for the four WHO-approved insecticide classes for ITN/IRS. The data were then mapped using Geographical Information Systems (GIS) and the IR mapper tool to provide spatial and temporal distribution data on insecticide resistance in An. gambiae sensu lato from Côte d'Ivoire between 1993 and 2014. RESULTS: Bioassay mortality decreased over time for all insecticide classes, though with significant spatiotemporal variation, such that stronger declines were observed in the southern ecological zone for DDT and pyrethroids than in the central zone, but with an apparently opposite effect for the carbamate and organophosphate. Variation in relative abundance of the molecular forms, coupled with dramatic increase in kdr 1014F frequency in M forms (An. coluzzii) seems likely to be a contributory factor to these patterns. Although records of resistance across insecticide classes have become more common, the number of classes tested in studies has also increased, precluding a conclusion that multiple resistance has also increased. CONCLUSION: Our analyses attempted synthesis of 22 years of bioassay data from Côte d'Ivoire, and despite a number of caveats and potentially confounding variables, suggest significant but spatially-variable temporal trends in insecticide resistance. In the light of such spatio-temporal dynamics, regular, systematic and spatially-expanded monitoring is warranted to provide accurate information on insecticide resistance for control programme management.

Efficacy of ICON® Maxx in the laboratory and against insecticide-resistant Anopheles gambiae in central Côte d'Ivoire.

BACKGROUND: Long-lasting treatment kits, designed to transform untreated nets into long-lasting insecticidal nets (LLINs), may facilitate high coverage with LLINs where non-treated nets are in place. In this study, the efficacy of ICON® Maxx (Syngenta) was evaluated under laboratory conditions and in an experimental hut trial in central Côte d'Ivoire, where Anopheles gambiae s.s. are resistant to pyrethroid insecticides. METHODS: In the laboratory, polyester and polyethylene net samples were treated with ICON® Maxx, washed up to 20 times and their efficacy determined in World Health Organization (WHO) cone assays against a susceptible laboratory An. gambiae s.s. colony. Over a 12-month period, the polyester nets were evaluated in a hut trial to determine mosquito deterrence, induced exophily, blood-feeding inhibition and mortality. RESULTS: In the laboratory, ICON® Maxx-treated polyethylene nets showed higher efficacy against pyrethroid-susceptible mosquitoes than polyester nets. After 20 washings, insecticidal efficacy in bioassays was 59.4% knockdown (KD) and 22.3% mortality for polyethylene, and 55.3% KD and 17.9% mortality for polyester nets. In experimental huts, treated nets showed strong deterrence, induced exophily and an over three-fold reduction in blood-fed mosquitoes. More than half (61.8%) of the mosquitoes entering the huts with treated nets were found dead the next morning despite high levels of KD resistance. After washing the treated nets, KD and mortality rates were close to or exceeded predefined WHO thresholds in cone bioassays. CONCLUSION: In contrast to previous laboratory investigation, ICON® Maxx-treated nets showed only moderate KD and mortality rates. However, under semi-field conditions, in an area where mosquitoes are resistant to pyrethroids, ICON® Maxx showed high deterrence, induced exophily and provided a significant reduction in blood-feeding rates; features that are likely to have a positive impact in reducing malaria transmission. The WHO cone test may not always be a good proxy for predicting product performance under field conditions.

Effect of repeated application of microbial larvicides on malaria transmission in central Côte d'Ivoire.

The effect of repeated applications of Bacillus thuringiensis var israeliensis (Bti) and B. sphaericus (Bs) on different entomologic parameters of malaria transmission was investigated in a village in central Côte d'Ivoire. The study area was monitored for potential mosquito breeding sites over a 7-month period. Microbial larvicides were applied once every 3 wk; first Bti (0.8 mg/liter), followed by Bs (10 mg/liter) 3-4 days later. Adult mosquitoes were collected inside and outside sentinel houses in 4 cross-sectional surveys using human landing catch. Repeated applications of Bti and Bs showed a decline in the biting rate of both Anopheles funestus and Anopheles gambiae. Moreover, the entomologic inoculation rate of An. funestus was significantly reduced (from 328 to 142, P = 0.005), whereas that of An. gambiae remained stable. In conclusion, microbial larvicides might play a role in an integrated approach for malaria control.

[Systematics and biology of Anopheles vectors of Plasmodium in Africa, recent data]. / Systématique et biologie des anophèles vecteurs de Plasmodium en Afrique, données récentes.

Renewed interest in research on Plasmodium vectors in Africa and development of genetic and molecular biology techniques has been spearheaded by the WHO and the PAL+ program of the French research ministry. New findings have led to a better understanding of the systematics and biology of the main vector groups. The purpose of this article is to describe the newest data on the Anopheles gambiae complex and the M and S forms of An. gambiae s.s., on species in the An. funestus group and genetic polymorphism of An. funestus, on the two probable species in the An. moucheti complex, and on An. mascarenesis.