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.

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.

High Levels of Admixture in Anopheles gambiae Populations from Côte d'Ivoire Revealed by Multilocus Genotyping.

Anopheles coluzzii and An. gambiae-the two most recently radiated species of the An. gambiae complex and the major Afrotropical malaria vector species-are identified by markers in the X-centromeric IGS rDNA region. Putative IGS-hybrids are rarely found in the field, except in restricted areas where genomic studies have led to the hypothesis that the observed IGS-patterns are due to cryptic taxa rather than to hybridization between the two species. We investigated the genome-wide levels of admixture in two villages in Côte d'Ivoire where high levels of IGS-hybrids have been detected, confirming unparalleled high frequencies in the coastal village. Genotyping of 24 Ancestry Informative Markers (AIMs) along the three chromosomes produced discordant results between the IGS-marker and the multilocus genotype obtained for AIMs across the whole genome (29%) as well as AIMs on chromosome-X (considered to be fundamental for species reproductive isolation) only (21%). Results highlight a complicated pattern of admixture that deserves deeper genomic analyses to understand better possible underlying causes (from extensive processes of hybridization to the existence of different cryptic taxa), and stress the need of developing advanced diagnostics for An. coluzzii, An. gambiae and putative new taxa, instrumental for assessing taxon-specific epidemiological characters.

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.

Effectiveness of Four Different Interventions Against Schistosoma haematobium in a Seasonal Transmission Setting of Côte d'Ivoire: A Cluster Randomized Trial.

BACKGROUND: Annual mass drug administration (MDA) using praziquantel is the cornerstone of schistosomiasis morbidity control but is not sufficient to interrupt transmission. We implemented a cluster-randomized trial to compare the effectiveness of 4 different intervention packages to interrupt transmission of Schistosoma haematobium in a seasonal transmission setting of Côte d'Ivoire. METHODS: Sixty-four localities with a S. haematobium prevalence in school children aged 13-14 years above 4% were randomly assigned to 1 of 4 intervention arms over a 3-year period: (1) the current standard strategy consisting of annual MDA before peak of transmission, (2) annual MDA after peak of transmission, (3) biannual MDA, and (4) standard MDA combined with snail control. The primary outcome was prevalence and intensity of S. haematobium infection in children aged 9-12 years 1 year after the final intervention, using urine filtration performed by experienced microscopists. RESULTS: By study end, we observed the lowest S. haematobium prevalence in the biannual MDA, compared to the standard treatment arm (0.6% vs 7.5%; odds ratio [OR] = 0.07, 95% confidence interval [CI] = .02 to .24). The prevalence in arms 2 and 4 was about 3.5%, which was not statistically significantly different from the standard strategy (both ORs 0.4, 95% CI = .1 to ~1.8). New cases of infection were still observed in all arms at study end. CONCLUSIONS: Biannual MDA was the only regimen that outperformed the standard treatment. All strategies resulted in decreased prevalence of infection; however, none of them was able to interrupt transmission of S. haematobium within a 3-year period. CLINICAL TRIALS REGISTRATION: ISRCTN10926858.

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.

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.

Effectiveness of school-based preventive chemotherapy strategies for sustaining the control of schistosomiasis in Côte d'Ivoire: Results of a 5-year cluster randomized trial.

BACKGROUND: Preventive chemotherapy using praziquantel is the mainstay for schistosomiasis control. However, there is little evidence on what is supposed to be the most effective school-based treatment strategy to sustain morbidity control. The aim of this study was to compare differences in Schistosoma mansoni prevalence and infection intensity between three different schedules of school-based preventive chemotherapy in an area with moderate prevalence of S. mansoni in Côte d'Ivoire. METHODOLOGY: Seventy-five schools were randomly assigned to one of three intervention arms: (i) annual school-based preventive chemotherapy with praziquantel (40 mg/kg) over four years; (ii) praziquantel treatment only in the first two years, followed by two years whithout treatment; and (iii) praziquantel treatment in years 1 and 3 without treatment in-between. Cross-sectional parasitologic surveys were carried out prior to each round of preventive chemotherapy. The difference in S. mansoni prevalence and infection intensity was assessed by multiple Kato-Katz thick smears, among children aged 9-12 years at the time of each survey. First-grade children, aged 5-8 years who had never received praziquantel, were also tested at baseline and at the end of the study. PRINCIPAL FINDINGS: Overall, 7,410 children aged 9-12 years were examined at baseline and 7,223 at the final survey. The baseline prevalence of S. mansoni was 17.4%, 20.2%, and 25.2% in arms 1, 2, and 3, respectively. In the final year, we observed the lowest prevalence of 10.4% in arm 1, compared to 18.2% in arm 2 and 17.5% in arm 3. The comparison between arms 1 and 2 estimated an odds ratio (OR) of 0.52 but the difference was not statistically significant (95% confidence interval (CI) = 0.23-1.16). Likewise the difference between arms 1 and 3 lacked statistical significance (OR = 0.55, 95% CI = 0.23-1.29). There was no noteworthy difference observed between arms 2 and 3 (OR = 1.06, 95% CI = 0.64-1.75). The lowest S. mansoni fecal egg counts in the final year survey were observed in arm 1 (7.9 eggs per gram of stool (EPG)). However, compared with 11.5 EPG in arm 2 and 15.4 EPG in arm 3, the difference lacked statistical significance. There were 4,812 first-grade children examined at baseline and 4,513 in the final survey. The overall prevalence of S. mansoni in these children slightly decreased in arms 1 (from 4.5% to 3.6%) and 2 (from 4.7% to 4.3%), but increased in arm 3 (from 6.8% to 7.9%). However, there was no significant difference in prevalence and infection intensity observed between study arms. CONCLUSIONS/SIGNIFICANCE: The three treatment schedules investigated led to a reduction in the prevalence and intensity of S. mansoni infection among children aged 9-12 years. Comparing intervention arms at the end of the study, no statistically significant differences were observed between annual treatement and the other two treatment schedules, neither in reduction of prevalence nor intensity of infection. It is important to combine our results with those of three sister trials conducted simultaneously in other African countries, before final recommendations can be drawn.

Role of Anopheles gambiae s.s. and Anopheles coluzzii (Diptera: Culicidae) in Human Malaria Transmission in Rural Areas of Bouaké, in Côte d'Ivoire.

Studies done in Bouaké (Côte d'Ivoire) about 20-yr ago reported that Anopheles gambiae s.l. Giles was the major malaria vector. The present study aimed to update these data and to identify the main vectors. Mosquitoes were collected in Allokokro and Petessou villages between June 2014 and December 2015 using the human landing catching method. Potential breeding sites of An. gambiae s.l. were identified in August and October 2014 and mapped using GPS. Anopheles species were morphologically and molecularly [polymerase chain reaction (PCR)] identified. Ovaries of female were dissected to determine the parity and infection with Plasmodium was detected in head and thorax by quantitative PCR. In Allokokro, the biting rate of An. gambiae s.s was significantly greater than Anopheles coluzzii, whereas, in Petessou, biting rates of both species were comparable. Plasmodium falciparum (Haemosporida: Plasmodiidae), Plasmodium malariae (Haemosporida: Plasmodiidae), and Plasmodium ovale (Haemosporida: Plasmodiidae) identified in both villages. The infection rates of An. gambiae s.s. and An. coluzzii were not significantly different. The entomological inoculation rate (EIR) of An. gambiae s.s. for P. falciparum was 9-fold greater than that of An. coluzzii in Allokokro; however, in Petessou, the EIRs of both species were comparable. In both village, An. gambiae s.s was responsible for P. falciparum and P. ovale transmission whereas An. coluzzii transmitted all three Plasmodium species.

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.

Evaluation of Human Exposure to Aedes Bites in Rubber and Palm Cultivations Using an Immunoepidemiological Biomarker.

Arbovirus infections, mainly transmitted by Aedes mosquito, are emerging in Africa. Efficient vector control requires an understanding of ecological factors which could impact on the risk of transmission, such as environmental changes linked to agricultural practices. The present study aims to assess the level of human exposure to Aedes mosquito bites in different agroecosystem area, using an immunological tool which quantifies human IgG antibody response to one Ae. aegypti salivary peptide. Specific IgG responses were assessed during dry and rainy seasons, in children living in different villages in Côte d'Ivoire: N'Zikro (rubber and oil palm exploitations), Ehania-V5 (oil palm), and Ayébo (without intensive agricultural activities). In the dry season, specific IgG levels were significantly lower in Ayébo compared to Ehania-V5 and N'Zikro and, interestingly, were similarly high in both villages with cultivations. In the rainy season, no difference of specific IgG was observed between villages. Specific IgG responses remained therefore high during both seasons in villages associated with intensive agricultural. The rubber and oil palm cultivations could maintain a high level of human exposure to Aedes mosquito bites during both dry and rainy seasons. These agricultural activities could represent a permanent risk factor of the transmission of arboviruses.

Dynamics of freshwater snails and Schistosoma infection prevalence in schoolchildren during the construction and operation of a multipurpose dam in central Côte d'Ivoire.

BACKGROUND: The construction and operation of small multipurpose dams in Africa have a history of altering the transmission of water-based diseases, including schistosomiasis. The current study was designed to investigate the abundance and dynamics of schistosomiasis intermediate host snails and Schistosoma infections in humans during the construction and the first years of operation of a small multipurpose dam in Côte d'Ivoire. METHODS: The study was carried out in Raffierkro and four neighbouring villages in central Côte d'Ivoire between 2007 and 2012. Snails were collected by two experienced investigators using scoops and forceps for 15 min at each site. Snails were identified at genera and, whenever possible, species level, and subjected to testing for cercarial shedding. Schoolchildren aged 6-15 years were examined once every year for Schistosoma haematobium and S. mansoni infection, using urine filtration and duplication Kato-Katz thick smears, respectively. Additionally, 551 adults were examined for Schistosoma infection before (June 2007) and 359 individuals 2 years after dam construction (June 2009). RESULTS: Overall, 1 700 snails belonging to nine different genera were collected from 19 sampling sites. Bulinus (potential intermediate host snails of S. haematobium) and Pila were the most common genera, whereas Biomphalaria (potential intermediate host snail of S. mansoni), Lymnaea, Physa and Melanoides were found in two villages. During the first-year sampling period, 65 snails were collected, of which 13 (20%) were schistosomiasis intermediate hosts. In subsequent years, out of 1 635 snails collected, 1 079 (66%) were identified as potential intermediate host for schistosomiasis, but none were shedding cercariae. The prevalence of S. mansoni among adults in the study area was low (0.4% in 2007 and 0.3% in 2009), whereas the prevalence of S. haematobium declined from 13.9% to 2.9% in this two-year period. CONCLUSIONS: The low prevalence of schistosomiasis in humans and the absence of infected intermediate host snails during the construction and early phase of operation of a small multipurpose dam suggest that there was no or only very little local transmission. However, the considerable increase in the number of intermediate host snails and their dispersion in irrigation canals call for rigorous surveillance, so that adequate public health measures can be taken in case of early signs of an outbreak.

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.