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.

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.

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.

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.

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.

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.