Differential insecticide resistance in Anopheles arabiensis populations in the seaside area of Mbour and its suburbs in Senegal.

Regular monitoring of insecticide resistance status is an important step in implementing appropriate and adapted insecticide-based strategies for vector control. In Senegal, Indoor Residual Spraying (IRS) and a national distribution campaign for long-lasting insecticide-treated net (LLIN) have been implemented since 2007 and 2009, respectively to prevent malaria transmission. To expand and ensure the sustainability of these strategies, we conducted a study on the status of insecticide resistance in malaria vectors in the seaside area of Mbour and its suburbs where no data were previously available. Anopheles larvae were sampled from four study sites (two in both coastal and inland areas) and reared to adulthood in the insectarium. Non-blood-fed females aged 3-5 days were then tested for susceptibility to permethrin, deltamethrin, lambdacyhalothrin, bendiocarb and pirimiphos-methyl. PCR amplification was used to identify sibling species of the An. gambiae complex and genotyping for the presence of resistance knockdown (kdr) L1014S, L1014F and Ace-1 G119S. Anopheles arabiensis was the only species present in the area. At all four sites, mosquitoes were resistant to deltamethrin, permethrin, and lambdacyhalothrin, and exhibited varying degrees of resistance to bendiocarb and pirimiphos-methyl. Overall, high levels of leucine-serine/phenylalanine substitutions at position 1014 (L1014S/L1014F) were observed, with frequencies ranging from 76.4 to 85.2 % for L1014F, and from 43.2 to 66.7 % for L1014S, compared to 8.1 to 28.3 for the Ace-1 G119S mutation. These results indicate a high level of phenotypic and genotypic resistance to insecticides, which is alarming, as it could have a significant impact on the operational effectiveness of current vector control tools that rely on pyrethroids. However, in the case of bendiocarb and pirimiphos-methyl, while some level of tolerance was observed, their potential use requires regular monitoring to prevent operational failure, as their deployment could potentially lead to an increase in resistance to them.

Host feeding preferences of malaria vectors in an area of low malaria transmission.

Studying the behaviour and trophic preferences of mosquitoes is an important step in understanding the exposure of vertebrate hosts to vector-borne diseases. In the case of human malaria, transmission increases when mosquitoes feed more on humans than on other animals. Therefore, understanding the spatio-temporal dynamics of vectors and their feeding preferences is essential for improving vector control measures. In this study, we investigated the feeding behaviour of Anopheles mosquitoes at two sites in the Sudanian areas of Senegal where transmission is low following the implementation of vector control measures. Blood-fed mosquitoes were collected monthly from July to November 2022 by pyrethrum spray catches in sleeping rooms of almost all houses in Dielmo and Ndiop villages, and blood meals were identified as from human, bovine, ovine, equine and chicken by ELISA. Species from the An. gambiae complex were identified by PCR. The types and numbers of potential domestic animal hosts were recorded in each village. The Human Blood Index (HBI) and the Manly Selection Ratio (MSR) were calculated to determine whether hosts were selected in proportion to their abundance. Spatio-temporal variation in HBI was examined using the Moran's index. A total of 1251 endophilic Anopheles females were collected in 115 bedrooms, including 864 blood fed females of 6 species. An. arabiensis and An. funestus were predominant in Dielmo and Ndiop, respectively. Of the 864 blood meals tested, 853 gave a single host positive result mainly on bovine, equine, human, ovine and chicken in decreasing order in both villages. Overall, these hosts were not selected in proportion to their abundance. The human host was under-selected, highlighting a marked zoophily for the vectors. Over time and space, the HBI were low with no obvious trend, with higher and lower values observed in each of the five months at different points in each village. These results highlight the zoophilic and exophagic behaviour of malaria vectors. This behaviour is likely to be a consequence of the distribution and use of LLINs in both villages and may increase risk of residual outdoor transmission. This underlines the need to study the feeding host profile of outdoor resting populations and how domestic animals may influence malaria epidemiology in order to tailor effective malaria vector control strategies in the two villages.

Evaluation of the residual efficacy and physical durability of five long-lasting insecticidal nets (LLINs) in Senegal.

BACKGROUND: The preventive and curative strategies of malaria are based on promoting the use of long-lasting insecticidal nets (LLINs) and treating confirmed cases with artemisinin-based combination therapy. These strategies have led to a sharp decline in the burden of malaria, which remains a significant public health problem in sub-Saharan countries. The objective of this study was to determine and compare the residual efficacy of LLINs recommended by the World Health Organization. METHODS: The study was conducted in six villages in two sites in Senegal located in the Sahelo-Sudanian area of the Thiès region, 70 km from Dakar and in Mbagame, a semi-urban zone in the Senegal River Valley. A census was conducted of all sleeping places in each household to be covered by LLINs. Five brands of LLIN were distributed, and every six months, retention rates, net use, maintenance, physical integrity, insecticide chemical content, and biological efficacy were examined for each type of LLIN. RESULTS: A total of 3012 LLINs were distributed in 1249 households in both sites, with an average coverage rate of 94% (95% CI 92.68-95.3). After 36 months, the average retention rate was 12.5% and this rate was respectively 20.5%, 15.1%, 10%, 7%, and 3% for Olyset Net®, Dawa Plus® 2.0, PermaNet® 2.0, NetProtect® and Life Net®, respectively. The proportion of LLINs with holes and the average number of holes per mosquito net increased significantly during each follow-up, with a large predominance of size 1 (small) holes for all types of LLINs distributed. During the three-year follow-up, bioassay mortality rates of a susceptible strain of insectary reared Anopheles coluzzii decreased in the following net types: in Dawa Plus® 2.0 (100% to 51.7%), PermaNet® 2.0 (96.6% to 83%), and Olyset Net® (96.6% to 33.3%). Mortality rates remained at 100% in Life Net® over the same time period. After 36 months, the average insecticide content per brand of LLIN decreased by 40.9% for Dawa Plus® 2.0, 31% for PermaNet® 2.0, 39.6% for NetProtect® and 51.9% for Olyset Net® and 40.1% for Life Net. CONCLUSIONS: Although some net types retained sufficient insecticidal activity, based on all durability parameters measured, none of the net types survived longer than 2 years.

Entomological Survey Confirms Changes in Mosquito Composition and Abundance in Senegal and Reveals Discrepancies among Results by Different Host-Seeking Female Traps.

Mosquitoes-borne diseases are major public health issues particularly in Africa. Vector control interventions and human-made environmental/climatic changes significantly affect the distribution and abundance of vector species. We carried out an entomological survey targeting host-seeking mosquitos in two different ecological contexts-coastal and inland-in Senegal, by CDC-light and BG-sentinel traps. Results show high predominance of Culex quinquefasciatus (90%) and of Anopheles arabiensis within malaria vectors (46%), with mean numbers of females/trap/nights =8 and <1, respectively, reinforcing previous evidence of changes in species composition and abundance, highlighting thus increasing risk of transmission of filariasis and emerging arboviruses in the Senegambia region. From the methodological perspective, results show a higher specificity of BG traps for Cx. quinquefasciatus and of CDC traps for An. gambiae s.l. and highlight that, despite both traps target the host-seeking fraction of the population, they provide different patterns of species abundance, temporal dynamics and host-seeking activity, leading to possible misinterpretation of the species bionomics. This draws attention to the need of taking into account trapping performance, in order to provide realistic quantification of the number of mosquitoes per units of space and time, the crucial parameter for evaluating vector-human contact, and estimating risk of pathogen transmission.

Insecticide resistance status and mechanisms in Aedes aegypti populations from Senegal.

Aedes aegypti is the main epidemic vector of arboviruses in Africa. In Senegal, control activities are mainly limited to mitigation of epidemics, with limited information available for Ae. aegypti populations. A better understanding of the current Ae. aegypti susceptibility status to various insecticides and relevant resistance mechanisms involved is needed for the implementation of effective vector control strategies. The present study focuses on the detection of insecticide resistance and reveals the related mechanisms in Ae. aegypti populations from Senegal. Bioassays were performed on Ae. aegypti adults from nine Senegalese localities (Matam, Louga, Barkedji, Ziguinchor, Mbour, Fatick, Dakar, Kédougou and Touba). Mosquitoes were exposed to four classes of insecticides using the standard WHO protocols. Resistance mechanisms were investigated by genotyping for pyrethroid target site resistance mutations (V1016G, V1016I, F1534C and S989P) and measuring gene expression levels of key detoxification genes (CYP6BB2, CYP9J26, CYP9J28, CYP9J32, CYP9M6, CCEae3a and GSTD4). All collected populations were resistant to DDT and carbamates except for the ones in Matam (Northern region). Resistance to permethrin was uniformly detected in mosquitoes from all areas. Except for Barkédji and Touba, all populations were characterized by a susceptibility to 0.75% Permethrin. Susceptibility to type II pyrethroids was detected only in the Southern regions (Kédougou and Ziguinchor). All mosquito populations were susceptible to 5% Malathion, but only Kédougou and Matam mosquitoes were susceptible to 0.8% Malathion. All populations were resistant to 0.05% Pirimiphos-methyl, whereas those from Louga, Mbour and Barkédji, also exhibited resistance to 1% Fenitrothion. None of the known target site pyrethroid resistance mutations was present in the mosquito samples included in the genotyping analysis (performed in > 1500 samples). In contrast, a remarkably high (20-70-fold) overexpression of major detoxification genes was observed, suggesting that insecticide resistance is mostly mediated through metabolic mechanisms. These data provide important evidence to support dengue vector control in Senegal.

Anopheles arabiensis seasonal densities and infection rates in relation to landscape classes and climatic parameters in a Sahelian area of Senegal.

BACKGROUND: The influence of environmental and climatic factors on malaria vector bionomics and transmission is an important topic in the context of climatic change particularly at macro-geographical level. Sahelian areas could be particularly affected due to heterogeneous features including high inter-annual variability in rainfall and others associated parameters. Therefore, baseline information on the impact of environmental and climatic factors on malaria transmission at micro-geographical level is required for vector risk management and implementation of control strategies. METHODS: Malaria vectors were collected indoors by pyrethrum spray catches in 14 villages belonging to 4 different landscape classes (wooded savanna, shrubby savanna, bare soils and steppe) in the sylvo-pastoral area of Senegal. Plasmodium falciparum infection rates were determined using an indirect enzyme-linked immunosorbent assay (ELISA). RESULTS: An. arabiensis was the predominant species in all landscape classes and was the only species collected at the end of the rainy season excepted in villages located in bare soils where it cohabited with An. coluzzii. Mean temperature and relative humidity showed similar variations in all the landscape classes covered whereas rainfall was more heterogeneous in terms of pattern, frequency and amount. The mean densities of An. arabiensis displayed high seasonal differences with peaks observed in August or September. A positive non-significant correlation was observed between An. arabiensis densities for rainfall and humidity whereas a negative non-significant correlation was reported for temperature. Plasmodium falciparum-infected mosquitoes were detected only in wooded savanna and bare soils villages. CONCLUSIONS: These observations suggest key roles played by landscape classes and rainfall in malaria vector densities, infection rates and malaria transmission that could be more pronounced in villages situated in wooded savanna and bare soils. Due to the close relationship between environmental and meteorological parameters in this Sahelian region, additional studies on the impact of these parameters are required to further ascertain their association with entomological parameters involved in malaria transmission. From the public health point of view, such information could be useful for human population settlements as well as for monitoring and modelling purposes giving early warning system for implementation of interventions in these unstable transmission zones.

Spatio-temporal analysis of host preferences and feeding patterns of malaria vectors in the sylvo-pastoral area of Senegal: impact of landscape classes.

BACKGROUND: The study of vector feeding behaviour is an important step in the understanding of the epidemiology of vector borne diseases. The main objective of this work was to study the spatio-temporal host preferences and blood-feeding patterns of malaria vectors in a pastoral area of Senegal where cattle breeding is the main human activity. METHODS: Malaria vectors were collected indoors by pyrethrum spray catch in 16 villages belonging to 4 different landscape classes (wooded savanna, shrubby savanna, bare soils and steppe). Blood meals sources were determined using a direct enzyme-linked immunosorbent assay (ELISA). RESULTS: The blood meal origins of 1886 freshly fed An. gambiae s.l. were determined. Among these blood meals, most were taken on a single host: 40.1% on human and 37.1% on animal. The range in proportions of blood meals taken from human were 25-62.4% in wooded savanna villages, 23.5-61.9% in shrubby savanna villages, 31.3-70% in bare soils villages and 57.7-68.7 in steppe villages. Blood meals taken from bovines were very heterogeneous with two clusters localized in the Northeast and Southwest axis of the study area that corresponds to the distribution of the main water ponds. Patent mixed blood meals taken from human and non-human were significantly higher than those taken from two animals, the highest proportions being observed in September (shrubby savanna, bare soils and steppe villages) or October (wooded savanna villages). CONCLUSIONS: These observations suggest that in this pastoral area, differences in feeding patterns of malaria vectors are merely linked to the specific localization of villages and are not influenced by landscape class distribution. In addition, the temporal variations in the anthropophilic rates are influenced by the presence of standing water in the study area.

Malaria transmission pattern in an area selected for clinical trials in the sudanian area of senegal (west Africa).

Malaria transmission pattern was studied in 3 villages (Toubanding, Daga Ndoup, and Keur Samba Guèye) situated within an area selected for clinical trials. The study was conducted in the rainy season from July to December 2011. The main objective of this work was to gather baseline data on malaria transmission intensity and other entomological parameters before the advent of clinical trials. Mosquitoes were collected by Human-Landing Collections (HLCs) and by pyrethrum spray catches (PSCs). Five anopheline species were collected, namely, An. arabiensis, An. gambiae, An. funestus, An. pharoensis, and An. rufipes, giving a heterogeneous distribution within the study area. The populations dynamics of the vectors varied temporarily in each village depending on the pattern of the rainy season. Transmission intensity estimated by the entomological inoculation rate (EIR) was measured in each of the three villages with the variations linked to the microecological differences between the villages. Measurements were calculated for August, September, and October and were found to vary between 4 and 30 infected bites per person over the study period with a peak intensity observed in September. These results indicate that epidemiological field trials on malaria could be conducted in this area on the basis of the differences observed with transmission intensity, micro-ecological variations, and the objectives of the trials.