Human and animal serological surveys suggest that West Nile virus (WNV) circulation is widely distributed in Madagascar. However, there are no reported West Nile fever outbreaks or epizootics in the country and only one fatal human case has been reported to date. Currently there is very limited information on the maintenance and the transmission of WNV in Madagascar and particularly on the mosquito species involved in transmission cycles. In 2014, we initiated a study to investigate mosquito species composition, relative abundance, and trophic behavior in Mitsinjo District close to Lake Kinkony, a WNV endemic area in north-western Madagascar. We collected a total of 2519 adult mosquitoes belonging to 21 different species. The most abundant species was Aedeomyia (Aedeomyia) madagascarica Brunhes, Boussès & da Cunha Ramos, which made up 83% of all the mosquitoes collected. Mosquito abundance was associated with proximity to the lake (Morafeno and Ankelimitondrotra). Additionally, a correlation was observed between the lake-side biotope and the abundance of mosquito vectors in Morafeno. WNV RNA was detected in one pool of Ae. madagascarica and one pool of Anopheles (Cellia) pauliani Grjebine, suggesting that these two species may be involved in the maintenance and/or transmission of WNV in Madagascar.

BackgroundMalaria is still a heavy public health concern in Madagascar. Few studies combining parasitology and entomology have been recently conducted despite the need for such information to design proper vector control measures. In a region of moderate to intense transmission of both Plasmodium falciparum and Plasmodium vivax, we conducted a combined parasitology and entomology survey in two nearby villages, across a malaria transmission season from December 2016 to April 2017.Methodology/Principal findingsCommunity-based surveys were conducted in the two close by villages at three time points during a single malaria transmission season. Plasmodium carriage in the human populations was determined by Rapid Diagnostic Tests (RDTs), microscopy and real-time PCR. Anthropophilic mosquitoes were captured by human landing captures and presence of Plasmodium sporozoites was assessed by robust Real Time PCR. Overall human malaria prevalence was 8.0% by RDT, 4.8% by microscopy and 11.9% by PCR, mainly due to P. falciparum detected in 92.2% of the PCR positive samples and Plasmodium vivax (5.7%). No significant differences in Plasmodium human carriage was observed between the 2 villages at any time point. Of the 1553 anopheline mosquitoes tested, 13 were found carrying Plasmodium sporozoites, the majority of them being captured outdoor. The mosquito sporozoite indices were not significantly different between the two villages. However, our entomological analysis revealed that Anopheles coustani was the main vector in one village, being responsible of 25.5 infective bites during the whole survey, whereas it was Anopheles arabiensis in the other village with 15 infective bites. In addition, we found a significant higher number of endophagic An. coustani and An. arabiensis in one village compared to the other.Conclusions/SignificanceDespite similar human malaria prevalence in two close by villages, the entomological survey demonstrated the contribution of two different mosquito species in each village, and importantly the role of a suspected secondary malaria vector, An. coustani, as the main vector in one village. This, along with its higher endophagic rate in that village, highlights the importance of combining parasitology and entomology surveys for better targeting the actual local malaria vector. Such study should contribute to the malaria pre-elimination goal established under the 2018-2022 National Malaria Strategic Plan.Author SummaryMalaria is still a major health concern in many countries in sub-Saharan Africa such as Madagascar. In this study, we determined the contribution of malaria vectors in the transmission of Plasmodium parasites in two nearby villages in an area of moderate to high malaria transmission in Madagascar. We collected, during a single malaria transmission season, parasitological data in the human population and entomological data in the mosquito population, in order to evaluate Plasmodium carriage in these two populations. The results showed that despite similarity in human malaria prevalence and in vector species diversity in each village, the contribution of vectors was different between the two villages. An. arabiensis was the major vector in Ambohitromby while it was An. coustani that played this role in Miarinarivo. Importantly, this study is the first that clearly demonstrates that An. coustani could act as a major local vector in Madagascar. Such study should help deploying adapted malaria vector control and contributing to the malaria pre-elimination goal established under the 2018-2022 National Malaria Strategic Plan.

Background: Malaria is still a heavy public health concern in Madagascar. Few studies combining parasitology and entomology have been conducted despite the need for accurate information to design effective vector control measures. In a Malagasy region of moderate to intense transmission of both Plasmodium falciparum and P. vivax, parasitology and entomology have been combined to survey malaria transmission in two nearby villages. Methods: Community-based surveys were conducted in the villages of Ambohitromby and Miarinarivo at three time points (T1, T2 and T3) during a single malaria transmission season. Human malaria prevalence was determined by rapid diagnostic tests (RDTs), microscopy and real-time PCR. Mosquitoes were collected by human landing catches and pyrethrum spray catches and the presence of Plasmodium sporozoites was assessed by TaqMan assay. Results: Malaria prevalence was not significantly different between villages, with an average of 8.0% by RDT, 4.8% by microscopy and 11.9% by PCR. This was mainly due to P. falciparum and to a lesser extent to P. vivax. However, there was a significantly higher prevalence rate as determined by PCR at T2 (χ 2 2 = 7.46, P = 0.025). Likewise, mosquitoes were significantly more abundant at T2 (χ 2 2 = 64.8, P < 0.001), especially in Ambohitromby. At T1 and T3 mosquito abundance was higher in Miarinarivo than in Ambohitromby (χ 2 2 = 14.92, P < 0.001). Of 1550 Anopheles mosquitoes tested, 28 (1.8%) were found carrying Plasmodium sporozoites. The entomological inoculation rate revealed that Anopheles coustani played a major contribution in malaria transmission in Miarinarivo, being responsible of 61.2

Introduction In Madagascar, malaria control relies on the countrywide use of long lasting insecticide treated bed nets (LLINs) and on indoor residual spraying (IRS) in the central highland area as well as a small area on the eastern coast. We tested insecticide resistance mechanisms of Anopheles funestus from Tsararano, a malaria endemic village in the coastal health district of Marovoay. Methods Insecticide susceptibility bioassays were done in July 2017 on first-generation Anopheles funestus (F1) to assess (i) the susceptibility to permethrin (0.05%), deltamethrin (0.05%), DDT (4%), malathion (5%), fenitrothion (1%), and bendiocarb (0.1%); (ii) the effect of preexposure to the piperonyl butoxide (PBO) synergist; and (iii) the enzymatic activities of cytochrome P450, esterases, and glutathione S-transferases (GST). Results Our results demonstrated that An. funestus was phenotypically resistant to pyrethroids and bendiocarb, with a mortality rate (MR) of 33.6% (95%CI: 24.5-43.7%) and 86% (95%CI: 77.6-92.1%), respectively. In contrast, An. funestus were 100% susceptible to DDT and organophosphates (malathion and fenitrothion). Preexposure of An. funestus to PBO synergist significantly restored the susceptibility to bendiocarb (MR=100%) and increased the MR in the pyrethroid group, from 96% (95%CI: 90.0-98.9%) to 100% for deltamethrin and permethrin, respectively (χ2 = 43, df = 3, P< 0.0001). Enzymatic activities of cytochrome P450 and α-esterases were significantly elevated among An. funestus compared with the IPM reference strain (Mann-Whitney U= 30, P<0.0001; U = 145.5, P <0.0001, respectively). No significant differences of β-esterases activities compared to the IPM reference strain were observed (Mann-Whitney U = 392.5, P = 0.08). Conclusion In Tsararano, despite the absence of an IRS programme, there is evidence of high levels of insecticide resistance to pyrethroids and bendiocarb in An.