Spatiotemporal multiple insecticide resistance in Aedes aegypti populations in French Guiana: need for alternative vector control.

BACKGROUND: Aedes aegypti is the sole vector of urban arboviruses in French Guiana. Overtime, the species has been responsible for the transmission of viruses during yellow fever, dengue, chikungunya and Zika outbreaks. Decades of vector control have produced resistant populations to deltamethrin, the sole molecule available to control adult mosquitoes in this French Territory. OBJECTIVES: Our surveillance aimed to provide public health authorities with data on insecticide resistance in Ae. aegypti populations and other species of interest in French Guiana. Monitoring resistance to the insecticide used for vector control and to other molecule is a key component to develop an insecticide resistance management plan. METHODS: In 2009, we started to monitor resistance phenotypes to deltamethrin and target-site mechanisms in Ae. aegypti populations across the territory using the WHO impregnated paper test and allelic discrimination assay. FINDINGS: Eight years surveillance revealed well-installed resistance and the dramatic increase of alleles on the sodium voltage-gated gene, known to confer resistance to pyrethroids (PY). In addition, we observed that populations were resistant to malathion (organophosphorous, OP) and alpha-cypermethrin (PY). Some resistance was also detected to molecules from the carbamate family. Finally, those populations somehow recovered susceptibility against fenitrothion (OP). In addition, other species distributed in urban areas revealed to be also resistant to pyrethroids. CONCLUSION: The resistance level can jeopardize the efficiency of chemical adult control in absence of other alternatives and conducts to strongly rely on larval control measures to reduce mosquito burden. Vector control strategies need to evolve to maintain or regain efficacy during epidemics.

High malaria transmission in a forested malaria focus in French Guiana: How can exophagic Anopheles darlingi thwart vector control and prevention measures?

In French Guiana, malaria vector control and prevention relies on indoor residual spraying and distribution of long lasting insecticidal nets. These measures are based on solid epidemiological evidence but reveal a poor understanding of the vector. The current study investigated the behaviour of both vectors and humans in relation to the ongoing prevention strategies. In 2012 and 2013, Anopheles mosquitoes were sampled outdoors at different seasons and in various time slots. The collected mosquitoes were identified and screened for Plasmodium infection. Data on human behaviour and malaria episodes were obtained from an interview. A total of 3,135 Anopheles mosquitoes were collected, of which Anopheles darlingi was the predominant species (96.2%). For the December 2012-February 2013 period, the Plasmodium vivax infection rate for An. darlingi was 7.8%, and the entomological inoculation rate was 35.7 infective bites per person per three-month span. In spite of high bednet usage (95.7%) in 2012 and 2013, 52.2% and 37.0% of the participants, respectively, had at least one malaria episode. An. darlingi displayed heterogeneous biting behaviour that peaked between 20:30 and 22:30; however, 27.6% of the inhabitants were not yet protected by bednets by 21:30. The use of additional individual and collective protective measures is required to limit exposure to infective mosquito bites and reduce vector densities.

Updated Checklist of the Mosquitoes (Diptera: Culicidae) of French Guiana.

The incredible mosquito species diversity in the Neotropics can provoke major confusion during vector control programs when precise identification is needed. This is especially true in French Guiana where studies on mosquito diversity practically ceased 35 yr ago. In order to fill this gap, we propose here an updated and comprehensive checklist of the mosquitoes of French Guiana, reflecting the latest changes in classification and geographical distribution and the recognition of current or erroneous synonymies. This work was undertaken in order to help ongoing and future research on mosquitoes in a broad range of disciplines such as ecology, biogeography, and medical entomology. Thirty-two valid species cited in older lists have been removed, and 24 species have been added including 12 species (comprising two new genera and three new subgenera) reported from French Guiana for the first time. New records are from collections conducted on various phytotelmata in French Guiana and include the following species: Onirion sp. cf Harbach and Peyton (2000), Sabethes (Peytonulus) hadrognathus Harbach, Sabethes (Peytonulus) paradoxus Harbach, Sabethes (Peytonulus) soperi Lane and Cerqueira, Sabethes (Sabethinus) idiogenes Harbach, Sabethes (Sabethes) quasicyaneus Peryassú, Runchomyia (Ctenogoeldia) magna (Theobald), Wyeomyia (Caenomyiella) sp. cf Harbach and Peyton (1990), Wyeomyia (Dendromyia) ypsipola Dyar, Wyeomyia (Hystatomyia) lamellata (Bonne-Wepster and Bonne), Wyeomyia (Miamyia) oblita (Lutz), and Toxorhynchites (Lynchiella) guadeloupensis (Dyar and Knab). At this time, the mosquitoes of French Guiana are represented by 235 species distributed across 22 genera, nine tribes, and two subfamilies.

Mosquito magnet® liberty plus trap baited with octenol confirmed best candidate for Anopheles surveillance and proved promising in predicting risk of malaria transmission in French Guiana.

BACKGROUND: In French Guiana, Mosquito Magnet® Liberty Plus trap baited with octenol (MMoct) has been proposed for sampling Anopheles darlingi after comparison with CDC light trap and Human landing catch (HLC). However, other available lures were not tested. The current study compared MMoct and MM baited with Lurex™ (MMlur) to HLC, and analysed entomological data from MMoct collection with malaria cases to facilitate malaria surveillance. METHODS: Two independent experiments were conducted during 2012 and 2013 in Saint-Georges town, French Guiana. The first experiment used Latin square design to compare MMoct and MMlur to HLC between 18:30 to 22:30 and 05:00 to 07:00. Parity rate was determined for An. darlingi from each sampling system. In the second experiment, a 24:00 hour collection was done for four consecutive days during the first week of each month and every four days for the rest of the month using MMoct. Portion of the 24 hour collection was dissected for parity rate. All anophelines were screened for Plasmodium infection by PCR. Data for number of malaria cases was analysed for association with density of An. darlingi. RESULTS: In the first experiment, 3,721 anopheline mosquitoes were collected over 21 nights. Of these, 95.7% was identified morphologically to five species and An. darlingi contributed 98.4%, mainly from HLC (75.1%, CI 95% [73.2-77.0]) than MMoct (14.1%, CI 95% [12.6-15.7]) and MMlur (10.8%, CI 95% [9.4-12.2]). Species richness was highest in HLC meanwhile species diversity index was greatest in MMoct. MMoct collected more parous An. darlingi than HLC (p<0.0001) and MMlur (p=0.0021). The second experiment amounted to 2035 females, 60.8% belonging to 10 species. Anopheles darlingi constituted 85.0% of the species and had parity rate of 52.3%. Specimens were uninfected with Plasmodium. Density of An. darlingi best correlated with malaria cases observed six weeks later (p=0.0016; r=0.4774). CONCLUSION: Though MMoct and MMlur performed well in sampling An. darlingi, MMoct captured more species and, therefore, would be useful for surveillance. Even if it collected mostly parous mosquitoes, MMoct proved useful in collecting entomological data required for predicting malaria emergence. It is a potential replacement for HLC.

Noninvasive detection of Zika virus in mosquito excreta sampled from wild mosquito populations in French Guiana.

Arboviruses can be difficult to detect in the field due to relatively low prevalence in mosquito populations. The discovery that infected mosquitoes can release viruses in both their saliva and excreta gave rise to low-cost methods for the detection of arboviruses during entomological surveillance. We implemented both saliva and excreta-based entomological surveillance during the emergence of Zika virus (ZIKV) in French Guiana in 2016 by trapping mosquitoes around households of symptomatic cases with confirmed ZIKV infection. ZIKV was detected in mosquito excreta and not in mosquito saliva in 1 trap collection out of 85 (1.2%). One female Ae. aegypti L. (Diptera: Culicidae) was found with a ZIKV systemic infection in the corresponding trap. The lag time between symptom onset in a ZIKV-infected individual living near the trap site and ZIKV detection in this mosquito was 1 wk. These results highlight the potential of detection in excreta from trapped mosquitoes as a sensitive and cost-effective method to non invasively detect arbovirus circulation.

Variation in Pyrethroid Resistance Phenotypes in Anopheles darlingi in an Area with Residual Malaria Transmission: Warning of Suspected Resistance in French Guiana.

Anopheles darlingi is the main vector of malaria in South America. In French Guiana, malaria transmission occurs inland and along the rivers with a regular reemergence in the lower Oyapock area. Control against malaria vectors includes indoor residual spraying of deltamethrin and the distribution of long-lasting impregnated bednets. In this context, the level of resistance to pyrethroids was monitored for 4 years using CDC bottle tests in An. darlingi populations. A loss of susceptibility to pyrethroids was recorded with 30-minute knock-down measured as low as 81%. However, no pyrethroid molecular resistance was found by sequencing a 170 base pair fragment of the S6 segment of domain II of the voltage-gated sodium channel gene. Fluctuation of resistance phenotypes may be influenced by the reintroduction of susceptible alleles from sylvatic populations or by other mechanisms of metabolic resistance.

Identification of Neotropical Culex Mosquitoes by MALDI-TOF MS Profiling.

The mosquito (Diptera: Culicidae) fauna of French Guiana encompasses 242 species, of which nearly half of them belong to the genus Culex. Whereas several species of Culex are important vectors of arboviruses, only a limited number of studies focus on them due to the difficulties to morphologically identify field-caught females. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been reported as a promising method for the identification of mosquitoes. Culex females collected in French Guiana were morphologically identified and dissected. Abdomens were used for molecular identification using the COI (cytochrome oxidase 1) gene. Legs and thorax of 169 specimens belonging to 13 Culex species, (i.e., Cx. declarator, Cx. nigripalpus, Cx. quinquefasciatus, Cx. usquatus, Cx. adamesi, Cx. dunni, Cx. eastor, Cx. idottus, Cx. pedroi, Cx. phlogistus, Cx. portesi, Cx. rabanicolus and Cx. spissipes) were then submitted to MALDI-TOF MS analysis. A high intra-species reproducibility and inter-species specificity of MS spectra for each mosquito body part tested were obtained. A corroboration of the specimen identification was revealed between MALDI-TOF MS, morphological and molecular results. MALDI-TOF MS protein profiling proves to be a suitable tool for identification of neotropical Culex species and will permit the enhancement of knowledge on this highly diverse genus.

Incrimination of Anopheles (Anopheles) intermedius Peryassú, An. (Nyssorhynchus) nuneztovari Gabaldón, An. (Nys.) oswaldoi Peryassú as natural vectors of Plasmodium falciparum in French Guiana.

Anopheles darlingi Root is the major vector of human malaria in the Neotropics and has been considered to be the sole malaria vector in French Guiana. The presence of other potential vectors suggests that malaria may be transmitted by other species under certain conditions. From 2006-2011, all anopheline specimens collected from 11 localities were assayed to determine if the Plasmodium circumsporozoite protein was present. In addition to An. darlingi, we found Anopheles oswaldoi, Anopheles intermedius and Anopheles nuneztovari specimens that were infected with Plasmodium sp. Further investigations on the behaviour and ecology of An. oswaldoi, An. intermedius and An. nuneztovari are necessary to determine their role in malaria transmission in French Guiana.

High susceptibility to Chikungunya virus of Aedes aegypti from the French West Indies and French Guiana.

OBJECTIVES: To estimate the vector competence of Aedes aegypti populations sampled from distinct anthropogenic environments in French Guiana, Guadeloupe and Martinique for the strain CHIKV 06.21. METHODS: F(1)/F(2) females were orally infected at titres of 10(6) and 10(7.5) pfu/ml in blood-meals. Disseminated infection rates (DIR) of mosquitoes were estimated using indirect fluorescent antibody assay on heads' squashes, 7 or 14 days post-infection (pi). RESULTS: At a titre of 10(7.5) pfu/ml, DIR ranged from 88.9% to 100.0% and were not significantly different whether assessed at day 7 or 14 pi. At a titre of 10(6) pfu/ml, DIR observed 7 days pi ranged from 37.6 to 62.0%. CONCLUSIONS: Ae. aegypti from French Guiana and French West Indies are highly competent to transmit CHIKV. An evaluation of DIR 7 days rather than 14 days pi is adequate to estimate vector competence. The titre of 10(6) pfu/ml allows us to distinguish Ae. aegypti populations originating from distinct environments (dense or diffuse housing) by their vector competence. This assessment is a prerequisite to better evaluate the potential risk of Chikungunya outbreaks once the virus is introduced from endemic regions.

Multiple insecticide resistance in Aedes aegypti (Diptera: Culicidae) populations compromises the effectiveness of dengue vector control in French Guiana.

In French Guiana, pyrethroids and organophosphates have been used for many years against Aedes aegypti. We aimed to establish both the resistance level of Ae. aegypti and the ultra low volume spray efficacy to provide mosquito control services with practical information to implement vector control and resistance management. Resistance to deltamethrin and fenitrothion was observed. In addition, the profound loss of efficacy of AquaK'othrine® and the moderate loss of efficacy of Paluthion® 500 were recorded. Fenitrothion remained the most effective candidate for spatial application in French Guiana until its removal in December 2010. Further investigation of the mechanism of resistance to deltamethrin demonstrated the involvement of mixed-function oxidases and, to a lesser extent, of carboxylesterases. However, these observations alone cannot explain the level of insecticide resistance we observed during tube and cage tests.

Annual variations in the number of malaria cases related to two different patterns of Anopheles darlingi transmission potential in the Maroni area of French Guiana.

BACKGROUND: With an Annual Parasite Incidence (API) of 132.1, in the high and moderate risks zones, the Maroni area of French Guiana has the second highest malaria incidence of South-America after Guyana (API = 183.54) and far above Brazil (API = 28.25). Malaria transmission is occurring despite strong medical assistance and active vector control, based on general WHO recommendations. This situation is generated by two main factors that are the social and cultural characteristics of this border area, where several ethnic groups are living, and the lack of understanding of transmission dynamics of the main mosquito vector, Anopheles darlingi. In this context, entomological data collected in two villages belonging to two different ethnic groups of the French border of the Maroni River, were retrospectively analysed to find out how the mosquito bionomics are related to the malaria transmission patterns. METHODS: Data were provided by human landing catches of mosquitoes carried out each month for two years in two villages belonging to two ethnic groups, the Amerindians Wayanas and the Aloukous of African origin. The mosquitoes were sorted by species, sex, date, hour and place of collection and processed for Plasmodium sp. parasite detection. The data were compiled to provide the following variables: human biting rates (HBR), parity rates (PR), numbers of infective bites (IB), entomological inoculation rates (EIR) and numbers of infected mosquitoes surviving enough to transmit (IMT). Spatial and temporal differences of variables between locations and during the night were tested by the Kruskall-Wallis analysis of variance to find out significant variations. RESULTS: The populations of the main mosquito vector An. darlingi showed significant variations in the spatial and temporal HBR/person/night and HBR/person/hour, IB/person/month and IB/person/hour, and IMT/village/night and IMT/village/hour. In the village of Loca (Aloukous), the IMT peaked from June to August with a very low transmission during the other months. The risks were higher during the first part of the night and an EIR of 10 infective bites per person and per year was estimated. In the village of Twenke (Wayanas), high level of transmission was reported all year with small peaks in March and October. The risk was higher during the second part of the night and an EIR of 5 infective bites per person and per year was estimated. CONCLUSION: For the first time in the past 40 years, the mosquito bionomics was related to the malaria transmission patterns in French Guiana. The peak of malaria cases reported from August to October in the Maroni region is concomitant with the significant peak of An. darlingi IMT, reported from the village of Loca where transmission is higher. However, the persistent number of cases reported all year long may also be related to the transmission in the Amerindian villages. The An. darlingi bionomics for these two close populations were found significantly different and may explain why a uniform vector control method is inadequate. Following these findings, malaria prevention measures adapted to the local conditions are needed. Finally, the question of the presence of An. darlingi sub-species is raised.

Redescription and placement of Wyeomyia rorotai Senevet, Chabelard amp; Abonnenc (Diptera: Culicidae) in the subgenus Decamyia based on morphological and molecular analyses.

Decamyia Dyar is a subgenus of Wyeomyia Theobald with three valid species. Wyeomyia rorotai Senevet, Chabelard Abonnenc, a species originally described rather briefly in the subgenus Dendromyia, is without subgeneric position in the genus. In the present work, we redescribe Wy. rorotai in all life stages and formally define its taxonomic placement in the subgenus Decamyia by combining morphological and molecular analyses based on the mitochondrial cytochrome c oxidase subunit I gene. We also show that Decamyia is a rather homogeneous group of four species, i.e. Wy. ulocoma (Theobald), Wy. pseudopecten Dyar Knab, Wy. felicia Dyar Núñez Tovar and Wy. rorotai, the immature stages of which almost exclusively inhabit the flower bracts of Heliconiaceae.

Anopheles darlingi bionomics and transmission of Plasmodium falciparum, Plasmodium vivax and Plasmodium malariae in Amerindian villages of the Upper-Maroni Amazonian forest, French Guiana.

French Guiana is one of the areas in South America most affected by malaria and where the disease has become a serious public health problem. In spite of this situation, little recent entomological data are available from the main localities where the disease occurs, even though they are crucial for development of an effective vector control strategy. A longitudinal entomological survey was carried out from March 2000-February 2002 in three Amerindian villages, namely Twenké, Taluène and Cayodé, located in the Amazonian forest of the Upper-Maroni area, to assess anopheline mosquitoes and malaria transmission dynamics. Anopheles darlingi (Diptera: Culicidae) was the most abundant mosquito species caught during the study. This efficient American malaria vector was active the entire year, but showed an evident peak of abundance during the main rainfall season, from April-June, with an average human biting rate of 255.5 bites per person per night. Parity rates were homogeneous all year, indicating no significant seasonal variability in female survival rates. Estimated vectorial capacity indices were higher during the rainy season, even though the risk of transmission was present throughout the year (VCI > 1). A total of 14 An. darlingi were found infected with Plasmodium falciparum, Plasmodium vivax or Plasmodium malariae. The annual circumsporozoite indices were 0.15, 0.14 and 0.05, and the entomological inoculation rates were 22.8, 27.4 and 14.4 infected bites per person per year in Twenké, Taluène and Cayodé, respectively. An. darlingiwas endo-exophagic and rather exophilic in these localities. The species was collected throughout the night but was more aggressive between 21:30-03:30 h and after 05:30 h. Parity rates were homogeneous during the entire night. Impregnated hammock and/or bed nets, coupled with the use of mosquito repellents, as well as the early treatment of malarial cases, appear to be the most suitable tools for fighting malaria in these Amerindian villages since the spraying of residual insecticides is inefficient because of vector biology and the housing structure.

Diversity and ecology of sand flies (Diptera: Psychodidae: Phlebotominae) in coastal French Guiana.

In French Guiana, at least five Leishmania species are known to be sympatically transmitted in sylvatic ecotopes. However, the previous surveys on the phlebotomine sand fly fauna were published 20 years ago. During that period, many ecological changes have occurred. Sand fly collections were conducted with CDC light traps in five stations representing the main ecotopes of French Guiana. A total of 817 sand flies belonging to 2 genera, 18 sub-genera, and 46 different species were identified. The species Lutzomyia umbratilis (16.6% of the collected specimens), Lu. infraspinosa (12.7%), Lu. ininii (8.0%), and Lu. flaviscutellata (6.1%) were the most common species. The stratification by height, activity period, and resting site preferences of the most abundant sand flies were analyzed. Population abundance and diversity were compared for each ecotope. The potential of certain sand fly species in leishmaniasis transmission is discussed.

Aedes aegypti survival and dengue transmission patterns in French Guiana.

Field survival of Aedes aegypti females is a key parameter for estimating the dengue transmission potential of a mosquito population. The objectives of this study were to explore the dynamics of these survival rates at different times of the year in French Guiana and to analyze the results from the perspective of dengue patterns. The mosquitoes were captured, marked, released, and recaptured during four consecutive days in six houses every month, for three to 24 months, from January 1997 to December 1998. Laboratory experiments showed no effects on female survival but some effect on the survival of males. Females' daily survival in the field varied from 0.525 to 1 but was mostly between 0.8 and 0.95 during the entire year, with a mean value of 0.913. The field survival of Ae. aegypti females in French Guiana was thus in agreement with the likely transmission of dengue and the dengue endemic patterns throughout the year. On the other hand, heavy rainfalls during this time were less favorable to Ae. aegypti survival, which may explain part of the El Niño effect on dengue epidemics in French Guiana. The methods and results on Ae. aegypti survival will be implemented in a global dengue surveillance network in French Guiana.

Anopheles darlingi (Diptera: Culicidae) Dynamics in Relation to Meteorological Data in a Cattle Farm Located in the Coastal Region of French Guiana: Advantage of Mosquito Magnet Trap.

Information on dynamics of anopheline mosquitoes is limited in the coastal zone of French Guiana compared with inland endemic areas. Importantly, improvement of surveillance techniques for assessing malaria transmission indicators and comprehension of impact of meteorological factors on Anopheles darlingi Root, the main malaria vector, are necessary. Anopheline mosquitoes were collected continuously during 2012 and 2013 using Mosquito Magnet traps baited with octenol and human landing catches. The two methods were compared based on trends in abundance and parity rate of An. darlingi. Impact of meteorological factors on An. darlingi density estimates was investigated using Spearman's correlation and by binomial negative regression analysis. In all, 11,928 anopheline mosquitoes were collected, and 90.7% (n = 10,815) were identified consisting of four species, with An. darlingi making up 94.9% (n = 10,264). An. darlingi specimens collected by the two methods were significantly correlated, and no difference in parity rate was observed. The abundance of this species peaked in September (dry season) and variations along the years were influenced by relative humidity, temperature, rainfall, and wind speed. Number of mosquitoes collected during peak aggression period was influenced by wind speed and rainfall. Data gathered in this study provide fundamental information about An. darlingi, which can facilitate the design of vector control strategies and construction of models for predicting malaria risk.

Spatiotemporal multiple insecticide resistance in Aedes aegypti populations in French Guiana: need for alternative vector control

BACKGROUND Aedes aegypti is the sole vector of urban arboviruses in French Guiana. Overtime, the species has been responsible for the transmission of viruses during yellow fever, dengue, chikungunya and Zika outbreaks. Decades of vector control have produced resistant populations to deltamethrin, the sole molecule available to control adult mosquitoes in this French Territory. OBJECTIVES Our surveillance aimed to provide public health authorities with data on insecticide resistance in Ae. aegypti populations and other species of interest in French Guiana. Monitoring resistance to the insecticide used for vector control and to other molecule is a key component to develop an insecticide resistance management plan. METHODS In 2009, we started to monitor resistance phenotypes to deltamethrin and target-site mechanisms in Ae. aegypti populations across the territory using the WHO impregnated paper test and allelic discrimination assay. FINDINGS Eight years surveillance revealed well-installed resistance and the dramatic increase of alleles on the sodium voltage-gated gene, known to confer resistance to pyrethroids (PY). In addition, we observed that populations were resistant to malathion (organophosphorous, OP) and alpha-cypermethrin (PY). Some resistance was also detected to molecules from the carbamate family. Finally, those populations somehow recovered susceptibility against fenitrothion (OP). In addition, other species distributed in urban areas revealed to be also resistant to pyrethroids. CONCLUSION The resistance level can jeopardize the efficiency of chemical adult control in absence of other alternatives and conducts to strongly rely on larval control measures to reduce mosquito burden. Vector control strategies need to evolve to maintain or regain efficacy during epidemics.

A survey of adult anophelines in French Guiana: enhanced descriptions of species distribution and biting responses.

In French Guiana, Anopheles darlingi is considered the main malaria vector. However, several reports have hypothesized the implication of other anopheline species in malaria transmission for the territory. Data on the ecology of these other potential vectors is rare or even unexplored in French Guiana. The aim of this study was to describe the biting habits of several anopheline species in multiple localities in French Guiana. Six sampling sites yielded 1,083 anopheline adults. Results indicated the presence of An. darlingi in all study locations and it was the only species to be collected inside villages. Other anophelines collected included An. aquasalis, An. braziliensis, An. intermedius, An. mediopunctatus, An. nuneztovari, An. oswaldoi, and An. triannulatus, all of which were associated with open areas and forests. The environment and time, at which biting behavior was recorded, varied for each species. It was noted that An. oswaldoi showed a daytime rhythm in open areas. This study is the first to report on the biting habits of a range of anophelines in French Guiana that may play a role in malaria transmission. This information is vital to fully describe the risk of malaria transmission and thereby design appropriate vector control measures and malaria prevention programs.

High malaria transmission in a forested malaria focus in French Guiana: How can exophagic Anopheles darlingi thwart vector control and prevention measures?

In French Guiana, malaria vector control and prevention relies on indoor residual spraying and distribution of long lasting insecticidal nets. These measures are based on solid epidemiological evidence but reveal a poor understanding of the vector. The current study investigated the behaviour of both vectors and humans in relation to the ongoing prevention strategies. In 2012 and 2013, Anopheles mosquitoes were sampled outdoors at different seasons and in various time slots. The collected mosquitoes were identified and screened for Plasmodium infection. Data on human behaviour and malaria episodes were obtained from an interview. A total of 3,135 Anopheles mosquitoes were collected, of which Anopheles darlingi was the predominant species (96.2%). For the December 2012-February 2013 period, the Plasmodium vivax infection rate for An. darlingi was 7.8%, and the entomological inoculation rate was 35.7 infective bites per person per three-month span. In spite of high bednet usage (95.7%) in 2012 and 2013, 52.2% and 37.0% of the participants, respectively, had at least one malaria episode. An. darlingi displayed heterogeneous biting behaviour that peaked between 20:30 and 22:30; however, 27.6% of the inhabitants were not yet protected by bednets by 21:30. The use of additional individual and collective protective measures is required to limit exposure to infective mosquito bites and reduce vector densities.