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Context: Recent publications on WHO recommended methods for estimating the survival of LLINs are good guidelines for assessing the performance of long-lasting insecticidal nets (LLINs). Thus, this field trial study was undertaken to evaluate the durability of the Yorkool® LN mosquito net distributed during the 2017 campaign in Benin. Methods: The monitoring of Yorkool® LN nets was carried out in two districts (Djougou III and Barienou) in Djougou, department of Donga, northern Benin from October 2017 to March 2019. A representative sample of 250 households that had received the Yorkool® LN polyester LLINs during the 2017 campaign was selected in the rural and urban areas of each district and monitored for 6, 12 and 18 months. An evaluation of the survival of Yorkool® LN nets was conducted based on the rate of loss and physical condition of the surviving nets as measured by the proportional hole index (pHI). Finally, the chemical efficacy of these LLINs during each period was determined using the WHO cone tests. Results: Survival of Yorkool® LN nets was similar in both rural and urban areas, although there was a difference in survival between the 6-month (95.3%), 12-month (89.7%), and 18-month follow-up periods (74.4%). A difference in survival was also observed between the NetCalc model (84%) compared to the Yorkool® LN nets of this study (74.4%). The attrition rate was 29.6% for LLINs at 18 months. Surprisingly, the physical integrity of the LLINs was minimally affected in the municipality. Indeed, the proportion of mosquito nets in good condition without a hole was 51.8% compared to 56.8% with a hole after 18 months. Only 7.8% of the LLINs in the two districts were damaged compared to 2.6% which needed to be replaced. The washing frequency, location of the LLINs and the frequency of use are some factors contributing to the appearance of the holes in LLINs. The bio-efficacy results of LLINs based on the cone test were good with mortality rates of 74%, 66%, 72% and 58% respectively after baseline, 6, 12 and 18 months of use. Conclusions: The observed differences in the survival of Yorkool® LN nets are due to community living conditions and movements and not to the equipment used to manufacture LLINs. However, the estimated median survival has shown that Yorkool® LN nets would have an average lifespan of 2 years 8 months despite their fairly good physical condition. These results may be useful to the National Malaria Control Program (NMCP) during the period of replacement of these nets on the field.

Malaria is the deadliest mosquito-borne disease and kills predominantly people in sub-Saharan Africa (SSA). The now widespread mosquito resistance to pyrethroids, with rapidly growing resistance to other insecticide classes recommended by the World Health Organization (WHO), may overturn the successes gained in mosquito control in recent years. It is of utmost importance to search for new, inexpensive, and safe alternatives, with new modes of action, that might improve the efficacy of current insecticides. The efficacy of a novel mechanical insecticidal mineral derived from volcanic rock, ImergardTMWP, was investigated to determine its efficacy as a stand-alone residual wall spray and as a mixture with deltamethrin (K-Othrine&reg, Polyzone) in experimental huts in Cove, Benin. The evaluation was conducted with susceptible (Kisumu) and wild-type Anopheles gambiae (s.l.). Deltamethrin applied alone demonstrated 40&ndash, 45% mortality (at 72 h post-exposure) during the first four months, which declined to 25% at six months for wild An. gambiae from Cove. ImergardTMWP alone and mixed with deltamethrin, under the same assay conditions, produced 79&ndash, 82% and 73&ndash, 81% mortality, respectively, during the same six-month period. ImergardTMWP met the 80% WHO bio-efficacy threshold for residual activity for the first five months with 78% residual activity at six months. ImergardTMWP can be used as a mixture with chemical insecticides or as a stand-alone pesticide for mosquito control in Africa.

Background Despite the success of indoor residual insecticide spraying (IRS) in Africa, particularly in Benin, some gaps of information need to be filled to optimize the effectiveness of this intervention in the perspective of the country’s effort to eliminate malaria. In anticipation to the 2018 IRS campaign in two targeted regions of northern Benin, this study aimed, to collect baseline information on vector composition, spatio-temporal variation and peak malaria transmission in the Alibori and Donga, two targeted regions of northern Benin. Information collected will help to better plan the implementation and later on the impact assessment of this IRS campaign. Methods The study was carried out in four districts of the two IRS targeted regions of northern Benin. Human landing catches and pyrethrum spray catches protocols were used to assess the biting rate (HBR) and, biting/resting behaviour of malaria vector populations. After morphological identification of collected Anopheles, the heads and thoraxes of Anopheles gambiae sensu lato (s.l.) were analysed by the ELISA CSP tests to estimate the sporozoite index (SI). The entomological inoculation rate was calculated as the product of mosquito biting rate (HBR) and the SI. Results The biting rates of An. gambiae s.l., the major vector in this study sites, varied significantly from region to region. It was higher: in rural than in urban areas, in rainy season than in dry season, indoors than outdoors. Overall, SI was comparable between sites. The highest EIRs were observed in the Donga region (16.84 infectious bites/man/month in Djougou district and 17.64 infectious bites/man/month in Copargo district) and the lowest in the Alibori region (10.74 infectious bites/man/month at Kandi district and 11.04 infectious bites/man/month at Gogounou district). Conclusion This study showed the heterogeneous and various nature of malaria epidemiology in Northern Benin. Indeed, the epidemiological profile of malaria transmission in the Alibori and Donga regions is made of a single season of transmission interrupted by a dry season. This period of transmission is relatively longer in Donga region than in Alibori. This information can be used to guide the extension of IRS in the Alibori and in the Donga, by primarily targeting areas with short periods of transmission, and easy to cover.

Background: Malaria transmission is based on four essential elements: the vector, the parasite, humans and the environment. However, of the four elements, the environment is not sufficiently exploited. Methods: In the research presence carried out in 6 localities in North Benin, we used a microscopic vision of the health geographer, focusing on certain components of geography, entomology and meteorology to show what to show spatial disparities in malaria transmission using Arcgis 10.4, Global Mapper and SPSS 21.0 for regression and correlation analysis. Results: The results of our research show that the slopes are unstable. Also, the lower the altitude, the lower the water kinetics and consequently a lot of water stagnation favourable for the development of mosquito breeding sites. The explanatory power of the regression model means that 54.3% of the variation in positive mosquito breeding is explained by human population density. Conclusions: Benin must make significant progress in the elimination of malaria using a new effort to understand the ecology of vector mosquitoes based on spatial disparities in the fight against malaria.

Anopheles gambiae, which is the main malaria vector in Benin has developed high level of resistance to pyrethroid insecticides. This raises serious concerns to the future use of long- lasting insecticidal nets (LLIN) and indoor residual spraying (IRS). It is therefore important to seek better and effective resistance management strategies which will use organophosphates or carbamates as alternatives against pyrethroid resistant malaria vectors in the field. Larvae and pupae of A. gambiae s.l. mosquitoes were collected from the breeding sites in Dassa-Zoume and Zogbodomey districts. WHO susceptibility tests were conducted on unfed female mosquitoes aged 2-5 days old. WHO bioassays were performed with impregnated papers with fenitrothion 1%, pirimiphos-methyl 0.25%, and bendiocarb 0.1%. Polymerase chain reaction (PCR) techniques were used to detect species and Ace-1 mutations. A. gambiae Dassa-Zoume populations were susceptible to bendiocarb 0.1% with mortality rate of 99%. A. gambiae Zogbodomey populations were susceptible to pirimiphos-methyl 0.25% and fenitrothion 1% with mortality rates of 98.96 and 99%, respectively. PCR assay revealed that 100% of mosquitoes tested were A. gambiae s.s. The frequencies of Ace-1R mutation in A. gambiae Dassa-Zoume and Zogbodomey were 0%. Carbamates (bendiocarb) and organophosphates (fenitrothion and pirimiphos-methyl) have maintained their efficiency against A. gambiae Dassa-Zoume and Zogbodomey populations. Carbamates (bendiocarb) and organophosphates (fenitrothion and pirimiphos-methyl) have proven to be powerful alternatives against pyrethroid resistant malaria vectors such as A. gambiae Dassa-Zoume and Zogbodomey populations. The use of any of these three compounds in the centre Benin would be successful in malaria vector control. Key words: Anopheles gambiae, Ace-1, resistance, fenitrothion, pirimiphos-methyl, bendiocarb, Benin.

OBJECTIVE: To investigate what kind of mosquito sample is necessary for the determination of insecticide susceptibility in malaria vectors. METHODS: Larvae and pupae of Anopheles gambiae s.l. (An. gambiae) mosquitoes were collected from the breeding sites in Littoral and Oueme departments. The Centers for Disease Control and Prevention (CDC) susceptibility tests were conducted on unfed male and female mosquitoes aged 2-5 days old. CDC susceptibility tests were also conducted on unfed, blood fed and gravid female mosquitoes aged 2-5 days old. These susceptibility tests were also conducted on unfed and blood fed female mosquitoes aged 2-5 days old and 20 days old. CDC biochemical assay using synergist was also carried out to detect any increase in the activity of enzyme typically involved in insecticide metabolism. RESULTS: Female An. gambiae Ladji and Sekandji populations were more susceptible than the males when they were unfed and aged 2-5 days old. The mortality rates of blood fed female An. gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when females were unfed. In addition, the mortality rates of gravid female An. gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when they were unfed. The mortality rate obtained when female An. gambiae Sekandji populations were unfed and aged 20 days old was higher than the one obtained when these populations were unfed and aged 2-5 days old. The results obtained after effects of synergist penicillin in beeswax on F1 progeny of An. gambiae Ladji populations resistant to permethrin showed that mono-oxygenases were involved in permethrin resistant F1 progeny from Ladji. CONCLUSIONS: The resistance is a hereditary and dynamic phenomenon which can be due to metabolic mechanisms like overproduction of detoxifying enzymes activity. Many factors influence vector susceptibility to insecticide. Among these factors, there are mosquito sex, mosquito age, its physiological status. Therefore, it is useful to respect the World Health Organization criteria in the assessment of insecticide susceptibility tests in malaria vectors. Otherwise, susceptibility testing is conducted using unfed female mosquitoes aged 3-5 days old. Tests should also be carried out at (25±2) °C and (80±10)% relative humidity.

As a result of the free-insecticide treated net distribution by National Malaria Control Programme (NMCP) in July 2011 throughout the entire country, it is useful to investigate the dynamics of insecticide resistance in Anopheles gambiae from Benin and the metabolic resistance mechanisms involved in this evolution of resistance. Larvae and pupae of A. gambiae s.l. mosquitoes were collected from the breeding sites in Littoral, Oueme and Zou departments. Centers for Disease Control and Prevention (CDC), bioassays were performed with permethrin (21.5 µg per bottle) and dichlorodiphenyltrichloroethane (DDT) (100 µg per bottle) whereas World Health Organization (WHO) susceptibility tests were performed with lambdacyhalothrin (0.05%). CDC biochemical assays using synergists piperonyl butoxide (PBO) and ethacrynic acid (ETAA) were also carried out. A. gambiae Akron, Suru-lere and Bame populations were resistant to permethrin and DDT in 2008 and 2013. A. gambiae Akron and Suru-lere were resistant to lambdacyhalothrin in 2013. A. gambiae Akron were susceptible to this product in 2008 whereas lambdacyhalothrin resistance status in A. gambiae Suru-lere populations in 2008 required further investigation. Mono-oxygenases were involved in resistance of A. gambiae Suru-lere to pyrethroids whereas glutathione S-transferases (GSTs) may play a role in A. gambiae Akron and Suru-lere resistant to DDT. This study shows that metabolic resistance conferred by detoxifying enzymes is an indication of phenotypic resistance to both DDT and pyrethroids in Southern Benin. Key words: Dynamics, piperonyl butoxide, ethacrynic acid, insecticide, vectors, resistance.

Background From 2008 to 2013, a prevention intervention against malaria based on indoor residual spraying (IRS) was implemented in Benin. From 2008 to 2012, Ficam M®, a bendiocarb-containing product was used for house spraying, in association with pirimiphos methyl EC (Actellic EC) in 2013. This operation aimed to strengthen the effectiveness of treated nets so as to expedite the achievement of Millennium Development Goals (MDGs): the reduction of morbidity and mortality due to malaria by 75 % from 2000 to 2015. Methods Monitoring and evaluation (M&E) was implemented in order to evaluate the impact of IRS intervention on malaria transmission. Anopheles gambiae s.l. populations were sampled by human landing catch. In addition, window exit traps and pyrethrum spray catches were performed to assess exophagic behaviour of Anopheles vectors the main malaria vector in the treated areas. The residual activity of insecticide in the treated walls was also assessed using WHO bioassay test. Results The purpose of this project was to draw attention to new challenges and future prospects for the success of IRS in Benin. The main strength of the intervention was a large-scale operation in which more than 80 % of the houses were treated due to the strong adhesion of population. In addition, a significant reduction of the EIR in areas under IRS were observed. However, there were many challenges including the high cost of IRS implementation and the identification of suitable areas to implement IRS. This was because of the low and short residual effect of the insecticides recommended for IRS and the management strategy for vector resistance to insecticides. This indicated that challenges are accompanied by suggested solutions. For the cost of IRS to be accessible to states, then local organizations need to be created in partnership with the National Malaria Control Programme (NMCP) in order to ensure relevant planning and implementation of IRS. Conclusion As an anticipatory measure against vector resistance, this paper proposes various methods, such as periodic IRS based on a combination of two or three insecticides of different classes used in rotation every two or three years.

Background Insecticides are widely used to control malaria vectors and have significantly contributed to the reduction of malaria-caused mortality. In addition, the same classes of insecticides were widely introduced and used in agriculture in Benin since 1980s. These factors probably contributed to the selection of insecticide resistance in malaria vector populations reported in several localities in Benin. This insecticide resistance represents a threat to vector control tool and should be monitored. The present study reveals observed insecticide resistance trends in Benin to help for a better management of insecticide resistance. Methods Mosquito larvae were collected in eight sites and reared in laboratory. Bioassays were conducted on the adult mosquitoes upon the four types of insecticide currently used in public health in Benin. Knock-down resistance, insensitive acetylcholinesterase-1 resistance, and metabolic resistance analysis were performed in the mosquito populations based on molecular and biochemical analysis. The data were mapped using Geographical Information Systems (GIS) with Arcgis software. Results Mortalities observed with Deltamethrin (pyrethroid class) were less than 90% in 5 locations, between 90-97% in 2 locations, and over 98% in one location. Bendiocarb (carbamate class) showed mortalities ranged 90-97% in 2 locations and were over 98% in the others locations. A complete susceptibility to Pirimiphos methyl and Fenitrothion (organophosphate class) was observed in all locations with 98-100% mortalities. Knock-down resistance frequencies were high (0.78-0.96) and similar between Anopheles coluzzii, Anopheles gambiae, Anopheles arabiensis, and Anopheles melas. Insensitive acetylcholinesterase-1 was rare (0.002-0.1) and only detected in Anopheles gambiae in concomitance with Knock-down resistance mutation. The maps showed a large distribution of Deltamethrin resistance, Knock-down mutation and metabolic resistance throughout the country, a suspected resistance to Bendiocarb and detection of insensitive acetylcholinesterase-1 from northern Benin, and a wide distribution of susceptible vectors to Pirimiphos methyl and Fenitrothion. Conclusion This study showed a widespread resistance of malaria vectors to pyrethroid previously located in southern Benin, an early emergence of carbamates resistance from northern Benin and a full susceptibility to organophosphates. Several resistance mechanisms were detected in vectors with a potential cross resistance to pyrethroids through Knock-down and metabolic resistance mechanisms.

Since the first evidence of pyrethroids resistance in 1999 in Benin, mutations have rapidly increased in mosquitoes and it is now difficult to design a study including a control area where malaria vectors are fully susceptible. Few studies have assessed the after effect of resistance on the success of pyrethroid based prevention methods in mosquito populations. We therefore assessed the impact of resistance on the effectiveness of pyrethroids based indoor residual spraying (IRS) in semi-field conditions and long lasting insecticidal nets (LLINs) in laboratory conditions. The results observed showed low repulsion and low toxicity of pyrethroids compounds in the test populations. The toxicity of pyrethroids used in IRS was significantly low with An. gambiae s.l (< 46%) but high for other predominant species such as Mansonia africana (93% to 97%). There were significant differences in terms of the repellent effect expressed as exophily and deterrence compared to the untreated huts (P

Background To better control malaria, the clear and urgent need is for improved data to inform decision makers, but in several African countries, there is a lack of baseline data on vectors and variation in the intensity of malaria transmission. This has resulted in the implementation of vector control efforts that ignore variation in vector behaviour and intensity of transmission, an approach that is most often not cost-effective. This study presents a detailed entomological description of mosquito distribution and variation in potentially transmissible contacts of Plasmodium falciparum following a south to north transect in Benin. Method The study was conducted in five locations where environmental parameters were different and malaria prevalence ranged between 14 and 51%. The locations represent the main eco-epidemiological malaria areas in Benin. Mosquitoes were collected using human landing catches, pyrethrum spray catches and windows traps. They were taxonomically and molecularly identified. Head-thoraces of Anopheles gambiae s.l. were tested by enzyme-linked immunosorbent assay. Entomological indicators were estimated following WHO guidelines. Results The results showed variation between location and period in distribution of Anopheles coluzzii, An. gambiae, and Anopheles arabiensis (p

Objective To determine the efficacy of WHO impregnated paper and CDC coated bottle based on number of storage days and number of times of consecutive use, in the assessment of insecticide vector susceptibility tests in laboratory and field conditions. Methods Larvae and pupae of Anopheles gambiae s.l. mosquitoes were collected from the breeding sites in Seme-Kpodji and Cotonou districts in Southern Benin in April 2013 during the first rainy season. Anopheles gambiae s.l. mosquitoes were also collected from the breeding sites in Parakou district in Northern Benin in May 2013 at the beginning of the rainy season. Susceptibility tests were done using impregnated paper with bendiocarb (0.1%) following WHO protocol and stock solutions of permethrin (21.5 μg per bottle) and deltamethrin (12.5 μg per bottle) following CDC protocol on unfed female mosquitoes aged 2-5 days old. These bioassays were repeated a certain number of times. The temperature and relative humidity were monitored and recorded during the susceptibility tests. Results This study showed that a WHO impregnated paper with bendiocarb could be used four times during four consecutive days in field conditions. Regarding a Wheaton coated bottle with permethrin or deltamethrin, they could be used at least three times during four consecutive days in laboratory conditions. Conclusions The day storage and the number of times that a WHO impregnated paper and a CDC coated bottle maintained their efficacy are useful in the assessment of insecticide vectors susceptibility tests.

Background: This study aims to research two areas, one with a resistant and the other with a susceptible profile of An. gambiae to deltamethrin in the region of Plateau (southern Benin). In each area, eight localities were sought. Both areas were needed for the assessment of the impact of malaria vector resistance to pyrethroids on the effectiveness of Long Lasting Insecticidal Nets (LLINs). The susceptible area of An. gambiae to deltamethrin was used as a control. Methods: In total, 119 localities in the region of Plateau were screened by sampling An. gambiae s.l larvae. Female mosquitoes resulting from these larvae were exposed to 0.05% deltamethrin following WHO standards. PCR was used to identify species and molecular forms of the dead and alive mosquitoes. Finally, we identified kdr mutations (1014 F and 1014S) using the HOLA technique. Results: Fifty-six out of 119 prospected localities tested positive for Anopheles gambae s.l breeding sites. The results showed that An. gambiae was resistant to deltamethrin in 39 localities and susceptible in only 2 localities; resistance to deltamethrin was suspected in 15 localities. The HOLA technique confirmed the presence of kdr 1014 F mutation and the absence of kdr 1014S mutation. The kdr 1014 F mutation was found in both M and S molecular forms at relatively high frequencies therefore confirming the susceptibility tests. Conclusion: We were unable to identify the eight susceptible areas due to the overall resistance of An. gambiae to deltamethrin in the region of Plateau. To implement the study, we kept two areas, one with high resistance (R+++) and the other with low resistance (R+) of An. gambiae to deltamethrin.

BACKGROUND Using the same insecticide for multiple successive indoor residual spraying (IRS) cycles is not recommended; instead, the National Malaria Control Program (NMCP) has decided to select another insecticide (insecticide B) in addition to bendiocarb for indoor residual spraying. METHODS An experimental hut trial comparing the effectiveness of three classes of insecticides (one carbamate [bendiocarb], two organophosphates [fenitrothion and pirimiphos methyl] and one pyrethroid [lambdacyalothrin]) was conducted in Malanville, northern Benin, against wild free entered resistant Anopheles gambiae s.l. population to pyrethroids. RESULTS Fenitrothion and pirimiphos methyl yielded the highest rate of deterrence. Their mean rates were respectively 46.6% and 35.4%. Regarding blood feeding inhibition, only fenitrothion has induced a significant inhibition rate (25.4% as mean rate). As regards the exophily rates, only lambdacyhalothrin has induced the highest rate (39.7%). Pirimiphos methyl showed the highest mortality rate and also induced a mortality rate of at least 80% in blood fed An. gambiae population after 24 h observation time. Furthermore, the huts treated with pirimiphos methyl showed the highest residual effect, followed by lambdacyhalothrin. CONCLUSION Benin NMCP has selected pirimiphos methyl as insecticide B to alternate or combine to bendiocarb (carbamate) because of the adverse effects of fenitrothion on the sleepers and its short residual effect on walls.

Background The detection of insecticide resistance in natural populations of Anopheles vectors is absolutely necessary for malaria control. In the African region, the WHO insecticide susceptibility test is the most common method for assessing resistance status. In order to search for a simple, rapid and more reliable technique in the assessment of insecticide resistance in malaria vectors, we compared the WHO tests with the CDC bottle bioassay in the Ouemé province of southern Benin where insecticide resistance has been widely reported. Methods Larvae and pupae of Anopheles gambiae s.l. mosquitoes were collected from the breeding sites in Ouemé. WHO and CDC susceptibility tests were conducted simultaneously on unfed female mosquitoes aged 2–5 days old. WHO bioassays were performed with impregnated papers of deltamethrin (0.05%) and bendiocarb (0.1%), whereas CDC bioassays were performed with stock solutions of deltamethrin (12.5 μg per bottle) and bendiocarb (12.5 μg per bottle). PCR techniques were used to detect species, Kdr and Ace-1 mutations. CDC biochemical assays using synergists were also conducted to assess the metabolic resistance. Results A slight decrease in mortality rates was observed with 97.95% and 98.33% obtained from CDC and WHO bioassays respectively in populations of mosquitoes from Adjara and Dangbo. PCR revealed that all specimens tested were Anopheles gambiae s.s. The Kdr mutation was found at high frequency in all populations and both the Kdr mutation and mono-oxygenase enzymes were implicated as mechanisms of pyrethroid resistance in An. gambiae from Misserete. Conclusion This study emphasizes that both WHO and CDC bioassays give similar results with regards to the susceptibility of mosquitoes to insecticides in southern Benin. There were complementarities between both methods, however, some specificity was noted for each of the two methods used. Both Kdr and metabolic mechanisms were implicated in the resistance.

Background Studies indicate that physical damage to long-lasting insecticide-treated nets (LLINs) occurs at a surprisingly rapid rate following net distribution. To what extent does such damage affect the impact of LLINs? Can vectors pass a compromised LLIN barrier to bite? Do more resistant vectors enter the insecticide-treated nets (ITNs) through holes? Methods The study was carried out in three geo-locations. Two types of LLINs (polyester and polyethylene) with ‘standardized’ physical damage were compared with similarly damaged, but non-insecticidal (control) nets. The proportionate Holes Index (pHI) of each net was 276. Mosquitoes were captured inside the nets, identified taxonomically, and subjected to molecular analysis to estimate Knock-down resistance (Kdr) frequency. Results The most commonly observed species was Anopheles gambiae, accounting for approximately 70% (1,076/1,550) of the total mosquitoes collected both in LLINs and non-insecticidal nets. When compared with controls, number of vectors captured in torn LLINs was significantly reduced. Nonetheless in a night, an average of 5 An. gambiae s.l could enter the damaged LLINs to bite. Similar numbers of resistant mosquitoes were collected in both LLINs and non-insecticidal (control) nets (p > 0.05). Conclusions At a pHI of 276, man-vector contact was observed in torn LLINs. The insecticide at the surface of LLINs could only reduce the number of vectors. Resistant mosquitoes have opportunity to enter both non-insecticidal (control) nets and LLINs to bite.

Background In many parts of Africa as in Benin, the main strategies of vector control are based on the scaling-up of Long Lasting Insecticide Treated Nets (LLITNs) and indoor residual spraying (IRS). The need to understand the biological implications of IRS in large scale and full coverage of LLITNs is paramount. It is in this context that the present study was conducted. It aims to evaluate the effect of a large scale IRS using a non-pyrethroid insecticide and full coverage of deltamethrin treated nets on the behavior of An. gambiae s.l. in the intervention areas compared to untreated areas used as controls. Methods Mosquitoes were collected using human landing catches, pyrethrum spray catches and window exit traps to assess reduction of entry rate, endophily rate, endophagy rate and overall mortality rate in natural populations of An. gambiae s.l. before IRS and LLITNs intervention (2007) and after in 2008 and 2010. Results In the IRS arm, endophily rate was 67.13% before intervention and 4.5% after intervention, whereas in the control arm it was stable at 51.67% (P > 0 .05). In the LLITN arm endophily rates also decreased after intervention. After the IRS, no gravid mosquitoes were collected from all treated localities, but LLITN performance was not that spectacular. The proportion of mosquitoes biting indoors in the IRS arm decreased from 67.09% before intervention to 42.85% after intervention, compared to a low but significant decrease (71.31% to 57. 46%) in the LLITN arm. The use of vector control tools and behavior of the host would be the main factors that modify the behavior of taking a human blood meal observed on An. gambiae s.l. inside human dwellings. Conclusion The impact on the behavior of An. gambiae s.l. observed with the bendiocarb used in IRS was highly effective compared with the free distribution of LLITNs in terms of mortality and the decrease of proportions of indoor feeding. Despite this efficacy, there is a need for complementary tools and research of alternative strategy oriented on effective health education, and the use of powerful tools such as IRS, LLITNs, larviciding and repellents.

Background LLIN distribution, every three years, is a key intervention of Benin’s malaria control strategy. However, data from the field indicate that LLIN lifespan appears to vary based on both intrinsic (to the LLIN) and extrinsic factors. Methods We monitored two indicators of LLIN durability, survivorship and integrity, to validate the three-year-serviceable-life assumption. Interviews with net owners were used to identify factors associated with loss of integrity. Results Observed survivorship, after 18 months, was significantly less (p

Background The dynamics of mosquito populations depends on availability of suitable surface water for oviposition. It is well known that suitable management of mosquito larval habitats in the sub-Saharan countries, particularly during droughts, could help to suppress vector densities and malaria transmission. We conducted a field survey to investigate the spatial and seasonal distribution of mosquito larval habitats and identify drought-refugia for anopheline larvae. Methods A GIS approach was used to identify, geo-reference and follow up longitudinally from May 2012 to May 2013, all mosquito breeding sites in two rural sites (Yondarou and Thui), one urban (Kossarou), and one peri-urban (Pèdè) site at Kandi, a municipality in northeastern Benin. In Kandi, droughts are excessive with no rain for nearly six months and a lot of sunshine. A comprehensive record of mosquito larval habitats was conducted periodically in all sites for the identification of drought-refugia of anopheline larval stages. With geospatialisation data, seasonal larval distribution maps were generated for each study site with the software ArcGIS version 10.2. Results Overall, 187 mosquito breeding sites were identified of which 29.95% were recorded during drought. In rural, peri-urban and urban sites, most of the drought-refugia of anopheline larvae were domestic in nature (61.54%). Moreover, in rural settings, anopheline larvae were also sampled in cisterns and wells (25% of larval habitats sampled during drought in Yondarou and 20% in Thui). The mapping showed a significant decrease in the spatial distribution of mosquito larval habitats in rural, peri-urban and urban sites during drought, except in Yondarou (rural) where the aridity did not seem to influence the distribution of larval habitats. Conclusion Our data showed that the main drought-refugia of anopheline larvae were of a domestic nature as well as wells and cisterns. A suitable management of mosquito larvae in sub-Saharan countries, particularly during droughts, should target such larval habitats for a meaningful impact on the dynamics of mosquito populations and malaria transmission.

Background Following a mass distribution of long-lasting insecticidal nets (LLINs) in Benin, we used WHO guidelines to develop an assessment tool which is described in this report. It involved assessment of the three WHO indicators: survivorship, integrity and bio-efficacy. Methods To evaluate the assessment tool, we selected four communities, two in the Southern part of the country, and two in the North. One of the two assessment communities in each geographic setting had ready access to water and a higher reported frequency of washing LLINs. It was assumed that nets in communities with greater washing frequencies would show greater loss of durability. If the tool was sensitive enough to detect such differences, the field testing would confirm its suitability for general use in different settings in Benin. While durability indicators of survival and fabric integrity were quantified using standard WHO methodology, bio-efficacy was assessed using a ‘new’ alternative (to the WHO bioassay test), involving gas chromatography. Additionally, data management used current internet technology for ‘real time’ analysis at a central monitoring location. Results While no difference in survivorship was observed between sites with ready access to water for washing, both in the North and the South, there was a significant difference in integrity. In the South and in the North, nets from sites near water (Kessounou and Malanville) showed greater damage to integrity than did the nets from Allada and Kandi (sites far from water). As expected, LLIN integrity was significantly lower when a community was near water (p

Background In arid settings, droughts usually lead to periods of very low or no malaria transmission. However, in rural Kandi (Sonsoro) in northeastern Benin, several malaria cases are often diagnosed during dry seasons. The underlying factors accounting for this phenomenon remain unknown. Methods The entomological profile of Sonsoro has been studied compared to a location in urban Kandi (Gansosso) for a period of one year. During this period, Anopheles larval habitats were investigated and populations of Anopheles gambiae s.l. were sampled by human landing catches in both areas. Enzyme-linked immunosorbent assays (ELISA) for Plasmodium falciparum circumsporozoite protein (CSP) were conducted on vector specimens and the entomological inoculation rates (EIR) were determined per season (wet versus dry) in each area. In addition, during the severe drought period, Rapid Diagnostic Tests (RDTs) were conducted on school children under the age 10 years in these areas to provide a global view of drought-malaria prevalence and to perform a crossing with entomological data in Kandi. Results Overall, An. gambiae s.l. was particularly abundant in rural Kandi compared to the urban area with a significant decrease of vector density in both sites during the dry season. In this period, larval sampling data identified household water sources as potential breeding sites in urban and rural Kandi. We also observed a significant seasonal variation of the infectivity rate in both areas but for each period (season), the EIR was higher in the rural site than in the urban. Data of P. falciparum detection was the reflection of entomological findings. The drought-malaria prevalence was 5.5 times higher in rural Kandi as compared to urban Kandi. The presence of a permanent water site and the low level of urbanization in rural Kandi were identified as a risk factor. Conclusion Our data showed a high level of malaria transmission in the municipality of Kandi. Household water source plays an important role in maintaining the breeding of anopheles larvae and the malaria transmission in Kandi. In rural settings, the proximity to permanent water sites could probably be the aggravating factor.

Background A dynamic study on the transmission of malaria was conducted in two areas (R+ area: Low resistance area; R+++ area: High resistance area) in the department of Plateau in South Eastern Benin, where the population is protected by Long Lasting Insecticidal Nets (LLINs). The aim of this study was to determine if the resistance of malaria vectors to insecticides has an impact on their behavior and on the effectiveness of LLINs in the reduction of malaria transmission. Methods Populations of Anopheles gambiae s.l. were sampled monthly by human landing catch in the two areas to evaluate human biting rates (HBR). Collected mosquitoes were identified morphologically and female Anopheles mosquitoes were tested for the presence of Plasmodium falciparum antigen as assessed using ELISA. The entomological inoculation rate (EIR) was also calculated (EIR = HBR x sporozoitic index [S]). We estimated the parity rate by dissecting the females of An. gambiae. Finally, window catch and spray catch were conducted in order to assess the blood feeding rate and the exophily rate of vectors. Results After 6 months of tracking the mosquito's behavior in contact with the LLINs (Olyset) in R+++ and R+ areas, the entomological indicators of the transmission of malaria (parity rate and sporozoitic index) were similar in the two areas. Also, An. gambiae populations showed the same susceptibility to P. falciparum in both R+ and R+++ areas. The EIR and the exophily rate are higher in R+ area than in R+++ area. But the blood-feeding rate is lower in R+ area comparing to R+++. Conclusion The highest entomological inoculation rate observed in R+ area is mostly due to the strong aggressive density of An. gambiae recorded in one of the study localities. On the other hand, the highest exophily rate and the low blood-feeding rate recorded in R+ area compared to R+++ area are not due to the resistance status of An. gambiae, but due to the differences in distribution and availability of breeding sites for Anopheles mosquitoes between areas. However, this phenomenon is not related to the resistance status, but is related to the environment instead.