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Background Long-lasting insecticidal nets (LLINs) are designed to survive and sustain their physical barrier for 3 years in household conditions. However, studies have shown that most of these nets are usually torn or no longer present in the households in less than 3 years. This study was initiated in Benin to compare the survivorship and physical integrity of seven types of LLINs in a same socio-geographic area. Methods In August 2017, 1890 households were selected in 9 villages in the municipality of Zagnanado in central Benin. Each one of the selected households received one of the seven LLIN products: Aspirational®, DawaPlus® 2.0, OlysetNet®, PermaNet® 2.0, PermaNet® 3.0, Royal Sentry® and Yorkool®. Overall, 270 LLINs of each type were freely distributed in Zagnanado, at a rate of 30 LLINs per type per village. These bed nets have been monitored and evaluated every 6 months to identify the most resilient and preferred LLINs in the community. Net survivorship was assessed using the rate of net loss and physical condition. Results The survivorship of all types of LLIN was estimated at 92% (95% CI 90.33–92.96) after 6 months and 70% (95% CI 67.25–71.81) after a year of use. At 12 months, all bed nets monitored were below the NetCalc model threshold of 92.8% for an LLIN with a lifespan of 3 years. Only 1.73% of all types of LLIN had a visible loss of integrity after 6 months with a median proportionate hole index (PHI) estimated at zero. The percentage significantly increased after 12 months with 10.41% of damaged nets (all types of LLINs). The median PHI for each brand of net was 23, 196, 141, 23, 23, 121 and 72, respectively for Aspirational®, DawaPlus® 2.0, OlysetNet®, PermaNet® 2.0, PermaNet® 3.0, Royal Sentry® and Yorkool®. A significant difference was noted between the PHI at 6 and 12 months (p ®2.0, OlysetNet® and Royal Sentry® suffered significantly more damage compared to the others (p Conclusion The results of this study showed that after a year of use, the survivorship of the 7 LLIN products in households was lower than expected. However, all the LLIN products successfully met WHO standards for physical integrity after 12 months of use. The monitoring continues. The next steps will help to identify the most sustainable LLINs.

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.

Pyrethroid resistance is widespread in sub-Saharan Africa. The objective of this study was to assess the insecticide resistance intensity in Anopheles gambiae s.l. (Diptera: Culicidae) in four districts of Benin in order to better understand how pyrethroid-only nets are likely to be effective. Thus, adult females of An. gambiae s.l., reared from field-collected larvae were used for assessing resistance intensity to permethrin and deltamethrin. They were tested at 1×, 5×, and 10× the diagnostic dose, using both WHO susceptibility tube testing and CDC bottle bioassays. Identification of molecular species, as well as of L1014F Kdr and Ace-1R mutations was performed using the PCR. The level of expression of biochemical enzymes was also evaluated. Overall, moderate to high resistance intensity to permethrin and deltamethrin was observed, irrespective of the testing method. While the L1014F Kdr frequency was high (>75%), Ace-1R was low (≤6%) in An. gambiaes.s. and Anopheles coluzzii, the two predominant species [52% (95% CI: 44.8–59.1) and 45% (95% CI: 38.0–52.2), respectively]. Anopheles arabiensis was found at very low frequency (3%, 95%CI: 1.1–6.4). For Biochemical analyses, α and β-esterases were over-expressed in all four districts, while mixed-function oxidases (MFOs) were over-expressed in only one. Overall, the two testing methods led to comparable conclusions, though there were a few inconsistencies between them. The moderate-high resistance intensity observed in the study area suggests that dual active-ingredient (AI) long-lasting insecticidal nets (LLINs) may provide better control of insecticide-resistant mosquitoes.

Monitoring vector resistance to insecticides is a major component of resistance management. Adults of Anopheles gambiae s.l. from larvae collected at thirteen sites were tested with papers impregnated with bendiocarb, permethrin; deltamethrin; pirimiphos-methyl and induced bottles of insecticide including bendiocarb; deltamethrin, permethrin and pirimiphos-methyl. Molecular analyses were performed and detoxification enzyme levels were determined for each study site. This study revealed a generalized distribution of resistance of An. gambiae s.l. to pyrethroids (permethrin and deltamethrin) and a clear resistance to carbamates (bendiocarb) especially in the northern region of the country. This vector is still susceptible to organophosphates, especially pirimiphos-methyl, but resistance is a dynamic phenomenon and it would be necessary to monitor the susceptibility of these anopheles to pyrimiphos-methyl and to find alternative vector control methods to slow down the spread of resistance genes.

Background Since 2008, Indoor Residual Spraying (IRS) has been performed in Benin in 19 districts, including 4 in southern Benin, 9 in Atacora, and 8 in Atacora, Alibori and Donga in northern Benin. However, Benin still struggles with questions about IRS cost–benefit and epidemiological impact. Lessons learned and challenges from 10 years of IRS in Benin to be shared with the stakeholders involved in vector control implementation for decision-making. Methods Entomological parameters have been assessed entomological parameters in IRS communes since 2008. In all IRS intervention communes, decreases in human biting rate (HBR) of Anopheles gambiae, blood feeding inhibition and entomological inoculation rate (EIR) as compared to control district have been measured. Results EIR was reduced by 80–90%, which is encouraging, but should be observed with caution because: (i) the reduction may be insufficient to decrease epidemiological indicators given that the residual EIR in IRS districts is still higher than it is in some regions of stable malaria; (ii) the reduction in EIR is based on comparisons with control communes, but it is difficult to select control areas with the same environmental characteristics as intervention areas; (iii) despite the reduction, half of all mosquitoes that entered IRS-treated houses succeeded in taking human blood meals. Further, there are behaviours among Benin’s population that limit IRS efficacy, including recent data showing that > 90% of people are not protected by IRS between 7 and 10 p.m. This is due to the fact that they remain outdoors and that most people are not protected from mosquito bites after 10 p.m. because they either sleep outdoors without IRS protection or indoors without an ITN. Moreover, people have large amounts of clothing hanging on walls where mosquitoes can rest instead of IRS-treated walls. Finally, other components are important to consider in implementing IRS among which: (i) Vector resistance management strategies are sometimes poorly understood; this is actually different from the need to replace one insecticide with another after the emergence of resistance; (ii) African countries should prepare to finance IRS themselves. Conclusion To curtail residual malaria transmission, additional interventions able to target vectors escaping IRS should be prioritized.

Background The current study shows the results of three years of IRS entomological monitoring (2016, before intervention; 2017 and 2018, after intervention) performed in Alibori and Donga, northern Benin. Methods Mosquito collections were performed on a monthly basis using human landing catches and pyrethrum spray catches in six districts including four treated with Actellic 300 CS (Kandi, Gogounou, Djougou and Copargo) and two untreated (Bembèrèkè and Kouandé) which served as control sites. Key transmission indicators of Anopheles gambiae (s.l.) as well as the residual activity of Actellic 300 CS assessed through WHO cone tests, were determined. Results The residual efficacy duration of Actellic 300 CS after the two IRS campaigns (2017 and 2018) was 4–5 months (May–September). The parity rate and the sporozoite index of An. gambiae (s.l.) were 36.62% and 0.71%, respectively, after the first spray round in treated areas compared to 57.24% and 3.7%, respectively, in the control areas (P vs 9.75 ib/m in the control areas, corresponding to a reduction of 84.61%. A decrease in the parity rate (46.26%), sporozoite index (85.75%) and EIR (87.27%) was observed for An. gambiae (s.l.) after the first round of IRS (June–October 2017) compared to the pre-intervention period (June–October 2016). The density of An. gambiae (s.l.) ranged between 0.38–0.48 per house in treated areas vs 1.53–1.76 An. gambiae (s.l.) per house respectively after the first and second IRS rounds. Conclusions This study showed the positive impact of IRS in reducing key entomological parameters of malaria transmission in Alibori and Donga. However, the considerable blood-feeding rate of An. gambiae (s.l.) in spray areas, stress the need for the population to sleep under long-lasting insecticidal nets (LLINs) in addition, to prevent from mosquito bites which did not succeed in resting on sprayed walls.

Background: Climate variability influence the diversity and abundance of malaria vectors and thereby on malaria transmission dynamics. Examine its effect on Anopheles parameters involved in transmission may predict the potential malaria hotspot as a right target for its control intervention strategies. Here, we investigated the influence of meteorological parameters on the aggressiveness and infectivity of Anopheles in two health districts zones where IRS has been extended in Northern Benin. Methods: Mosquito collections were carried out using human landing catches to evaluate rates of aggression and infectivity in twelve villages. Concomitantly, meteorological data from synoptic stations of Benin and neighbouring countries were collected in 2016-2017. Results: The spatial distribution of infective bites of An. gambiae is characterized by an intense aggression in the rural villages of the study area. Analysis of variances showed significant HBR difference according to the period but not according to the locality. However, the same analysis carried out with the infectivity rate shows no significant difference according to the period and the locality. In addition, the number of infective bites per man per month is higher in August and October, and the climatic parameters that have mainly favoured aggression are wind speed, humidity, sunshine and temperature. Indeed, the peak of wind speed is concentrated around 1.2 km / h and in September (5 km / h) whereas the aggressiveness score of Anopheles in the region is greater than 10 infective bites per man a year. Conclusion Malaria transmission by Anopheles is influenced by climatic factors. The climate observed in the districts where IRS was extended in northern Benin has a real impact on Anopheles density and weakens current and future vector control strategies. This could lead to a series of modifications observed in anopheline populations just after IRS implementation ranging from a tendency to exophagy, from a decrease in the rate of blood-feeding to changes in the time, and change in aggressiveness. These phenomena most likely contribute to the sustainability of malaria transmission despite vector control measures. Keywords: Infectivity, aggression, Climate, Anopheles gambiae ( s.l. ), IRS, Benin.

Background This study aims to provide baseline data on the resistance status to insecticides, the frequency of mechanisms involved and the impact of the association with the synergist piperonyl butoxide (PBO) on resistant Anopheles gambiae (s.l.) populations in two regions of northern Benin, prior to an indoor residual spraying campaign and introduction of next generation long-lasting insecticidal nets (LLINs) incorporating PBO. Methods Adult Anopheles gambiae (s.l.) originating from larvae collected in two study regions (Alibori within the Kandi-Gogounou-Segbana districts and Donga within the Djougou-Copargo-Ouake districts) were tested with impregnated papers (bendiocarb 0.1%, pirimiphos-methyl 0.25%, permethrin 0.75% and deltamethrin 0.05%). The synergist PBO was used to check for the involvement of detoxification enzymes in pyrethroid resistant populations. Molecular analyses were performed for the identification of species within the Anopheles gambiae (s.l.) complex and kdr L1014F and G119S Ace-1 mutations. Biochemical assays assessed the activity of detoxification enzymes. Results Anopheles gambiae (s.l.) was resistant to pyrethroids, with a mortality range of 25–83% with deltamethrin and 6–55% with permethrin. A significant increase in mortality was observed after pre-exposure to PBO for both deltamethrin (63–99%) and permethrin (56–99%). With bendiocarb, An. gambiae (s.l.) were susceptible in Kandi (99% mortality), with possible resistance (92–95%) recorded in Djougou, Copargo, Gogounou, Ouake and Segbana. All study populations were fully susceptible to pirimiphos-methyl. The frequencies of resistant mutations varied according to species and sites: 0.67–0.88 for L1014F kdr and 0–0.06 for G119S Ace-1. Three study locations (Djougou, Gogounou and Kandi) showed high oxidase activity and four sites (Djougou, Ouake, Copargo and Kandi) showed elevated esterase activity. Conclusions This study confirms resistance to pyrethroids and suggests emerging bendiocarb resistance in An. gambiae (s.l.) populations in northern Benin. However, recovery of susceptibility to pyrethroids after PBO exposure, and susceptibility to organophosphates in the An. gambiae (s.l.) populations indicate that next generation LLINs incorporating PBO synergist combined with an indoor residual spraying (IRS) campaign with organophosphate insecticides may be regarded as alternative control tools.

Long-lasting insecticidal nets (LLINs) are an essential tool in the fight against malaria. Physical integrity, durability and bio-effectiveness are key variables in the effectiveness of LLINs. The objective of this study was to identify the main factors affecting the survival of three brands of LLINs with different physical characteristics and to assess their bio-effectiveness. A cohort consisting of 1500 LLINs (500 of each) of the brands: DawaPlus®2.0 (polyester, 150 denier, 40 g/m2 fabric weight), PermaNet®2.0 and Yorkool® (polyester, 75 denier, alternating knit pattern with 85 g/m2 fabric weight) was monitored every 6 months in the communes of Ketou, Dogbo and Djougou (from October 2017 to September 2019) based on attrition and integrity measures and median survival in years. We also determined bio-efficacy using the WHO cone test. The physical presence rate was 26.4%, 21.4% and 48.6% respectively for DawaPlus®2.0, PermaNet®2.0 and Yorkool®. The main cause of loss of the three LLINs was displacement, 43.6% (in rural areas) versus 43.2% (in urban areas) with no significant difference (p ˃ 0.05). The median proportional hole index (pHI) ranged from 578 (IQR: 219-843) at 6 months to 196 (IQR: 46-524.5). After 24 months of use, 86.1% were in good condition (0≤pHI

Background: Climate variability influence the diversity and abundance of malaria vectors and thereby on malaria transmission dynamics. Examine its effect on Anopheles parameters involved in transmission may predict the potential malaria hotspot as a right target for its control intervention strategies. Here, we investigated the influence of meteorological parameters on the aggressiveness and infectivity of Anopheles in two health districts zones where IRS has been extended in Northern Benin. Mosquito collections were carried out using human landing catches to evaluate rates of aggression and infectivity in twelve villages. Concomitantly, meteorological data from synoptic stations of Benin and neighbouring countries were collected in 2016-2017. The spatial distribution of infective bites of An. gambiae is characterized by an intense aggression in the rural villages of the study area. Analysis of variances showed significant HBR difference according to the period but not according to the locality. However, the same analysis carried out with the infectivity rate shows no significant difference according to the period and the locality. In addition, the number of infective bites per man per month is higher in August and October, and the climatic parameters that have mainly favoured aggression are wind speed, humidity, sunshine and temperature. Indeed, the peak of wind speed is concentrated around 1.2 km / h and in September (5 km / h) whereas the aggressiveness score of Anopheles in the region is greater than 10 infective bites per man a year. Malaria transmission by Anopheles is influenced by climatic factors. The climate observed in the districts where IRS was extended in northern Benin has a real impact on Anopheles density and weakens current and future vector control strategies. This could lead to a series of modifications observed in anopheline populations just after IRS implementation ranging from a tendency to exophagy, from a decrease in the rate of blood-feeding to changes in the time, and change in aggressiveness. These phenomena most likely contribute to the sustainability of malaria transmission despite vector control measures.

Background The complex Anopheles gambiae includes the most effective vectors of malaria in the world. The identification of these vector species and their seasonal and temporal distribution are essential for effective control of malaria. In Benin, this complex Anopheles gambiae is represented by Anopheles coluzzii, Anopheles gambiae, Anopheles arabiensis and Anopheles melas which occur near the lakes and lagoons of southern Benin. The present study provides information on the seasonal and temporal variation of Anopheles melas, the predominant malaria vector in Djegbadji, a coastal lagoon town of Benin. Methods Sampling of adult mosquitoes was performed at Djegbadji by night inside and outside of homes using human landing catches (from May 2013 to February 2015) and morning indoor residual spray catches (from January 2014 to December 2014). The collected mosquitoes were identified morphologically and Anopheles melas specimens were separated from other vectors by the palps biometrics technique. Results The results showed that the collected mosquitoes belonged to 5 genera and 8 species. It included: Anopheles gambiae sensu lato, Aedes aegypti, Culex quinquefasciatus, Culex decens, Culex nebulosus, Culex fatigans and Mansonia africana. Among the different species identified, Anopheles gambiae sensu lato had the highest abundance. At Djegbadji, Anopheles melas was the predominant malaria vector in dry season (90%), in rainy season (73.43%) and during flood periods (76.47%). In times of recession, there is a reduction of Anopheles melas density by 30.76% with a predominance of Anopheles gambiae sensu stricto (69.24%). The two methods used for the collection of Anopheles melas gave similar results (p = 0.35). Conclusion This study allowed us to update the data on malaria vector species in coastal lagoon areas of Benin. It also demonstrated that the predominant species of malaria vectors in this area was Anopheles melas. These findings will guide planning strategies against malaria vectors in the coastal lagoon areas of Benin