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Abstract Background Pyrethroid insecticides, carbamate and organophosphate are the classes of insecticides commonly used in agriculture for crop protection in Benin. Pyrethroids remain the only class of insecticides recommended by the WHO for impregnation of bed nets. Unfortunately, the high level of pyrethroid resistance in Anopheles gambiae s.l., threatens to undermine the success of pyrethroid treated nets. This study focuses on the investigation of agricultural practices in cotton growing areas, and their direct impact on larval populations of An. gambiae in surrounding breeding sites. Methods The protocol was based on the collection of agro-sociological data where farmers were subjected to semi-structured questionnaires based on the strategies used for crop protection. This was complemented by bioassay tests to assess the susceptibility of malaria vectors to various insecticides. Molecular analysis was performed to characterize the resistance genes and the molecular forms of An. gambiae. Insecticide residues in soil samples from breeding sites were investigated to determine major factors that can inhibit the normal growth of mosquito larvae by exposing susceptible and resistant laboratory strains. Results There is a common use by local farmers of mineral fertilizer NPK at 200 kg/ha and urea at 50 kg/hectare following insecticide treatments in both the Calendar Control Program (CCP) and the Targeted Intermittent Control Program (TICP). By contrast, no chemicals are involved in Biological Program (BP) where farmers use organic and natural fertilizers which include animal excreta. Susceptibility test results confirmed a high resistance to DDT. Mean mortality of An. gambiae collected from the farms practicing CCP, TICP and BP methods were 33%, 42% and 65% respectively. An. gambiae populations from areas using the CCP and TICP programs showed resistance to permethrin with mortality of 50% and 58% respectively. By contrast, bioassay test results of An. gambiae from BP areas gave a high level of susceptibility to permethrin with an average mortality of 94%. Molecular analysis identified An. gambiae s.s, and An. arabiensis with a high predominance of An. gambiae s.s (90%). The two molecular forms, M and S, were also determined with a high frequency of the S form (96%). The Kdr gene seemed the main target- site resistance mechanism detected in CCP, TICP, and BP areas at the rates ranging from 32 to 78%. The frequency of ace-1R gene was very low (< 0.1). The presence of inhibiting factors in soil samples under insecticide treatments were found and affected negatively in delaying the development of An. gambiae larval populations. Conclusions This research shows that Kdr has spread widely in An. gambiae, mainly in CCP and TICP areas where pyrethroids are extensively used. To reduce the negative impact of pesticides use in cotton crop protection, the application of BP-like programs, which do not appear to select for vector resistance would be useful. These results could serve as scientific evidence of the spread of resistance due to a massive agricultural use of insecticides and contribute to the management of pesticides usage on cotton crops hence reducing the selection pressure of insecticides on An. gambiae populations.

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. [ABSTRACT FROM PUBLISHER]

Background: This study investigates the effectiveness of new-generation mosquito nets, like Olyset® Plus and PermaNet® 3.0, and dual-action nets such as Interceptor® G2, against pyrethroid-resistant Anopheles gambiae mosquitoes following the 2023 mass distribution of long-lasting insecticidal nets in Benin. Methods: We tested wild mosquito populations from six communes in Benin against various pyrethroid (permethrin 0.75%, alphacypermethrin 0.05%, and deltamethrin 0.05%) using WHO tube tests. Additionally, we exposed mosquitoes to chlorfenapyr 100 µg/ml using the CDC bottle bioassay method. A subset of mosquitoes underwent biochemical and PCR tests to check the overexpression of metabolic enzymes and the Kdr L1014F mutation. We evaluated the effectiveness of Olyset® Plus, PermaNet® 3.0, and Interceptor® G2 nets using cone and tunnel tests on both laboratory and field populations of An. gambiae. Results: Overall, the highest mortality rate was 60% with pyrethroid and 98 to100% with chlorfenapyr. In cone tests, all three types of nets induced mortality rates above 80% in the susceptible laboratory strain of An. gambiae. Notably, Olyset® Plus showed the highest mortality rates for pyrethroid-resistant mosquitoes in cone tests, ranging from 81.03% (95% CI: 68.59–90.13) in Djougou to 96.08% (95% CI: 86.54–99.52) in Akpro-Missérété. PermaNet® 3.0 had variable rates, from 42.5% (95% CI: 27.04–59.11) in Djougou to 58.54% (95% CI: 42.11–73.68) in Porto-Novo. However, revealed good results for Interceptor® G2, with 94% (95% CI: 87.40–97.77) mortality and 89.09% blood sampling inhibition in local populations of An. gambiae. In comparison, Interceptor® had lower rates of 17% (95% CI: 10.23–25.82) and 60%, respectively. Conclusion: These results suggest that tunnel tests are effective for evaluating dual-active ingredient nets. Additionally, Interceptor® G2 and PBO nets like Olyset® Plus could be considered as alternatives against pyrethroid-resistant mosquitoes. [ABSTRACT FROM AUTHOR]

Background: The purpose of this report is to provide information on Culicidae diversity; biting behavior and spatio-seasonal variation of abundance of Anopheles gambiae s.l. and Culex quinquefasciatus in rural and urban settings of the Alibori and Donga regions, Northern Benin, where an indoor residual spraying (IRS) campaign to control malaria is planned. Methods: Both human landing catches, associated with pyrethrum spray catches were used to monitor the mosquito populations in 12 sites with 1 urban and 1 rural located in each of the 6 districts randomly selected in the two targeted regions. After morphological identification of all mosquito specimens, biting behavior and density of An. gambiae s.l. and Cx quinquefasciatus were studied. PCR was also performed on An. gambiae s.l., to identify sibling species and its seasonal variation. Results: A total of 10,367 mosquitoes were captured, related to 14 species of the genera, Anopheles , Aedes , Culex and Mansonia . Of the total species collection, 40.39% were An. gambiae s.l. and 56.85% were Cx. quinquefasciatus. An. gambiae s.l. was more abundant in Donga (2521 specimens) compared with Alibori (1666 specimens). The opposite trend was observed with Cx. quinquefasciatus (2162 specimens in Donga against 4028 in Alibori). An. gambiae s.l. was predominant and displayed a higher blood feeding rate in rural areas, whereas Cx. quinquefasciatus was in majority in urban areas. An. gambiae s.l. was more endophagic, whereas Cx. quinquefasciatus showed similar indoor and outdoor biting behavior. An. gambiae s.l. was composed of An. coluzzii found in majority in the drought, and An. gambiae , which was predominant in the rainy season. Conclusion: The predominance of the malaria vector, An. gambiae s.l. and their higher blood feeding rate and their significantly high endophagy in rural areas indicate that these areas should be primarily targeted with the IRS operations to have a substantial impact on malaria transmission. Endophagy, characteristic of An. gambiae s.l. in our study area, suggests that IRS will have a positive impact on vector control if implemented 1 week before June that is the onset of the rainy season.