Boulais Yovogan

The Plasmodium transmission-blocking endosymbiont Microsporidia MB was previously identified in Anopheles gambiae s.l., but its association with the carria

Background: Insecticide resistance amongst vector populations is a major challenge, exacerbated by the continued use of the same active ingredients. The present study assessed the impact of long-lasting insecticidal nets (LLINs) bi-treated with chlorfenapyr-alphacypermethrin (PY-CFP LLIN) or pyriproxyfen-alphacypermethrin (PY-PPF LLIN) on the genetic structure of resistant populations of Anopheles gambiae in 60 clusters divided into three arms from three districts in southern Benin. Methods: The study was conducted between September 2019 and October 2021 in 123 villages grouped in 60 clusters. Mosquitoes were collected indoors and outdoors using human landing catches (HLCs) in 4 households in each cluster every 3 months. After morphological identification, a subsample of An. gambiae sensu lato (s.l.) was analysed by PCR to detect the molecular species and the presence of L1014F vgsc-kdr and G119S-ace-1 mutations. Results: Anopheles coluzzii (56.9%) and An. gambiae sensu stricto (s.s.) (42.8%), with a few hybrids (0.2%), were identified within 4242 samples of An. gambiae tested. The frequency of L1014F vgsc-kdr decreased in An. coluzzii collected both indoors and outdoors locations in the PY-CFP LLIN and PY-PPF LLIN arms post-intervention compared to baseline. In An. gambiae, the frequency of the L1014F allele decreased in year one but increased above baseline in year 2. In both species, the allelic frequency of G119S-ace-1 was < 10%. For L1014F vgsc-kdr, the fixation index was positive (F _IS > 0) in both species. However, it was negative (F _IS < 0) for the presence of G119S-ace-1. Weak genetic differentiation, especially in the PY-PPF LLIN and PY-CFP LLIN arms (F _ST ≤ 0.05), was observed in An. gambiae s.s. populations with L1014F vgsc-kdr, while it was generally higher for both species with G119S-ace-1. Conclusion: The frequency of the L1014F vgsc-kdr resistance allele was high, while that of the G119S-ace-1 allele was low throughout the study period. Consistent changes in allele frequencies were not observed in any of the treatment arms suggesting that the pyrethroid component of dual AI (active ingredients) nets continues to select for the resistant allele and there is little if any evidence that the non-pyrethroid insecticide selects for the wild-type kdr allele.

The present cluster-randomised control trial aims to assess the entomological efficacy of pyrethroid-pyriproxyfen and pyrethroid-chlorfenapyr LLINs compared to the standard pyrethroid-only LLINs, in their third year of community usage. Adult mosquito collections were performed every 3 months, in 4 randomly selected houses in each of the 60 trial clusters, using human landing catches. Adult mosquitoes were morphologically identified and Anopheles vectors were molecularly speciated and screened for the presence of the L1014F kdr mutation using PCR. Plasmodium falciparum sporozoite infection was assessed using ELISA. A subset of An. gambiae s.l. was also dissected to examine parity and fertility rates across study arms. There was no evidence of a significant reduction in indoor vector density and entomological inoculation rate by the pyrethroid-pyriproxyfen [DR 0.94 (95% CI 0.46–1.88), p = 0.8527; and RR 1.10 (95% CI 0.44–2.72), p = 0.8380], and pyrethroid-chlorfenapyr [DR 0.74 (95% CI 0.37–1.48), p = 0.3946; and RR 1.00 (95% CI 0.40–2.50), p = 0.9957] LLINs, respectively. The same trend was observed outdoors. Frequencies of the L1014F kdr mutation, as well as parous and fertility rates, were similar between study arms. In the third year after net distribution, entomological indicators show that the two dual active-ingredients nets performed similarly to the standard pyrethroid-only LLIN. To maintain malaria gains, it is crucial that net distribution cycles fit with their operational lifespan.

Background: Long-lasting insecticidal nets (LLINs) may have different impacts on distinct mosquito vector species. We assessed the efficacy of pyrethroid-pyriproxyfen and pyrethroid-chlorfenapyr LLINs on the density of Anopheles gambiae s.s. and An. coluzzii compared to pyrethroid-only nets in a three-arm cluster randomised control trial in Benin. Methods: Indoor and outdoor collections of adult mosquitoes took place in 60 clusters using human landing catches at baseline and every 3 months for 2 years. After morphological identification, around 15% of randomly selected samples of An. gambiae s.l. were dissected to determine parity, species (using PCR). Results: Overall, a total of 46,613 mosquito specimens were collected at baseline and 259,250 in the eight quarterly collections post-net distribution. Post-net distribution, approximately 70% of the specimens of An. gambiae s.l. speciated were An. coluzzii, while the rest were mostly composed of An. gambiae s.s. with a small proportion (< 1%) of hybrids (An. gambiae/coluzzii). There was no evidence of a significant reduction in vector density indoors in either primary vector species [An. coluzzii: DR (density ratio) = 0.62 (95% CI 0.21–1.77), p = 0.3683 for the pyrethroid-pyriproxyfen LLIN and DR = 0.56 (95% CI 0.19–1.62), p = 0.2866 for the pyrethroid-chlorfenapyr LLIN, An. gambiae s.s.: DR = 0.52 (95% CI 0.18–1.46), p = 0.2192 for the pyrethroid-pyriproxyfen LLIN and DR = 0.53 (95% CI 0.19–1.46), p = 0.2222 for the pyrethroid-chlorfenapyr]. The same trend was observed outdoors. Parity rates of An. gambiae s.l. were also similar across study arms. Conclusions: Compared with pyrethroid-only LLINs, pyrethroid-chlorfenapyr LLINs and pyrethroid-pyriproxyfen LLINs performed similarly against the two primary mosquito species An. gambiae s.s. and An. coluzzii in Benin. Graphical

Selection of mosquito collection methods is of crucial importance to evaluate the impact of vector control tools on entomological outcomes. During a cluster randomised control trial evaluating the relative efficacy of two dual-active ingredient (a.i.) long-lasting insecticidal nets (LLINs) compared to pyrethroid-only LLINs, we assessed the performance of different mosquito collection methods: Human landing catches (HLC), Centers for Disease Control and Prevention (CDC) light traps, and pyrethrum spray catches (PSC). Anopheles mosquitoes were collected using three collection methods in 4 houses, in each of the 60 trial clusters at baseline and every quarter for 24 months using PSCs and HLCs, while CDC light traps were performed during two quarters only. Mean density of vectors collected per method per night was the highest with HLCs (15.9), followed by CDC light traps (6.8); with PSCs (1.1) collecting 10 times less mosquitoes than HLCs. All three collection methods collected fewer mosquitoes in the Interceptor G2^® dual a.i. arm, compared to the other trial arms, although only HLCs and PSCs demonstrated strong evidence of this due to a greater number of collection rounds undertaken, than CDC light traps. The broadly similar results regarding the differential impact of the two dual a.i. LLINs showed by the three collection methods suggest that the more ethically acceptable, cheaper, and logistically simpler methods such as CDC light traps could be prioritised for use in large community trials for measuring the efficacy of vector control tools.

The efficacy of a vector control tool in reducing mosquito biting is crucial for its acceptability. The present study compared the vector density of Culex spp. And Mansonia spp. across clusters, which received two dual-active ingredient (a.i.) long-lasting insecticidal nets (LLINs) and a standard pyrethroid-only LLIN, and assessed the seasonality of these mosquito genera. A total of 85,723 Culex spp. and 144,025 Mansonia spp. were caught over the study period. The density of Culex and Mansonia was reduced in all three arms over the study period. There was no evidence of a significant reduction in the indoor or outdoor density of Culex spp. in either dual-a.i. LLIN arm as compared to the standard pyrethroid-only net arm. A similar trend was observed with Mansonia spp. A high density of Culex spp. was found both in rainy and dry seasons, while for Mansonia spp., this was mainly observed during the rainy season. These results suggest that the novel insecticides in the dual-a.i. LLINs did not have an additional impact on these species and that pyrethroids might still be effective on them. Further work is required to determine whether these species of mosquitoes have resistance to the insecticides tested in this trial.

: Malaria remains the main cause of morbidity and mortality in Benin despite the scale-up of long-lasting insecticidal nets (LLINs), indoor residual spraying, and malaria case management. This study aimed to determine the malaria burden and its associated risk factors in a rural area of Benin characterized by high net coverage and pyrethroid-resistant mosquito vectors. A community-based cross-sectional survey was conducted in three districts in southern Benin. Approximately 4,320 randomly selected participants of all ages were tested for malaria using rapid diagnostic tests within 60 clusters. Risk factors for malaria infection were evaluated using mixed-effect logistic regression models. Despite high population net use (96%), malaria infection prevalence was 43.5% (cluster range: 15.1–72.7%). Children (58.7%) were more likely to be infected than adults (31.2%), with a higher malaria prevalence among older children (5–10 years: 69.1%; 10–15 years: 67.9%) compared with young children (< 5 years: 42.1%); however, young children were more likely to be symptomatic. High household density, low socioeconomic status, young age (< 15 years), poor net conditions, and low net usage during the previous week were significantly associated with malaria infection. Malaria prevalence remains high in this area of intense pyrethroid resistance despite high net use. New classes of LLINs effective against resistant vectors are therefore crucial to further reduce malaria in this area.

To increase the effectiveness of insecticide-treated nets (ITN) in areas of high resistance, new long-lasting insecticidal nets (LLINs) called new-generation nets have been developed. These nets are treated with the piperonyl butoxide (PBO) synergist which inhibit the action of detoxification enzymes. The effectiveness of the new-generation nets has been proven in some studies, but their specific effect on mosquitoes carrying detoxification enzymes and those carrying both detoxification enzymes and the knock-down resistance gene in Benin is not well known. Thus, the objective of this study is to evaluate the efficacy of LLINs treated with PBO on multi-resistant Anopheles gambiae s.l. The study occurred in seven cities in Benin, Abomey, Cotonou, Porto-Novo, Zangnanado, Parakou, Malanville and Tanguiéta, and included ten locations selected on a north–south transect. Mosquito larvae were collected from these sites, and adult females from these larvae were exposed to single-pyrethroid-treated nets (LifeNet, PermaNet 2.0, Olyset Net) and bi-treated nets (PermaNet 3.0 and Olyset Plus) based on their level of resistance and using WHO cone tests following WHO guidelines. The different LLINs showed 100% mortality of the susceptible laboratory strain Kisumu and the resistant strain Ace-1R Kisumu. However, with the resistant laboratory strain kdr-Kisumu, mortality was low (16–32%) for all LLINs except PermaNet 3.0 (82.9%). The mortality of local strains carrying only the kdr mechanism varied from 0 to 47% for the single-pyrethroid-treated LLINs and 9 to 86% for bi-treated LLINs. With local strains carrying several mechanisms of resistance (kdr + detoxification enzymes), the observed mortality with different LLINs was also low except for PermaNet 3.0, which induced significantly higher mortality, usually greater than 75% (p < 0.001), with multi-resistant strains. The inhibition of the mortalities induced by the LLINs (11–96%) on multi-resistant field populations was similar to the inhibition observed with the laboratory strain carrying only the knock-down resistance mechanism (kdr-Kisumu) (p > 0.05). This study showed that the new-generation LLINs treated with pyrethroids and PBO showed better efficacy compared to conventional LLINs. Although the addition of PBO significantly increased the mortality of mosquitoes, the significant role of the kdr resistance gene in the low efficacy of LLINs calls for LLIN technology innovation that specifically targets this mechanism.