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Background The World Health Organization (WHO) recommends that pregnant women in sub-Saharan Africa (SSA) receive a free long-lasting insecticidal net (LLIN) during their first antenatal care (ANC) visit to prevent malaria. This study, conducted in Benin, evaluates the distribution and utilization rates of LLINs provided at the first ANC visit among pregnant women. Methods Data were collected from 14 public and private health centers located in urban and rural areas across Southern, Central, and Northern Benin. Pregnant women were enrolled in the study during their initial ANC visit and were subsequently visited at home twice, where a questionnaire was administered. The study assessed the distribution and use of LLINs during the first ANC visit. After the second home visit, the LLIN found on the pregnant women’s sleeping unit was collected to evaluate its physical integrity and bio-efficacy. Chi-square tests were used to compare each indicator across three variables: region, urban/rural setting, and public/private status of health centers. Results A total of 718 pregnant women were included in the study. LLIN ownership and usage before the first ANC visit were 94% [89-97%] and 93% [85-97%], respectively. During the first ANC visit, 63% [40-80%] of the pregnant women received an LLIN, but only 11% [7-22%] installed it on their sleeping area. During the pregnancy period, 72% [64-78%] of the LLINs in use were found to be either physically damaged or not bio-effective. Conclusion The distribution of LLINs to pregnant women during their first ANC visit was inadequate, with only a small fraction of recipients actively using the net. This shortfall leads to suboptimal protection for this vulnerable population during pregnancy.

This study provides detailed characteristics of vector populations in preparation for a three-arm cluster randomized controlled trial (RCT) aiming to compare the community impact of dual active-ingredient (AI) long-lasting insecticidal nets (LLINs) that combine two novel insecticide classes-chlorfenapyr or pyriproxifen-with alpha-cypermethrin to improve the prevention of malaria transmitted by insecticide-resistant vectors compared to standard pyrethroid LLINs. The study was carried out in 60 villages across Cove, Zangnanando and Ouinhi districts, southern Benin. Mosquito collections were performed using human landing catches (HLCs). After morphological identification, a sub-sample of Anopheles gambiae s.l. were dissected for parity, analyzed by PCR for species and presence of L1014F kdr mutation and by ELISA-CSP to identify Plasmodium falciparum sporozoite infection. WHO susceptibility tube tests were performed by exposing adult An. gambiae s.l., collected as larvae from each district, to 0.05% alphacypermethrin, 0.75% permethrin, 0.1% bendiocarb and 0.25% pirimiphos-methyl. Synergist assays were also conducted with exposure first to 4% PBO followed by alpha-cypermethrin. An. gambiae s.l. (n = 10807) was the main malaria vector complex found followed by Anopheles funestus s.l. (n = 397) and Anopheles nili (n = 82). An. gambiae s.l. was comprised of An. coluzzii (53.9%) and An. gambiae s.s. (46.1%), both displaying a frequency of the L1014F kdr mutation >80%. Although more than 80% of people slept under standard LLIN, human biting rate (HBR) in An. gambiae s.l. was higher indoors [26.5 bite/person/night (95% CI: 25.2-27.9)] than outdoors [18.5 b/p/n (95% CI: 17.4-19.6)], as were the trends for sporozoite rate (SR) [2.9% (95% CI: 1.7-4.8) vs 1.8% (95% CI: 0.6-3.8)] and entomological inoculation rate (EIR) [21.6 infected bites/person/month (95% CI: 20.4-22.8) vs 5.4 (95% CI: 4.8-6.0)]. Parous rate was 81.6% (95%CI: 75.4-88.4). An. gambiae s.l. was resistant to alpha-cypermethrin and permethrin but, fully susceptible to bendiocarb and pirimiphos-methyl. PBO pre-exposure followed by alpha-cypermethrin treatment induced a higher 24 hours mortality compared to alphacypermethrin alone but not exceeding 40%. Despite a high usage of standard pyrethroid LLINs, the study area is characterized by intense malaria transmission. The main vectors An. coluzzii and An. gambiae s.s. were both highly resistant to pyrethroids and displayed multiple resistance mechanisms, L1014F kdr mutation and mixed function oxidases. These conditions of the study area make it an appropriate site to conduct the trial that aims to assess the effect of novel dual-AI LLINs on malaria transmitted by insecticide-resistant vectors.

Background In 2023, an estimated 36 million pregnancies occurred in malaria endemic sub-Saharan Africa, but only 44% received the WHO recommended ≥ 3 doses of intermittent preventive treatment (IPTp3). Group Antenatal Care (G-ANC) is a service delivery model associated with higher quality of and greater retention in ANC, in which pregnant women are enrolled into groups at their first ANC visit and subsequent care is provided in groups. A cluster-randomized controlled trial was conducted in Atlantique Department, Benin, to assess whether G-ANC improved ANC retention and IPTp3 uptake at community level. Methods Forty purposively selected health facilities (HF) were randomized 1:1 to control (individual ANC) or G-ANC. Cross-sectional household surveys to measure uptake of ANC and IPTp were conducted in each HF catchment area before and after implementation among randomly selected women who had given birth in the previous 12 months. Changes in coverage were assessed using a difference-in-difference approach, adjusting for HF clustering. Results At baseline (N = 1259), coverage of at least 4 ANC visits (ANC4) and IPTp3 was 52.8% and 48.0%, respectively, in the intervention catchment, and 44.9% and 49.4% in the control catchment. Coverage of ANC4 improved in both arms by endline (N = 1280), to 56.7% in the intervention and 46.1% in the control, but the difference in the increase was not significant between arms (p = 0.51). Coverage of IPTp3 increased non-significantly (p = 0.26), to 53.2% (intervention) and 49.7% (control). Overall, only 140 (10.6%) surveyed women reported participating in G-ANC. Participation improved coverage of both ANC4 (65.0% vs 50.5%, p = 0.002; odds ratio (OR) 1.9, 95% CI 1.4–2.5) and IPTp3 (64.0 vs 50.6%, p = 0.004; OR = 1.8, 95% CI 1.2–2.6). Conclusions G-ANC increased ANC attendance and IPTp3 uptake among women who participated, but participation was limited. Understanding and addressing the barriers to participation is critical if G-ANC is to be used more widely to increase IPTp coverage. Trial Registration : PACTR202405487752509

Background Long-lasting insecticidal nets (LLINs) are currently the primary method of malaria control in sub-Saharan Africa and have contributed to a significant reduction in malaria burden over the past 15 years. However, this progress is threatened by the wide-scale selection of insecticide-resistant malaria vectors. It is, therefore, important to accelerate the generation of evidence for new classes of LLINs. Methods This protocol presents a three-arm superiority, single-blinded, cluster randomized controlled trial to evaluate the impact of 2 novel dual-active ingredient LLINs on epidemiological and entomological outcomes in Benin, a malaria-endemic area with highly pyrethroid-resistant vector populations. The study arms consist of (i) Royal Guard® LLIN, a net combining a pyrethroid (alpha-cypermethrin) plus an insect growth regulator (pyriproxyfen), which in the adult female is known to disrupt reproduction and egg fertility; (ii) Interceptor G2® LLIN, a net incorporating two adulticides (alpha-cypermethrin and chlorfenapyr) with different modes of action; and (iii) the control arm, Interceptor® LLIN, a pyrethroid (alpha-cypermethrin) only LLIN. In all arms, one net for every 2 people will be distributed to each household. Sixty clusters were identified and randomised 1:1:1 to each study arm. The primary outcome is malaria case incidence measured over 24 months through active case detection in a cohort of 25 children aged 6 months to 10 years, randomly selected from each cluster. Secondary outcomes include 1) malaria infection prevalence (all ages) and prevalence of moderate to severe anaemia in children under 5 years old, measured at 6 and 18 months post-intervention; 2) entomological indices measured every 3 months using human landing catches over 24 months. Insecticide resistance intensity will also be monitored over the study period. Discussion This study is the second cluster randomised controlled trial to evaluate the efficacy of these next-generation LLINs to control malaria transmitted by insecticide-resistant mosquitoes. The results of this study will form part of the WHO evidence-based review to support potential public health recommendations of these nets and shape malaria control strategies of sub-Saharan Africa for the next decade. Trial registration ClinicalTrials.gov, NCT03931473 , registered on 30 April 2019.

Symptomatic and asymptomatic malaria patients are considered as the reservoirs of human Plasmodium. In the present study, we have evaluated the Plasmodium falciparum merozoite surface protein-1 (Pfmsp1) and protein-2 (Pfmsp2) genetic diversity among the symptomatic and asymptomatic malaria infection from health facilities in Cotonou, Benin Republic. A cross-sectional study recruited 158 individuals, including 77 from the asymptomatic and 81 from the symptomatic groups. The parasites were genotyped using Nested Polymerase Chain Reaction. Samples identified as Plasmodium falciparum were genotyped for their genetic diversity. No significant difference was observed in the overall multiplicity of infection (MOI) between the asymptomatic and symptomatic groups. In the symptomatic group, the overall frequency of K1, MAD20, and RO33 allelic family was more predominant (98.5%) followed by 3D7 (87.3%) and FC27 (83.1%). However, in asymptomatic group, the K1 alleles were the most prevalent (100%) followed by FC27 (89.9%), 3D7 (76.8%), MAD20 (60.5%), and RO33 (35.5%). The frequency of multiple allelic types (K1+MAD20+RO33) at the Pfmsp1 loci in the symptomatic infections was significantly higher when compared to that of the asymptomatic ones (97% vs. 34%, p < 0.05), whereas no difference was observed in the frequency of multiple allelic types (3D7 and FC27) at the Pfmsp2 loci between the two groups. The high presence of msp1 multiple infections in the symptomatic group compared to asymptomatic ones suggests an association between the genetic diversity and the onset of malaria symptoms. These data can provide valuable information in the development of a vaccine that could reduce the symptomatic disease.

Background Insecticide-based interventions have averted more than 500 million malaria cases since 2000, but insecticide resistance in mosquitoes could bring about a rebound in disease and mortality. This study investigated whether insecticide resistance was associated with increased incidence of clinical malaria. Methods In an area of southern Benin with insecticide resistance and high use of insecticide-treated nets (ITNs), malaria morbidity and insecticide resistance were measured simultaneously in 30 clusters (villages or collections of villages) multiple times over the course of 2 years. Insecticide resistance frequencies were measured using the standard World Health Organization bioassay test. Malaria morbidity was measured by cases recorded at health facilities both in the whole population using routinely collected data and in a passively followed cohort of children under 5 years old. Results There was no evidence that incidence of malaria from routinely collected data was higher in clusters with resistance frequencies above the median, either in children aged under 5 (RR = 1.27 (95% CI 0.81–2.00) p = 0.276) or in individuals aged 5 or over (RR = 1.74 (95% CI 0.91–3.34) p = 0.093). There was also no evidence that incidence was higher in clusters with resistance frequencies above the median in the passively followed cohort (RR = 1.11 (0.52–2.35) p = 0.777). Conclusions This study found no association between frequency of resistance and incidence of clinical malaria in an area where ITNs are the principal form of vector control. This may be because, as other studies have shown, ITNs continue to offer some protection from malaria even in the presence of insecticide resistance. Irrespective of resistance, nets provide only partial protection so the development of improved or supplementary vector control tools is required to reduce Africa’s unacceptably high malaria burden.

Background In Benin, malaria vector control mostly relies on long-lasting, insecticidal-treated bed nets (LLINs) and indoor residual spraying (IRS) operations. From 2011 to 2016, an IRS programme has been implemented in Atacora region. However, in 2017 the programme was withdrawn from two other regions in the northern part of the country, with hopes that gains would be relatively sustained because of the seasonality of malaria transmission. What would be the vulnerability of populations to malaria after the withdrawal of IRS? Methods Monthly mosquito collections were performed through human landing captures (HLCs) for 24 months (from January to December 2016 during the last IRS campaign, and from January to December 2018, 2 years after the withdrawal of IRS). Vector mosquitoes biting density was sampled by HLC and was tested for presence of Plasmodium falciparum sporozoites. The carcass of these mosquitoes (abdomens, wing, legs) were subjected to molecular species identification using polymerase chain reaction (PCR) assays. Results It is noticed a drastic increase (~ 3 times higher) of vector abundance after the withdrawal of IRS. Mosquito biting rates in the 3 survey districts increased significantly after IRS was withdrawn. In 2018, after IRS cessation a significant increase of entomological inoculation rate was recorded, where each inhabitant received an average of 94.9 infected bites/year to 129.21 infected bites/year against an average of 17.15 infected bites/year to 24.82 infected bites/year in 2016. Conclusion It is obvious that the withdrawal of IRS confers a vulnerability of the population with regard to the malaria transmission. Robust monitoring is needed to better understand when and where IRS should be most adequate, or can be safely withdrawn. In case of withdrawal, adapted accompanying measures should be proposed according to the context not only to maintain the gains capitalized with IRS, but also to avoid any rebound of transmission.

Background Malaria control is heavily reliant on insecticides, especially pyrethroids. Resistance of mosquitoes to insecticides may threaten the effectiveness of insecticide-based vector control and lead to a resurgence of malaria in Africa. Methods In 21 villages in Southern Benin with high levels of insecticide resistance, the resistance status of local vectors was measured at the same time as the prevalence of malaria infection in resident children. Results Children who used LLINs had lower levels of malaria infection [odds ratio = 0.76 (95% CI 0.59, 0.98, p = 0.033)]. There was no evidence that the effectiveness of nets was different in high and low resistance locations (p = 0.513). There was no association between village level resistance and village level malaria prevalence (p = 0.999). Conclusions LLINs continue to offer individual protection against malaria infection in an area of high resistance. Insecticide resistance is not a reason to stop efforts to increase coverage of LLINs in Africa.

With global malaria cases on the rise, the WHO has placed increased emphasis on National Malaria Programmes to tailor interventions to country and programmatic needs. This paper presents the Plus Project’s experience of applying a codesign approach to design country-specific models of perennial malaria chemoprevention (PMC), a chemoprevention intervention aimed at reducing morbidity and mortality due to malaria and anaemia in children. Codesign workshops were held in each of the project’s focus countries (Benin, Cameroon, Côte d'Ivoire and Mozambique) with the primary objective of designing the country-specific PMC model. The three-and-a-half-day workshops were adapted to each country’s context and included stakeholders from national and subnational malaria, immunisation and child health programmes, as well as national and international development partners and research institutions. The meetings were iterative and collaborative, harnessing a variety of participatory methods including journey mapping and surveys to reach group consensus on the PMC models best suited to each country’s specific context. The Plus Project’s codesign approach resulted in four different PMC strategies, with a range from four to eight contact points and different codelivery interventions, each taking advantage of country-specific health system delivery platforms, operational logistics and political contexts. This collaborative, codesign process also helped gather additional programmatic insights to aid PMC implementation while providing an opportunity to increase stakeholder buy-in. With an emphasis on collaborative decision-making, the learnings collected through these workshops can be applied to a variety of programmatic applications, extending beyond malaria.

Background Increasing pyrethroid resistance has been an undesirable correlate of the rapid increase in coverage of insecticide-treated nets (ITNs) since 2000. Whilst monitoring of resistance levels has increased markedly over this period, longitudinal monitoring is still lacking, meaning the temporal and spatial dynamics of phenotypic resistance in the context of increasing ITN coverage are unclear. Methods As part of a large WHO-co-ordinated epidemiological study investigating the impact of resistance on malaria infection, longitudinal monitoring of phenotypic resistance to pyrethroids was undertaken in 290 clusters across Benin, Cameroon, India, Kenya and Sudan. Mortality in response to pyrethroids in the major anopheline vectors in each location was recorded during consecutive years using standard WHO test procedures. Trends in mosquito mortality were examined using generalised linear mixed-effect models. Results Insecticide resistance (using the WHO definition of mortality < 90%) was detected in clusters in all countries across the study period. The highest mosquito mortality (lowest resistance frequency) was consistently reported from India, in an area where ITNs had only recently been introduced. Substantial temporal and spatial variation was evident in mortality measures in all countries. Overall, a trend of decreasing mosquito mortality (increasing resistance frequency) was recorded (Odds Ratio per year: 0.79 per year (95% CI: 0.79–0.81, P < 0.001). There was also evidence that higher net usage was associated with lower mosquito mortality in some countries. Discussion Pyrethroid resistance increased over the study duration in four out of five countries. Insecticide-based vector control may be compromised as a result of ever higher resistance frequencies.