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The source of malaria vector populations that re-establish at the beginning of the rainy season is still unclear yet knowledge of mosquito behaviour is required to effectively institute control measures. Alternative hypotheses like aestivation, local refugia, migration between neighbouring sites, and long-distance migration (LDM) are stipulated to support mosquito persistence. This work assessed the malaria vector persistence dynamics and examined various studies done on vector survival  via these hypotheses; aestivation, local refugia, local or long-distance migration across sub-Saharan Africa, explored a range of methods used, ecological parameters and highlighted the knowledge trends and gaps. The results about a particular persistence mechanism that supports the re-establishment of Anopheles gambiae , Anopheles coluzzii or Anopheles arabiensis in sub-Saharan Africa were not conclusive given that each method used had its limitations. For example, the Mark-Release-Recapture (MRR) method whose challenge is a low recapture rate that affects its accuracy, and the use of time series analysis through field collections whose challenge is the uncertainty about whether not finding mosquitoes during the dry season is a weakness of the conventional sampling methods used or because of hidden shelters. This, therefore, calls for further investigations emphasizing the use of ecological experiments under controlled conditions in the laboratory or semi-field, and genetic approaches, as they are known to complement each other. This review, therefore, unveils and assesses the uncertainties that influence the different malaria vector persistence mechanisms and provides recommendations for future studies.

Macroinvertebrate predators such as backswimmers (Heteroptera: Notonectidae), dragonflies (Odonata: Aeshnidae), and predatory diving beetles (Coleoptera: Dytiscidae) naturally inhabit aquatic ecosystems. Some aquatic ecosystems inhabited by these macroinvertebrate predator taxa equally form malaria vector larval habitats. The presence of these predators in malaria vector larval habitats can negatively impact on development, adult body size, fecundity, and longevity of the malaria vectors, which form important determinants of their fitness and future vectorial capacity. These potential negative impacts caused by aquatic macroinvertebrate predators on malaria vectors warrant their consideration as biocontrol agents in an integrated program to combat malaria. However, the use of these macroinvertebrate predators in malaria biocontrol is currently constrained by technical bottlenecks linked to their generalist predatory tendencies and often long life cycles, demanding complex rearing systems. We reviewed the literature on the use of aquatic macroinvertebrate predators for biocontrol of malaria vectors from the An. gambiae s.l. complex. The available information from laboratory and semi-field studies has shown that aquatic macroinvertebrates have the potential to consume large numbers of mosquito larvae and could thus offer an additional approaches in integrated malaria vector management strategies. The growing number of semi-field structures available in East and West Africa provides an opportunity to conduct ecological experimental studies to reconsider the potential of using aquatic macroinvertebrate predators as a biocontrol tool. To achieve a more sustainable approach to controlling malaria vector populations, additional, non-chemical interventions could provide a more sustainable approach, in comparison with the failing chemical control tools, and should be urgently considered for integration with the current mosquito vector control campaigns. Graphical Abstract

Background Members of the Anopheles gambiae complex are major malaria vectors in sub-Saharan Africa. Their larval stages inhabit a variety of aquatic habitats in which, under natural circumstances, they are preyed upon by different taxa of aquatic macroinvertebrate predators. Understanding the potential impact of predators on malaria vector larval population dynamics is important for enabling integrated local mosquito control programmes with a stronger emphasis on biocontrol approaches. This study experimentally evaluated the predation efficacy and foraging strategy of three common aquatic macroinvertebrate predators of An. gambiae , diving beetles (Coleoptera), backswimmers (Hemiptera), and dragonfly nymphs (Odonata) in a semi-field system in South-Eastern Tanzania. Methods An array of alternating small and large basins used as aquatic habitats was created in two compartments of a semi-field system and filled with well water. Field-collected adult diving beetles, backswimmers or dragonfly nymphs were randomly assigned to these habitats and Anopheles arabiensis larvae were added as prey in half of the habitats. The number of mosquito larvae consumed, predator mobility across habitats and mortality were recorded at 24, 48 and 72 h. Results The presence of An. gambiae larvae in habitats significantly increased the survival of backswimmer and dragonfly nymphs, which are not mobile. In contrast, diving beetles survived well under any initial condition by preferentially flying away from habitats without prey to nearby larger habitats with prey. The larval predation rates of predacious diving beetle, backswimmer and dragonfly nymphs were stable over time at a mean of 3.2, 7.0 and 9.6 larvae consumed each day. Conclusion This study demonstrates that aquatic macroinvertebrate predators display adaptive foraging behaviour in response to prey presence and aquatic habitat size. It also confirms the ability of these predators to significantly reduce An. gambiae larval densities in aquatic habitats, thus their potential for consideration as additional biocontrol tools for mosquito population reduction.

Sustainable reductions in African malaria transmission require innovative tools for mosquito control. One proposal involves the use of low-threshold gene drive in Anopheles vector species, where a ‘causal pathway’ would be initiated by (i) the release of a gene drive system in target mosquito vector species, leading to (ii) its transmission to subsequent generations, (iii) its increase in frequency and spread in target mosquito populations, (iv) its simultaneous propagation of a linked genetic trait aimed at reducing vectorial capacity for Plasmodium , and (v) reduced vectorial capacity for parasites in target mosquito populations as the gene drive system reaches fixation in target mosquito populations, causing (vi) decreased malaria incidence and prevalence. Here the scope, objectives, trial design elements, and approaches to monitoring for initial field releases of such gene dive systems are considered, informed by the successful implementation of field trials of biological control agents, as well as other vector control tools, including insecticides, Wolbachia, larvicides, and attractive-toxic sugar bait systems. Specific research questions to be addressed in initial gene drive field trials are identified, and adaptive trial design is explored as a potentially constructive and flexible approach to facilitate testing of the causal pathway. A fundamental question for decision-makers for the first field trials will be whether there should be a selective focus on earlier points of the pathway, such as genetic efficacy via measurement of the increase in frequency and spread of the gene drive system in target populations, or on wider interrogation of the entire pathway including entomological and epidemiological efficacy. How and when epidemiological efficacy will eventually be assessed will be an essential consideration before decisions on any field trial protocols are finalized and implemented, regardless of whether initial field trials focus exclusively on the measurement of genetic efficacy, or on broader aspects of the causal pathway. Statistical and modelling tools are currently under active development and will inform such decisions on initial trial design, locations, and endpoints. Collectively, the considerations here advance the realization of developer ambitions for the first field trials of low-threshold gene drive for malaria vector control within the next 5 years.

Background Malaria is often persistent in communities surrounded by mosquito breeding habitats. Anopheles gambiae sensu lato exploit a variety of aquatic habitats, but the biotic determinants of its preferences are poorly understood. This study aimed to identify and quantify macroinvertebrates in different habitat types with determined water physico-chemical parameters to establish those preferred by An. gambiae s.l. larvae as well as their predators and competitors. Methods A field survey was conducted in Kibuye and Kayonjo villages located in the vicinity of the River Sezibwa, north-eastern Uganda to identify Anopheline larval habitats shared by aquatic insects. Habitats were geo-recorded and as streams, ponds, temporary pools and roadside ditches. From October to December 2017, random microhabitats/quadrats were selected from each habitat type, their water physico-chemical parameters (electrical conductivity, total dissolved solids, temperature and pH) were measured, and they were sampled for macroinvertebrates using standard dippers. All collected arthropod macroinvertebrates were then morphologically identified to family level and enumerated. Results Principal component analysis showed that the four larval habitat types were characterized by distinct physico-chemical parameter profiles. Ponds and streams had the highest number and diversity of macroinvertebrate insect taxa and sustained few An. gambiae s.l. larvae. Anopheles gambiae s.l. were more common in roadside ditches and particularly abundant in temporary pools which it commonly shared with Dytiscidae (predaceous diving beetles) and Culex spp. Cluster correlation analysis conducted on the abundance of these taxa within quadrats suggested that An. gambiae s.l. and Dytiscidae have the most similar patterns of microhabitat use, followed by Cybaeidae (water spiders). Whilst Culex spp. co-occurred with An. gambiae s.l. in some habitats, there was only partial niche overlap and no clear evidence of competition between the two mosquito taxa. Conclusions Ponds and streams are habitats that host the largest diversity and abundance of aquatic insect taxa. Anopheles gambiae s.l. larvae distinctively preferred temporary pools and roadside ditches, where they were exposed to few predators and no apparent competition by Culex spp. Further studies should aim to test the impact of Dytiscidae and Cybaeidae on An. gambiae s.l. dynamics experimentally. Graphical Abstract

Background Members of the Anopheles gambiae complex, such as Anopheles gambiae sensu stricto ( An. gambiae s.s.), Anopheles coluzzii and Anopheles arabiensis , are among the key malaria vectors in sub-Saharan Africa. These species are often abundant in areas of intense rice and maize farming with temporary water pools reflecting the dependence of their larvae on the pollen shed in such pools as food. In this study we explored the oviposition preference of wild-caught gravid An. arabiensis in response to maize and rice pollen in artificial aquatic habitats in a semi-field system. Methods Twelve experimental breeding habitats were established in each of the two large compartments of a semi-field system. Rice or maize pollen was added into eight randomly selected habitats in eachcompartment; the remaining four habitats of each compartment were used as control habitats without pollens. In the first experiment, 40 gravid An. arabiensis were released in each compartment and left overnight to choose egg-laying habitats, following which the eggs were sampled and counted. The second experiment differed from the first experiment only in that the counted eggs were returned to the respective habitat where the development of the resultant larvae was monitored and recorded until pupation. Results Pollen types strongly affected the oviposition behaviour of gravid An. arabiensis . Females preferred to lay eggs in habitats with rice pollen on the water surface over those with maize pollen, and in habitats with maize pollen on the water surface over pollen-less controls. The development of larvae was significantly affected by the type of pollen in the habitats. The highest total number of An. arabiensis offspring were produced in habitats with rice pollen compared to those with maize pollen and no pollen. However, larval development success was comparatively lower in habitats containing rice pollen than those with maize pollen and no pollen, suggesting that the habitats with rice pollen were overcrowded. Conclusion This study demonstrates that pollen types on the surface of aquatic habitats influence the oviposition site selection behaviour of gravid An. arabiensis and has carry-over effect on the developmental success of their offspring. Graphical abstract

Background Anopheles gambiae continues to be widespread and an important malaria vector species complex in Uganda. New approaches to malaria vector control are being explored including population suppression through swarm reductions and genetic modification involving gene drives. Designing and evaluating these new interventions require good understanding of the biology of the target vectors. Anopheles mosquito swarms have historically been hard to locate in Uganda and therefore have remained poorly characterized. In this study we sought to identify and characterize An. gambiae s.l mosquito swarms in three study sites of high An. gambiae s.l prevalence within Central Uganda. Methods Nine sampling visits were made to three villages over a 2-year period. Sampling targeted both wet and dry seasons and was done for 2 days per village during each trip, using sweep nets. All swarm data were analysed using the JMP 14 software (SAS Institute, Inc., Cary, NC, USA), parametrically or non-parametrically as appropriate. Results Most of the An. gambiae s.s. swarms sampled during this study were single-species swarms. However, some mixed An. gambiae s.s. and Culex spp. mosquito swarms were also observed. Swarms were larger in the wet season than in the dry season. Mean swarm height ranged from 2.16 m to 3.13 m off the ground and only varied between villages but not by season. Anopheles gambiae mosquitoes were present in all three villages, preferred to swarm over bare ground markers, and could be effectively sampled by field samplers. Conclusions This study demonstrated that An. gambiae s.l swarms could be effectively located and sampled in South Central Uganda and provided in-depth descriptions of hitherto poorly understood aspects of An. gambiae local swarm characteristics. Swarms were found close to inhabited households and were greater in size and number during the rainy season. Anopheles gambiae s.s swarms were significantly associated with bare ground markers and were sometimes at heights over 4 m above the ground, showing a necessity to develop tools suitable for swarm sampling at these heights. While mixed species swarms have been reported before elsewhere, this is the first documented instance of mixed genus swarms found in Uganda and should be studied further as it could have implications for swarm sampling explorations where multiple species of mosquitoes exist. Graphical Abstract

Background Currently, formal mechanisms for bioinformatics support are limited. The H3Africa Bioinformatics Network has implemented a public and freely available Helpdesk (HD), which provides generic bioinformatics support to researchers through an online ticketing platform. The following article reports on the H3ABioNet HD (H3A-HD)‘s development, outlining its design, management, usage and evaluation framework, as well as the lessons learned through implementation. Results The H3A-HD evaluated using automatically generated usage logs, user feedback and qualitative ticket evaluation. Evaluation revealed that communication methods, ticketing strategies and the technical platforms used are some of the primary factors which may influence the effectivity of HD. Conclusion To continuously improve the H3A-HD services, the resource should be regularly monitored and evaluated. The H3A-HD design, implementation and evaluation framework could be easily adapted for use by interested stakeholders within the Bioinformatics community and beyond.

Documenting isolation is notoriously difficult for species with vast polymorphic populations. High proportions of shared variation impede estimation of connectivity, even despite leveraging information from many genetic markers. We overcome these impediments by combining classical analysis of neutral variation with assays of the structure of selected variation, demonstrated using populations of the principal African malaria vector Anopheles gambiae. Accurate estimation of mosquito migration is crucial for efforts to combat malaria. Modeling and cage experiments suggest that mosquito gene drive systems will enable malaria eradication, but establishing safety and efficacy requires identification of isolated populations in which to conduct field testing. We assess Lake Victoria islands as candidate sites, finding one island 30 km offshore is as differentiated from mainland samples as populations from across the continent. Collectively, our results suggest sufficient contemporary isolation of these islands to warrant consideration as field‐testing locations and illustrate shared adaptive variation as a useful proxy for connectivity in highly polymorphic species.