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Background: The efficacy of vector control tools depends on the behavior of the vector species. Many studies have sought to determine the feeding behavior of Anopheles mosquitoes in different settings of Ethiopia. We have performed a systematic review aimed to generate pooled evidence on the overall and species-specific blood meal sources of Anopheles mosquitoes in Ethiopia. Methods: A search for relevant articles was performed in two electronic databases (PubMed and Science Direct) and three search engines (Google Scholar, Research Gate and Google) between 11 March and 2 April 2024. Following the initial identification of articles, we used EndNote X8 software and removed duplicate articles and screened the remaining articles by careful reading of their titles and abstracts. The full text of articles that passed this screening phase was retrieved, read and evaluated against predetermined selection criteria. The final decision for inclusion in the systematic review was made after a methodological quality check using the JBI critical appraisal checklist. All relevant data were extracted from tables, figures and texts of the included articles using a premade template in Excel, and the data were analyzed using Stata version 14 software. Results: Of the 2431 studies identified, 27 met the inclusion criteria; all were published between 1997 and 2024. At 215 data points (frequency of tests of each Anopheles species by location and method of mosquito collections), 18,771 Anopheles mosquitoes belonging to 23 species or species complexes were tested for blood meal sources. The commonest sources of blood meals for Anopheles mosquitoes were bovine (36.0%, n = 6758) and human (29.4%, n = 5520). Among the tested anophelines, Anopheles (An.) arabiensis accounted for 67.9% (n = 12,741), followed by An. pharoensis, An. demeilloni and An. stephensi at 10.0%, 5.6% and 4.4%, respectively. Overall, there was no difference in the mean proportion of An. arabiensis detected with domestic animal blood (33.4%, 95% confidence interval [CI] 32.4–34.4%) and those detected with human blood (31.8%, 95% CI 30.9–32.8%). However, a greater proportion of the outdoor collected An. arabiensis were found to feed on bovines (47.9%, 95% CI 35.3–60.6) compared to humans (12.9%, 95% CI 0.8–24.9, P < 0.01). The foraging ratio (FR), which accounts for host availability, was greater for bovines (FR = 0.7) than for humans (FR = 0.2) for An. arabiensis, indicating preferential feeding on bovine hosts. This host preference was supported by the host preference index (human:bovine = 0.4). Anopheles pharoensis was detected with a slightly higher human blood index (53.5%, n = 1005) compared to bovine blood index (45.2%, n = 849). In contrast, An. demeilloni, An. coustani and An. marshalli were detected with a higher bovine blood index. Recently invaded urban malaria vector, An. stephensi was found with a higher ovine blood index. Conclusions: Bovine and human hosts are common sources of a blood meal for Anopheles mosquitoes. In terms of host availability, An. arabiensis showed preferential feeding on bovines/cattle. Targeting domestic animals, bovines and ovines with endectocides could supplement current vector control interventions. Study registration: The protocol of this study was registered on the International Prospective Register of Systematic Reviews, registration no. CRD42024515725. [ABSTRACT FROM AUTHOR]

Abstract Background The efficacy of vector control tools depends on the behavior of the vector species. Many studies have sought to determine the feeding behavior of Anopheles mosquitoes in different settings of Ethiopia. We have performed a systematic review aimed to generate pooled evidence on the overall and species-specific blood meal sources of Anopheles mosquitoes in Ethiopia. Methods A search for relevant articles was performed in two electronic databases (PubMed and Science Direct) and three search engines (Google Scholar, Research Gate and Google) between 11 March and 2 April 2024. Following the initial identification of articles, we used EndNote X8 software and removed duplicate articles and screened the remaining articles by careful reading of their titles and abstracts. The full text of articles that passed this screening phase was retrieved, read and evaluated against predetermined selection criteria. The final decision for inclusion in the systematic review was made after a methodological quality check using the JBI critical appraisal checklist. All relevant data were extracted from tables, figures and texts of the included articles using a premade template in Excel, and the data were analyzed using Stata version 14 software. Results Of the 2431 studies identified, 27 met the inclusion criteria; all were published between 1997 and 2024. At 215 data points (frequency of tests of each Anopheles species by location and method of mosquito collections), 18,771 Anopheles mosquitoes belonging to 23 species or species complexes were tested for blood meal sources. The commonest sources of blood meals for Anopheles mosquitoes were bovine (36.0%, n = 6758) and human (29.4%, n = 5520). Among the tested anophelines, Anopheles (An.) arabiensis accounted for 67.9% (n = 12,741), followed by An. pharoensis, An. demeilloni and An. stephensi at 10.0%, 5.6% and 4.4%, respectively. Overall, there was no difference in the mean proportion of An. arabiensis detected with domestic animal blood (33.4%, 95% confidence interval [CI] 32.4–34.4%) and those detected with human blood (31.8%, 95% CI 30.9–32.8%). However, a greater proportion of the outdoor collected An. arabiensis were found to feed on bovines (47.9%, 95% CI 35.3–60.6) compared to humans (12.9%, 95% CI 0.8–24.9, P

Beginning in 2023, we observed increased Plasmodium vivax malaria cases at an institution in Los Angeles, California, USA. Most cases were among migrants from China who traveled to the United States through South and Central America. US clinicians should be aware of possible P. vivax malaria among immigrants from China.

Abstract Background “Regeneration time” (RT) denotes the time required to obtain a stable mortality rate for mosquitoes exposed to insecticide-treated nets (ITNs) after three consecutive washes of a net in a day. The RT informs the wash interval used to artificially age ITNs to simulate their lifetime performance under user conditions (20 washes). RT was estimated following World Health Organization (WHO) longitudinal method (LM) procedures. Longitudinal evaluation may introduce heterogeneity due to mosquito batch variability, complicating RT determination. To overcome this, nets at each stage of regeneration (i.e., 1, 2, 3, 5 and 7 days post wash) were prepared in advance and refrigerated; then, a complete regeneration series was tested with a single mosquito batch on 1 testing day, completing four series over 4 days. This study compared the complete series method (CSM) against the LM. Methods The overall heterogeneity in the methods for estimating RT of one incorporated alpha-cypermethrin and piperonyl butoxide (PBO) and one incorporated permethrin with PBO ITNs was determined using laboratory-reared resistant Anopheles arabiensis under standard laboratory conditions. LM methods and CSM were compared in two experiments with refrigerated nets acclimated for (i) 2 h (test 1) and (ii) 3 h (test 2). Four regeneration replicates per day were tested per ITN product with 50 mosquitoes exposed per replicate (equivalent sample size to LM). The heterogeneity from these methods was compared descriptively. Results The intra-method variability for unwashed pieces was minimal, with variance of 1.26 for CSM and 1.18 for LM. For unwashed nets, LM had substantially greater variance and ratio of LM:CSM was 2.66 in test 1 and 2.49 in test 2. The magnitude of mortality measured in bioassays depended on sample acclimation after refrigeration. Conclusions The CSM is a convenient method for determining the regeneration times. ITNs are prepared in advance, reducing pressure to prepare all samples to start on a single day. A complete regeneration series of samples is removed from the refrigerator, defrosted and evaluated on a single day with one mosquito batch reducing the influence of mosquito batch heterogeneity on results. Replicates can be conducted over several days but do not have to be conducted on consecutive days, allowing easy facility scheduling. Graphical Abstract

Background: "Regeneration time" (RT) denotes the time required to obtain a stable mortality rate for mosquitoes exposed to insecticide-treated nets (ITNs) after three consecutive washes of a net in a day. The RT informs the wash interval used to artificially age ITNs to simulate their lifetime performance under user conditions (20 washes). RT was estimated following World Health Organization (WHO) longitudinal method (LM) procedures. Longitudinal evaluation may introduce heterogeneity due to mosquito batch variability, complicating RT determination. To overcome this, nets at each stage of regeneration (i.e., 1, 2, 3, 5 and 7 days post wash) were prepared in advance and refrigerated; then, a complete regeneration series was tested with a single mosquito batch on 1 testing day, completing four series over 4 days. This study compared the complete series method (CSM) against the LM. Methods: The overall heterogeneity in the methods for estimating RT of one incorporated alpha-cypermethrin and piperonyl butoxide (PBO) and one incorporated permethrin with PBO ITNs was determined using laboratory-reared resistant Anopheles arabiensis under standard laboratory conditions. LM methods and CSM were compared in two experiments with refrigerated nets acclimated for (i) 2 h (test 1) and (ii) 3 h (test 2). Four regeneration replicates per day were tested per ITN product with 50 mosquitoes exposed per replicate (equivalent sample size to LM). The heterogeneity from these methods was compared descriptively. Results: The intra-method variability for unwashed pieces was minimal, with variance of 1.26 for CSM and 1.18 for LM. For unwashed nets, LM had substantially greater variance and ratio of LM:CSM was 2.66 in test 1 and 2.49 in test 2. The magnitude of mortality measured in bioassays depended on sample acclimation after refrigeration. Conclusions: The CSM is a convenient method for determining the regeneration times. ITNs are prepared in advance, reducing pressure to prepare all samples to start on a single day. A complete regeneration series of samples is removed from the refrigerator, defrosted and evaluated on a single day with one mosquito batch reducing the influence of mosquito batch heterogeneity on results. Replicates can be conducted over several days but do not have to be conducted on consecutive days, allowing easy facility scheduling. [ABSTRACT FROM AUTHOR]

BACKGROUND: The potential reappearance and/or expansion of vector‐borne diseases is one of the terrifying issues awaiting humanity in the context of climate change. The presence of competent Anopheles vectors, as well as suitable environmental circumstances, may result in the re‐emergence of autochthonous Malaria, after years of absence. In Morocco, international travel and migration movements from Malaria‐endemic areas have recently increased the number of imported cases, raising awareness of Malaria's possible reintroduction. Using machine learning we developed model predictions, under current and future (2050) climate, for the prospective distribution of Anopheles claviger, Anopheles labranchiae, Anopheles multicolor, and Anopheles sergentii implicated or incriminated in Malaria transmission. RESULTS: All modelled species are expected to find suitable habitats and have the potential to become established in the northern and central parts of the country, under present‐day conditions. Distinct changes in the distributions of the four mosquitoes are to be expected under climate change. Even under the most optimistic scenario, all investigated species are likely to acquire new habitats that are now unsuitable, placing further populations in danger. We also observed a northward and altitudinal shift in their distribution towards higher altitudes. CONCLUSION: Climate change is expected to expand the potential range of malaria vectors in Morocco. Our maps and predictions offer a way to intelligently focus efforts on surveillance and control programmes. To reduce the threat of human infection, it is crucial for public health authorities, entomological surveillance teams, and control initiatives to collaborate and intensify their actions, continuously monitoring areas at risk. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Abstract Background Malaria remains an enduring public health concern in Indonesia, exacerbated by its equatorial climate that fosters the proliferation of Anopheles mosquitoes. This study seeks to assess the performance of the malaria elimination programme comprehensively. Methods Between May and August 2022, a qualitative study was conducted in Muara Enim Regency, South Sumatra Province, involving 22 healthcare professionals from diverse backgrounds. These informants were strategically chosen for their pivotal roles in providing profound insights into various facets of the malaria elimination programme. This encompasses inputs such as human resources, budgetary allocation, and infrastructural support; processes like case identification and management, capacity enhancement, epidemiological surveillance, prevention measures, outbreak control, and enhanced communication and educational initiatives; and, notably, the programme’s outcomes. Data were collected through 3-h Focus Group Discussions (FGDs) divided into two groups, each with 12 participants: healthcare professionals and programme managers. Additionally, in-depth interviews (IDIs) were conducted with ten informants. Employing the Input-Process-Output (IPO) model, this study meticulously analysed the healthcare system dynamics and the interventions’ efficacy. Results The study unveiled many challenges during the input phase, including the absence of entomologists and a shortage of diagnostic tools. Despite these obstacles, it documented remarkable accomplishments in the output domain, marked by significant advancements in the distribution of mosquito nets and the successful implementation of the Early Warning System (EWS). Despite the adversities, the programme has made substantial strides towards malaria elimination. Conclusions Urgent action is imperative to bolster the effectiveness of the malaria elimination programme. Key measures encompass augmenting the entomologist workforce, optimizing resource allocation, and ensuring stringent adherence to regional regulations. Addressing these concerns will enhance programme efficacy, yielding enduring public health benefits. This research substantially contributes to Indonesia’s ongoing malaria elimination endeavours, furnishing actionable insights for programme enhancement. Consequently, this research holds significant importance for the malaria elimination drive.

Abstract Background Insecticide resistance is reducing the efficacy of vector control interventions, consequently threatening efforts to control vector-borne diseases, including malaria. Investigating the prevalence of molecular markers of resistance is a useful tool for monitoring the spread of insecticide resistance in disease vectors. The Bijagós Archipelago (Bijagós) in Guinea-Bissau is a region of stable malaria transmission where insecticide-treated nets are the mainstay for malaria control. However, the prevalence of molecular markers of insecticide resistance in malaria vectors is not well understood. Methods A total of 214 Anopheles mosquitoes were analysed from 13 islands across the Bijagós. These mosquitoes were collected using CDC light traps in November 2019, during the peak malaria transmission season. High-throughput multiplex amplicon sequencing was used to investigate the prevalence of 17 different molecular markers associated with insecticide resistance in four genes: vgsc, rdl, ace1 and gste2. Results Of the 17 screened mutations, four were identified in mosquitoes from the Bijagós: vgsc L995F (12.2%), N1570Y (6.2%) and A1746S (0.7%) and rdl A269G (1.1%). This study is the first to report the L995F knock-down resistance (kdr)-west allele in Anopheles melas on the Archipelago. An additional eight non-synonymous single-nucleotide polymorphisms were identified across the four genes which have not been described previously. The prevalences of the vgsc L995F and N1570Y mutations were higher on Bubaque Island than on the other islands in this study; Bubaque is the most populous island in the archipelago, with the greatest population mobility and connection to continental Guinea-Bissau. Conclusions This study provides the first surveillance data for genetic markers present in malaria vectors from islands across the Bijagós Archipelago. Overall prevalence of insecticide resistance mutations was found to be low. However, the identification of the vgsc L995F and N1570Y mutations associated with pyrethroid resistance warrants further monitoring. This is particularly important as the mainstay of malaria control on the islands is the use of pyrethroid insecticide-treated nets. Graphical Abstract

Background: Malaria remains an enduring public health concern in Indonesia, exacerbated by its equatorial climate that fosters the proliferation of Anopheles mosquitoes. This study seeks to assess the performance of the malaria elimination programme comprehensively. Methods: Between May and August 2022, a qualitative study was conducted in Muara Enim Regency, South Sumatra Province, involving 22 healthcare professionals from diverse backgrounds. These informants were strategically chosen for their pivotal roles in providing profound insights into various facets of the malaria elimination programme. This encompasses inputs such as human resources, budgetary allocation, and infrastructural support; processes like case identification and management, capacity enhancement, epidemiological surveillance, prevention measures, outbreak control, and enhanced communication and educational initiatives; and, notably, the programme's outcomes. Data were collected through 3-h Focus Group Discussions (FGDs) divided into two groups, each with 12 participants: healthcare professionals and programme managers. Additionally, in-depth interviews (IDIs) were conducted with ten informants. Employing the Input-Process-Output (IPO) model, this study meticulously analysed the healthcare system dynamics and the interventions' efficacy. Results: The study unveiled many challenges during the input phase, including the absence of entomologists and a shortage of diagnostic tools. Despite these obstacles, it documented remarkable accomplishments in the output domain, marked by significant advancements in the distribution of mosquito nets and the successful implementation of the Early Warning System (EWS). Despite the adversities, the programme has made substantial strides towards malaria elimination. Conclusions: Urgent action is imperative to bolster the effectiveness of the malaria elimination programme. Key measures encompass augmenting the entomologist workforce, optimizing resource allocation, and ensuring stringent adherence to regional regulations. Addressing these concerns will enhance programme efficacy, yielding enduring public health benefits. This research substantially contributes to Indonesia's ongoing malaria elimination endeavours, furnishing actionable insights for programme enhancement. Consequently, this research holds significant importance for the malaria elimination drive. [ABSTRACT FROM AUTHOR]

Background: Insecticide resistance is reducing the efficacy of vector control interventions, consequently threatening efforts to control vector-borne diseases, including malaria. Investigating the prevalence of molecular markers of resistance is a useful tool for monitoring the spread of insecticide resistance in disease vectors. The Bijagós Archipelago (Bijagós) in Guinea-Bissau is a region of stable malaria transmission where insecticide-treated nets are the mainstay for malaria control. However, the prevalence of molecular markers of insecticide resistance in malaria vectors is not well understood. Methods: A total of 214 Anopheles mosquitoes were analysed from 13 islands across the Bijagós. These mosquitoes were collected using CDC light traps in November 2019, during the peak malaria transmission season. High-throughput multiplex amplicon sequencing was used to investigate the prevalence of 17 different molecular markers associated with insecticide resistance in four genes: vgsc, rdl, ace1 and gste2. Results: Of the 17 screened mutations, four were identified in mosquitoes from the Bijagós: vgsc L995F (12.2%), N1570Y (6.2%) and A1746S (0.7%) and rdl A269G (1.1%). This study is the first to report the L995F knock-down resistance (kdr)-west allele in Anopheles melas on the Archipelago. An additional eight non-synonymous single-nucleotide polymorphisms were identified across the four genes which have not been described previously. The prevalences of the vgsc L995F and N1570Y mutations were higher on Bubaque Island than on the other islands in this study; Bubaque is the most populous island in the archipelago, with the greatest population mobility and connection to continental Guinea-Bissau. Conclusions: This study provides the first surveillance data for genetic markers present in malaria vectors from islands across the Bijagós Archipelago. Overall prevalence of insecticide resistance mutations was found to be low. However, the identification of the vgsc L995F and N1570Y mutations associated with pyrethroid resistance warrants further monitoring. This is particularly important as the mainstay of malaria control on the islands is the use of pyrethroid insecticide-treated nets. [ABSTRACT FROM AUTHOR]

Malaria and lymphatic filariasis (LF) are two serious public health challenges in sub-Saharan Africa, and both diseases are transmitted by Anopheles mosquitoes. Successful control of both diseases requires detailed information on transmission dynamics; thus, this study investigated malaria and LF transmission indices in two (2) communities (Jidawa and Kargo) in North-West Nigeria. Anopheles mosquitoes were sampled from twenty-five (25) randomly selected houses from each of the two communities using pyrethrum spray collection (PSC). The samples were identified morphologically and molecularly characterised using polymerase chain reaction (PCR). Human biting rate (HBR), indoor resting density (IRD), sporozoite rate (SR) and entomological inoculation rate (EIR) were calculated using standard formulae. The thorax region of the collected samples were dissected and smeared; then, Plasmodium and Wuchereria bancrofti parasites were identified using microscopy. A total of 2417 Anopheles mosquitoes were collected, and all were identified morphologically as An. gambiae s.l. Further molecular identification of sibling species revealed that An. gambiae and An. arabiensis were the only sibling species present. A total of 818 Anopheles mosquitoes were screened for Plasmodium and Wuchereria bancrofti parasites. A total of 180 samples were positive for Plasmodium parasites (Jidawa = 151; Kargo = 29), and none was positive for W. bancrofti (0%). Result of entomological indices for malaria transmission showed that indoor resting density was higher in Jidawa (10 mosquitoes/room/night) while human biting rate (2.07 bites/person/night), sporozoite rate (29.3%) and entomological inoculation rate (0.61) were higher in Kargo. In total, 35.2% of the samples were blood-fed while 67.4% were parous. There is active transmission of malaria in the two communities but not LF, suggesting the effectiveness of mass drug administration for LF. Concerted efforts should be focused on malaria control as transmission of the disease persists. [ABSTRACT FROM AUTHOR]

Malaria, caused by Plasmodium parasites and transmitted through Anopheles mosquitoes, remains a formidable global health challenge. This abstract provides an overview of the intricate molecular mechanisms underlying malaria pathogenesis, explores current therapeutic approaches, and highlights emerging insights that may shape future strategies for malaria control. The intricate dance between Plasmodium parasites and their human hosts begins with the mosquito's bite, leading to the invasion of erythrocytes by Plasmodium species. We delve into the molecular mechanisms governing parasite entry and subsequent replication within host cells, shedding light on key factors such as erythrocyte surface receptors and parasite-encoded proteins critical to invasion and survival. While malaria treatment has relied heavily on antimalarial drugs, the emergence of drug resistance necessitates ongoing exploration of novel therapeutic strategies. This abstract reviews current antimalarial drug classes, their mechanisms of action, and the challenges posed by drug resistance. We also highlight promising drug candidates and innovative approaches in the pipeline, including the use of advanced molecular techniques and immunotherapies. Emerging insights from genomics, proteomics, and transcriptomics have deepened our understanding of Plasmodium biology and host-parasite interactions. We discuss the potential of these omics approaches in identifying new drug targets, understanding drug resistance mechanisms, and developing vaccines. Additionally, we examine the role of human genetics in influencing susceptibility to malaria and response to treatment. Vector control remains a critical component of malaria prevention. We touch upon emerging strategies, such as genetically modified mosquitoes and novel insecticides, in the context of integrated vector management programs. Finally, we emphasize the importance of a multifaceted approach to malaria control, combining advances in molecular biology, drug development, vector control, and public health interventions. [ABSTRACT FROM AUTHOR]

Abstract Background Measuring risk of malaria transmission is complex, especially in case of Plasmodium vivax. This may be overcome using membrane feeding assays in the field where P. vivax is endemic. However, mosquito-feeding assays are affected by a number of human, parasite and mosquito factors. Here, this study identified the contributions of Duffy blood group status of P. vivax-infected patients as a risk of parasite transmission to mosquitoes. Methods A membrane feeding assay was conducted on a total of 44 conveniently recruited P. vivax infected patients in Adama city and its surroundings in East Shewa Zone, Oromia region, Ethiopia from October, 2019 to January, 2021. The assay was performed in Adama City administration. Mosquito infection rates were determined by midgut dissections at seven to 8 days post-infection. Duffy genotyping was defined for each of the 44 P. vivax infected patients. Results The infection rate of Anopheles mosquitoes was 32.6% (296/907) with 77.3% proportion of infectious participants (34/44). Infectiousness of participants to Anopheles mosquitoes appeared to be higher among individuals with homozygous Duffy positive blood group (TCT/TCT) than heterozygous (TCT/CCT), but the difference was not statistically significant. The mean oocyst density was significantly higher among mosquitoes fed on blood of participants with FY*B/FY*BES than other genotypes (P = 0.001). Conclusion Duffy antigen polymorphisms appears to contribute to transmissibility difference of P. vivax gametocytes to Anopheles mosquitoes, but further studies are required.

BackgroundDespite extensive irrigation development in Ethiopia, limited studies assessed the impact of irrigation on malaria vector mosquito composition, abundance and seasonality. This study aimed to evaluate the impact of sugarcane irrigation on species composition, abundance and seasonality of malaria vectors.MethodsAdult Anopheles mosquitoes were collected using CDC light traps from three irrigated and three non-irrigated clusters in and around Arjo-Didessa sugarcane irrigation scheme in southwestern Ethiopia. Mosquitoes were surveyed in four seasons: two wet and two dry, in 2018 and 2019. Mosquito species composition, abundance and seasonality were compared between irrigated and non-irrigated clusters. Anopheles mosquitoes were sorted out to species using morphological keys and molecular techniques. Chi square was used to test the relationships between Anopheles species occurrence, and environmental and seasonal parameters.ResultsOverall, 2108 female Anopheles mosquitoes comprising of six species were collected. Of these, 92.7% (n = 1954) were from irrigated clusters and 7.3% (n = 154) from the non-irrigated. The Anopheles gambiae complex was the most abundant (67.3%) followed by Anopheles coustani complex (25.3%) and Anopheles pharoensis (5.7%). PCR-based identification revealed that 74.7% (n = 168) of the An. gambiae complex were Anopheles arabiensis and 22.7% (n = 51) Anopheles amharicus. The density of An. gambiae complex (both indoor and outdoor) was higher in irrigated than non-irrigated clusters. The overall anopheline mosquito abundance during the wet seasons (87.2%; n = 1837) was higher than the dry seasons (12.8%; n = 271).ConclusionThe ongoing sugarcane irrigation activities in Arjo-Didessa created conditions suitable for malaria transmitting Anopheles species diversity and abundance. This could drive malaria transmission in Arjo-Didessa and its environs in both dry and wet seasons. Currently practiced malaria vector interventions need to be strengthened by including larval source management to reduce vector abundance in the irrigated areas.

Abstract Background Insecticide resistance is a key barrier to long-term malaria control, and it may be exacerbated by poor agricultural pesticide use. Current practices, however, do not link public health and agricultural pesticide use. This study investigated the perspectives of farmers and other stakeholders regarding the integration of agricultural and public health measures to address resistance. Additionally, the feasibility of participatory workshops to increase the farmers’ understanding and participation in pesticide stewardship was assessed. Methods Four themes were investigated: pesticide awareness, practices, and opinions of; insecticide resistance in malaria vectors; the effectiveness of current malaria prevention tools; and the links between agricultural and public health pesticide usage. Participatory workshops and field training were held with entomologists, farmers, and agricultural specialists, focusing on agro-ecosystem practices related to pest control; and local farmers were involved in live-testing for insecticides resistance of local Anopheles mosquitoes. Results Most farmers (94%) considered pesticides effective, and nearly half of them (n = 198, 46.4%) could identify and name crop pests and diseases, mostly using local names. Three quarters were unaware of mosquito larvae in their fields, and only 7% considered their fields as potential sources of mosquitoes. Two thirds were uninformed of any effects that agricultural pesticides may have on mosquitoes, and three quarters had never heard of resistance in malaria mosquitoes. Experts from various sectors acknowledged that agricultural pesticides might impact malaria control through increasing resistance. They did, however, emphasize the importance of crop protection and advocated for the use of pesticides sparingly and non-chemical approaches. Farmers learnt how to discriminate between malaria vectors and non-vectors, identify agricultural pests and diseases, choose and use pesticides effectively, and conduct resistance tests during the participatory workshops. Conclusion This study emphasizes the significance of enhancing subsistence farmers’ awareness of mosquito ecology as well as merging public health and agricultural pest management measures. Participatory techniques have the potential to raise stakeholder awareness and engagement, resulting in more effective resistance management.

Background: Measuring risk of malaria transmission is complex, especially in case of Plasmodium vivax. This may be overcome using membrane feeding assays in the field where P. vivax is endemic. However, mosquito-feeding assays are affected by a number of human, parasite and mosquito factors. Here, this study identified the contributions of Duffy blood group status of P. vivax-infected patients as a risk of parasite transmission to mosquitoes. Methods: A membrane feeding assay was conducted on a total of 44 conveniently recruited P. vivax infected patients in Adama city and its surroundings in East Shewa Zone, Oromia region, Ethiopia from October, 2019 to January, 2021. The assay was performed in Adama City administration. Mosquito infection rates were determined by midgut dissections at seven to 8 days post-infection. Duffy genotyping was defined for each of the 44 P. vivax infected patients. Results: The infection rate of Anopheles mosquitoes was 32.6% (296/907) with 77.3% proportion of infectious participants (34/44). Infectiousness of participants to Anopheles mosquitoes appeared to be higher among individuals with homozygous Duffy positive blood group (TCT/TCT) than heterozygous (TCT/CCT), but the difference was not statistically significant. The mean oocyst density was significantly higher among mosquitoes fed on blood of participants with FY*B/FY*BES than other genotypes (P = 0.001). Conclusion: Duffy antigen polymorphisms appears to contribute to transmissibility difference of P. vivax gametocytes to Anopheles mosquitoes, but further studies are required. [ABSTRACT FROM AUTHOR]

This study evaluated the impact of combining house screens with long-lasting insecticidal nets (LLINs) on mosquito host-seeking, resting, and biting behavior. Intervention houses received house screens and LLINs, while control houses received only LLINs. Centre for Disease Control light traps, pyrethrum spray collections and human landing catches were used to assess the densities of indoor and outdoor host-seeking, indoor resting, and biting behavior of malaria vectors in 15 sentinel houses per study arm per sampling method. The protective efficacy of screens and LLINs was estimated through entomological inoculation rates (EIRs). There were 68% fewer indoor host-seeking Anopheles funestus (RR = 0.32, 95% CI 0.20–0.51, p < 0.05) and 63% fewer An. arabiensis (RR = 0.37, 95% CI 0.22–0.61, p < 0.05) in screened houses than unscreened houses. There was a significantly higher indoor biting rate for unscreened houses (6.75 bites/person/h [b/p/h]) than for screened houses (0 b/p/h) (χ2 = 6.67, df = 1, p < 0.05). The estimated indoor EIR in unscreened houses was 2.91 infectious bites/person/six months, higher than that in screened houses (1.88 infectious bites/person/six months). Closing eaves and screening doors and windows has the potential to reduce indoor densities of malaria vectors and malaria transmission.

Background: Insecticide resistance is a key barrier to long-term malaria control, and it may be exacerbated by poor agricultural pesticide use. Current practices, however, do not link public health and agricultural pesticide use. This study investigated the perspectives of farmers and other stakeholders regarding the integration of agricultural and public health measures to address resistance. Additionally, the feasibility of participatory workshops to increase the farmers' understanding and participation in pesticide stewardship was assessed. Methods: Four themes were investigated: pesticide awareness, practices, and opinions of; insecticide resistance in malaria vectors; the effectiveness of current malaria prevention tools; and the links between agricultural and public health pesticide usage. Participatory workshops and field training were held with entomologists, farmers, and agricultural specialists, focusing on agro-ecosystem practices related to pest control; and local farmers were involved in live-testing for insecticides resistance of local Anopheles mosquitoes. Results: Most farmers (94%) considered pesticides effective, and nearly half of them (n = 198, 46.4%) could identify and name crop pests and diseases, mostly using local names. Three quarters were unaware of mosquito larvae in their fields, and only 7% considered their fields as potential sources of mosquitoes. Two thirds were uninformed of any effects that agricultural pesticides may have on mosquitoes, and three quarters had never heard of resistance in malaria mosquitoes. Experts from various sectors acknowledged that agricultural pesticides might impact malaria control through increasing resistance. They did, however, emphasize the importance of crop protection and advocated for the use of pesticides sparingly and non-chemical approaches. Farmers learnt how to discriminate between malaria vectors and non-vectors, identify agricultural pests and diseases, choose and use pesticides effectively, and conduct resistance tests during the participatory workshops. Conclusion: This study emphasizes the significance of enhancing subsistence farmers' awareness of mosquito ecology as well as merging public health and agricultural pest management measures. Participatory techniques have the potential to raise stakeholder awareness and engagement, resulting in more effective resistance management. [ABSTRACT FROM AUTHOR]

In recent days, in tropical and subtropical regions, secondary vectors of Anopheles mosquitoes are becoming more important in transmitting diseases to humans as primary vectors. Various molecular techniques have separated closely related Anopheles subpictus and Anopheles vagus mosquitoes based on their diversity with other mosquito species. Despite their widespread distribution, the An. subpictus and An. vagus mosquitoes, which carry Plasmodium in their salivary glands, were not considered primary malaria vectors in India. An. vagus mosquitoes are zoophilic and physically similar to An. subpictus. We intend to identify An. subpictus and An. vagus mosquito's sister species based on their Interspaced Transcribed Region-2 (ITS2). We isolated the midgut gDNA from each mosquito and used ITS2-PCR and Sanger sequencing to characterize the mosquito species. BioEdit software aligned the sequences, and MEGA7 built a phylogenetic tree from them. According to this study, the information gathered from these mosquito samples fits the An. subpictus species A form and the An. vagus Indian form. Furthermore, gut microbiome plays an important role in providing nutrients, immunity, and food processing, whereas mosquitoes' midgut microbiota changes their hosts and spreads illnesses. So, we used the Illumina sequencer to look at the gut microbiome diversity of An. subpictus and An. vagus mosquitoes using 16S rRNA-based metagenomic sequencing. Both mosquito species had an abundant phylum of Pseudomonadota (Proteobacteria), Bacillota, Bacteroidota, and Actinomycetota in their gut microbiomes. Notably, both mosquito species had the genus Serratia in their gut. In the subpictus midgut, the genus of Haematosprillum bacteria was dominant, whereas in the vagus mosquito, the genus of Salmonella was dominant. Notably, current research has observed the Sodalis spp. Bacterial genus for the first time., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Recent experiments have produced several Anopheles gambiae homing gene drives that disrupt female fertility genes, thereby eventually inducing population collapse. Such drives may be highly effective tools to combat malaria. One such homing drive, based on the zpg promoter driving CRISPR/Cas9, was able to eliminate a cage population of mosquitoes. A second version, purportedly improved upon the first by incorporating an X-shredder element (which biases inheritance towards male offspring), was similarly successful. Here, we analyze experimental data from each of these gene drives to extract their characteristics and performance parameters and compare these to previous interpretations of their experimental performance. We assess each suppression drive within an individual-based simulation framework that models mosquito population dynamics in continuous space. We find that the combined homing/X-shredder drive is actually less effective at population suppression within the context of our mosquito population model. In particular, the combined drive often fails to completely suppress the population, instead resulting in an unstable equilibrium between drive and wild-type alleles. By contrast, otherwise similar drives based on the nos promoter may prove to be more promising candidates for future development than originally thought.

Malaria is a widespread vector-borne disease in the tropics and subtropics causing nearly half a million deaths every year. Malaria vector control intervention mainly rely on the control of adults using Indoor residual sprayings (IRS) and long lasting insecticidal nets (LLINs). The purpose of this study was to assess the species composition of Anopheles mosquitoes and determine the environmental and physicochemical parameters of their breeding habitats in Bambasi district, Benshangul Gumuz regional state, northwestern Ethiopia. Three major Anopheles breeding habitats were identified in three Kebeles namely; drainage ditch (Keshmando), swamp (Amba 46), and stagnant water (Amba 47). Anopheles mosquito larvae were sampled twice a month from September 2020 to November 2020. A total of 2185 Anopheles mosquito larvae were collected. Of those collected larvae three Anopheles species (Anopheles gambiae s.l. An. funestus and An. coustani complex) were identified. Anopheles gambiae s.l was the most abundant whereas An. funestus and An. coustani were the least in all the study kebeles. Of the three kebeles, Amba 47 was found the most productive for Anopheles followed by Amba 46 and Keshmando. The highest mean density of larvae per dip was sampled in September in all the study sites. The three sampling sites varied in physicochemical characteristics. The findings of this study showed that dissolved oxygen (DO) was highest (7.07 ± 0.55 mg/L) in the swamps and lowest (0.32 ± 0.04 mg/L) in the drainage ditches. Conductivity across different habitats showed wide variations. There were slight variations in temperature between different habitats. Higher total dissolved solids (TDS) 12.19 ± 0.26 mg/L was recorded from the drainage ditches; whereas TDS 9.49 ± 1.62 mg/L was recorded from the swamp. Salinity in the drainage ditches and stagnant water was 5.54 ± 1.00PSU and 3.30 ± 0.97 PSU respectively. There were negative strong correlation between the larval density with temperature and EC but positive correlation between larval density with salinity. However, there was no significant correlation between Anopheles larval density with TDS and DO. In conclusion this study suggested that environmental and physicochemical factors could play an important role in the development of mosquito larvae. Therefore, characterizing mosquito larval habitats is important for targeted control of malaria vectors in Ethiopia. [ABSTRACT FROM AUTHOR]

Wolbachia infection in Anopheles albimanus mosquitoes can render mosquitoes less capable of spreading malaria. We developed and analyzed a mechanistic compartmental ordinary differential equation model to evaluate the effectiveness of Wolbachia-based vector control strategies among wild Anopheles mosquitoes in Haiti. The model tracks the mosquito life stages, including egg, larva, and adult (male and female). It also accounts for critical biological effects, such as the maternal transmission of Wolbachia through infected females and cytoplasmic incompatibility, which effectively sterilizes uninfected females when they mate with infected males. We derive and interpret dimensionless numbers, including the basic reproductive number and next-generation numbers. The proposed system presents a backward bifurcation, which indicates a threshold infection that needs to be exceeded to establish a stable Wolbachia infection. The sensitivity analysis ranks the relative importance of the epidemiological parameters at baseline. We simulate different intervention scenarios, including prerelease mitigation using larviciding and thermal fogging before the release, multiple releases of infected populations, and different release times of the year. Our simulations show that the most efficient approach to establishing Wolbachia is to release all the infected mosquitoes immediately after the prerelease mitigation process. Moreover, the model predicts that it is more efficient to release during the dry season than the wet season.

Understanding the temporal dynamics of mosquito populations underlying vector-borne disease transmission is key to optimizing control strategies. Many questions remain surrounding the drivers of these dynamics and how they vary between species—questions rarely answerable from individual entomological studies (that typically focus on a single location or species). We develop a novel statistical framework enabling identification and classification of time series with similar temporal properties, and use this framework to systematically explore variation in population dynamics and seasonality in anopheline mosquito time series catch data spanning seven species, 40 years and 117 locations across mainland India. Our analyses reveal pronounced variation in dynamics across locations and between species in the extent of seasonality and timing of seasonal peaks. However, we show that these diverse dynamics can be clustered into four 'dynamical archetypes', each characterized by distinct temporal properties and associated with a largely unique set of environmental factors. Our results highlight that a range of environmental factors including rainfall, temperature, proximity to static water bodies and patterns of land use (particularly urbanicity) shape the dynamics and seasonality of mosquito populations, and provide a generically applicable framework to better identify and understand patterns of seasonal variation in vectors relevant to public health. [ABSTRACT FROM AUTHOR]

Abstract Background Despite extensive irrigation development in Ethiopia, limited studies assessed the impact of irrigation on malaria vector mosquito composition, abundance and seasonality. This study aimed to evaluate the impact of sugarcane irrigation on species composition, abundance and seasonality of malaria vectors. Methods Adult Anopheles mosquitoes were collected using CDC light traps from three irrigated and three non-irrigated clusters in and around Arjo-Didessa sugarcane irrigation scheme in southwestern Ethiopia. Mosquitoes were surveyed in four seasons: two wet and two dry, in 2018 and 2019. Mosquito species composition, abundance and seasonality were compared between irrigated and non-irrigated clusters. Anopheles mosquitoes were sorted out to species using morphological keys and molecular techniques. Chi square was used to test the relationships between Anopheles species occurrence, and environmental and seasonal parameters. Results Overall, 2108 female Anopheles mosquitoes comprising of six species were collected. Of these, 92.7% (n = 1954) were from irrigated clusters and 7.3% (n = 154) from the non-irrigated. The Anopheles gambiae complex was the most abundant (67.3%) followed by Anopheles coustani complex (25.3%) and Anopheles pharoensis (5.7%). PCR-based identification revealed that 74.7% (n = 168) of the An. gambiae complex were Anopheles arabiensis and 22.7% (n = 51) Anopheles amharicus. The density of An. gambiae complex (both indoor and outdoor) was higher in irrigated than non-irrigated clusters. The overall anopheline mosquito abundance during the wet seasons (87.2%; n = 1837) was higher than the dry seasons (12.8%; n = 271). Conclusion The ongoing sugarcane irrigation activities in Arjo-Didessa created conditions suitable for malaria transmitting Anopheles species diversity and abundance. This could drive malaria transmission in Arjo-Didessa and its environs in both dry and wet seasons. Currently practiced malaria vector interventions need to be strengthened by including larval source management to reduce vector abundance in the irrigated areas.

Abstract Background The success of indoor interventions that target mosquitoes for malaria control is partially dependent on early evening and outdoor biting behaviours of mosquito vectors. In southwest Ethiopia, people and cattle live in proximity, which calls to investigate whether the presence of cattle increase or decrease bites from malaria mosquito vectors. This study assessed both host-seeking and overnight activity of malaria mosquito vectors given the presence or absence of cattle in Chano Mille village, Arba Minch district, Ethiopia. Methods Anopheles species density and activity time was compared when a calf was: (i) placed inside; (ii) 1 m away from; or (iii) absent from a tent with a human volunteer resting insides using hourly human landing catches (HLC) conducted from 18:00–0:00 h for 3 months. This trial was performed close to the shore of the Lake Abaya to minimize the interference of other animals on mosquito movement. The overnight activity of malaria vectors was assessed within a Chano village from 18:00–6:00 h with collections carried out both indoors and outdoors by HLC. Generalized estimating equations were used to statistically assess differences. Results Anopheles pharoensis was significantly more prevalent when a calf was present either inside (42%, P

Beginning in 2023, we observed increased Plasmodium vivax malaria cases at an institution in Los Angeles, California, USA. Most cases were among migrants from China who traveled to the United States through South and Central America. US clinicians should be aware of possible P. vivax malaria among immigrants from China.

Entomological data gathering is essential for monitoring malaria vector disease risks and selection of appropriate interventions for the protection of exposed human populations. This study assessed the relative abundance, species composition, and sporozoite infection rates of indoor resting An. gambiae s.l. malaria vectors in six communities across 3 Local Government Areas in Kwara State, Nigeria. Total number of mosquitoes collected by Pyrethrum Spray Catch method over a period of eighth months were correlated with rainfall values in the area. Plasmodium falciparum sporozoite infection rates and sibling species identification of collected An. gambiae s.l. mosquito samples were determined by ELISA and PCR respectively. Results showed a positive correlation (r = 0.639, p = 0.08) between rainfall and numbers of Anopheles mosquitoes in the study areas. The overall composition of the An. gambiae s.l sibling species in the collected samples from all the six communities showed the predominance of An. gambiae s.s 298 (75.3%) compared to An. coluzzii 94(23.7%) and An. arabiensis 4(1.0%). However, the sporozoite infection rate of An. coluzzii (22.3%) was higher compared to An. gambiae s.s (12.8%) and An. arabiensis (0%). Mean numbers of An. gambiae s.l mosquitoes were significantly higher in Ilorin west LGA compared to Asa (F = 17.81, P < 0.001) and Ilorin East LGAs (F = 22.81, P < 0.001). Sporozoite rates of both An. gambiae s.s and An. coluzzii sibling species were higher in Ilorin West communities (Aiyede 21%, Ogundele 32%) compared to Asa (Idi Emi 11.1%, Lasoju 5.1%) and Ilorin East (Oke Oyi 2.4%, Ote-efan 0%) communities. Prevalence of sporozoite-infected An. gambiae s.s and An. coluzzii indoors highlight the need for effective insecticide treated bed-nets interventions to protect the residents from malaria risks. Higher numbers of An. coluzzii in the swampy rice marshed Ilorin West LGA communities require larval source management as an additional strategy for effective malaria vector con [ABSTRACT FROM AUTHOR]

BACKGROUND፡ Malaria is one of the main parasitic diseases and a major health issue in some countries. This study aims to determine the rate and type of infections of Anopheles mosquitoes with malaria parasites using the molecular LAMP method in the Southeastern Iran. METHODS: In this study, 400 Anopheles mosquitoes were collected by the Zahedan Medical Insecticide Center in Nikshahr City, a high-risk area of malaria transmission in Sistan- Baluchestan Province. The mosquitoes were caught manually (by hand) in domestic (humans and animals), natural, and artificial outdoor places (Shelter pits). After DNA extraction, the LAMP method was used, which was compared with Multiplex Nested- PCR as a standard method. RESULTS: Out of 400 samples collected from Nikshahr City, 6 samples (1.5%) were infected with Plasmodium vivax. No Plasmodium falciparum or a mix (Plasmodium vivax and Plasmodium falciparum) was detected in this study. CONCLUSIONS: The results of this study indicate that in places with transmission of both species, i.e. Plasmodium vivax and Plasmodium falciparum, detection of malaria parasites by the LAMP method could be very useful in spotting infections in the field. Thus, molecular epidemiological studies could be conducted annually to monitor malaria in endemic regions. The results of this research show that contamination with mosquito malaria vectors is increasing in Nikshahr City, and it seems that more studies will be required to eliminate malaria until 2026. [ABSTRACT FROM AUTHOR]

Abstract Background Elimination of malaria depends on mastering transmission and understanding the biological basis of Plasmodium infection in the vector. The first mosquito organ to interact with the parasite is the midgut and its transcriptomic characterization during infection can reveal effective antiplasmodial responses able to limit the survival of the parasite. The vector response to Plasmodium vivax is not fully characterized, and its specificities when compared with other malaria parasites can be of fundamental interest for specific control measures. Methods Experimental infections were performed using a membrane-feeding device. Three groups were used: P. vivax-blood-fed, blood-fed on inactivated gametocytes, and unfed mosquitoes. Twenty-four hours after feeding, the mosquitoes were dissected and the midgut collected for transcriptomic analysis using RNAseq. Nine cDNA libraries were generated and sequenced on an Illumina HiSeq2500. Readings were checked for quality control and analysed using the Trinity platform for de novo transcriptome assembly. Transcript quantification was performed and the transcriptome was functionally annotated. Differential expression gene analysis was carried out. The role of the identified mechanisms was further explored using functional approaches. Results Forty-nine genes were identified as being differentially expressed with P. vivax infection: 34 were upregulated and 15 were downregulated. Half of the P. vivax-related differentially expressed genes could be related to autophagy; therefore, the effect of the known inhibitor (wortmannin) and activator (spermidine) was tested on the infection outcome. Autophagic activation significantly reduced the intensity and prevalence of infection. This was associated with transcription alterations of the autophagy regulating genes Beclin, DRAM and Apg8. Conclusions Our data indicate that P. vivax invasion of An. aquasalis midgut epithelium triggers an autophagic response and its activation reduces infection. This suggests a novel mechanism that mosquitoes can use to fight Plasmodium infection.

Abstract Malaria is a serious global health burden, affecting more than 200 million people each year in over 90 countries, predominantly in Africa, Asia and the Americas. Since the year 2000, a concerted effort to combat malaria has reduced its incidence by more than 40%, primarily due to the use of insecticide-treated bednets, indoor residual spraying and artemisinin-based combination drug therapies. Nevertheless, the cost of control is expected to nearly triple over the next decade and the current downward trend in disease transmission is threatened by the rise of resistance to drugs and insecticides. Novel strategies that are sustainable and cost-effective are needed to help usher in an era of malaria elimination. The most effective strategies thus far have focussed on control of the mosquito vector. The sterile insect technique (SIT) is a potentially powerful strategy that aims to suppress mosquito populations through the unproductive mating of wild female mosquitoes with sterile males that are released en masse. The technique and its derivatives are currently not appropriate for malaria control because it is difficult to sterilise males without compromising their ability to mate, and because anopheline males cannot be easily separated from females, which if released, could contribute to disease transmission. Advances in genome sequencing technologies and the development of transgenic techniques provide the tools necessary to produce mosquito sexing strains, which promise to improve current malaria-control programs and pave the way for new ones. In this review, the progress made in the development of transgenic sexing strains for the control of Anopheles gambiae, a major vector of human malaria, is discussed.

Mentha piperita (peppermint) essential oil nanoemulsion was prepared by low energy spontaneous emulsification method. GC-MS analysis revealed the composition of peppermint essential oil and menthol (45.2%) was the major bioactive compound along with menthone (15.39%), neomethol (8.1%), menthyl acetate (7.7%) and isomenthone (7.4%). Optimization of the nanoemulsion preparation process was done by Response Surface Methodology (RSM) with Central Composite Design (CCD). The droplet diameter and polydispersity index at optimized conditions (15% oil concentration, 25% surfactant concentration and Tween80 as surfactant) were 39.2 nm and 0.22 respectively. Optimized peppermint oil nanoemulsion (OPNE) was optically transparent, spherical in morphology and was stable for 6 months. OPNE formulation demonstrated dose, time and size-dependent larvicidal activity against malaria vectors with LC50 value of 48 ppm and 123 ppm against Anopheles culicifacies and Anopheles stephensi respectively. The LC50 values were 90 ppm and 163 ppm against Anopheles culicifacies and Anopheles stephensi correspondingly for the bigger droplet size formulation (PNE, droplet diameter: 129.6 nm) confirming the droplet size-dependent larvicidal activity of the nanoemulsion. The results of this study propose that peppermint oil-based nanoemulsion possibly be used as an eco-friendly larvicide for mosquito vector control strategies. [ABSTRACT FROM AUTHOR]

Malaria vectors are found virtually in all communities of Nigeria. Understanding the transmitting potential of these vectors in relationship with the malaria parasite (MP) infection is essential to design a sustainable community base control programme. This study aimed at determining this relationship in six communities in Ekiti State, Nigeria. The communities were randomly selected through a multistage sampling. Blood samples were screened for MP infection among human volunteers during dry and rainy seasons in each community. Adult mosquitoes were collected indoors for twelve months in the communities using light traps. Female Anopheles mosquitoes collected were characterized by Polymerase Chain Reactions and circumsporozoite proteins of the infected mosquitoes were detected by Enzyme-Linked Immunosorbent Assay. Prevalence of MP infection, human biting rates (HBR), sporozoite rates and entomological inoculation rates (EIRs) were determined using appropriate formulae. Overall prevalence of MP infection was 26% in dry season and 38% in rainy season (P=0.001). Female Anopheles mosquitoes collected were 428. Strong relationship existed between HBR and MP infection in rainy season (R2= 0.671) but the relationship was weak (R2= 0.209) in dry season. Sporozoites were detected in 2.3% of the female Anopheles mosquitoes collected. EIRs were between 0.100 - 0.252 infective bites/person/night. Transmission of malaria parasites occurred throughout the year particularly during rainy season when there were recorded EIRs. Malaria control interventions will be more appropriate during raining season in this region. [ABSTRACT FROM AUTHOR]

Among the complex microbial community living in the mosquito midgut, some bacteria (e.g., Enterobacter spp.) can deliver effector molecules with anti-Plasmodium effects suppressing the development of malaria parasites (Plasmodium falciparum) before the öokinete can penetrate the mosquito midgut epithelium. Despite knowledge of this phenomenon, only a few studies have defined the diversity of microbiota in wild-caught adult Anopheles species. The objective of this study was to analyze and compare the bacterial microbiota in different Anopheles species, including representatives of the primary malaria vectors in western Thailand. Wild female Anopheles species were sampled from malaria-endemic areas in Tak and Mae Hong Son provinces near the Thai-Myanmar border. Midgut/abdominal bacterial diversity was assessed by examining the 16S rRNA gene, V3 hypervariable region, using PCR-Temporal Temperature Gel Electrophoresis (PCR-TTGE) profiling and sequence analysis. A total of 24 bacterial genera were identified from eight Anopheles species. Five bacterial genera were newly reported in Anopheles mosquitoes (Ferrimonas, Megasphaera, Pectobacterium, Shimwellia, and Trabulsiella). Five genera, including Megasphaera, were detected exclusively in a single-malaria (Plasmodium vivax) infected Anopheles minimus and not observed in other non-infected mosquitoes. The use of PCR-TTGE provides the first characterization of the midgut bacterial microbiome present in wild adult Anopheles in Thailand. Evidence that microbiota might impact pathogen development (suppression) in Anopheles and thereby reduce the risk of pathogen transmission deserves more studies to describe the presence and better understand the biological role of bacteria in natural mosquito populations.

Abstract Background Plasmodium falciparum malaria remains a leading cause of death in tropical regions of the world. Despite efforts to reduce transmission, rebounds associated with the persistence of malaria vectors have remained a major impediment to local elimination. One area that remains poorly understood is how Anopheles populations survive long dry seasons to re-emerge following the onset of the rains. Methods We developed a suite of mathematical models to explore the impact of different dry-season mosquito survival strategies on the dynamics of vector populations. We fitted these models to an Anopheles population data set from Mali to estimate the model parameters and evaluate whether incorporating aestivation improved the fit of the model to the observed seasonal dynamics. We used the fitted models to explore the impact of intervention strategies that target aestivating mosquitoes in addition to targeting active mosquitoes and larvae. Results Including aestivation in the model significantly improved our ability to reproduce the observed seasonal dynamics of vector populations as judged by the deviance information criterion (DIC). Furthermore, such a model resulted in more biologically plausible active mosquito survival times (for A. coluzzii median wet season survival time of 10.9 days, 95% credible interval (CrI): 10.0–14.5 days in a model with aestivation versus 38.1 days, 95% CrI: 35.8–42.5 days in a model without aestivation; similar patterns were observed for A. arabiensis). Aestivation also generated enhanced persistence of the vector population over a wider range of both survival times and fecundity levels. Adding vector control interventions that target the aestivating mosquito population is shown to have the potential to enhance the impact of existing vector control. Conclusions Dry season survival attributes appear to drive vector population persistence and therefore have implications for vector control. Further research is therefore needed to better understand these mechanisms and to evaluate the additional benefit of vector control strategies that specifically target dormant mosquitoes.

Malaria parasites are transmitted in nature through the bites of female Anopheles mosquitoes. However, the vector distribution varies in time and space. This study aimed to determine the species composition, abundance and climatic factors influencing Anopheles mosquitoes in Anambra East Local Government Area of Anambra State, Nigeria from October 2016 to September 2017. Adult Anopheles mosquitoes were collected from indoor and outdoor locations using Pyrethrum Knockdown Collection (PKC) and Human Landing Catch (HLC) Methods respectively. Anopheles mosquito larvae were collected using dipping method. Morphological identification of Anopheles species was carried out using standard identification keys. Climatic data was obtained from Nigerian Meteorological Agency Office in Anambra State. A total of 8181 female Anopheles mosquitoes which comprised 4127 (50.4%) larvae and 4054 (49.6%) adults were collected (P > 0.05) in the study. Four Anopheles species: An. gambiae s. l (70.1%), An. funestus group (18.2%), An. moucheti (6.3%) and An. nili (5.4%) were identified (P < 0.05). In the study area, 2608 (31.9%), 3025 (37.0%) and 2548 (31.1%) Anopheles mosquitoes were collected from Aguleri, Igbariam and Nsugbe respectively; with 100% species overlap. In each selected selected town, the Simpson’s index of diversity was ~2 and Shannon-wiener diversity was ~1. Only An. gambiae s. l showed varied seasonal abundance with wet season contributing 67.9% and dry season 32.1% of the overall An. gambiae s. l collection (P < 0.05). The correlation between rainfall and An. gambiae s. l. abundance was significantly strong (r = 0.66; P < 0.05). No significant correlation was found between Anopheles species abundance and temperature as well as relative humidity. The study revealed the preponderance of four Anopheles species: An. gambiae s. l., An. funestus, An. moucheti and An. nili; and rainfall is the only climatic factor that causes temporal change in the abundance of one of the species, An. gambiae s. l. in the study area. [ABSTRACT FROM AUTHOR]

Background: The success of indoor interventions that target mosquitoes for malaria control is partially dependent on early evening and outdoor biting behaviours of mosquito vectors. In southwest Ethiopia, people and cattle live in proximity, which calls to investigate whether the presence of cattle increase or decrease bites from malaria mosquito vectors. This study assessed both host-seeking and overnight activity of malaria mosquito vectors given the presence or absence of cattle in Chano Mille village, Arba Minch district, Ethiopia. Methods: Anopheles species density and activity time was compared when a calf was: (i) placed inside; (ii) 1 m away from; or (iii) absent from a tent with a human volunteer resting insides using hourly human landing catches (HLC) conducted from 18:00–0:00 h for 3 months. This trial was performed close to the shore of the Lake Abaya to minimize the interference of other animals on mosquito movement. The overnight activity of malaria vectors was assessed within a Chano village from 18:00–6:00 h with collections carried out both indoors and outdoors by HLC. Generalized estimating equations were used to statistically assess differences. Results: Anopheles pharoensis was significantly more prevalent when a calf was present either inside (42%, P < 0.001), or adjacent to (46%, P = 0.002) a tent relative to a tent without a calf present. The presence of a calf did not affect densities of the primarily anthropophilic species A. gambiae (s.l.), or An. tenebrosus. Anopheles gambiae (s.l.) (P < 0.001) and An. pharoensis (P = 0.015) both had a tendency for early evening biting between 19:00 h and 20:00 h. Anopheles gambiae (s.l.) was mainly biting humans outdoors in the village. Conclusions: The presence of calves within and close to human dwellings acts to draw malaria mosquitoes toward the human occupant with the potential to increase their risk of malaria. Hence, deployment of cattle far from human residence could be recommended to reduce human exposure. Outdoor and early evening biting could threaten the success of current indoor-based interventions. Hence, tools could be designed to reduce this threat. [ABSTRACT FROM AUTHOR]

Among the complex microbial community living in the mosquito midgut, some bacteria (e.g., Enterobacter spp.) can deliver effector molecules with anti- Plasmodium effects suppressing the development of malaria parasites (Plasmodium falciparum) before the öokinete can penetrate the mosquito midgut epithelium. Despite knowledge of this phenomenon, only a few studies have defined the diversity of microbiota in wild-caught adult Anopheles species. The objective of this study was to analyze and compare the bacterial microbiota in different Anopheles species, including representatives of the primary malaria vectors in western Thailand. Wild female Anopheles species were sampled from malaria-endemic areas in Tak and Mae Hong Son provinces near the Thai-Myanmar border. Midgut/abdominal bacterial diversity was assessed by examining the 16S rRNA gene, V3 hypervariable region, using PCR-Temporal Temperature Gel Electrophoresis (PCR-TTGE) profiling and sequence analysis. A total of 24 bacterial genera were identified from eight Anopheles species. Five bacterial genera were newly reported in Anopheles mosquitoes (Ferrimonas , Megasphaera , Pectobacterium , Shimwellia , and Trabulsiella). Five genera, including Megasphaera , were detected exclusively in a single-malaria (Plasmodium vivax) infected Anopheles minimus and not observed in other non-infected mosquitoes. The use of PCR-TTGE provides the first characterization of the midgut bacterial microbiome present in wild adult Anopheles in Thailand. Evidence that microbiota might impact pathogen development (suppression) in Anopheles and thereby reduce the risk of pathogen transmission deserves more studies to describe the presence and better understand the biological role of bacteria in natural mosquito populations. [ABSTRACT FROM AUTHOR]

Despite the implementation of preventive measures in airports and aircrafts, the risk of importing Plasmodium spp. infected mosquitoes is still present in malaria-free countries. Evidence suggests that mosquitoes have found a new alliance with the globalization of trade and climate change, leading to an upsurge of malaria parasite transmission around airports. The resulting locally acquired form of malaria is called Airport malaria. However, piecemeal information is available, regarding its epidemiological and entomological patterns, as well as the challenges in the diagnosis, treatment, and prevention. Understanding these issues is a critical step towards a better implementation of control strategies. To cross reference this information, we conducted a systematic review on 135 research articles published between 1969 (when the first cases of malaria in airports were reported) and 2020 (i.e., 51 years later). It appears that the risk of malaria transmission by local mosquito vectors in so called malaria-free countries is not zero; this risk is more likely to be fostered by infected vectors coming from endemic countries by air or by sea. Furthermore, there is ample evidence that airport malaria is increasing in these countries. From 2010 to 2020, the number of cases in Europe was 7.4 times higher than that recorded during the 2000–2009 decade. This increase may be associated with climate change, increased international trade, the decline of aircraft disinsection, as well as delays in case diagnosis and treatment. More critically, current interventions are weakened by biological and operational challenges, such as drug resistance in malaria parasites and vector resistance to insecticides, and logistic constraints. Therefore, there is a need to strengthen malaria prevention and treatment for people at risk of airport malaria, and implement a rigorous routine entomological and epidemiological surveillance in and around airports.

Abstract Background Due to outdoor and residual transmission and insecticide resistance, long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) will be insufficient as stand-alone malaria vector control interventions in many settings as programmes shift toward malaria elimination. Combining additional vector control interventions as part of an integrated strategy would potentially overcome these challenges. Larval source management (LSM) and structural house improvements (HI) are appealing as additional components of an integrated vector management plan because of their long histories of use, evidence on effectiveness in appropriate settings, and unique modes of action compared to LLINs and IRS. Implementation of LSM and HI through a community-based approach could provide a path for rolling-out these interventions sustainably and on a large scale. Methods/design We will implement community-based LSM and HI, as additional interventions to the current national malaria control strategies, using a randomised block, 2 × 2 factorial, cluster-randomised design in rural, southern Malawi. These interventions will be continued for two years. The trial catchment area covers about 25,000 people living in 65 villages. Community participation is encouraged by training community volunteers as health animators, and supporting the organisation of village-level committees in collaboration with The Hunger Project, a non-governmental organisation. Household-level cross-sectional surveys, including parasitological and entomological sampling, will be conducted on a rolling, 2-monthly schedule to measure outcomes over two years (2016 to 2018). Coverage of LSM and HI will also be assessed throughout the trial area. Discussion Combining LSM and/or HI together with the interventions currently implemented by the Malawi National Malaria Control Programme is anticipated to reduce malaria transmission below the level reached by current interventions alone. Implementation of LSM and HI through a community-based approach provides an opportunity for optimum adaptation to the local ecological and social setting, and enhances the potential for sustainability. Trial Registration Registered with The Pan African Clinical Trials Registry on 3 March 2016, trial number PACTR201604001501493.

This study evaluated the impact of combining house screens with long-lasting insecticidal nets (LLINs) on mosquito host-seeking, resting, and biting behavior. Intervention houses received house screens and LLINs, while control houses received only LLINs. Centre for Disease Control light traps, pyrethrum spray collections and human landing catches were used to assess the densities of indoor and outdoor host-seeking, indoor resting, and biting behavior of malaria vectors in 15 sentinel houses per study arm per sampling method. The protective efficacy of screens and LLINs was estimated through entomological inoculation rates (EIRs). There were 68% fewer indoor host-seeking Anopheles funestus (RR = 0.32, 95% CI 0.20-0.51, p < 0.05) and 63% fewer An. arabiensis (RR = 0.37, 95% CI 0.22-0.61, p < 0.05) in screened houses than unscreened houses. There was a significantly higher indoor biting rate for unscreened houses (6.75 bites/person/h [b/p/h]) than for screened houses (0 b/p/h) (χ 2 = 6.67, df = 1, p < 0.05). The estimated indoor EIR in unscreened houses was 2.91 infectious bites/person/six months, higher than that in screened houses (1.88 infectious bites/person/six months). Closing eaves and screening doors and windows has the potential to reduce indoor densities of malaria vectors and malaria transmission.

Malaria molecular surveillance remains critical in detecting and tracking emerging parasite resistance to anti-malarial drugs. The current study employed molecular techniques to determine Plasmodium species prevalence and characterize the genetic diversity of Plasmodium falciparum and Plasmodium malariae molecular markers of sulfadoxine-pyrimethamine resistance in humans and wild Anopheles mosquito populations in Cameroon. Anopheles mosquito collections and parasitological survey were conducted in villages to determine Plasmodium species infection, and genomic phenotyping of anti-folate resistance was accomplished by sequencing the dihydrofolate-reductase ( dhfr ) and dihydropteroate-synthase ( dhps ) genes of naturally circulating P. falciparum and P. malariae isolates. The malaria prevalence in Elende was 73.5% with the 5-15 years age group harboring significant P. falciparum (27%) and P. falciparum + P. malariae (19%) infections. The polymorphism breadth of the pyrimethamine-associated Pfdhfr marker revealed a near fixation (94%) of the triple-mutant -A 16 I 51 R 59 N 108 I 164 . The Pfdhps backbone mediating sulfadoxine resistance reveals a high frequency of the V 431 A 436 G 437 K 540 A 581 A 613 alleles (20.8%). Similarly, the Pmdhfr N 50 K 55 L 57 R 58 S 59 S 114 F 168 I 170 haplotype (78.4%) was predominantly detected in the asexual blood stage. In contrast, the Pmdhps - S 436 A 437 occured at 37.2% frequency. The combined quadruple N 50 K 55 L 57 R 58 S 59 S 114 F 168 I 170 &#95; S 436 G 437 K 540 A 581 A 613 (31.9%) was the major circulating haplotype with similar frequency in humans and mosquitoes. This study highlights the increasing frequency of the P. malariae parasite mostly common in asymptomatic individuals with apparent P. falciparum infection. Interventions directed at reducing malaria transmission such as the scaling-up of SP are favoring the emergence and spread of multiple drug-resistant alleles between the human and mosquito host systems., Competing Interests: The authors declare no conflict of interest.

Malaria and lymphatic filariasis (LF) are two serious public health challenges in sub-Saharan Africa, and both diseases are transmitted by Anopheles mosquitoes. Successful control of both diseases requires detailed information on transmission dynamics; thus, this study investigated malaria and LF transmission indices in two (2) communities (Jidawa and Kargo) in North-West Nigeria. Anopheles mosquitoes were sampled from twenty-five (25) randomly selected houses from each of the two communities using pyrethrum spray collection (PSC). The samples were identified morphologically and molecularly characterised using polymerase chain reaction (PCR). Human biting rate (HBR), indoor resting density (IRD), sporozoite rate (SR) and entomological inoculation rate (EIR) were calculated using standard formulae. The thorax region of the collected samples were dissected and smeared; then, Plasmodium and Wuchereria bancrofti parasites were identified using microscopy. A total of 2417 Anopheles mosquitoes were collected, and all were identified morphologically as An. gambiae s.l. Further molecular identification of sibling species revealed that An. gambiae and An. arabiensis were the only sibling species present. A total of 818 Anopheles mosquitoes were screened for Plasmodium and Wuchereria bancrofti parasites. A total of 180 samples were positive for Plasmodium parasites (Jidawa = 151; Kargo = 29), and none was positive for W. bancrofti (0%). Result of entomological indices for malaria transmission showed that indoor resting density was higher in Jidawa (10 mosquitoes/room/night) while human biting rate (2.07 bites/person/night), sporozoite rate (29.3%) and entomological inoculation rate (0.61) were higher in Kargo. In total, 35.2% of the samples were blood-fed while 67.4% were parous. There is active transmission of malaria in the two communities but not LF, suggesting the effectiveness of mass drug administration for LF. Concerted efforts should be focused on malaria control as transmission of the disease persists., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Malaria is endemic and stable, being a major cause of morbidity and mortality in Nigeria. Malaria prevalence and mosquito dynamics were studied in the rural villages of Alulu-Nike Community, Enugu East Local Government Area of Enugu State for a period of 12 months. Malaria was diagnosed with microscopy method. Mosquito collection was done using pyrethroid knockdown collection method for indoor biting and resting adult mosquitoes; human bait method for outdoor adult mosquito and larval collection from different breeding sites. Mosquito identification was done using morphological characteristics and PCR. A total of 1,440 people were screened for malaria and 476(32.4%) were infected. The overall mean malaria intensity was 545 parasites per microlitre of blood (p/μl). Nnegbune Village had the highest malaria prevalence 86 (41.5%) which was significant (p<0.05) and the highest mean malaria intensity 741 p/μl. Age 0-10 years had the highest malaria prevalence 229(44.7) and mean intensity 991 p/μl. The age specific malaria prevalence was statistically significant (p<0.05). Malaria intensity was not significant in all categories. Malaria prevalence and intensity were not dependent on sex and occupation of the participants (p>0.05). The primary education group 204(38.6) and the nonformal education group 63(34.2) were mostly infected while the tertiary education group were the least infected 68(26.6) which was significant (p<0.05). PCR characterization of the 990 indoor biting and resting adult mosquitoes revealed Anopheles gambiae ss and A. arabiensis as the sibling species of A. gambiae sl. A. gambiae ss, A. arabiensis and A. funestus (malaria vectors) constituted 764 (77.1%) while the Culicines accounted for 226(22.8%) of the total indoor collection. The highest malaria vectors as well as the highest mosquitoes were collected from Nnegbune village. Of the 694 outdoor biting adult mosquitoes collected. A. gambiae sl 7(1.0%) was the only malaria vector collected while other species of culicine mosquitoes accounted for 687(98.99%) of the outdoor collections. A total of 1,162 mosquito larvae were collected from different villages of Alulu-Nike Community out of which A. gambiae yielded 45(3.9%) while culicine mosquitoes yielded 1,117(96.1%) of the total larval sampling. The highest collection of larvae was from used and discarded tyres 822(70.7%). Mosquitoes were more abundant in wet months than in dry months. The distribution of indoor and outdoor mosquitoes and larvae in different villages and in different months showed significant results (p<0.05). Entomological study of mosquitoes is important for planning and optimizing malaria control in both time and space. [ABSTRACT FROM AUTHOR]

The behavioural effect of tobacco smoke on endophilic Anopheles mosquitoes was observed in two North Central Nigerian villages. Pyrethrum spray and exit trap collections of mosquitoes were conducted, between April and November 2014, in two different villages where tobacco smoking was common among the residents. Anopheles mosquito samples collected were identified to sibling species level using species-specific polymerase chain reaction. Presence of human blood in the blood-fed mosquitoes was determined by Enzyme-linked immunosorbent assay. Numbers of mosquitoes collected from the rooms inhabited by tobacco smokers and non-smokers were compared using Student’s t-test (p

The determination of the trophic preferences of the Anopheles gambiae Giles (Diptera: Culicidae) is a decisive parameter for the monitoring and the prevention of malaria risk transmission. Currently, arthropod blood feeding sources are identified using immunological or molecular biology traditional techniques. Despite the effectiveness of these methods, they present several limitations, and notably, they are time-consuming and costly techniques. A recent study demonstrated that MALDI-TOF MS could be a useful tool for the identification of blood meal origins in freshly engorged mosquitoes. However, the limited number of blood vertebrate species tested to date, did not allow an assessment of the efficiency of MALDI-TOF MS in distinguishing blood MS spectra among close host species, such as humans versus primates. Therefore, in the present study, blood from ten distinct vertebrate host species, including four domestic species, four wild species, and two primates, was selected to control the reliability of MALDI-TOF MS based identification. Host blood species-specific MS profiles, up to 24 h post-feeding in engorged Anopheles abdomens, were confirmed. Blind tests underlined the high specificity of MS spectra for the recognition of each host species, preventing misidentification. Nevertheless, an accurate analysis of the results from MS spectra queried against the MS database revealed that the reliability of identification is directly linked to the comprehensiveness of the MS reference database. Finally, the rapidity, the low-cost reagents, the simplicity of data analysis, and the accuracy of the tool for blood origin determination, make this proteomic strategy a promising complementary method for the elucidation of host/vector interactions. [ABSTRACT FROM AUTHOR]

The behavioural effect of tobacco smoke on endophilic Anopheles mosquitoes was observed in two North Central Nigerian villages. Pyrethrum spray and exit trap collections of mosquitoes were conducted, between April and November 2014, in two different villages where tobacco smoking was common among the residents. Anopheles mosquito samples collected were identified to sibling species level using species-specific polymerase chain reaction. Presence of human blood in the blood-fed mosquitoes was determined by Enzyme-linked immunosorbent assay. Numbers of mosquitoes collected from the rooms inhabited by tobacco smokers and non-smokers were compared using Student's t-test (p<0.05). At least four times higher exit rates, five times less gravid females and significantly lower numbers of An. gambiae s.l mosquitoes were found in rooms inhabited by tobacco smokers compared to equal sized rooms occupied by non-smokers in each village. Blood feeding rates (77-88%) and human blood indices (0.82-0.87) of mosquitoes were generally high in the habitations of both tobacco smokers and non-smokers. There was predominance of An. gambiae (≥ 86%) species compared to An. coluzzii (≤ 14%) in both villages. Reduced mosquito endophily was observed in the habitations of tobacco smokers compared to the non-smokers. Widespread tobacco smoking habits should be investigated for possible contributions to mosquito behavioural resistance and residual malaria parasite transmission within rural communities. [ABSTRACT FROM AUTHOR]

The detection of insecticides resistance status in a natural population of Anopheles vectors is a vital tool for malaria control intervention strategy against Anopheles gambiae sensu lato, which is the main malaria vector in Nigeria. This study was conducted to determine the susceptibility status of the female Anopheles mosquitoes to Propoxor (0.1%) (Carbamate) and Alphacympermetrin (0.5%) (Pyrethroid) insecticides in Keffi LGA of Nasarawa State, Nigeria, between March and June, 2015. The larvae were collected from natural water bodies and other potential breeding sites from five different sites in Keffi LGA namely; Angwan Lambu, High Court, Angwan Kwara, Antau Bridge and Pyanku they were raised in the insectary located at Nasarawa State University, Keffi. WHO bioassay were performed using 2-3 days old, glucose-fed but non-blood fed female Anopheles mosquitoes. The knock down effect of Propoxur in the five locations in Keffi LGA was more pronounced with mortalities ranging from 84%-98% (p<0.05) and Alphacympermethrin recording a higher knock down in Angwan Kwara with 95% and with mortalities ranging from 78%-85% (p<0.05). The result of the bioassay showed reduced efficacy of Alphacypermethrin with resistance in some locations. However, Propoxur recorded higher mortality (100%, 100%, 95%, 100%, and 100%) respectively in the five sampled areas. Using chi-square statistical analysis; there was a significant difference between knockdown times with respect to Propoxur (p<0.05). Insecticide based intervention strategy remain the principal vector control measure in malaria endemic countries prompting the recommendation of Propoxur in the future vector control programs because high mortalities of the vectors were noticed in the assay run by the Carbamate class of insectide (Propoxur). [ABSTRACT FROM AUTHOR]

Background: Due to outdoor and residual transmission and insecticide resistance, long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) will be insufficient as stand-alone malaria vector control interventions in many settings as programmes shift toward malaria elimination. Combining additional vector control interventions as part of an integrated strategy would potentially overcome these challenges. Larval source management (LSM) and structural house improvements (HI) are appealing as additional components of an integrated vector management plan because of their long histories of use, evidence on effectiveness in appropriate settings, and unique modes of action compared to LLINs and IRS. Implementation of LSM and HI through a community-based approach could provide a path for rolling-out these interventions sustainably and on a large scale.Methods/design: We will implement community-based LSM and HI, as additional interventions to the current national malaria control strategies, using a randomised block, 2 × 2 factorial, cluster-randomised design in rural, southern Malawi. These interventions will be continued for two years. The trial catchment area covers about 25,000 people living in 65 villages. Community participation is encouraged by training community volunteers as health animators, and supporting the organisation of village-level committees in collaboration with The Hunger Project, a non-governmental organisation. Household-level cross-sectional surveys, including parasitological and entomological sampling, will be conducted on a rolling, 2-monthly schedule to measure outcomes over two years (2016 to 2018). Coverage of LSM and HI will also be assessed throughout the trial area.Discussion: Combining LSM and/or HI together with the interventions currently implemented by the Malawi National Malaria Control Programme is anticipated to reduce malaria transmission below the level reached by current interventions alone. Implementation of LSM and HI through a community-based approach provides an opportunity for optimum adaptation to the local ecological and social setting, and enhances the potential for sustainability.Trial Registration: Registered with The Pan African Clinical Trials Registry on 3 March 2016, trial number PACTR201604001501493. [ABSTRACT FROM AUTHOR]