Intense malarial transmission during the dry season in irrigated rice-growing areas: a case study in Sakassou, Côte d'Ivoire.

The health district of Sakassou is one of the 83 health districts in Côte d'Ivoire, located in a zone with very high malarial transmission rates, with an incidence rate of ≥40% Therefore, to guide vector control methods more effectively, it was crucial to have a good understanding of the vectors in the area. This study aimed to determine the level of malarial transmission during the dry season in Sakassou, Côte d'Ivoire. Female Anopheles mosquitoes were sampled using human landing catches (HLCs) and pyrethrum spraying catches (PSCs). The larvae were collected using the 'dipping' method. A total of 10,875 adult female mosquitoes of Anopheles gambiae were collected. The PCR analysis revealed that all individuals were Anopheles coluzzii. The geographical distribution of potential breeding sites of Anopheles showed the presence of An. coluzzii in all the wetlands of the city of Sakassou. During the dry season, the human-biting rate of An. coluzzii was 139.1 bites/person/night. An exophagic trend was displayed by an adult female of An. coluzzii. The entomological inoculation rate during the dry season was 1.49 infectious bites/person/night. This study demonstrated that An. coluzzii was the main vector of malarial transmission in Sakassou, and the intensity of transmission remains high throughout the dry season.

Geospatial joint modeling of vector and parasite serology to microstratify malaria transmission.

The World Health Organization identifies a strong surveillance system for malaria and its mosquito vector as an essential pillar of the malaria elimination agenda. Anopheles salivary antibodies are emerging biomarkers of exposure to mosquito bites that potentially overcome sensitivity and logistical constraints of traditional entomological surveys. Using samples collected by a village health volunteer network in 104 villages in Southeast Myanmar during routine surveillance, the present study employs a Bayesian geostatistical modeling framework, incorporating climatic and environmental variables together with Anopheles salivary antigen serology, to generate spatially continuous predictive maps of Anopheles biting exposure. Our maps quantify fine-scale spatial and temporal heterogeneity in Anopheles salivary antibody seroprevalence (ranging from 9 to 99%) that serves as a proxy of exposure to Anopheles bites and advances current static maps of only Anopheles occurrence. We also developed an innovative framework to perform surveillance of malaria transmission. By incorporating antibodies against the vector and the transmissible form of malaria (sporozoite) in a joint Bayesian geostatistical model, we predict several foci of ongoing transmission. In our study, we demonstrate that antibodies specific for Anopheles salivary and sporozoite antigens are a logistically feasible metric with which to quantify and characterize heterogeneity in exposure to vector bites and malaria transmission. These approaches could readily be scaled up into existing village health volunteer surveillance networks to identify foci of residual malaria transmission, which could be targeted with supplementary interventions to accelerate progress toward elimination.

Insufficient duration of insecticidal efficacy of Yahe® insecticide-treated nets in Papua New Guinea.

BACKGROUND: Insecticide-treated nets (ITNs) are the backbone of anti-malarial vector control in Papua New Guinea (PNG). Over recent years the quality and performance of ITNs delivered to PNG decreased, which has likely contributed to the stagnation in the malaria control effort in the country. The present study reports results from the first 24 months of a durability study with the ITN product Yahe LN® in PNG. METHODS: The durability study was conducted in four villages on the northern coast of PNG, in an area with high malaria parasite transmission, following WHO-recommended methodology adapted to the local scenario. A cohort of n = 500 individually identifiable Yahe® ITNs was distributed by the PNG National Malaria Control Programme from October to December 2021. Insecticidal efficacy of the ITNs was tested using cone bioassays with fully pyrethroid susceptible Anopheles farauti colony mosquitoes at baseline and at 6 months intervals, alongside evaluation of physical integrity and the proportion of ITNs lost to follow-up. A questionnaire was used to collect information on ITN end user behaviour, such as the frequency of use and washing. The observations from the durability study were augmented with simulated laboratory wash assays. RESULTS: Gradual uptake and replacement of previous campaign nets by the communities was observed, such that at 6 months 45% of all newly distributed nets were in use in their designated households. Insecticidal efficacy of the Yahe® nets, expressed as the percent 24 h mortality in cone bioassays decreased from 91 to 45% within the first 6 months of distribution, even though > 90% of study nets had never been washed. Insecticidal efficacy decreased further to < 20% after 24 months. ITNs accumulated physical damage (holes) at a rate similar to previous studies, and 35% were classified as 'too torn' by proportional hole index after 24 months. ITNs were lost to follow-up such that 61% of cohort nets were still present after 24 months. Laboratory wash assays indicated a rapid reduction in insecticidal performance with each consecutive wash such that average 24 h mortality was below 20% after 10 washes. CONCLUSION: Yahe® ITNs are not performing as per label claim in an area with fully pyrethroid susceptible vectors, and should be investigated more comprehensively and in other settings for compliance with currently recommended durability and efficacy thresholds. The mass distribution of low quality ITN products with variable performance is one of the major ongoing challenges for global malaria control in the last decade.

[The mosquitoes (Diptera: Culicidae) of Djibouti: bibliographical review (1970-2023)]. / Les moustiques (Diptera : Culicidae) de Djibouti : revue bibliographique (1970-2023).

Background and justification: The Republic of Djibouti is located in the Horn of Africa, on the Gulf of Aden and the Bab-el-Mandeb detroit, at the southern entrance to the Red Sea. Prior to its independence in 1977, the Republic of Djibouti was known by two names: "Côte française des Somalis" until 1967, then "Territoire Français de Afars et Issas". As part of our doctoral research on the ecology of mosquitoes in Djibouti, we noted a lack of information on the species encountered, and felt it essential to draw up a list of species before embarking on ecological monitoring. The aim of this work is to survey publications on mosquitoes in Djibouti and to synthesize data from this scientific literature in order to update the national inventory of Culicidae. Materials and methods: An exhaustive search of electronic bibliographic databases (PubMed, Scopus, HAL Open Archive, Science Direct and Google Scholar) was carried out. Reference lists were filtered to access additional articles in order to obtain more data. Two keywords were used: "Djibouti" and "French Territory of Afars and Issas". A selection of scientific publications on Djibouti mosquitoes and/or diseases transmitted by mosquito vectors was made. Researches were conducted in articles selected. The names of the species listed were checked and validated by referring to the site Mosquito Taxonomic Inventory. Results: A total of 13 studies, published between 1970 and 2023, were found. Over the years, the composition of the Culicidae fauna has become well known. In part, the movement of people traveling to and from neighboring countries has been linked to the detection of new species and the reappearance of mosquito species in Djibouti. Numerous studies have been carried out over the years, including purely taxonomic studies and others focusing on the incrimination of mosquito vectors and the characterization of the pathogens they transmit. A total of 37 species, belonging to two subfamilies (Anophelinae and Culicinae), of mosquitoes divided between 7 genera (Aedes, Anopheles, Culex, Culiseta, Lutzia, Mimomyia and Uranotaenia) have been mentioned across the country. The number of species per genus is distributed as follows: 5 species of Aedes including 1 subspecies, 14 species of Anopheles including two subspecies, 12 species of Culex including 1 subspecies, 1 species for each of the genera Culiseta and Lutzia and finally 2 species respectively for the genera Mimomiya and Uranotaenia. Five species have been incriminated as vectors of diseases such as malaria, dengue fever, yellow fever, West Nile virus and chikungunya. Others are known for their potential role in pathogen transmission, including Zika and Rift Valley virus. Discussion - Conclusion: The bibliographical research enabled us to summarize the research carried out over more than half a century in the history of Djibouti, and to update the inventory of the country's mosquitoes, which now includes 37 species. Species names were reviewed and updated, and the case of Anopheles gambiae was also addressed. Two species mentioned as part of the Culicidae fauna of Djibouti appeared to be doubtful and are up for discussion. These results provide a useful information base for defining vector control priorities in Djibouti. They will also inform, guide and facilitate future consultations of our database. In addition, this study will help to identify research ways on mosquitoes in Djibouti.

A cell atlas of the adult female Aedes aegypti midgut revealed by single-cell RNA sequencing.

Aedes aegypti is a primary vector for transmitting various arboviruses, including Yellow fever, dengue and Zika virus. The mosquito midgut is the principal organ for blood meal digestion, nutrient absorption and the initial site of arbovirus infection. Although a previous study delineated midgut's transcriptome of Ae. aegypti at the single-nucleus resolution, there still lacks an established protocol for isolating and RNA sequencing of single cells of Ae. aegypti midgut, which is required for investigating arbovirus-midgut interaction at the single-cell level. Here, we established an atlas of the midgut cells for Ae. aegypti by single-cell RNA sequencing. We annotated the cell clusters including intestinal stem cells/enteroblasts (ISC/EB), cardia cells (Cardia), enterocytes (EC, EC-like), enteroendocrine cells (EE), visceral muscle (VM), fat body cells (FBC) and hemocyte cells (HC). This study will provide a foundation for further studies of arbovirus infection in mosquito midgut at the single-cell level.

Altered thermal preferences of infected or immune-challenged Aedes aegypti and Aedes japonicus mosquitoes.

Temperature is a critical factor shaping physiology, life cycle, and behaviour of ectothermic vector insects, as well as the development and multiplication of pathogens within them. However, the influence of pathogen infections on thermal preferences (behavioural thermoregulation) is not well-understood. The present study examined the thermal preferences of mosquitoes (Aedes aegypti and Ae. japonicus) infected with either Sindbis virus (SINV) or Dirofilaria immitis over 12 days post exposure (p.e.) or injected with a non-pathogenic Sephadex bead over 24 h in a thermal gradient (15-30 °C). SINV-infected Ae. aegypti preferred 5 °C warmer temperatures than non-infected ones at day 6 p.e., probably the time of highest innate immune response. In contrast, D. immitis-infected Ae. japonicus preferred 4 °C cooler temperatures than non-infected ones at day 9 p.e., presumably a stress response during the migration of third instar larvae from their development site to the proboscis. Sephadex bead injection also induced a cold preference in the mosquitoes but to a level that did not differ from control-injections. The cold preference thus might be a strategy to escape the risk of desiccation caused by the wound created by piercing the thorax. Further research is needed to uncover the genetic and physiological mechanisms underlying these behaviours.

Efficacy of pyrethroid-pyriproxyfen and pyrethroid-chlorfenapyr nets on entomological indicators of malaria transmission: third year of a randomised controlled trial in Benin.

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

Mosquitocidal effect of ivermectin-treated nettings and sprayed walls on Anopheles gambiae s.s.

Ivermectin (IVM) has been proposed as a new tool for malaria control as it is toxic on vectors feeding on treated humans or cattle. Nevertheless, IVM may have a direct mosquitocidal effect when applied on bed nets or sprayed walls. The potential for IVM application as a new insecticide for long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) was tested in this proof-of-concept study in a laboratory and semi-field environment. Laboratory-reared, insecticide-susceptible Kisumu Anopheles gambiae were exposed to IVM on impregnated netting materials and sprayed plastered- and mud walls using cone bioassays. The results showed a direct mosquitocidal effect of IVM on this mosquito strain as all mosquitoes died by 24 h after exposure to IVM. The effect was slower on the IVM-sprayed walls compared to the treated nettings. Further work to evaluate possibility of IVM as a new insecticide formulation in LLINs and IRS will be required.

Sampling efficiency and screening of Aedes albopictus for yellow fever virus in Niger Delta region of Nigeria.

Introduction: Aedes albopictus, like Aedes aegypti, is a virulent vector of arboviruses especially the well-documented spread of yellow fever around the world. Although yellow fever is prevalent in Nigeria, there is a paucity of information in the Niger Delta region on the distribution of Aedes mosquito vectors and molecular detection of the virus in infected mosquitoes. This study sampled Aedes mosquitoes around houses associated with farms from four communities (Otolokpo, Ute-Okpu, Umunede, and Ute Alohen) in Ika North-East Local Government Area of Delta State, Nigeria. Methods: various sampling methods were used in Aedes mosquito collection to test their efficacy in the survey. Mosquitoes in holding cages were killed by freezing and morphologically identified. A pool of 15 mosquitoes per Eppendorf tube was preserved in RNAi later for yellow fever virus screening. Two samples were molecularly screened for each location. Results: seven hundred and twenty-five (725) mosquitoes were obtained from the various traps. The mean abundance of the mosquitoes was highest in m-HLC (42.9) compared to the mosquitoes sampled using other techniques (p<0.0001). The mean abundance of mosquitoes was lowest in Center for Disease Control (CDC) light traps without attractant (0.29). No yellow fever virus strain was detected in all the mosquitoes sampled at the four locations. Conclusion: this study suggests that Aedes albopictus are the mosquitoes commonly biting around houses associated with farms. More so, yellow fever virus was not detected in the mosquitoes probably due to the mass vaccination exercise that was carried out the previous year in the study area. More studies are required using the m-HLC to determine the infection rate in this endemic area.

A First Report on the Isolation and Characterization of a Highly Potential Indigenous Mosquitocidal Bacterium (Bacillus thuringiensis Subspecies Israelensis VCRC B647) from Red Soil, India.

BACKGROUND: To control mosquito vectors causing human diseases, bacterial biopesticides are currently in use. Indeed, the recent development of resistance to these bacterial agents has impeded its applications. Under these circumstances, the search for novel bacterial agents with mosquitocidal activity is unavoidable. In this study, a novel mosquitocidal bacterium was isolated from red soils of agricultural field. OBJECTIVES: The objective of this study was to isolate and identify new mosquitocidal bacteria from the natural environment. MATERIALS AND METHODS: Soil samples were collected during 2021-2022 from Tirupathur district of Tamil Nadu, South India. The samples were bioprocessed for culturing the bacterial colony in a suitable culture medium (Nutrient Yeast Salt Medium), and after 72 h, the bacterial cell mass was removed and lyophilized. Bioassays (mosquito toxicity assays) were carried out to screen the bacterial colonies for mosquitocidal effect. The potential colony was further analyzed, and identified for its application in mosquito control. RESULTS: The new isolate screened from red soil was identified as Bacillus thuringiensis subspecies israelensis (VCRC B647) as per the ilvD gene sequence analysis. The strain was found to be potentially effective in controlling mosquito larvae, and further biochemical analyses, bacterial growth, biomass, and protein content were investigated. The new isolate did not show any toxic effect on nontarget aquatic organisms. CONCLUSION: It is significant to depict that the mosquitocidal action of this new isolate (Bti) is highly significant than the reference strain of Bti-H14. It is concluded that this is the first report that an indigenous strain of Bti VCRC B647 is very effective in mosquito control.

Tiger prowling: Distribution modelling for northward-expanding Aedes albopictus (Diptera: Culicidae) in Japan.

The Asian tiger mosquito, Aedes albopictus, is a significant public health concern owing to its expanding habitat and vector competence. Disease outbreaks attributed to this species have been reported in areas under its invasion, and its northward expansion in Japan has caused concern because of the potential for dengue virus infection in newly populated areas. Accurate prediction of Ae. albopictus distribution is crucial to prevent the spread of the disease. However, limited studies have focused on the prediction of Ae. albopictus distribution in Japan. Herein, we used the random forest model, a machine learning approach, to predict the current and potential future habitat ranges of Ae. albopictus in Japan. The model revealed that these mosquitoes prefer urban areas over forests in Japan on the current map. Under predictions for the future, the species will expand its range to the surrounding areas and eventually reach many areas of northeastern Kanto, Tohoku District, and Hokkaido, with a few variations in different scenarios. However, the affected human population is predicted to decrease owing to the declining birth rate. Anthropogenic and climatic factors contribute to range expansion, and urban size and population have profound impacts. This prediction map can guide responses to the introduction of this species in new areas, advance the spatial knowledge of diseases vectored by it, and mitigate the possible disease burden. To our knowledge, this is the first distribution-modelling prediction for Ae. albopictus with a focus on Japan.

Potential of MALDI-TOF MS biotyping to detect deltamethrin resistance in the dengue vector Aedes aegypti.

Insecticide resistance in mosquitoes is spreading worldwide and represents a growing threat to vector control. Insecticide resistance is caused by different mechanisms including higher metabolic detoxication, target-site modification, reduced penetration and behavioral changes that are not easily detectable with simple diagnostic methods. Indeed, most molecular resistance diagnostic tools are costly and labor intensive and then difficult to use for routine monitoring of insecticide resistance. The present study aims to determine whether mosquito susceptibility status against the pyrethroid insecticides (mostly used for mosquito control) could be established by the protein signatures of legs and/or thoraxes submitted to MALDI-TOF Mass Spectrometry (MS). The quality of MS spectra for both body parts was controlled to avoid any bias due to unconformity protein profiling. The comparison of MS profiles from three inbreeds Ae. aegypti lines from French Guiana (IRF, IR03, IR13), with distinct deltamethrin resistance genotype / phenotype and the susceptible reference laboratory line BORA (French Polynesia), showed different protein signatures. On both body parts, the analysis of whole protein profiles revealed a singularity of BORA line compared to the three inbreeding lines from French Guiana origin, suggesting that the first criteria of differentiation is the geographical origin and/or the breeding history rather than the insecticide susceptibility profile. However, a deeper analysis of the protein profiles allowed to identify 10 and 11 discriminating peaks from leg and thorax spectra, respectively. Among them, a specific peak around 4870 Da was detected in legs and thoraxes of pyrethroid resistant lines compared to the susceptible counterparts hence suggesting that MS profiling may be promising to rapidly distinguish resistant and susceptible phenotypes. Further work is needed to confirm the nature of this peak as a deltamethrin resistant marker and to validate the routine use of MS profiling to track insecticide resistance in Ae. aegypti field populations.

Predicting the age of field Anopheles mosquitoes using mass spectrometry and deep learning.

Mosquito-borne diseases like malaria are rising globally, and improved mosquito vector surveillance is needed. Survival of Anopheles mosquitoes is key for epidemiological monitoring of malaria transmission and evaluation of vector control strategies targeting mosquito longevity, as the risk of pathogen transmission increases with mosquito age. However, the available tools to estimate field mosquito age are often approximate and time-consuming. Here, we show a rapid method that combines matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry with deep learning for mosquito age prediction. Using 2763 mass spectra from the head, legs, and thorax of 251 field-collected Anopheles arabiensis mosquitoes, we developed deep learning models that achieved a best mean absolute error of 1.74 days. We also demonstrate consistent performance at two ecological sites in Senegal, supported by age-related protein changes. Our approach is promising for malaria control and the field of vector biology, benefiting other disease vectors like Aedes mosquitoes.

Evaluating the mosquito vector range for two orthobunyaviruses: Oya virus and Ebinur Lake virus.

BACKGROUND: Mosquito-borne viruses cause various infectious diseases in humans and animals. Oya virus (OYAV) and Ebinur Lake virus (EBIV), belonging to the genus Orthobunyavirus within the family Peribunyaviridae, are recognized as neglected viruses with the potential to pose threats to animal or public health. The evaluation of vector competence is essential for predicting the arbovirus transmission risk. METHODS: To investigate the range of mosquito vectors for OYAV (strain SZC50) and EBIV (strain Cu20-XJ), the susceptibility of four mosquito species (Culex pipiens pallens, Cx. quinquefasciatus, Aedes albopictus, and Ae. aegypti) was measured through artificial oral infection. Then, mosquito species with a high infection rate (IR) were chosen to further evaluate the dissemination rate (DR), transmission rate (TR), and transmission efficiency. The viral RNA in each mosquito sample was determined by RT-qPCR. RESULTS: The results revealed that for OYAV, Cx. pipiens pallens had the highest IR (up to 40.0%) among the four species, but the DR and TR were 4.8% and 0.0%, respectively. For EBIV, Cx. pipiens pallens and Cx. quinquefasciatus had higher IR compared to Ae. albopictus (1.7%). However, the EBIV RNA and infectious virus were detected in Cx. pipiens pallens, with a TR of up to 15.4% and a transmission efficiency of 3.3%. CONCLUSIONS: The findings indicate that Cx. pipiens pallens was susceptible to OYAV but had an extremely low risk of transmitting the virus. Culex pipiens pallens and Cx. quinquefasciatus were susceptible to EBIV, and Cx. pipiens pallens had a higher transmission risk to EBIV than Cx. quinquefasciatus.

Trends of Mansonia (Diptera, Culicidae, Mansoniini) in Porto Velho: Seasonal patterns and meteorological influences.

Entomological research is vital for shaping strategies to control mosquito vectors. Its significance also reaches into environmental management, aiming to prevent inconveniences caused by non-vector mosquitoes like the Mansonia Blanchard, 1901 mosquito. In this study, we carried out a five-year (2019-2023) monitoring of these mosquitoes at ten sites in Porto Velho, Rondônia, using SkeeterVac SV3100 automatic traps positioned between the two hydroelectric complexes on the Madeira River. Throughout this period, we sampled 153,125 mosquitoes, of which the Mansonia genus accounted for 54% of the total, indicating its prevalence in the region. ARIMA analysis revealed seasonal patterns of Mansonia spp., highlighting periods of peak density. Notably, a significant decreasing trend in local abundance was observed from July 2021 (25th epidemiological week) until the end of the study. Wind speed was observed to be the most relevant meteorological factor influencing the abundance of Mansonia spp. especially in the Joana D'Arc settlement, although additional investigation is needed to comprehensively analyze other local events and gain a deeper understanding of the ecological patterns of this genus in the Amazon region.

Factors associated with mosquito control among construction workers: A systematic review.

INTRODUCTION: Workers in the construction industry frequently work in construction sites with numerous areas that can potentially accumulate water, such as tanks, wet cement surfaces, or water puddles. These water collection sites become ideal breeding grounds for mosquito infestation, which leads to a higher prevalence of mosquito-borne diseases, especially malaria and dengue among construction workers. Despite that numerous factors have been identified in controlling vector-borne diseases, the specific factors that influence mosquito control at construction sites have yet to be explored. AIMS: This systematic review aims to determine the factors associated with mosquito control among construction workers. METHODS: Primarily, articles related to factors associated with mosquito control among construction workers were collected from two different online databases (ScienceDirect and EBSCOhost). Two independent reviewers were assigned to screen the titles and abstracts of the collected data, stored in Microsoft Excel, against the inclusion and exclusion criteria. Afterwards, the quality of the included articles was critically assessed using the Mixed Method Appraisal Tool (MMAT). Of the 171 articles identified, 4 were included in the final review. RESULTS: Based on the thorough evaluation, mosquito-related knowledge, practical mosquito prevention measures, and Larval Source Management (LSM) were identified as vital factors associated with mosquito control among construction workers. The significant association between mosquito-related knowledge and control practices indicates higher knowledge linked to effective practices, particularly among female workers and those who were recently infected with malaria. Concurrently, there were notable challenges regarding sustainable preventive measures and larval control methods in construction settings. CONCLUSION: Implementing effective mosquito control, including knowledge and practice on mosquito control together with vector control, is highly required to suppress the expanding mosquito population. It is recommended that employers provide continuous mosquito control education and training to their employees and reward them with incentives, while employees should comply with the guidelines set by their employers to ensure successful mosquito control and reduce the spread of mosquito-borne diseases in the construction industry.

Mosquito Microbiomes of Rwanda: Characterizing Mosquito Host and Microbial Communities in the Land of a Thousand Hills.

Mosquitoes are a complex nuisance around the world and tropical countries bear the brunt of the burden of mosquito-borne diseases. Rwanda has had success in reducing malaria and some arboviral diseases over the last few years, but still faces challenges to elimination. By building our understanding of in situ mosquito communities in Rwanda at a disturbed, human-occupied site and at a natural, preserved site, we can build our understanding of natural mosquito microbiomes toward the goal of implementing novel microbial control methods. Here, we examined the composition of collected mosquitoes and their microbiomes at two diverse sites using Cytochrome c Oxidase I sequencing and 16S V4 high-throughput sequencing. The majority (36 of 40 species) of mosquitoes captured and characterized in this study are the first-known record of their species for Rwanda but have been characterized in other nations in East Africa. We found significant differences among mosquito genera and among species, but not between mosquito sexes or catch method. Bacteria of interest for arbovirus control, Asaia, Serratia, and Wolbachia, were found in abundance at both sites and varied greatly by species.

Zika emergence, persistence, and transmission rate in Colombia: a nationwide application of a space-time Markov switching model.

Zika, a viral disease transmitted to humans by Aedes mosquitoes, emerged in the Americas in 2015, causing large-scale epidemics. Colombia alone reported over 72,000 Zika cases between 2015 and 2016. Using national surveillance data from 1121 municipalities over 70 weeks, we identified sociodemographic and environmental factors associated with Zika's emergence, re-emergence, persistence, and transmission intensity in Colombia. We fitted a zero-state Markov-switching model under the Bayesian framework, assuming Zika switched between periods of presence and absence according to spatially and temporally varying probabilities of emergence/re-emergence (from absence to presence) and persistence (from presence to presence). These probabilities were assumed to follow a series of mixed multiple logistic regressions. When Zika was present, assuming that the cases follow a negative binomial distribution, we estimated the transmission intensity rate. Our results indicate that Zika emerged/re-emerged sooner and that transmission was intensified in municipalities that were more densely populated, at lower altitudes and/or with less vegetation cover. Warmer temperatures and less weekly-accumulated rain were also associated with Zika emergence. Zika cases persisted for longer in more densely populated areas with more cases reported in the previous week. Overall, population density, elevation, and temperature were identified as the main contributors to the first Zika epidemic in Colombia. We also estimated the probability of Zika presence by municipality and week, and the results suggest that the disease circulated undetected by the surveillance system on many occasions. Our results offer insights into priority areas for public health interventions against emerging and re-emerging Aedes-borne diseases.

A simple, field-applicable method to increase the infectivity of wild isolates of Plasmodium falciparum to mosquito vectors.

BACKGROUND: The direct membrane feeding assay (DMFA), whereby gametocyte-infected blood is collected from human donors and from which mosquitoes feed through a membrane, is proving essential for assessing parameters influencing Plasmodium transmission potential in endemic countries. The success of DMFAs is closely tied to gametocyte density in the blood, with relatively high gametocytaemia ensuring optimal infection levels in mosquitoes. As transmission intensity declines with control efforts, the occurrence of asymptomatic individuals with low gametocyte densities, who can significantly contribute to the infectious reservoir, is increasing. This poses a limitation to studies relying on the experimental infection of large numbers of mosquitoes with natural isolates of Plasmodium. A simple, field-applicable method is presented for improving parasite infectivity by concentrating Plasmodium falciparum gametocytes. METHODS: Anopheles gambiae received one of the following 5 blood treatments through DMFA: (i) whole blood (WB) samples from naturally-infected donors; (ii) donor blood whose plasma was replaced with the same volume of Plasmodium-naive AB + serum (1:1 control); (iii) plasma replaced with a volume of malaria-naïve AB + serum equivalent to half (1:1/2), or to a quarter (1:1/4), of the initial plasma volume; and (v) donor blood whose plasma was fully removed (RBC). The experiment was repeated 4 times using 4 distinct wild parasite isolates. Seven days post-infection, a total of 1,095 midguts were examined for oocyst presence. RESULTS: Substituting plasma with reduced amounts (1:1/2 and 1:1/4) of Plasmodium-naive AB + serum led to a 31% and 17% increase of the mosquito infection rate and to a 85% and 308% increase in infection intensity compared to the 1:1 control, respectively. The full removal of plasma (RBC) reduced the infection rate by 58% and the intensity by 64% compared to the 1:1 control. Reducing serum volumes (1:1/2; 1:1/4 and RBC) had no impact on mosquito feeding rate and survival when compared to the 1:1 control. CONCLUSIONS: Concentrating gametocytic blood by replacing natural plasma by lower amount of naive serum can enhance the success of mosquito infection. In an area with low gametocyte density, this simple and practical method of parasite concentration can facilitate studies on human-to-mosquito transmission such as the evaluation of transmission-blocking interventions.

Vegetation structure drives mosquito community composition in UK's largest managed lowland wetland.

PURPOSE: The rising burden of mosquito-borne diseases in Europe extends beyond urban areas, encompassing rural and semi-urban regions near managed and natural wetlands evidenced by recent outbreaks of Usutu and West Nile viruses. While wetland management policies focus on biodiversity and ecosystem services, few studies explore the impact on mosquito vectors. METHODS: Our research addresses this gap, examining juvenile mosquito and aquatic predator communities in 67 ditch sites within a South England coastal marsh subjected to different wetland management tiers. Using joint distribution models, we analyse how mosquito communities respond to abiotic and biotic factors influenced by wetland management. RESULTS: Of the 12 mosquito species identified, Culiseta annulata (Usutu virus vector) and Culex pipiens (Usutu and West Nile virus vector) constitute 47% of 6825 larval mosquitoes. Abundant predators include Coleoptera (water beetles) adults, Corixidae (water boatmen) and Zygoptera (Damselfy) larvae. Models reveal that tier 3 management sites (higher winter water levels, lower agricultural intensity) associated with shade and less floating vegetation are preferred by specific mosquito species. All mosquito species except Anopheles maculipennis s.l., are negatively impacted by potential predators. Culiseta annulata shows positive associations with shaded and turbid water, contrary to preferences of Corixidae predators. CONCLUSIONS: Tier 3 areas managed for biodiversity, characterised by higher seasonal water levels and reduced livestock grazing intensity, provide favourable habitats for key mosquito species that are known vectors of arboviruses, such as Usutu and West Nile. Our findings emphasise the impact of biodiversity-focused wetland management, altering mosquito breeding site vegetation to enhance vector suitability. Further exploration of these trade-offs is crucial for comprehending the broader implications of wetland management.