Search

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

The mosquito Aedes aegypti is an important vector of diseases including dengue, Zika, chikungunya, and yellow fever. Olfaction is a critical modality for mosquitoes enabling them to locate hosts, sources of nectar, and sites for oviposition. GABA is an essential neurotransmitter in olfactory processing in the insect brain, including the primary olfactory center, the antennal lobe. Previous work with Ae. aegypti has suggested that antennal lobe inhibition via GABA may be involved in the processing of odors. However, little is known about GABA receptor expression in the mosquito brain, or how they may be involved in odor attraction. In this context, generating mutants that target the mosquitos olfactory responses, and particularly the GABAergic system, is essential to achieve a better understanding of these diverse processes and olfactory coding in these disease vectors. Here we demonstrate the potential of a transgenic line using the QF2 transcription factor, GABA-B1QF2-ECFP, as a new neurogenetic tool to investigate the neural basis of olfaction in Ae. aegypti. Our results show that the gene insertion has a moderate impact on mosquito fitness. Moreover, the line presented here was crossed with a QUAS reporter line expressing the green fluorescent protein and used to determine the location of the metabotropic GABA-B1 receptor expression. We find high receptor expression in the antennal lobes, especially the cell bodies surrounding the antennal lobes. In the mushroom bodies, receptor expression was high in the Kenyon cells, but had low expression in the mushroom body lobes. Behavioral experiments testing the fruit odor attractants showed that the mutants lost their behavioral attraction. Together, these results show that the GABA-B1QF2-ECFP line provides a new tool to characterize GABAergic systems in the mosquito nervous system.

Mosquitoes transmit pathogens that pose a threat to millions of people globally. Unfortunately, widespread insecticide resistance makes it difficult to control these public health pests. General mechanisms of resistance, such as target site mutations or increased metabolic activity, are well established. However, many questions regarding the dynamics of these adaptations in the context of developmental and environmental conditions require additional exploration. One aspect of resistance that deserves further study is the role of heat shock proteins (HSPs) in insecticide tolerance. Studies show that mosquitoes experiencing heat stress before insecticide exposure demonstrate decreased mortality. This is similar to the observed reciprocal reduction in mortality in mosquitoes exposed to insecticide prior to heat stress. The environmental shifts associated with climate change will result in mosquitoes occupying environments with higher ambient temperatures, which could enhance existing insecticide resistance phenotypes. This physiological relationship adds a new dimension to the problem of insecticide resistance and further complicates the challenges that vector control and public health personnel face. This article reviews studies illustrating the relationship between insecticide resistance and HSPs or hsp genes as well as the intersection of thermotolerance and insecticide resistance. Further study of HSPs and insecticide resistance could lead to a deeper understanding of how environmental factors modulate the physiology of these important disease vectors to prepare for changing climatic conditions and the development of novel strategies to prevent vector-borne disease transmission.

Background: Most insects are poikilotherms and ectotherms, so their body temperature is predicated by environmental temperature. With climate change, insect body temperature is rising, which affects how insects develop, survive, and respond to infection. Aging also affects insect physiology by deteriorating body condition and weakening immune proficiency via senescence. Aging is usually considered in terms of time, or chronological age, but it can also be conceptualized in terms of body function, or physiological age. We hypothesized that warmer temperature decouples chronological and physiological age in insects by accelerating senescence. To investigate this, we reared the African malaria mosquito, Anopheles gambiae, at 27 °C, 30 °C and 32 °C, and measured survival starting at 1-, 5-, 10- and 15-days of adulthood after no manipulation, injury, or a hemocoelic infection with Escherichia coli or Micrococcus luteus. Then, we measured the intensity of an E. coli infection to determine how the interaction between environmental temperature and aging shapes a mosquito's response to infection. Results: We demonstrate that longevity declines when a mosquito is infected with bacteria, mosquitoes have shorter lifespans when the temperature is warmer, older mosquitoes are more likely to die, and warmer temperature marginally accelerates the aging-dependent decline in survival. Furthermore, we discovered that E. coli infection intensity increases when the temperature is warmer and with aging, and that warmer temperature accelerates the aging-dependent increase in infection intensity. Finally, we uncovered that warmer temperature affects both bacterial and mosquito physiology. Conclusions: Warmer environmental temperature accelerates aging in mosquitoes, negatively affecting both longevity and infection outcomes. These findings have implications for how insects will serve as pollinators, agricultural pests, and disease vectors in our warming world. [ABSTRACT FROM AUTHOR]

Background: Fibrinogen-related protein 1 (frep1) is a member of the pattern-recognizing receptor family (PRR) which generates an innate immune response after recognizing the pattern associated molecular pattern (PAMP) that occurs on the surface of microorganisms. The main objective of this study is to characterize frep1 and its in-silico analysis in Anopheles stephensi. Methods and result: The DNA was extracted from female Anopheles stephensi. PCR was performed for complete analysis of frep1 using specific primers. The gene sequence of frep1 was identified by Sanger sequencing. The bioinformatics structure analysis approach revealed the presence of 3 exons and 4 introns in the frep1. The sequence of frep1 was submitted to NCBI GeneBank with accession number ON817187.1. Quantitative real-time PCR was performed to analyze frep1 expression. At the developmental stage, frep1 is highly expressed in the L1 stage, egg, and adult female mosquito. In addition, frep1 is highly expressed in the tissue fat body, midgut, and salivary gland. After blood-fed, an upregulation of frep1 at 48 h in the midgut, and downregulation in fat body were observed at different time intervals. Conclusion: The genomic data of frep1 is encoded by 12,443 bp. The frep1 has a significant role in the early metamorphosis. Its expression in fat body and midgut suggests it could be important for fat metabolism and post-blood digestion. The conserved domain could be targeted for vector control. Further study is required to elucidate its function against malaria parasites to confirm its agonist role in malaria transmission. [ABSTRACT FROM AUTHOR]

Our knowledge of alphavirus genetic diversity is mainly based on viruses isolated from anthropophilic mosquito species, humans, and livestock during outbreaks. Studies on alphaviruses from sylvatic amplification cycles in sub-Saharan Africa have been conducted less often than from epizootic environments. To gain insight into alphavirus diversity in enzootic transmission cycles, we collected over 23,000 mosquitoes in lowland rainforest and savannah gallery forest in southwestern Uganda and tested them for alphavirus infections. We detected Sindbis virus (SINV) in a Culex Culex sp. mosquito and Middelburg virus (MIDV) in Eretmapodites intermedius and Mansonia africana. MIDV is a mosquito-borne alphavirus that causes febrile illness in sheep, goats, and horses and was previously not known to occur in Uganda. SINV, also a mosquito-borne alphavirus, causes mild infections in humans. Full genomes of SINV and MIDV were sequenced, showing a nucleotide identity of 99% to related strains. Both isolates replicated to high titres in a wide variety of vertebrate cells. Our data suggest endemic circulation of SINV and MIDV in Uganda. [ABSTRACT FROM AUTHOR]

Larval habitat availability, Typha phytotelmata use and the relation of environmental factors to annual dynamics of the mosquito Wyeomyia leucostigma Lutz, 1904 (larvae and females) were studied in Costanera Sur Ecological Reserve at Buenos Aires, Argentina. Bimonthly, 1520 plants were examined, detecting W. leucostigma in 80.8% of surveys. Larval habitat availability was associated with temperature, but not with rainfall. In addition, the high temperature was associated with greater biting rate. Dynamics of host-seeking were recorded over 10 months of the year and were associated with temperature as well. Furthermore, an accidental fire led to an increase of W. leucostigma abundance likely based on an increase of the Thypha marshes productivity that occurred after the fire. [ABSTRACT FROM AUTHOR]

The present study was carried out for the first time in Morocco to valorise plant species of the Artemisia genus. Its aim was to evaluate the larvicidal activity of essential oils from six Artemisia plants (A. herba alba, A. absinthium, A. campestris L., A. negrei, A. pontica, and A. arborescens) against Culex pipiens mosquitoes, which are recognised as potential vectors of West Nile virus. Bioinsecticides derived from EOs have shown promise as effective mosquito control agents. The EOs were analysed using GC–MS and GC‐FID and were obtained by hydrodistillation from the aerial parts of the plants. Besides, the experiment was carried out according to the standard WHO protocol. Thujone and camphor were identified as the predominant compounds in all six Artemisia plants. A. herba alba demonstrated the highest larvicidal activity with LC50 and LC90 values of 4.85 and 15.39 μg/mL, respectively. Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA) were employed to explore the correlation between major EO compounds and larvicidal efficacy. The results suggest that these findings could pave the way for the development of tailored biocides that specifically target important disease vectors. [ABSTRACT FROM AUTHOR]

Mosquitoes are known to choose oviposition habitats actively based on their suitability for survival of their offspring. They thus avoid laying eggs in dangerous habitats with aquatic predators through detection of their cues. Visual, tactile, and chemical cues emitted by certain predators have been shown to reduce oviposition in some mosquitoes, which in turn may affect their adult population sizes. However, those tests tend to be limited to species from temperate regions. Moreover, mosquito oviposition response to predator cues have been shown to be predator specific, but this has not been investigated for many potential predators worldwide. In the African subtropics, temporary ponds can be important mosquito breeding habitats and the same habitats are also used by a group of specialized killifish that can survive dry periods using drought resistant eggs. Although they are reported as mosquito predators, it is unknown whether mosquitoes actively avoid laying eggs in ponds with annual killifish. We investigated this using the mosquito Culex quinquefasciatus and the free and caged killifish Nothobranchius neumanni in the outdoor mesocosm experiments in Tanzania. We found that Cx. quinquefasciatus had 71% lower oviposition in mesocosms with free swimming killifish and 68% lower oviposition in mesocosms with caged killifish. We conclude that predator-released chemical cues alone are enough to explain the observed responses. If the cues can be isolated and chemically identified can be developed into a biological mosquito control strategy and used to safeguard public health in areas where mosquito borne diseases are endemic. [ABSTRACT FROM AUTHOR]

Background: Mosquito host feeding patterns are an important factor of the species-specific vector capacity determining pathogen transmission routes. Culex pipiens s.s./Cx. torrentium are competent vectors of several arboviruses, such as West Nile virus and Usutu virus. However, studies on host feeding patterns rarely differentiate the morphologically indistinguishable females. Methods: We analyzed the host feeding attraction of Cx. pipiens and Cx. torrentium in host-choice studies for bird, mouse, and a human lure. In addition, we summarized published and unpublished data on host feeding patterns of field-collected specimens from Germany, Iran, and Moldova from 2012 to 2022, genetically identified as Cx. pipiens biotype pipiens, Cx. pipiens biotype molestus, Cx. pipiens hybrid biotype pipiens × molestus, and Cx. torrentium, and finally put the data in context with similar data found in a systematic literature search. Results: In the host-choice experiments, we did not find a significant attraction to bird, mouse, and human lure for Cx. pipiens pipiens and Cx. torrentium. Hosts of 992 field-collected specimens were identified for Germany, Iran, and Moldova, with the majority determined as Cx. pipiens pipiens, increasing the data available from studies known from the literature by two-thirds. All four Culex pipiens s.s./Cx. torrentium taxa had fed with significant proportions on birds, humans, and nonhuman mammals. Merged with the data from the literature from 23 different studies showing a high prevalence of blood meals from birds, more than 50% of the blood meals of Cx. pipiens s.s. were identified as birds, while up to 39% were human and nonhuman mammalian hosts. Culex torrentium fed half on birds and half on mammals. However, there were considerable geographical differences in the host feeding patterns. Conclusions: In the light of these results, the clear characterization of the Cx. pipiens s.s./Cx. torrentium taxa as ornithophilic/-phagic or mammalophilic/-phagic needs to be reconsidered. Given their broad host ranges, all four Culex taxa could potentially serve as enzootic and bridge vectors. [ABSTRACT FROM AUTHOR]

Background: Aedes aegypti, the primary vector of dengue, chikungunya, and Zika viruses, poses a significant public health threat worldwide. Traditional control methods using insecticides are increasingly challenged by resistance and environmental concerns. The sterile insect technique (SIT) offers an eco-friendly alternative that has been successfully applied to other insect pests. This article aims to briefly review Ae. aegypti management in Cuba, highlighting the accomplishments, challenges, and future directions of the SIT. Main body: Here we provide a brief summary of the extensive history of Ae. aegypti control efforts in Cuba. After a successful eradication campaign in the 1980s, a resurgence of dengue cases has been observed in recent years, suggesting that traditional control methods may have limited effectiveness under current conditions. In response, Cuba initiated a phased approach to develop and evaluate the feasibility of SIT for Ae. aegypti control, starting in 2008. Initial research focused on Ae. aegypti mating behavior and sterilization methods, followed by successful laboratory and semi-field trials that demonstrated population suppression. The first open-field trial in 2020 confirmed the efficacy of the SIT in reducing Ae. aegypti populations under real-world conditions. Currently, the research is in a phase involving a cluster-randomized superiority-controlled trial. This planned trial will compare the standard vector control program with the same program augmented by the SIT, aiming to assess the impact of the SIT on dengue incidence as the primary outcome. Implementing robust epidemiological trials to evaluate the effectiveness of the SIT is complex due to potential spillover effects from mosquito and human movement across study areas. Additionally, conducting the SIT requires significant development and operational investments. Despite these challenges, the ongoing Cuban trial holds promise for establishing the SIT as an effective and sustainable tool for Ae. aegypti control and for reducing the burden of mosquito-borne diseases. Conclusions: The phased evaluation conducted in Cuba confirms the efficacy of the SIT against Ae. aegypti, highlighting its potential for sustainable mosquito-borne disease management. The effective implementation of multi-site trials will be crucial in providing evidence of the potential of the sterile insect technique as part of a strategy to reduce the incidence of arboviral diseases. [ABSTRACT FROM AUTHOR]

Background: Rift Valley fever virus (RVFV) is a zoonotic mosquito-borne virus with serious implications for livestock health, human health, and the economy in Africa, and is suspected to be endemic in north-eastern KwaZulu-Natal (KZN), South Africa. The vectors of RVFV in this area are poorly known, although several species, such as Aedes (Neomelaniconion) mcintoshi, Aedes (Neomelaniconion) circumluteolus, Aedes (Aedimorphus) durbanensis, and Culex (Lasioconops) poicilipes may be involved. The aim of the study was to determine the vertebrate blood meal sources of potential RVFV mosquito vectors in north-eastern KZN and to characterize the host-biting network. Methods: Blood-fed mosquitoes were collected monthly from November 2019 to February 2023 using a backpack aspirator, CO2-baited Centers for Disease Control and Prevention (CDC) miniature light traps and tent traps, in the vicinity of water bodies and livestock farming households. The mosquitoes were morphologically identified. DNA was extracted from individual mosquitoes and used as templates to amplify the vertebrate cytochrome c oxidase I (COI) and cytochrome b (cytb) genes using conventional polymerase chain reaction (PCR). Amplicons were sequenced and queried in GenBank and the Barcode of Life Data systems to identify the vertebrate blood meal sources and confirm mosquito identifications. All mosquitoes were screened for RVFV using real time reverse transcription (RT)-PCR. Results: We identified the mammalian (88.8%) and avian (11.3%) blood meal sources from 409 blood-fed mosquitoes. Aedes circumluteolus (n = 128) made up the largest proportion of collected mosquitoes. Cattle (n = 195) and nyala (n = 61) were the most frequent domestic and wild hosts, respectively. Bipartite network analysis showed that the rural network consisted of more host-biting interactions than the reserve network. All mosquitoes tested negative for RVFV. Conclusions: Several mosquito species, including Ae. circumluteolus, and vertebrate host species, including cattle and nyala, could play a central role in RVFV transmission. Future research in this region should focus on these species to better understand RVFV amplification. [ABSTRACT FROM AUTHOR]

Introduction: This research aimed to assess housing design consideration features and architects' perceptions of their effectiveness in malaria prevention. Previous studies have highlighted various architectural design elements that can reduce mosquito entry into homes, thus contributing to malaria prevention. This study aims to build on these insights by evaluating specific design features and gathering professional opinions on their effectiveness. Methods: The methodological approach adopted was quantitative, with data collected using a structured questionnaire containing closed-ended questions and a checklist. The observation checklist was used to ascertain mosquito entry reduction features incorporated in 208 systematically selected houses in Garneton East, Kitwe, Zambia. Additionally, a structured questionnaire containing fourteen key design features identified from the literature was administered to 76 architects purposively sampled to establish their perceptions of the effectiveness of these design factors in malaria prevention. Data collected was analyzed using descriptive statistics and exploratory factor analysis. Results: The findings revealed that most houses needed to incorporate design features to reduce mosquito entry. The common features incorporated were ceilings (86.80%), screened air ventilation (59.80%), and window screens (54.40%). The use of window and door screens, as well as mosquito-repellent paint, were perceived as the most effective measures by the architects. Discussion: The results suggest a keen awareness among architects regarding the efficacy of certain design features in malaria prevention. Despite this awareness, the implementation of these features in actual housing designs needs to be improved. This indicates a need for greater effort and adaptation in building regulations to promote the use of mosquito entry reduction features. Enhancing housing designs with these features can significantly reduce malaria morbidity and mortality. Therefore, it is recommended that building regulations be updated to support and encourage the incorporation of mosquito entry reduction features in housing designs. [ABSTRACT FROM AUTHOR]

In the absence of an effective vaccine, integrated vector control (IVC) is the key method for controlling dengue cases. The COVID-19 pandemic lockdown significantly weakened the surveillance, preventive insecticide application, and vector control effectiveness. Therefore, the Department of Health issued a memorandum urging barangays to keep implementing interventions to prevent dengue cases from growing in number. This study aimed to describe the status of the implementation of the IVC component of the National Dengue Prevention and Control Program by the Barangay Health Stations (BHSs) of District 5, Manila City, the Philippines, before the COVID-19 pandemic in 2019 and during the pandemic in 2020. The pandemic affected numerous government programs, so the IVC status before and amid the pandemic is worth describing. The study used a cross-sectional descriptive design with a remote survey being conducted with 144 BHS from 08-23 Feb 2021 using a self-administered questionnaire. Results showed that the BHS conducted larviciding (84.02%), adulticiding either through indoor residual spraying [IRS (86.81%)] or space spraying (77.08%) in 2019. Distributions of insecticide-treated nets [ITN (27.78%)], insecticide-treated curtains [ITC (17.36%)], smoke repellents (54.17%), and skin repellents (50.00%) were also implemented. In 2020 amid the pandemic, 81.94% of BHS implemented larviciding, whereas 83.33% and 77.08% conducted IRS and space spraying, respectively. In addition, ITNs (24.31%), ITCs (17.36%), smoke repellents (52.08%), and skin repellents (44.44%) were also distributed in 2020. Despite the pandemic, most barangays in District 5 continued to implement IVC strategies, which contributed to vector density and possibly prevented an increase in mosquito density that could have led to dengue outbreaks. [ABSTRACT FROM AUTHOR]

Temperature is commonly acknowledged as one of the primary forces driving ectotherm vector populations, most notably by influencing metabolic rates and survival. Although numerous experiments have shown this for a wide variety of organisms, the vast majority has been conducted at constant temperatures and changes therein, while temperature is far from constant in nature, and includes seasonal and diurnal cycles. As fluctuating temperatures have been described to affect metabolic processes at (sub)cellular level, this calls for studies evaluating the relative importance of temperature fluctuations and the changes therein. To gain insight in the effects of temperature fluctuations on ectotherm development, survival, and sex ratio, we developed an inexpensive, easily reproducible, and open‐source, Arduino‐based temperature control system, which emulates natural sinusoidal fluctuations around the average temperature. We used this novel setup to compare the effects of constant (mean) temperatures, most commonly used in experiments, block schemes, and natural sinusoidal fluctuations as well as an extreme variant with twice its amplitude using the cosmopolitan mosquito species Culex pipiens s.l. as a study organism. Our system accurately replicated the preprogrammed temperature treatments under outdoor conditions, even more accurately than traditional methods. While no effects were detected on survival and sex ratio within the ranges of variation evaluated, development was sped up considerably by including temperature fluctuations, especially during pupation, where development under constant temperatures took almost a week (30%) longer than under natural fluctuations. Doubling the amplitude further decreased development time by 1.5 days. These results highlight the importance of including (natural) oscillations in experiments on ectotherm organisms – both aquatic and terrestrial – that use temperature as a variable. Ultimately, these results have major repercussions for downstream effects at larger scales that may be studied with applications such as ecological niche models, disease risk models, and assessing ecosystem services that rely on ectotherm organisms. [ABSTRACT FROM AUTHOR]

Dengue is an arbovirus transmitted by the Aedes spp. mosquito. Approximately 390 million infections occur annually per World Health Organization estimates, with significant increases in infections throughout the last decade. The disease is endemic in warmer climates throughout the world, though cases may also be imported to non-endemic regions by returning travelers. Patients experience a wide variety of symptoms ranging from asymptomatic infection to severe disease requiring critical care. Emergency clinicians should consider the diagnosis of dengue in patients from endemic areas presenting with a flu-like illness, rash, and evidence of bleeding. [ABSTRACT FROM AUTHOR]

Modeling biological datasets represents an essential step in processing and exploiting biological information. Selecting features and improving modeling quality are critical in building a high-performance predictive model. In this article, we have presented and applied a novel approach to select features and to improve the modeling quality using the presence/absence data of three mosquito species in Morocco. This approach uses a recursive search of feature subsets conditioned on improving the modeling quality compared to an initially chosen solution. It has led to a significant improvement in the modeling quality compared to another study carried out on the same dataset, where the accuracy of the models improved with a range varying between 0.062 and 0.198. The relevance of this approach also extends to the search for solutions that achieve the same performance with different subsets, known as multiple solutions. These solutions demonstrate that various combinations of explanatory features can explain the target feature, leading to categorizing them according to their impact on the modeling. This work has provided a good explanation of the distribution of mosquito species thanks to the improved modeling quality, opening up the possibility of having relevant solutions and discovering new explanatory modes for the features. [ABSTRACT FROM AUTHOR]

Mosquitoes are vectors of many important human diseases. The prolonged and widespread use of insecticides has led to the development of mosquito resistance to these insecticides. The gut microbiota is considered the master of host development and physiology; it influences mosquito biology, disease pathogen transmission, and resistance to insecticides. Understanding the role and mechanisms of mosquito gut microbiota in mosquito insecticide resistance is useful for developing new strategies for tackling mosquito insecticide resistance. We searched online databases, including PubMed, MEDLINE, SciELO, Web of Science, and the Chinese Science Citation Database. We searched all terms, including microbiota and mosquitoes, or any specific genera or species of mosquitoes. We reviewed the relationships between microbiota and mosquito growth, development, survival, reproduction, and disease pathogen transmission, as well as the interactions between microbiota and mosquito insecticide resistance. Overall, 429 studies were included in this review after filtering 8139 search results. Mosquito gut microbiota show a complex community structure with rich species diversity, dynamic changes in the species composition over time (season) and across space (environmental setting), and variation among mosquito species and mosquito developmental stages (larval vs. adult). The community composition of the microbiota plays profound roles in mosquito development, survival, and reproduction. There was a reciprocal interaction between the mosquito midgut microbiota and virus infection in mosquitoes. Wolbachia, Asaia, and Serratia are the three most studied bacteria that influence disease pathogen transmission. The insecticide resistance or exposure led to the enrichment or reduction in certain microorganisms in the resistant mosquitoes while enhancing the abundance of other microorganisms in insect-susceptible mosquitoes, and they involved many different species/genera/families of microorganisms. Conversely, microbiota can promote insecticide resistance in their hosts by isolating and degrading insecticidal compounds or altering the expression of host genes and metabolic detoxification enzymes. Currently, knowledge is scarce about the community structure of mosquito gut microbiota and its functionality in relation to mosquito pathogen transmission and insecticide resistance. The new multi-omics techniques should be adopted to find the links among environment, mosquito, and host and bring mosquito microbiota studies to the next level. [ABSTRACT FROM AUTHOR]

The mosquito, Aedes aegypti, is the principal vector for several arboviruses. The mosquito midgut is the initial tissue that gets infected with an arbovirus acquired along with a blood meal from a vertebrate host. Blood meal ingestion leads to midgut tissue distention thereby increasing the pore size of the surrounding basal lamina. This allows newly synthesized virions to exit the midgut by traversing the distended basal lamina to infect secondary tissues of the mosquito. We conducted a quantitative label‐free proteomic time course analysis with saline meal‐fed Ae. aegypti females to identify host factors involved in midgut tissue distention. Around 2000 proteins were detected during each of the seven sampling time points and 164 of those were uniquely expressed. Forty‐five of 97 differentially expressed proteins were upregulated during the 96‐h time course and most of those were involved in cytoskeleton modulation, metabolic activity, and vesicle/vacuole formation. The F‐actin‐modulating Ae. aegypti (Aa)‐gelsolin was selected for further functional studies. Stable knockout of Aa‐gelsolin resulted in a mosquito line, which showed distorted actin filaments in midgut‐associated tissues likely due to diminished F‐actin processing by gelsolin. Zika virus dissemination from the midgut of these mosquitoes was diminished and delayed. The loss of Aa‐gelsolin function was associated with an increased induction of apoptosis in midgut tissue indicating an involvement of Aa‐gelsolin in apoptotic signaling in mosquitoes. Here, we used proteomics to discover a novel host factor, Aa‐gelsolin, which affects the midgut escape barrier for arboviruses in mosquitoes and apoptotic signaling in the midgut. [ABSTRACT FROM AUTHOR]

Background: Analyses of the temporal distribution of mosquitoes are presented in statistical charts, but it is difficult to prove in statistics whether differences in peak periods exist among different years or habitats. This study aimed to investigate the application of circular statistics in determining the peak period and a comparison of differences. Methods: Surveillance of adult mosquitoes was conducted twice a month by light traps in five different habitats from March to November for 3 years (2021–2023) in Qingdao, Shandong Province, China. The Kruskal–Wallis test was performed to determine the differences in mosquito density among different years and habitats. Circular statistics and line charts were employed to determine the peak period and a comparison of differences. Results: Among a total of 14,834 adult mosquitoes comprising five mosquito species from four genera, Culex pipiens pallens was dominant and accounted for 89.6% of the specimens identified. Aedes albopictus, Armigeres subalbatus, and Anopheles sinensis made up 5.7%, 4.2%, and 0.5%, respectively. Culex tritaeniorhynchus accounted for less than 0.1%. The mean mosquito density (females/trap night) for the trapping period was 10.3 in 2021, 5.6 in 2022, and 3.6 in 2023. Among five habitats, the highest mosquito density was 8.9 in livestock sheds, followed by 6.8 in parks, 5.9 in rural dwellings, 5.5 in urban dwellings, and 5.4 in hospitals. No statistically significant differences were found among different years (H = 1.96, d.f. 2, P = 0.376) and habitats (H = 0.45, d.f. 4, P = 0.978). Overall, the peak period of mosquito activity fell in the months from June to September. The peak period among 3 years differed significantly (F(2,7022) = 119.17, P < 0.01), but there were no statistically significant differences in peak period among different habitats (F(4,7020) = −159.09, P > 0.05). Conclusion: Circular statistics could be effectively combined with statistical charts to elucidate the peak period of mosquitoes and determine the differences in statistics among different years and habitats. These findings will provide valuable information for mosquito control and public health management. [ABSTRACT FROM AUTHOR]

Wolbachia continue to be reported in species previously thought to lack them, particularly Aedes aegypti mosquitoes. The presence of Wolbachia in this arbovirus vector is considered important because releases of mosquitoes with transinfected Wolbachia are being used around the world to suppress pathogen transmission and these efforts depend on a lack of Wolbachia in natural populations of this species. We previously assessed papers reporting Wolbachia in natural populations of Ae. aegypti and found little evidence that seemed convincing. However, since our review, more and more papers are emerging on Wolbachia detections in this species. Our purpose here is to evaluate these papers within the context of criteria we previously established but also new criteria that include the absence of releases of transinfections within the local areas being sampled which has contaminated natural populations in at least one case where novel detections have been reported. We also address the broader issue of Wolbachia detection in other insects where similar issues may arise which can affect overall estimates of this endosymbiont more generally. We note continuing shortcomings in papers purporting to find natural Wolbachia in Ae. aegypti which are applicable to other insects as well. [ABSTRACT FROM AUTHOR]

Malaria drug research and development efforts have resurged in the last decade following the decelerating rate of mortality and malaria cases in endemic regions. The inefficiency of malaria interventions is largely driven by the spreading resistance of the Plasmodium falciparum parasite to current drug regimens and that of the malaria vector, the Anopheles mosquito, to insecticides. In response to the new eradication agenda, drugs that act by breaking the malaria transmission cycle (transmission-blocking drugs), which has been recognized as an important and additional target for intervention, are being developed. These drugs take advantage of the susceptibility of Plasmodium during population bottlenecks before transmission (gametocytes) and in the mosquito vector (gametes, zygotes, ookinetes, oocysts, sporozoites). To date, compounds targeting stage V gametocytes predominate in the chemical library of transmission-blocking drugs, and some of them have entered clinical trials. The targeting of Plasmodium mosquito stages has recently renewed interest in the development of innovative malaria control tools, which hold promise for the application of compounds effective at these stages. In this review, we highlight the major achievements and provide an update on the research of transmission-blocking drugs, with a particular focus on their chemical scaffolds, antiplasmodial activity, and transmission-blocking potential. [ABSTRACT FROM AUTHOR]

Mosquitoes are prime examples of vectors for various diseases such as malaria, the West Nile virus, elephantiasis, dengue fever, and yellow fever. The repeated use of chemical pesticides has created numerous obstacles and environmental risks including mosquito resistance to insecticides. This work aimed at assessing the larvicidal effect of chlorophyllin (a water-soluble substance obtained after removing the phytol tail from chlorophyll) and coumarin as photosensitizers, compared to the microbial insecticide Bacillus thuringiensis israelensis (Bti) and permethrin, a chemical insecticide, against the third larval instar of Culex pipiens larvae monitoring its mode of action and its genotoxic effect. Photosensitizers exposed to sunlight generate reactive oxygen species (ROS), with singlet oxygen (¹O2) capable of killing parasitic organisms primarily in aquatic systems. Our experiments demonstrated a high potential activity of Na-Cu chlorophyllin against mosquito larvae after at least 8 hours of incubation in the dark. The LC50 values were 0.22x10-3, 0.96x10-2, 13.7x10-2, and 4.59x10-1 mg/ l after 24 hours of exposure to chlorophyllin, Bti, permethrin, and coumarin, respectively. The molecular changes after treatment with chlorophyllin were tracked using RAPD-PCR with six arbitrary DNA primers. Results confirmed no significant changes or genetic damage after treatment with chlorophyll derivatives. [ABSTRACT FROM AUTHOR]

Simple Summary: This study investigates the factors influencing local populations of Aedes albopictus, a highly invasive mosquito species known for its role in spreading vector-borne diseases. Aimed at enhancing vector control effectiveness, the research focused on understanding the interplay between meteorological elements, human activity, and pest management efforts in determining mosquito abundance at local scales. Conducted over a study season at a botanical garden, the investigation utilized weekly BG-Trap data to observe adult Ae. albopictus dynamics. Results indicated a significant seasonal trend affected by temperature and modulated by rainfall events responsible for the observed bimodal abundance pattern. Additionally, nearby stagnant water and human presence were positively linked to mosquito abundance, while larvicide effectiveness varied with the preceding rainfall levels and application timing. Given the potential for global warming to shift weather patterns and increase the frequency of extreme events, these findings underscore the importance of integrating fine-scale ecological parameters with broader climatic trends in mosquito abatement strategies. This comprehensive approach could enhance the efficacy of interventions and mitigate the impact of mosquito-borne illnesses. Mosquito species, including the Asian tiger mosquito, can transmit disease-causing pathogens such as dengue, Zika, and chikungunya, with their population dynamics influenced by a variety of factors including climate shifts, human activity, and local environmental conditions. Understanding these dynamics is vital for effective control measures. Our study, conducted in Jardí Botanic Marimurtra from May to November 2021, monitored Ae. albopictus activity using BG-Traps and investigated larval control effects. We employed Generalized Linear Mixed Models to analyze variables like weather, human presence, and larvicidal control on adult mosquito abundance. Adults of Ae. albopictus exhibited a seasonal pattern influenced by temperature but with bimodal peaks linked to cumulative rainfall. Proximity to stagnant water and visitor influx directly affected mosquito captures. Additionally, the effectiveness of larvicide treatments depended on interactions between preceding rainfall levels and treatment timing. Our research emphasizes the significance of studying vector ecology at local scales to enhance the efficacy of control programs and address the escalating burden of vector-borne diseases. Considering the impacts of extreme weather events and climate shifts is essential for the development of robust vector control strategies. Furthermore, our distinct findings serve as a prime illustration of utilizing statistical modeling to gain mechanistic insights into ecological patterns and processes. [ABSTRACT FROM AUTHOR]

Simple Summary: The present study examined whether extracts of hemp leaves were toxic to Aedes aegypti larvae and determined which compound(s) were responsible for the toxicity. We found that larvae, from both insecticide-resistant and -susceptible strains were killed by hemp leaf extract within 48 h of exposure. Furthermore, we found that an abundant cannabinoid (cannabidiol) within the extract was the primary active compound. This study suggests that hemp extracts and cannabidiol are potentially valuable sources for developing biopesticides to control mosquitoes. To mitigate pyrethroid resistance in mosquito vectors of emerging and re-emerging human pathogens, there is an urgent need to discover insecticides with novel modes of action. Natural alternatives, such as extracts derived from plants, may serve as substitutes for traditional synthetic insecticides if they prove to be sustainable, cost-effective, and safe for non-target organisms. Hemp (Cannabis sativa) is a sustainable plant known to produce various secondary metabolites with insecticidal properties, including terpenoids and flavonoids. The goal of this study was to assess the larvicidal activity of hemp leaf extract on mosquito larvae from both pyrethroid-susceptible (PS) and pyrethroid-resistant (PR) strains of Aedes aegypti. Another goal was to identify which components of the extract were responsible for any observed larvicidal activity. We found that a methanol extract of hemp leaves induced similar concentration-dependent larvicidal activity against PS (LC50: 4.4 ppm) and PR (LC50: 4.3 ppm) strains within 48 h. Partitioning of the leaf extract between methanol and hexane fractions revealed that full larvicidal activity was restricted to the methanol fraction. Analysis of this fraction by gas chromatography–mass spectrometry and nuclear magnetic resonance showed it to be dominated by cannabidiol (CBD). Larvicidal assays using authentic CBD confirmed this compound was primarily responsible for the toxicity of the hemp leaf extract against both strains. We conclude that hemp leaf extracts and CBD have the potential to serve as viable sources for the development of novel mosquito larvicides. [ABSTRACT FROM AUTHOR]

Introduction: Widespread transmission of Japanese encephalitis virus (JEV) genotype four (GIV) occurred across mainland Australia in 2022. This resulted in forty-five human cases, including seven deaths, and the identification of JEV infection in over 80 commercial piggeries. Materials and Methods: We collected mosquitoes which were trapped using CO2-baited light traps deployed near piggeries reporting disease or in regions linked to human cases in the Wide Bay region in the state of Queensland. Mosquitoes from four traps yielded JEV RNA by real-time RT-PCR. Pools containing RNA positive mosquitoes were inoculated onto mosquito cell monolayers. Discussion: A single isolate of JEV was obtained from a pool of mixed mosquito species. Near whole genome sequencing and phylogenetic analysis of the JEV isolate demonstrated its high genomic relatedness with JEV GIV pig sequences sampled from Queensland and the state of New South Wales in 2022. Conclusion: We report the first isolation of JEV GIV from mosquitoes collected in Australia. With only a few JEV GIV isolates available globally, the isolate we report will be essential for future research of JEV host interactions, evolution and disease markers, and development of effective therapies, vaccines, diagnostic assays, and mosquito control strategies. [ABSTRACT FROM AUTHOR]

In a previous study to understand how the chikungunya virus (CHIKV) E1 glycoprotein β-strand c functions, we identified several attenuating variants at E1 residue V80 and the emergence of second-site mutations in the fusion loop (E1-M88L) and hinge region (E1-N20Y) with the V80 variants in vivo. The emergence of these mutations led us to question how changes in E1 may contribute to CHIKV infection at the molecular level. Here, we use molecular dynamics to understand how changes in the E1 glycoprotein may influence the CHIKV glycoprotein E1-E2 complex. We found that E1 domain II variants lead to E2 conformational changes, allowing us to hypothesize that emerging variants E1-M88L and E1-N20Y could also change E2 conformation and function. We characterized CHIKV E1-M88L and E1-N20Y in vitro and in vivo to understand how these regions of the E1 glycoprotein contribute to host-specific infection. We found that CHIKV E1-N20Y enhanced infectivity in mosquito cells, while the CHIKV E1-M88L variant enhanced infectivity in both BHK-21 and C6/36 cells and led to changes in viral cholesterol-dependence. Moreover, we found that E1-M88L and E1-N20Y changed E2 conformation, heparin binding, and interactions with the receptor Mxra8. Interestingly, the CHIKV E1-M88L variant increased replication in Mxra8-deficient mice compared to WT CHIKV, yet was attenuated in mouse fibroblasts, suggesting that residue E1-M88 may function in a cell-type-dependent entry. Taken together, these studies show that key residues in the CHIKV E1 domain II and hinge region function through changes in E1-E2 dynamics to facilitate cell- and host-dependent entry. IMPORTANCE Arboviruses are significant global public health threats, and their continued emergence around the world highlights the need to understand how these viruses replicate at the molecular level. The alphavirus glycoproteins are critical for virus entry in mosquitoes and mammals, yet how these proteins function is not completely understood. Therefore, it is critical to dissect how distinct glycoprotein domains function in vitro and in vivo to address these gaps in our knowledge. Here, we show that changes in the CHIKV E1 domain II and hinge alter E2 conformations leading to changes in virus-receptor and -glycosaminoglycan interactions and cell-specific infection. These results highlight that adaptive changes in E1 can have a major effect on virus attachment and entry, furthering our knowledge of how alphaviruses infect mammals and insects. [ABSTRACT FROM AUTHOR]

Machine learning methods have seen increased application to geospatial environmental problems, such as precipitation nowcasting, haze forecasting, and crop yield prediction. However, many of the machine learning methods applied to mosquito population and disease forecasting do not inherently take into account the underlying spatial structure of the given data. In our work, we apply a spatially aware graph neural network model consisting of GraphSAGE layers to forecast the presence of West Nile virus in Illinois, to aid mosquito surveillance and abatement efforts within the state. More generally, we show that graph neural networks applied to irregularly sampled geospatial data can exceed the performance of a range of baseline methods including logistic regression, XGBoost, and fully‐connected neural networks. Plain Language Summary: Many machine learning methods have been applied to geospatial data, that is data that has a spatial dimension and corresponds to particular regions on Earth. However, many of these methods do not account for the fact that data recorded at nearby locations will be correlated. We apply a deep learning method called a graph neural network to overcome this issue. Our application to West Nile virus forecasting in Illinois could aid mosquito surveillance and abatement efforts within the state. This shows that graph neural networks could be a good option for other geospatial data, since they outperform a range of baseline methods in our particular problem. Key Points: Many machine learning methods applied to environmental problems do not account for spatial dependenceWe apply a spatially aware graph neural network model to forecast West Nile virusGraph neural networks applied to irregularly sampled geospatial data can outperform a range of baseline methods [ABSTRACT FROM AUTHOR]

Abstract Background Mosquito host feeding patterns are an important factor of the species-specific vector capacity determining pathogen transmission routes. Culex pipiens s.s./Cx. torrentium are competent vectors of several arboviruses, such as West Nile virus and Usutu virus. However, studies on host feeding patterns rarely differentiate the morphologically indistinguishable females. Methods We analyzed the host feeding attraction of Cx. pipiens and Cx. torrentium in host-choice studies for bird, mouse, and a human lure. In addition, we summarized published and unpublished data on host feeding patterns of field-collected specimens from Germany, Iran, and Moldova from 2012 to 2022, genetically identified as Cx. pipiens biotype pipiens, Cx. pipiens biotype molestus, Cx. pipiens hybrid biotype pipiens × molestus, and Cx. torrentium, and finally put the data in context with similar data found in a systematic literature search. Results In the host-choice experiments, we did not find a significant attraction to bird, mouse, and human lure for Cx. pipiens pipiens and Cx. torrentium. Hosts of 992 field-collected specimens were identified for Germany, Iran, and Moldova, with the majority determined as Cx. pipiens pipiens, increasing the data available from studies known from the literature by two-thirds. All four Culex pipiens s.s./Cx. torrentium taxa had fed with significant proportions on birds, humans, and nonhuman mammals. Merged with the data from the literature from 23 different studies showing a high prevalence of blood meals from birds, more than 50% of the blood meals of Cx. pipiens s.s. were identified as birds, while up to 39% were human and nonhuman mammalian hosts. Culex torrentium fed half on birds and half on mammals. However, there were considerable geographical differences in the host feeding patterns. Conclusions In the light of these results, the clear characterization of the Cx. pipiens s.s./Cx. torrentium taxa as ornithophilic/-phagic or mammalophilic/-phagic needs to be reconsidered. Given their broad host ranges, all four Culex taxa could potentially serve as enzootic and bridge vectors. Graphical Abstract

Abstract Background Aedes aegypti, the primary vector of dengue, chikungunya, and Zika viruses, poses a significant public health threat worldwide. Traditional control methods using insecticides are increasingly challenged by resistance and environmental concerns. The sterile insect technique (SIT) offers an eco-friendly alternative that has been successfully applied to other insect pests. This article aims to briefly review Ae. aegypti management in Cuba, highlighting the accomplishments, challenges, and future directions of the SIT. Main body Here we provide a brief summary of the extensive history of Ae. aegypti control efforts in Cuba. After a successful eradication campaign in the 1980s, a resurgence of dengue cases has been observed in recent years, suggesting that traditional control methods may have limited effectiveness under current conditions. In response, Cuba initiated a phased approach to develop and evaluate the feasibility of SIT for Ae. aegypti control, starting in 2008. Initial research focused on Ae. aegypti mating behavior and sterilization methods, followed by successful laboratory and semi-field trials that demonstrated population suppression. The first open-field trial in 2020 confirmed the efficacy of the SIT in reducing Ae. aegypti populations under real-world conditions. Currently, the research is in a phase involving a cluster-randomized superiority-controlled trial. This planned trial will compare the standard vector control program with the same program augmented by the SIT, aiming to assess the impact of the SIT on dengue incidence as the primary outcome. Implementing robust epidemiological trials to evaluate the effectiveness of the SIT is complex due to potential spillover effects from mosquito and human movement across study areas. Additionally, conducting the SIT requires significant development and operational investments. Despite these challenges, the ongoing Cuban trial holds promise for establishing the SIT as an effective and sustainable tool for Ae. aegypti control and for reducing the burden of mosquito-borne diseases. Conclusions The phased evaluation conducted in Cuba confirms the efficacy of the SIT against Ae. aegypti, highlighting its potential for sustainable mosquito-borne disease management. The effective implementation of multi-site trials will be crucial in providing evidence of the potential of the sterile insect technique as part of a strategy to reduce the incidence of arboviral diseases. Graphical Abstract

Abstract Background Rift Valley fever virus (RVFV) is a zoonotic mosquito-borne virus with serious implications for livestock health, human health, and the economy in Africa, and is suspected to be endemic in north-eastern KwaZulu-Natal (KZN), South Africa. The vectors of RVFV in this area are poorly known, although several species, such as Aedes (Neomelaniconion) mcintoshi, Aedes (Neomelaniconion) circumluteolus, Aedes (Aedimorphus) durbanensis, and Culex (Lasioconops) poicilipes may be involved. The aim of the study was to determine the vertebrate blood meal sources of potential RVFV mosquito vectors in north-eastern KZN and to characterize the host-biting network. Methods Blood-fed mosquitoes were collected monthly from November 2019 to February 2023 using a backpack aspirator, CO2-baited Centers for Disease Control and Prevention (CDC) miniature light traps and tent traps, in the vicinity of water bodies and livestock farming households. The mosquitoes were morphologically identified. DNA was extracted from individual mosquitoes and used as templates to amplify the vertebrate cytochrome c oxidase I (COI) and cytochrome b (cytb) genes using conventional polymerase chain reaction (PCR). Amplicons were sequenced and queried in GenBank and the Barcode of Life Data systems to identify the vertebrate blood meal sources and confirm mosquito identifications. All mosquitoes were screened for RVFV using real time reverse transcription (RT)-PCR. Results We identified the mammalian (88.8%) and avian (11.3%) blood meal sources from 409 blood-fed mosquitoes. Aedes circumluteolus (n = 128) made up the largest proportion of collected mosquitoes. Cattle (n = 195) and nyala (n = 61) were the most frequent domestic and wild hosts, respectively. Bipartite network analysis showed that the rural network consisted of more host-biting interactions than the reserve network. All mosquitoes tested negative for RVFV. Conclusions Several mosquito species, including Ae. circumluteolus, and vertebrate host species, including cattle and nyala, could play a central role in RVFV transmission. Future research in this region should focus on these species to better understand RVFV amplification. Graphical Abstract

ObjectiveTo evaluate the field mosquito surveillance efficacy between the black box method and the light trap method.MethodsEight light traps and eight black boxes were placed in each of the eight field sites in Baoshan District of Shanghai, with a minimum distance of 10 meters between each traps. Surveillance was conducted twice a month from May to September 2023.ResultsThe total number of mosquitoes (273) and female mosquitoes (228) captured by the black box method were 0.795 and 0.774 times higher than those caught by the light trap method, respectively. There was no statistically significant difference (P>0.05) between the mosquito density (t=1.723 5, P=0.100 2) and the female mosquito density (t=1.805 9, P=0.101 1), and there was a correlation (P0.05) and female mosquitoes(χ2=0.394 4, P>0.05) captured by black box and light trap methods. In four different habitats, the composition ratios of mosquito species captured by the black box and light trap methods were: hospitals (38.10% and 38.77%) > rural housing areas (36.26% and 37.61%) > parks (14.65% and 12.54%)> urban housing areas (10.99% and 11.08%), and the composition ratios of female mosquitoes captured by the black box and light trap methods were: rural housing areas (40.35% and 42.52%) > hospitals (37.72% and 36.05%) > parks (13.16% and 13.27%) > urban housing areas (8.77% and 8.16%). In different habitats, there was no significant difference in the composition ratio of mosquitoes captured by two monitoring methods(χ2=0.5987, P>0.05), and there was no significant difference in the composition ratio of female mosquitoes(χ2=0.2993, P>0.05).ConclusionWith widen application and fewer requirements for equipment, operation skills, and use cost the black box method can be interchanged with the light trap method for monitoring mosquito density, especially in disaster areas or remote areas with difficult in having access to electricity.

Abstract Background Analyses of the temporal distribution of mosquitoes are presented in statistical charts, but it is difficult to prove in statistics whether differences in peak periods exist among different years or habitats. This study aimed to investigate the application of circular statistics in determining the peak period and a comparison of differences. Methods Surveillance of adult mosquitoes was conducted twice a month by light traps in five different habitats from March to November for 3 years (2021–2023) in Qingdao, Shandong Province, China. The Kruskal–Wallis test was performed to determine the differences in mosquito density among different years and habitats. Circular statistics and line charts were employed to determine the peak period and a comparison of differences. Results Among a total of 14,834 adult mosquitoes comprising five mosquito species from four genera, Culex pipiens pallens was dominant and accounted for 89.6% of the specimens identified. Aedes albopictus, Armigeres subalbatus, and Anopheles sinensis made up 5.7%, 4.2%, and 0.5%, respectively. Culex tritaeniorhynchus accounted for less than 0.1%. The mean mosquito density (females/trap night) for the trapping period was 10.3 in 2021, 5.6 in 2022, and 3.6 in 2023. Among five habitats, the highest mosquito density was 8.9 in livestock sheds, followed by 6.8 in parks, 5.9 in rural dwellings, 5.5 in urban dwellings, and 5.4 in hospitals. No statistically significant differences were found among different years (H = 1.96, d.f. 2, P = 0.376) and habitats (H = 0.45, d.f. 4, P = 0.978). Overall, the peak period of mosquito activity fell in the months from June to September. The peak period among 3 years differed significantly (F (2,7022) = 119.17, P 0.05). Conclusion Circular statistics could be effectively combined with statistical charts to elucidate the peak period of mosquitoes and determine the differences in statistics among different years and habitats. These findings will provide valuable information for mosquito control and public health management. Graphical Abstract

Knowledge of insect dispersal is relevant to the control of agricultural pests, vector-borne transmission of human and veterinary pathogens, and insect biodiversity. Previous studies in a malaria endemic area of the Sahel region in West Africa revealed high-altitude, long-distance migration of insects and various mosquito species. The objective of the current study was to assess whether similar behavior is exhibited by mosquitoes and other insects around the Lake Victoria basin region of Kenya in East Africa. Insects were sampled monthly from dusk to dawn over 1 year using sticky nets suspended on a tethered helium-filled balloon. A total of 17,883 insects were caught on nets tethered at 90, 120, and 160 m above ground level; 818 insects were caught in control nets. Small insects (0.5 cm, n = 2,334) and mosquitoes (n = 299). Seven orders were identified; dipteran was the most common. Barcoding molecular assays of 184 mosquitoes identified 7 genera, with Culex being the most common (65.8%) and Anopheles being the least common (5.4%). The survival rate of mosquitoes, experimentally exposed to high-altitude overnight, was significantly lower than controls maintained in the laboratory (19% vs. 85%). There were no significant differences in mosquito survival and oviposition rate according to capture height. These data suggest that windborne dispersal activity of mosquito vectors of malaria and other diseases occurs on a broad scale in sub-Saharan Africa.

Abstract Background Female Aedes aegypti mosquitoes can spread disease-causing pathogens when they bite humans to obtain blood nutrients required for egg production. Following a complete blood meal, host-seeking is suppressed until eggs are laid. Neuropeptide Y-like receptor 7 (NPYLR7) plays a role in endogenous host-seeking suppression and previous work identified small-molecule NPYLR7 agonists that inhibit host-seeking and blood-feeding when fed to mosquitoes at high micromolar doses. Methods Using structure–activity relationship analysis and structure-guided design we synthesized 128 compounds with similarity to known NPYLR7 agonists. Results Although in vitro potency (EC50) was not strictly predictive of in vivo effect, we identified three compounds that reduced blood-feeding from a live host when fed to mosquitoes at a dose of 1 μM—a 100-fold improvement over the original reference compound. Conclusions Exogenous activation of NPYLR7 represents an innovative vector control strategy to block mosquito biting behavior and prevent mosquito–human host interactions that lead to pathogen transmission. Graphical Abstract

Abstract Background Cache Valley virus (CVV) is an understudied Orthobunyavirus with a high spillover transmission potential due to its wide geographical distribution and large number of associated hosts and vectors. Although CVV is known to be widely distributed throughout North America, no studies have explored its geography or employed computational methods to explore the mammal and mosquito species likely participating in the CVV sylvatic cycle. Methods We used a literature review and online databases to compile locality data for CVV and its potential vectors and hosts. We linked location data points with climatic data via ecological niche modeling to estimate the geographical range of CVV and hotspots of transmission risk. We used background similarity tests to identify likely CVV mosquito vectors and mammal hosts to detect ecological signals from CVV sylvatic transmission. Results CVV distribution maps revealed a widespread potential viral occurrence throughout North America. Ecological niche models identified areas with climate, vectors, and hosts suitable to maintain CVV transmission. Our background similarity tests identified Aedes vexans, Culiseta inornata, and Culex tarsalis as the most likely vectors and Odocoileus virginianus (white-tailed deer) as the most likely host sustaining sylvatic transmission. Conclusions CVV has a continental-level, widespread transmission potential. Large areas of North America have suitable climate, vectors, and hosts for CVV emergence, establishment, and spread. We identified geographical hotspots that have no confirmed CVV reports to date and, in view of CVV misdiagnosis or underreporting, can guide future surveillance to specific localities and species. Graphical Abstract

Abstract Background Salinity, exacerbated by rising sea levels, is a critical environmental cue affecting freshwater ecosystems. Predicting ecosystem structure in response to such changes and their implications for the geographical distribution of arthropod disease vectors requires further insights into the plasticity and adaptability of lower trophic level species in freshwater systems. Our study investigated whether populations of the mosquito Culex pipiens, typically considered sensitive to salt, have adapted due to gradual exposure. Methods Mesocosm experiments were conducted to evaluate responses in life history traits to increasing levels of salinity in three populations along a gradient perpendicular to the North Sea coast. Salt concentrations up to the brackish–marine transition zone (8 g/l chloride) were used, upon which no survival was expected. To determine how this process affects oviposition, a colonization experiment was performed by exposing the coastal population to the same concentrations. Results While concentrations up to the currently described median lethal dose (LD50) (4 g/l) were surprisingly favored during egg laying, even the treatment with the highest salt concentration was incidentally colonized. Differences in development rates among populations were observed, but the influence of salinity was evident only at 4 g/l and higher, resulting in only a 1-day delay. Mortality rates were lower than expected, reaching only 20% for coastal and inland populations and 41% for the intermediate population at the highest salinity. Sex ratios remained unaffected across the tested range. Conclusions The high tolerance to salinity for all key life history parameters across populations suggests that Cx. pipiens is unlikely to shift its distribution in the foreseeable future, with potential implications for the disease risk of associated pathogens. Graphical Abstract

Abstract Background Aedes aegypti is the primary mosquito vector for several arboviruses, such as dengue, chikungunya and Zika viruses, which cause frequent outbreaks of human disease in tropical and subtropical regions. Control of these outbreaks relies on vector control, commonly in the form of insecticide sprays that target adult female mosquitoes. However, the spatial coverage and frequency of sprays needed to optimize effectiveness are unclear. In this study, we characterize the effect of ultra-low-volume (ULV) indoor spraying of pyrethroid insecticides on Ae. aegypti abundance within households. We also evaluate the effects of spray events during recent time periods or in neighboring households. Improved understanding of the duration and distance of the impact of a spray intervention on Ae. aegypti populations can inform vector control interventions, in addition to modeling efforts that contrast vector control strategies. Methods This project analyzes data from two large-scale experiments that involved six cycles of indoor pyrethroid spray applications in 2 years in the Amazonian city of Iquitos, Peru. We developed spatial multi-level models to disentangle the reduction in Ae. aegypti abundance that resulted from (i) recent ULV treatment within households and (ii) ULV treatment of adjacent or nearby households. We compared fits of models across a range of candidate weighting schemes for the spray effect, based on different temporal and spatial decay functions to understand lagged ULV effects. Results Our results suggested that the reduction of Ae. aegypti in a household was mainly due to spray events occurring within the same household, with no additional effect of sprays that occurred in neighboring households. Effectiveness of a spray intervention should be measured based on time since the most recent spray event, as we found no cumulative effect of sequential sprays. Based on our model, we estimated the spray effect is reduced by 50% approximately 28 days after the spray event. Conclusions The reduction of Ae. aegypti in a household was mainly determined by the number of days since the last spray intervention in that same household, highlighting the importance of spray coverage in high-risk areas with a spray frequency determined by local viral transmission dynamics. Graphical abstract

Abstract 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. Graphical Abstract