Search

Mosquitoes are the most common disease vectors worldwide. In coastal cities, the spread, activity, and longevity of vector mosquitoes are influenced by environmental factors such as temperature, humidity, and rainfall, which affect their geographic distribution, biting rates, and lifespan. We examined mosquito abundance and species composition before and after Hurricane Irma in Miami, Dade County, Florida, and identified which mosquito species predominated post-Hurricane Irma. Our results showed that mosquito populations increased post-Hurricane Irma: 7.3 and 8.0 times more mosquitoes were captured in 2017 than at baseline, 2016 and 2018 respectively. Warmer temperatures accelerated larval development, resulting in faster emergence of adult mosquitoes. In BG-Sentinel traps, primary species like Ae. tortills, Cx. nigripalpus, and Cx. quinquefasciatus dominated the post-Hurricane Irma period. Secondary vectors that dominated post-Hurricane Irma include An. atropos, An. crucians, An. quadrimaculatus, Cx. erraticus, and Ps. columbiae. After Hurricane Irma, the surge in mosquito populations in Miami, Florida heightened disease risk. To mitigate and prevent future risks, we must enhance surveillance, raise public awareness, and implement targeted vector control measures., (© 2024. The Author(s).)

Mosquito vectors of pathogens (e.g., Aedes, Anopheles, and Culex spp. which transmit dengue, Zika, chikungunya, West Nile, malaria, and others) are of increasing concern for global public health. These vectors are geographically shifting under climate and other anthropogenic changes. As small-bodied ectotherms, mosquitoes are strongly affected by temperature, which causes unimodal responses in mosquito life history traits (e.g., biting rate, adult mortality rate, mosquito development rate, and probability of egg-to-adult survival) that exhibit upper and lower thermal limits and intermediate thermal optima in laboratory studies. However, it remains unknown how mosquito thermal responses measured in laboratory experiments relate to the realized thermal responses of mosquitoes in the field. To address this gap, we leverage thousands of global mosquito occurrences and geospatial satellite data at high spatial resolution to construct machine-learning based species distribution models, from which vector thermal responses are estimated. We apply methods to restrict models to the relevant mosquito activity season and to conduct ecologically plausible spatial background sampling centered around ecoregions for comparison to mosquito occurrence records. We found that thermal minima estimated from laboratory studies were highly correlated with those from the species distributions (r = 0.87). The thermal optima were less strongly correlated (r = 0.69). For most species, we did not detect thermal maxima from their observed distributions so were unable to compare to laboratory-based estimates. The results suggest that laboratory studies have the potential to be highly transportable to predicting lower thermal limits and thermal optima of mosquitoes in the field. At the same time, lab-based models likely capture physiological limits on mosquito persistence at high temperatures that are not apparent from field-based observational studies but may critically determine mosquito responses to climate warming. Our results indicate that lab-based and field-based studies are highly complementary; performing the analyses in concert can help to more comprehensively understand vector response to climate change., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Athni et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

In the last 2 decades, there has been an increase in the geographic range and frequency of vector-borne diseases. Management of mosquito populations has become challenging due to increasing rates of resistance to existing insecticidal products and formulations. Several alternative tools have emerged to suppress or replace mosquito populations. One of these tools is the In2Care Mosquito Station (In2Care station). This dual-action station contains the insect growth regulator pyriproxyfen which disrupts the development of immatures and the entomopathogenic fungus Beauveria bassiana (B. bassiana) strain GHA which kills exposed adult mosquitoes. The In2Care stations have previously been shown to effectively control Aedes aegypti in field settings at a density of 6 stations/acre rather than the label-recommended 10 stations/acre. To further test the efficacy of low station density deployment, we deployed In2Care stations in the Pleasant Street Historic District of Gainesville, Florida, at a density of 3 stations/acre over a period of 2 years in the presence or absence of ground larvicidal applications. The deployment of stations resulted in no measurable impact on Ae. aegypti and Culex quinquefasciatus adult or immature abundance suggesting that the low-density deployment of In2Care stations is insufficient to reduce Ae. aegypti and Cu. quinquefasciatus abundance within treatment areas., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The northern house mosquito, Culex pipiens, employs diapause as an essential survival strategy during winter, inducing important phenotypic changes such as enhanced stress tolerance, lipid accumulation, and extended longevity. During diapause, the cessation of reproductive development represents another distinctive phenotypic change, underlining the need for adjusted modulation of gene expressions within the ovary. Although considerable advancements in screening gene expression profiles in diapausing and non-diapausing mosquitoes, there remains a gap in tissue-specific transcriptomic profiling that could elucidate the complicated formation of diverse diapause features in Cx. pipiens. Here, we filled this gap by utilizing RNA sequencing, providing a detailed examination of gene expression patterns in the fat body and ovary during diapause compared to non-diapause conditions. Functional annotation of upregulated genes identified associations with carbohydrate metabolism, stress tolerance, immunity, and epigenetic regulation. The validation of candidate genes using quantitative real-time PCR verified the differentially expressed genes identified in diapausing mosquitoes. Our findings contribute novel insights into potential regulators during diapause in Cx. pipiens, thereby opening possible avenues for developing innovative vector control strategies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The synthesized pyrazolopyrimidine derivatives conjugated with selenium nanoparticles were prepared via a reaction of pyrazolone 1 with aryl-aldehyde and malononitrile or 3-oxo-3-phenylpropanenitrile in the presence ammonium acetate or pipridine using an ultrasonic bath as a modified method in the organic synthesis for such materials. The structure of the synthesized compounds was elucidated through various techniques. All the synthesized pyrazolopyrimidines were used in the synthesis of selenium nanoparticles (SeNPs). These nanoparticles were confirmed using UV-spectra, Dynamic Light scattering and (TEM) techniques. The larvicidal efficiency;of the synthesized;compounds; was investigated against some strains such as Culex pipiens;and Musca domestica larvae. Bioassay test showed pyrazolopyrimide derivatives to exhibit an acceptable larvicidal;bio-efficacy. The derivative (3) exhibited;the highest;efficiency for more than; lab strains of both species. Moreover, C. pipiens larvae were more sensitive towards the examined compounds than M. domestica. The field;strain displayed lower affinity for the 2 folds compounds. Some biochemical changes were tracked through analysis of insect main metabolites (protein, lipid and carbohydrate), in addition to measuring the changes in seven enzymes after treatment. Generally, there was a reduction in the protein, lipids and carbohydrates after treatment with all tested compounds. Moreover, a decrement was noticed for acetylcholine esterase and glutathione;S-transferase; enzymes. There was an increment in the acid;phosphatase; and alkaline phosphatase. In addition, there was elevation in Phenoloxidase level but it noticed the declination in both Cytochrome P450 and Ascorbate peroxidase activity after treatment both flies with derivatives of selenium-nanoparticles in both lab and field strain. Generally, the experiments carried out indicate that antioxidant and detoxification enzymes may play a significant role in mechanism of action of our novel nanocompounds. The cytotoxicity of the synthesized compounds and conjugated with SeNPs showed enhanced compatibility with human normal fibroblast cell line (BJ1) with no toxic effect., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Insects use seasonal diapause as an alternative strategy to endure adverse seasons. This developmental trajectory is induced by environmental cues like short-day lengths in late summer and early fall, but how insects measure day length is unknown. The circadian clock has been implicated in regulating photoperiodic or seasonal responses in many insects, including the Northern house mosquito, Culex pipiens, which enters adult diapause. To investigate the potential control of diapause by circadian control, we employed ChIP-sequencing to identify the downstream targets of a circadian transcription factor, PAR domain protein 1 (PDP1), that contribute to the hallmark features of diapause. We identified the nearest genes in a 10 kb region of the anticipated PDP1 binding sites, listed prospective targets and searched for PDP1-specific binding sites. By examining the functional relevance to diapause-specific behaviours and modifications such as metabolic pathways, lifespan extension, cell cycle regulation and stress tolerance, eight genes were selected as targets and validated using ChIP-qPCR. In addition, qRT-PCR demonstrated that the mRNA abundance of PDP1 targets increased in the heads of diapausing females during the middle of the scotophase (ZT17) compared with the early photophase (ZT1), in agreement with the peak and trough of PDP1 abundance. Thus, our investigation uncovered the mechanism by which PDP1 might generate a diapause phenotype in insects., (© 2024 The Author(s). Insect Molecular Biology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.)

Globalisation, climate change and international trade are the factors contributing to the spread of Aedes albopictus (Diptera: Culicidae) and Ae. aegypti into new areas. In newly invaded habitats, these non-native species can serve as arbovirus disease vectors or increase the risk of disease spill over. These mosquitoes continue to emerge in new areas where they have or will have overlapping ranges with other resident mosquito species. The study investigates how invasive Aedes mosquitoes compete with the native Culex pipiens in Türkiye, which might affect the overall mosquito population dynamics and disease transmission risks. Both Aedes species exhibited contrasting responses to interspecific competition with Cx. pipiens. While Ae. albopictus suffers reduced emergence primarily in larger containers with abundant food, Ae. aegypti surprisingly thrives in mixed cultures under all food conditions. Adult Cx. pipiens emergence drops by half against Ae. albopictus and under specific conditions with Ae. aegypti. Competition influences mosquito size differently across species and life stages. Culex pipiens females grow larger when competing with Ae. aegypti, potentially indicating resource advantage or compensatory strategies. However, Ae. albopictus size shows more nuanced responses, suggesting complex interactions at play. Understanding how invasive and native mosquitoes interact with each other can provide insights into how they adapt and coexist in shared habitats. This knowledge can inform effective control strategies. The study highlights the differential responses of invasive Aedes species and the potential for managing populations based on their competitive interactions with the native Cx. pipiens. It can contribute to improved monitoring and prediction systems for the spread of invasive mosquitoes and the associated disease risks., (© 2024 The Author(s). Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.)

Mosquitoes, one of the deadliest animals on the planet, cause millions of fatalities each year by transmitting several human illnesses. Synthetic pesticides were previously used to prevent the spread of diseases by mosquitoes, which was effective in protecting humans but caused serious human health problems, environmental damage, and developed mosquito pesticide resistance. This research focuses on exploring new, more effective, safer, and environmentally friendly compounds to improve mosquito vector management. Phytochemicals are possible biological agents for controlling pests and many are target-specific, rapidly biodegradable, and eco-friendly. The potential of extracts of Lantana camara, Melia azedarach, Nerium oleander, Ricinus communis, and Withania somnifera against 3rd instar Culex pipiens (Common house mosquito) larvae was evaluated. Methanol extracts had more toxic effects against Cx. pipiens larvae (95-100%, 24 h post-treatment) than aqueous extracts (63-91%, 24 h post-treatment). The methanol extracts of Nerium oleander (LC 50  = 158.92 ppm) and Ricinus communis (LC 50  = 175.04 ppm) were very effective at killing mosquito larvae, 24 h after treatment. N. oleander (LC 50  = 373.29 ppm) showed high efficacy in aqueous plant extracts. Among the different extracts of the five plants screened, the methanol extract of R. communis recorded the highest ovicidal activity of 5% at 800 ppm concentration. Total developmental duration and growth index were highly affected by R. communis and M. azedarach methanol extracts. In field tests it was clear that plant extracts decreased mosquito larval density, especially when mixed with mosquito Bti briquette, with stability up to seven days for N. oleander. GC-MS results showed that the methanol extract had a higher number of chemical compounds, particularly with more terpene compounds. A high-performance liquid chromatography (HPLC) technique was used to detect the existence of non-volatile polyphenols and flavonoids. All five methanol extracts showed high concentrations of active ingredients such as gallic acid, chlorogenic acid (more than 100 μg/ml) and the rosmarinic acid was also found in all the five extracts in addition to 17 active polyphenols and flavonoids presented at moderate to low concentrations. Molecular modeling of 18 active ingredients detected by the HPLC were performed to the vicinity of one of the fatty acid binding proteins of lm-FABP (PDB code: 2FLJ). Rutin, Caffeic acid, coumaric acid and rosmarinic acid which presented densely in R. communis and N. oleander showed multiple and stable intermolecular hydrogen bonding and π-π stacking interactions. The inhibition ability of the fatty acid binding protein, FABP4, was evaluated with remarkable receptor inhibition evident, especially with R. communis and N. oleander having inhibitory concentrations of IC 50  = 0.425 and 0.599 µg/mL, respectively. The active phytochemical compounds in the plants suggest promising larvicidal and ovicidal activity, and have potential as a safe and effective alternative to synthetic insecticides., (© 2024. The Author(s).)

Failure of conventional mosquito control strategies to curb the population of vectors have made the humans vulnerable to serious medical problems transmitted by them. This effect has been compounded by global climate change enabling the mosquitoes to cross geographical boundaries and cause trouble in regions where they were initially not found. As such, the scientific community has been compelled to devise alternative and innovative strategies of mosquito control that can be integrated with the conventional practices to implement multi-phasic approach of vector management. Culex quinquefasciatus is one such mosquito species that is reported to be one of the primary vectors of lymphatic filariasis and many other diseases of global health concern. However, not much is known about its breeding habitat ecology and microbial properties that have enabled the species to achieve reproductive success in urbanized habitats. The current investigation was carried out at Digha, West Bengal, India. The region, despite being endemic for lymphatic filariasis, has rarely been explored for its mosquito diversity and/or their breeding habitat characteristics. Therefore, these were attempted. For survey and sampling, seven villages were chosen, namely, Duttapur, Jatimati, Champabani, Padima, Gobindabasan, Bhagibaharampur and Palsandapur. The study showed that Cx. quinquefasciatus is the dominant mosquito species at the sampling sites with the highest density of their larvae being recorded from man-made structures like drains and pools close to human habitations and livestock. The study was, therefore, restricted to Cx. quinquefasciatus. Seasonal abundance showed that they were most prevalent in the monsoon followed by summer. The physicochemical characterization showed their larvae to prefer almost neutral pH (6.9 to 7.3), low chloride concentration (98 to 258 ppm) and turbidity. As far as other parameters are concerned, they were tolerant towards a wide range allowing them to adapt varied habitats in the study areas. The bacterial profiling of their natural habitat waters revealed the presence of Paenibacillus nanensis DGX1(OQ690670), Bacillus cereus DGX2(OQ690675), Bacillus sp. DGX3(OQ690700) and Escherichia coli DGX4(OQ690701). Bacillus cereus was found to have high oviposition attractant properties in oviposition assays. Bacillus cereus was also obtained from the midgut of third instar larvae indicating that they had entered from the surrounding medium and colonized the larval gut. Subsequent tests exhibited the roles of B. cereus in larval development. Numerous plant products have been reported either as insecticides for killing larvae or adult mosquitoes or as repellents for mosquito biting and the best alternatives for mosquito control. Larvicidal potential of emulsified neem oil formulation against the field collected 3rd instar larvae of Culex quinquefasciatus mosquito under laboratory conditions was also evaluated. The information thus obtained can be pooled to generate larval source markers and larval source management practices by altering their habitats that cannot be removed. Furthermore, the time of implementation of these strategies can also be planned., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

A researcher may have many reasons for wanting to establish new laboratory colonies from field-collected mosquitoes. In particular, the ability to study the diversity found within and among natural populations in a controlled laboratory environment opens up a wide range of possibilities for understanding how and why burdens of vector-borne disease vary over space and time. However, field-collected mosquitoes are often more difficult to work with than established laboratory strains, and considerable logistical challenges are involved in safely transporting field-collected mosquitoes into the laboratory. Here, we provide advice for researchers working with Aedes aegypti , Anopheles gambiae , and Culex pipiens, as well as notes on other closely related species. We provide guidance on each stage of the life cycle and highlight the life stages for which it is easiest to initiate new laboratory colonies for each species. In accompanying protocols, we provide methods detailing Ae. aegypti egg collection and hatching as well as how to transport larvae and pupae from the field., (© 2024 Cold Spring Harbor Laboratory Press.)

The common house mosquito ( Culex pipiens ) is a native vector for West Nile virus (WNV). Invasive species like the tiger mosquito ( Aedes albopictus ) and Asian bush mosquito ( Aedes japonicus ) are rapidly spreading through Europe, posing a major threat as vectors for dengue, chikungunya (CHIKV), and Japanese encephalitis virus (JEV). These mosquitoes share a similar ecological niche as larvae, but the carry-over effects of aquatic larval interactions to the terrestrial adult stage remain largely unknown and their medical relevance requires further investigation. This study examines the context dependency of larval interactions among Aedes albopictus , Aedes japonicus , and Culex pipiens . The survival, development time, growth, and energetic storage were measured in different European populations within density-response (intraspecific) experiments and replacement (interspecific) experiments at 20 °C and 26 °C. Overall, Ae. japonicus was the weakest competitor, while competition between Ae. albopictus and Cx. pipiens varied with temperature. Adults emerging from this larval competition were infected as follows: Culex pipiens with WNV, Ae. albopictus with CHIKV, and Ae. japonicus with JEV. While no JEV infection was observed, mosquitoes experiencing interspecific interactions during their larval stages exhibited higher infection rates and viral RNA titers for CHIKV and WNV. This increased susceptibility to viral infection after larval competition suggests a higher risk of arbovirus transmission in co-occurring populations.

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 (LD 50 ) (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., (© 2024. The Author(s).)

This study aims to determine the phenological characteristics of thermal responses in the larvae of a Culex pipiens complex field population at the individual level under the influence of thermal regime of its habitat. The analysis is based on a structured population model quantifying the thermal responses of development time and survival under variable conditions and characterising the variety between the larvae (interindividual variety). During the study performed in Turkish Thrace on a monthly basis between May 2021 and June 2022, a total of 3744 larvae were reared as peer larval cohorts and 2330 larvae as siblings in artificial containers to be fully exposed to the natural thermal condition that was recorded hourly. The development process of larvae was monitored daily from egg to adult. As a result, a total of 4788 adult mosquitoes emerged, with a development period ranging from 8 to 52 days in the females and 7 to 50 days in the males, and the survival rate was found to range from 0% to 100%. Both parameters varied by month and individuals, and the variations manifested itself, particularly in the colder periods. The results indicate that the variation between the individuals in terms of thermal response in the larvae of C. pipiens, along with the thermal acclimation ability, appears to be fate determinant in resisting fluctuating thermal regimes, surviving in concurrent climate change and adapting to new conditions with modifications in the seasonal phenology, such as maintaining reproductive dynamics throughout the winter thanks to global warming., (© 2024 Royal Entomological Society.)

The field of bio-fabricated noble metallic nanoparticles (NPs) has gained significant attention in applied research due to their eco-friendly and biocompatible nature. This study focuses on employing a green synthesis method to produce silver and gold nanoparticles (bio-fabricated) using a Mangrove plant extract and assessing their insecticidal and growth-inhibitory effects for environmentally friendly pest control. The resulting NPs underwent comprehensive characterization through various spectroscopy techniques. The morphology of both silver and gold mediated nanoparticles of Avicennia marina leaf extract displayed a spherical shape, with average sizes measuring around 70-80 nm and 95-100 nm, respectively. Regarding cytotoxicity, the inhibitory effects of silver nanoparticles were less than that observed by the extract alone while gold nanoparticles showed stronger cell growth inhibitory effects on splenic cells. The hepatic toxicity of silver and gold nanoparticles showed significant toxic effects as compared to A. marina extract alone. Notably, as prepared silver nanoparticles exhibited substantial larvicidal toxicity as compared to gold nanoparticles, when tested against fourth instar Culex pipiens larvae. These biocompatible silver and gold nanoparticles prepared from A. marina leaf extract hold promise for future applications as larvicides to effectively control mosquito species.

This study investigated the pathogenic potential of Pseudomonas protegens on mosquito larvae of the two species Culex pipiens and Aedes albopictus, representing major threats for disease transmission in the Mediterranean area and worldwide. The bacterium achieved to kill over 90% of the mosquito larvae within 72 h after exposition to a bacterial concentration of 100 million CFU/ml. These lethal effects were concentration dependent and a significantly higher susceptibility was associated with younger larvae of both mosquito species. Significant slowdown of immature (larval and pupal) development and decrease in adult emergence rate after treatment with sub-lethal doses of the bacterium were also detected. This study reports for the first time the insecticidal activity of a root-associated biocontrol bacterium against aquatic mosquito larvae., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Culex quinquefasciatus is a cosmopolitan species widely distributed in the tropical and subtropical areas of the world. Due to its long history of close association with humans, the transmission of arboviruses and parasites have an important role in veterinary and public health. Adult females feed mainly on birds although they can also feed on humans and other mammals. On the other hand, larvae are able to feed on a great diversity of microorganisms, including microalgae, present in natural or artificial breeding sites with a high organic load. These two particularities, mentioned above, are some of the reasons why this mosquito is so successful in the environment. In this work, we report the identification of a microalga found during field sampling in artificial breeding sites, in a group of discarded tires with accumulated rainwater. Surprisingly, only one of them had a bright green culture without mosquito larvae while the other surrounding tires contained a large number of mosquito larvae. We isolated and identified this microorganism as Neochloris aquatica, and it was evaluated as a potential biological control agent against Cx. quinquefasciatus. The oviposition site preference in the presence of the alga by gravid females, and the effects on larval development were analyzed. Additionally, microalga effect on Cx. quinquefasciatus wild type, naturally infected with the endosymbiotic bacterium Wolbachia (w+) and Wolbachia free (w-) laboratory lines was explored. According to our results, even though it is chosen by gravid females to lay their eggs, the microalga had a negative effect on the development of larvae from both populations. Additionally, when the larvae were fed with a culture of alga supplemented with balanced fish food used as control diet, they were not able to reverse its effect, and were unable to complete development until adulthood. Here, N. aquatica is described as a biological agent, and as a potential source of bioactive compounds for the control of mosquito populations important in veterinary and human health., Competing Interests: The authors have declared that no competing interests exist.

Urbanization processes are increasing globally. Anthropogenic alterations in the environment have profound effects on biodiversity. Decreased biodiversity due to biotic homogenization processes as a consequence of urbanization often result in increased levels of mosquito vector species and vector-borne pathogen transmission. Understanding how anthropogenic alterations in the environment will affect the abundance, richness, and composition of vector mosquito species is crucial for the implementation of effective and targeted mosquito control strategies. We hypothesized that anthropogenic alterations in the environment are responsible for increasing the abundance of mosquito species that are adapted to urban environments such as Aedes aegypti and Culex quinquefasciatus. Therefore, our objective was to survey mosquito relative abundance, richness, and community composition in Miami-Dade County, Florida, in areas with different levels of urbanization. We selected 24 areas, 16 remote areas comprised of natural and rural areas, and 8 urban areas comprised of residential and touristic areas in Miami-Dade County, Florida. Mosquitoes were collected weekly in each area for 24 h for 5 consecutive weeks from August to October 2020 using BG-Sentinel traps baited with dry ice. A total of 36,645 mosquitoes were collected, from which 34,048 were collected in the remote areas and 2,597 in the urban areas. Our results show a clear and well-defined pattern of abundance, richness, and community composition according to anthropogenic modifications in land use and land cover. The more urbanized a given area the fewer species were found and those were primary vectors of arboviruses, Ae. aegypti and Cx. quinquefasciatus., (© 2021. The Author(s).)

Culex quinquefasciatus, a member of the Culex pipiens complex, is widespread in Saudi Arabia and other parts of the world. It is a vector for lymphatic filariasis, Rift Valley fever, and West Nile virus. Studies have shown the deleterious effect of RNA interference (RNAi)-mediated knockdown of various lethal genes in model and agricultural pest insects. RNAi was proposed as a tool for mosquito control with a focus on Aedes aegypti and Anopheles gambiae. In this study, we examined the effect of RNAi of selected target genes on both larval mortality and adult emergence of Cx. quinquefasciatus through two delivery methods: soaking and nanoparticles. Ten candidate genes were selected for RNAi based on their known lethal effect in other insects. Disruption of three genes, chitin synthase-1, inhibitor of apoptosis 1, and vacuolar adenosine triphosphatase, resulted in the highest mortality among the selected genes using the two treatment methods. Silencing the other seven genes resulted in a medium to low mortality in both assays. These three genes are also active against a wide range of insects and could be used for RNAi-based mosquito control in the future., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Culex longitubus Somboon, Namgay & Harbach is described as a new species of the Mimeticus Subgroup of the subgenus Culex. The larva is most similar to the larva of Cx. tianpingensis Chen from China, but is distinguished by the length of the siphon and the anal papillae, the form of the comb scales and pecten spines, and the development of setae 7-P, 13-T, 1-X, and 4-X. The adults have wing markings and male genitalia similar to those of species of the Mimeticus Complex. Phylogenetic analysis of COI sequences revealed that the new species is closely related to Cx. murrelli Lien of the Mimulus Complex. The immature stages of the new species were found in stagnant pools and marshes at high altitudes in several districts of Bhutan., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Culex pipiens pallens is an important vector of lymphatic filariasis and epidemic encephalitis. Mosquito control is the main strategy used for the prevention of mosquito-borne diseases. Bacillus thuringiensis israelensis (Bti) is an entomopathogenic bacterium widely used in mosquito control. In this study, we profiled the microbiota and transcriptional response of the larvae of Cx. pipiens pallens exposed to different concentrations of Bti. The results demonstrated that Bti induced a significant effect on both the microbiota and gene expression of Cx. pipiens pallens. Compared to the control group, the predominant bacteria changed from Actinobacteria to Firmicutes, and with increase in the concentration of Bti, the abundance of Actinobacteria was gradually reduced. Similar changes were also detected at the genus level, where Bacillus replaced Microbacterium, becoming the predominant genus in Bti-exposed groups. Furthermore, alpha diversity analysis indicated that Bti exposure changed the diversity of the microbota, possibly because the dysbiosis caused by the Bti infection inhibits some bacteria and provides opportunities to other opportunistic taxa. Pathway analysis revealed significant enhancement for processes associated with sphingolipid metabolism, glutathione metabolism and glycerophospholipid metabolism between all Bti-exposed groups and control group. Additionally, genes associated with the Toll and Imd signaling pathway were found to be notably upregulated. Bti infection significantly changed the bacterial community of larvae of Cx. pipiens pallens., (© 2021. The Author(s).)

Culex quinquefasciatus Say is a mosquito distributed in both tropical and subtropical regions of the world. It is a night-active, opportunistic blood-feeder and vectors many animal and human diseases, including West Nile Virus and avian malaria. Current vector control methods (e.g. physical/chemical) are increasingly ineffective; use of insecticides also imposes hazards to both human and ecosystem health. Advances in genome editing have allowed the development of genetic insect control methods, which are species-specific and, theoretically, highly effective. CRISPR/Cas9 is a bacteria-derived programmable gene editing tool that is functional in a range of species. We describe the first successful germline gene knock-in by homology dependent repair in C. quinquefasciatus. Using CRISPR/Cas9, we integrated an sgRNA expression cassette and marker gene encoding a fluorescent protein fluorophore (Hr5/IE1-DsRed, Cq7SK-sgRNA) into the kynurenine 3-monooxygenase (kmo) gene. We achieved a minimum transformation rate of 2.8%, similar to rates in other mosquito species. Precise knock-in at the intended locus was confirmed. Insertion homozygotes displayed a white eye phenotype in early-mid larvae and a recessive lethal phenotype by pupation. This work provides an efficient method for engineering C. quinquefasciatus, providing a new tool for developing genetic control tools for this vector., (© 2021. The Author(s).)

The bio-efficacy of crude ethyl acetate extract, fractions and a compound phenyl acetic acid from the ethyl acetate extract of Streptomyces collinus was evaluated on Culex quinquefasciatus Say and Aedes aegypti L. mosquitoes (Diptera: Culicidae). The larvae were exposed to concentrations of 2.5, 5.0, 7.5 and 10.0 ppm for fractions and 0.5, 1.0, 1.5 and 2.0 ppm for compound. After 24 h, the larval mortality was assessed and the LC₅₀ and LC 90 values were calculated. Similarly, per cent ovicidal activity was calculated for eggs after 120 h post treatment for phenyl acetic acid. Among the eleven fractions screened, fraction 7 from the ethyl acetate extract of Streptomyces collinus exhibited good larvicidal activity against both mosquito species. The LC 50 and LC 90 values of fraction 7 were 4.42, 6.23 ppm against Cx. quinquefasciatus larvae and 5.13, 14.51 ppm against Ae. aegypti larvae, respectively. Further, the isolated compound, phenyl acetic acid from fraction 7 recorded 100% larvicidal activity at 2 ppm concentration with LC 50 and LC 90 values of 2.07, 4.87 ppm on Cx. quinquefasciatus larvae and 3.81, 9.87 ppm on Ae. aegypti larvae, respectively. Phenyl acetic acid presented 50.3% and 42.0% ovicidal activity against Cx. quinquefasciatus and Ae. aegypti eggs at 2 ppm concentration after 120 h post treatment. The compound, phenyl acetic acid could be used in mosquito control programme., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: Clusters of sex-specific loci are predicted to shape the boundaries of the M/m sex-determination locus of the dengue vector mosquito Aedes aegypti, but the identities of these genes are not known. Identification and characterization of these loci could promote a better understanding of mosquito sex chromosome evolution and lead to the elucidation of new strategies for male mosquito sex separation, a requirement for several emerging mosquito population control strategies that are dependent on the mass rearing and release of male mosquitoes. This investigation revealed that the methylthioribulose-1-phosphate dehydratase (MtnB) gene, which resides adjacent to the M/m locus and encodes an evolutionarily conserved component of the methionine salvage pathway, is required for survival of female larvae., Results: Larval consumption of Saccharomyces cerevisiae (yeast) strains engineered to express interfering RNA corresponding to MtnB resulted in target gene silencing and significant female death, yet had no impact on A. aegypti male survival or fitness. Integration of the yeast larvicides into mass culturing protocols permitted scaled production of fit adult male mosquitoes. Moreover, silencing MtnB orthologs in Aedes albopictus, Anopheles gambiae, and Culex quinquefasciatus revealed a conserved female-specific larval requirement for MtnB among different species of mosquitoes., Conclusions: The results of this investigation, which may have important implications for the study of mosquito sex chromosome evolution, indicate that silencing MtnB can facilitate sex separation in multiple species of disease vector insects.

An extensive network of storm water conveyance systems in urban areas, often referred to as the "underground storm drain system" (USDS), serves as significant production habitats for mosquitoes. Knowledge of whether USDS habitats are suitable for newly introduced dengue vectors Aedes aegypti and Ae. albopictus will help guide surveillance and control efforts. To determine whether the USDS functions as a suitable larval habitat for Culex, Ae. aegypti and Ae. albopictus in southern California, we examined mosquito habitat utilization and larval survivorship using laboratory microcosm studies. The data showed that USDS constituted 4.1% of sampled larval habitats for Ae. aegypti and Ae. albopictus, and 22.0% for Cx. quinquefasciatus. Furthermore, USDS water collected in the summer completely inhibited Aedes larval development, but yielded a 15.0% pupation rate for Cx. quinquefasciatus. Food supplementation in the microcosms suggests that nutrient deficiency, toxins and other factors in the USDS water led to low success or complete failure of larval development. These results suggest that USDS habitats are currently not major productive larval habitats for Aedes mosquitoes in southern California. Our findings prompt inclusion of assessments of pupal productivity in USDS habitats and adult mosquito resting sites in the mosquito surveillance program.

The aim of the present study was to describe the morphology of the eggs of Culex (Culex) saltanensis Dyar that occurs in the Neotropical region. Eggs of the Cx. (Cux.) saltanensis were collected at the Mata Atlântica FIOCRUZ campus, fixed in 1% osmium tetroxide, prepared for mounting on metal supports, observed under a scanning electron microscope, and described morphologically. The eggs had a coniform shape with a length of approximately 0.5 mm (505-510 µm) and a width in the median portion of 117 µm (113-123 µm). Upper portion is lined with tubers of irregular shape and varying sizes (0.64-1.31 µm), located on a cross-linked matrix forming bands observed under optical microscopy. The micropyle is encased in a necklace of approximately 6.6-µm plates arranged in a flower-like shape. Comparing Cx. (Cux.) saltanensis eggs with several species of different genera, important divergent characteristics can be observed. However, this study points to the need for new descriptions of eggs of species belonging to the same subgenus in order to analyze if there will be differences between them. Culex (Cux.) saltanensis eggs have particular characteristics not observed in eggs of other Culicidae genera., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Insecticides, especially pyrethroids, are the most important in the insect pest control and preventing insect vector-borne human diseases. However, insect pests, including mosquitoes, have developed resistance in the insecticides that used against them. Cytochrome P450s are associated with insecticide resistance through overexpression and detoxification mechanisms in insect species. In this study, we utilized a powerful tool, the RNAi technique, to determine the roles of key P450 genes overexpressed in permethrin resistant mosquitoes that confer insecticide resistance to unravel the molecular basis of resistance mechanisms in the mosquito Culex quinquefasciatus. The results showed that knockdown of 8 key P450 genes using RNAi techniques significantly decreased resistance to permethrin in resistant mosquitoes. In silico modeling and docking analysis further revealed the potential metabolic function of overexpressed P450 genes in the development of insecticide resistance in mosquitoes. These findings not only highlighted the functional importance of these P450 genes in insecticide resistance, but also revealed that overexpression of multiple P450 genes was responsible for the high levels of insecticide resistance in a mosquito population of Culex quinquefasciatus.

Infections associated with the emergence of multidrug resistance and mosquito-borne diseases have resulted in serious crises associated with high mortality and left behind a huge socioeconomic burden. The chemical investigation of Lavandula coronopifolia aerial parts extract using HPLC-MS/MS led to the tentative identification of 46 compounds belonging to phenolic acids, flavonoids and their glycosides, and biflavonoids. The extract displayed larvicidal activity against Culex pipiens larvae (LC 50 = 29.08 µg/mL at 72 h). It significantly inhibited cytochrome P-450 monooxygenase (CYP450), acetylcholinesterase (AChE), and carboxylesterase (CarE) enzymes with the comparable pattern to the control group, which could explain the mode of larvae toxification. The extract also inhibited the biofilm formation of Pseudomonas aeruginosa by 17-38% at different Minimum Inhibitory Concentrations (MICs) (0.5-0.125 mg/mL) while the activity was doubled when combined with ciprofloxacin (ratio = 1:1 v : v ). In conclusion, the wild plant, L. coronopifolia, can be considered a promising natural source against resistant bacteria and infectious carriers.

Pesticide resistance in arthropod vectors of disease agents is a growing issue globally. Despite the importance of resistance monitoring to inform mosquito control programs, no regional monitoring programs exist in the United States. The Northeastern Regional Center for Excellence in Vector-Borne Diseases (NEVBD) is a consortium of researchers and public health practitioners with a primary goal of supporting regional vector control activities. NEVBD initiated a pesticide resistance monitoring program to detect resistant mosquito populations throughout the northeastern United States. A regionwide survey was distributed to vector control agencies to determine needs and refine program development and in response, a specimen submission system was established, allowing agencies to submit Culex pipiens (L.) (Diptera:Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae) for pesticide resistance testing. NEVBD also established larvicide resistance diagnostics for Bacillus thuringiensis israelensis (Bti) and methoprene. Additional diagnostics were developed for Cx. pipiens resistance to Lysinibacillus sphaericus. We received 58 survey responses, representing at least one agency from each of the 13 northeastern U.S. states. Results indicated that larvicides were deployed more frequently than adulticides, but rarely paired with resistance monitoring. Over 18,000 mosquitoes were tested from six states. Widespread low-level (1 × LC-99) methoprene resistance was detected in Cx. pipiens, but not in Ae. albopictus. No resistance to Bti or L. sphaericus was detected. Resistance to pyrethroids was detected in many locations for both species. Our results highlight the need for increased pesticide resistance testing in the United States and we provide guidance for building a centralized pesticide resistance testing program., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Predator-released kairomones indicating the presence of predatory fish are known to alter the behaviour and life-history traits of several aquatic herbivores. Culex mosquitoes (Diptera: Culicidae) respond to such cues by altering oviposition habitat selection and larval development time. These responses differ among fish species indicating composition differences among fish-released cues, but the recognition pattern is not clear. This study tested the dependence of fish recognition to co-evolution and the level of threat to larvae. We compared Culex responses to three fish, the invasive larvivorous Gambusia affinis (Cyprinodontiformes: Poeciliidae), the native larvivorous Aphanius mento (Cyprinodontiformes: Cyprinodontidae) and the native algivorous Garra rufa (Cypriniformes: Cyprinidae). The effects of cues released by the three fish on Culex oviposition and life-history shifts were compared in field mesocosms and lab tests. Our results showed that ovipositing females were significantly repelled only by cues originated from G. affinis, while developing larvae response was a more general one. Our results support the idea that fish-released kairomones differ in composition or quantities among different species., (© 2020 The Royal Entomological Society.)

Populations of Culex stigmatosoma and Cx. thriambus have been documented in the southwestern USA with a southward range extension to northern South America and Central America, respectively. Studies conducted in California indicate both species are potential vectors of West Nile virus. However, vector competence studies are lacking for other parts of the USA. During a multicounty regional surveillance study west of San Antonio, Texas, multiple errors were observed in the Texas distributional literature of these species. These errors involved incorrect distributional information in Texas and US publications. Evidence to correct these errant records was found upon further analysis of Texas literature and curated specimens. Therefore, the aims of this study were to present that evidence and then combine the corrected records with additional records from the Texas Department of State Health Services and from larval collections made during other Texas surveillance studies., (Copyright © 2021 by The American Mosquito Control Association, Inc.)

Background: Most female mosquitoes are anautogenous and must blood feed on a vertebrate host to produce eggs. Prior studies show that the number of eggs females lay per clutch correlates with the volume of blood ingested and that protein is the most important macronutrient for egg formation. In contrast, how whole blood, blood fractions and specific blood proteins from different vertebrates affect egg formation is less clear. Since egg formation is best understood in Aedes aegypti, we examined how blood and blood components from different vertebrates affect this species and two others: the malaria vector Anopheles gambiae and arbovirus vector Culex quinquefasciatus., Methods: Adult female mosquitoes were fed blood, blood fractions and purified major blood proteins from different vertebrate hosts. Markers of reproductive response including ovary ecdysteroidogenesis, yolk deposition into oocytes and number of mature eggs produced were measured., Results: Ae. aegypti, An. gambiae and C. quinquefasciatus responded differently to meals of whole blood, plasma or blood cells from human, rat, chicken and turkey hosts. We observed more similarities between the anthropophiles Ae. aegypti and An. gambiae than the ornithophile C. quinquefasciatus. Focusing on Ae. aegypti, the major plasma-derived proteins (serum albumin, fibrinogen and globulins) differentially stimulated egg formation as a function of vertebrate host source. The major blood cell protein, hemoglobin, stimulated yolk deposition when from pigs but not humans, cows or sheep. Serum albumins from different vertebrates also variably affected egg formation. Bovine serum albumin (BSA) stimulated ovary ecdysteroidogenesis, but more weakly induced digestive enzyme activities than whole blood. In contrast, BSA-derived peptides and free amino acids had no stimulatory effects on ecdysteroidogenesis or yolk deposition into oocytes., Conclusions: Whole blood, blood fractions and specific blood proteins supported egg formation in three species of anautogenous mosquitoes but specific responses varied with the vertebrate source of the blood components tested.

Background: Aerial applications of insecticides that target adult mosquitoes are widely used to reduce transmission of West Nile virus to humans during periods of epidemic risk. However, estimates of the reduction in abundance following these treatments typically focus on single events, rely on pre-defined, untreated control sites and can vary widely due to stochastic variation in population dynamics and trapping success unrelated to the treatment., Methods: To overcome these limitations, we developed generalized additive models fitted to mosquito surveillance data collected from CO 2 -baited traps in Sacramento and Yolo counties, California from 2006 to 2017. The models accounted for the expected spatial and temporal trends in the abundance of adult female Culex (Cx.) tarsalis and Cx. pipiens in the absence of aerial spraying. Estimates for the magnitude of deviation from baseline abundance following aerial spray events were obtained from the models., Results: At 1-week post-treatment with full spatial coverage of the trapping area by pyrethroid or pyrethrin products, Cx. pipiens abundance was reduced by a mean of 52.4% (95% confidence intrval [CI] - 65.6, - 36.5%) while the use of at least one organophosphate pesticide resulted in a mean reduction of 76.2% (95% CI - 82.8, - 67.9%). For Cx. tarsalis, at 1-week post-treatment with full coverage there was a reduction in abundance of 30.7% (95% CI - 54.5, 2.5%). Pesticide class was not a significant factor contributing to the reduction. In comparison, repetition of spraying over three to four consecutive weeks resulted in similar estimates for Cx. pipiens and estimates of somewhat smaller magnitude for Cx. tarsalis., Conclusions: Aerial adulticides are effective for achieving a rapid short-term reduction of the abundance of the primary West Nile virus vectors, Cx. tarsalis and Cx. pipiens. A larger magnitude of reduction was estimated in Cx. pipiens, possibly due to the species' focal distribution. Effects of aerial sprays on Cx. tarsalis populations are likely modulated by the species' large dispersal ability, population sizes and vast productive larval habitat present in the study area. Our modeling approach provides a new way to estimate effects of public health pesticides on vector populations using routinely collected observational data and accounting for spatio-temporal trends and contextual factors like weather and habitat. This approach does not require pre-selected control sites and expands upon past studies that have focused on the effects of individual aerial treatment events.

Larval mosquitoes have a more limited home range and lower resistance to adverse environment than adults, thus can be ideal targets for vector control in some cases. Coagulation-flocculation technology, which could be used for water treatment in breeding sites of several vector mosquito species, can significantly change both the distribution of organic particles and surface sediment characteristics in water environment. The aim of this study was to explore the effect, principle and possibility of using coagulation-flocculation technology in immature mosquitoes killing. In this study, dechlorinated water was treated with Poly Aluminum Chloride (PACl, sewage treatment using), and we observed the impacts of PACl treatment on the development and survival of immature Culex pipiens pallens mosquitoes. When exposed to PACl treatment, physical effect is believed to be a main reason of coagulation-flocculation caused high larvae mortality: Ⅰ) alum floc layer increases the difficulty of larvae foraging, leads larvae starving to death; (Ⅱ) the little floc particles could attach to the lateral hair of larvae, which impede floatation process and then surface respiration by larval mosquitoes. The alum floc layer had a good killing effect on the mosquito larvae, presented the half lethal time (LT 50 ) of 2d, the 90% lethal time (LT 90 ) of 8.7±7.3 ∼ 14±4.5 d, and the pupation rate of 0 ∼ (6.5±0.5)%, respectively. Our results indicates alum floc, produced by PACl coagulation-flocculation, was shown to be highly active against 1 st ∼2 nd instar larvae, the high mortality rate of immature mosquitoes as a result of physical effect. The observations suggest that coagulation-flocculation technology offers a novel potential approach to a sustainable and low-impact mosquito control method., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Advances in technology have greatly stimulated the understanding of insect-specific viruses (ISVs). Unfortunately, most of these findings are based on sequencing technology, and laboratory data are scarce on the transmission dynamics of ISVs in nature and the potential effects of these viruses on arboviruses. Mesonivirus is a class of ISVs with a wide geographical distribution. Recently, our laboratory reported the isolation of a novel strain of mesonivirus, Yichang virus (YCV), from Culex mosquitoes, China. In this study, the experimental infection of YCV by the oral route for adult and larvae mosquitoes, and the vertical transmission has been conducted, which suggests that YCV could adopt a mixed-mode transmission. Controlled experiments showed that the infectivity of YCV depends on the mosquito species, virus dose, and infection route. The proliferation curve and tissue distribution of YCV in Cx. quinquefasciatus and Ae. albopictus showed that YCV is more susceptible to Ae. albopictus and is located in the midgut. Furthermore, we also assessed the interference of YCV with flaviviruses both in vitro and in vivo. YCV significantly inhibited the proliferation of DENV-2 and ZIKV, in cell culture, and reduced transmission rate of DENV-2 in Ae. albopictus. Our work provides insights into the transmission of ISVs in different mosquito species during ontogeny and their potential ability to interact with mosquito-borne viruses., Competing Interests: The authors have declared that no competing interests exist.

Predation is a critical factor that mediates population stability, community structure, and ecosystem function. Predatory natural enemies can contribute to the regulation of disease vector groups such as mosquitoes, particularly where they naturally co-occur across landscapes. However, we must understand inter-population variation in predatory efficiency if we are to enhance vector control. The present study thus employs a functional response (FR; resource use under different densities) approach to quantify and compare predatory interaction strengths among six populations of a predatory temporary pond specialist copepod, Lovenula raynerae, from the Eastern Cape of South Africa preying on second instar Culex pipiens complex mosquito larvae. All individuals from the sampled populations were predatory and drove significant mortality through per capita predation rates of 0.75-1.10 mosquitoes/h at maximum densities over a 5-h feeding time. Individuals from all copepod populations exhibited Type II FRs with no significant differences in attack rates. On the other hand, there were significant differences in handling times, and therefore also maximum feeding rates (maximum experimental prey density: 32), suggesting possible genetic differences among populations that influenced predation. Owing to a widespread distribution in arid landscapes, we propose that predatory calanoid copepods such as L. raynerae play a key regulatory role at the landscape scale in the control of disease vector mosquito populations. We propose that these ecosystems and their specialist biota should thus be conserved and enhanced (e.g., via selective breeding) owing to the ecosystem services they provide in the context of public health., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The use of essential oils as ecofriendly tools for vector management is one of the mainstreams for biopesticide research. We evaluated the larvicidal properties of Commiphora erythraea (opoponax) essential oil and its fractions against Culex restuans Theobald, Culex pipiens L., and Aedes aegypti L. The use of bio-based amylose-N-1-hexadecylammonium chloride inclusion complex (Hex-Am) and amylose-sodium palmitate inclusion complex (Na-Palm) as emulsifiers for C. erythraea essential oil was also investigated. Bisabolene was the most abundant chemical constituent in the whole essential oil (33.9%), fraction 2 (62.5%), and fraction 4 (23.8%) while curzerene (32.6%) and α-santalene (30.1%) were the dominant chemical constituents in fractions 1 and 3, respectively. LC50 values for the whole essential oil were 19.05 ppm for Cx. restuans, 22.61 ppm for Cx. pipiens, and 29.83 ppm for Ae. aegypti and differed significantly. None of the four C. erythraea essential oil fractions were active against mosquito larvae. Two CYP450 genes (CYP6M11 and CYP6N12) and one GST gene (GST-2) were significantly upregulated in Ae. aegypti larvae exposed to C. erythraea essential oil suggesting their potential involvement in metabolic pathways for C. erythraea essential oil. Essential oil emulsions produced with Hex-Am were more toxic than the whole essential oil while those produced with Na-Palm had similar toxicity as the whole essential oil. These findings demonstrate that C. erythraea essential oil is a promising source of mosquito larvicide and that the use of Hex-Am as an emulsifier can enhance the insecticidal properties of C. erythraea essential oil., (Published by Oxford University Press on behalf of Entomological Society of America 2020.)

The impact of increasing resistance of mosquitoes to conventional pesticides has led to investigate various unique tools and pest control strategies. Herein, we assessed the potency of flupyradifurone, a novel pesticide, on fourth instar larvae of Culex quinquefasciatus Say. Further, we evaluated the synergistic action of piperonyl butoxide (PBO) and the octopamine receptor agonists (OR agonists) chlordimeform (CDM) and amitraz (AMZ) on the toxicity of flupyradifurone in comparison with sulfoxaflor and nitenpyram to increase their toxicity on Cx. quinquefasciatus. Results demonstrated that flupyradifurone was the most potent pesticide followed by sulfoxaflor and nitenpyram. Further, the synergetic effect of PBO, CDM, and AMZ was significant for all selected pesticides especially flupyradifurone. However, AMZ had the most significant effect in combination with the selected pesticides followed by CDM and PBO. The toxicity of the pesticides was time-dependent and increased over time from 24, 48, to 72 h of exposure in all experiments. The results indicate that flupyradifurone is a promising component in future mosquito control programs., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Four different tests showed the effectiveness of Azolla pinnata plant extracts against Aedes aegypti and Aedes albopictus mosquitoes. In the adulticidal test, there was a significant increase in mortality as test concentration increases and A. pinnata extracts showed LC 50 and LC 95 values of 2572.45 and 6100.74 ppm, respectively, against Ae. aegypti and LC 50 and LC 95 values of 2329.34 and 5315.86 ppm, respectively, against Ae. albopictus. The ovicidal test showed 100% eggs mortality for both species tested for all the concentrations tested at 1500 ppm, 1000 ppm, 500 ppm, 250 ppm and 125 ppm. Both tested samples of Ae. aegypti and Ae. albopictus did not lay any eggs in the plastic cups filled with the A. pinnata extract but instead opted to lay eggs in the plastic cups filled with water during the oviposition deterrence test. Similarly, the non-choice test of Ae. aegypti mosquitoes laid eggs on the sucrose solution meant for the nutrient source of the mosquitoes instead of in the plastic cup that was designed to facilitate oviposition filled with the extract. This clearly indicates the presence of bioactive compounds which are responsible in adulticidal and ovicidal activity in Aedes mosquitoes and at the same time inducing repellence towards the mosquitoes. The LC-MS results showed mainly three important chemical compounds from A. pinnata extracts such as 1-(O-alpha-D-glucopyranosyl)-(1,3R,25R)-hexacosanetriol, Pyridate and Nicotinamide N-oxide. All these chemicals have been used for various applications such as both emulsion and non-emulsion type of cosmetics, against mosquito vector such as Culex pipens and Anopheles spp. Finally, the overall view of these chemical components from A. pinnata extracts has shown the potential for developing natural product against dengue vectors.

Mosquitoes (Diptera: Culicidae) in the Culex pipiens complex play a key role in the transmission and therefore epidemiology of a number of human and animal pathogens globally. These mosquitoes, and sympatric species of the genus Culex Linnaeus that are not within the Cx. pipiens complex, are often considered 'impossible' to distinguish by morphology in the adult female stage. In the United States, this is particularly true for Culex pipiens s.l. and Culex restuans Theobald, both of which are competent vectors of West Nile virus, but likely play different roles in the transmission cycle. Therefore, we undertook an in-depth morphological evaluation of matched larval exuviae and adult specimens that revealed five useful morphological characters that are informative to distinguish Cx. pipiens s.l. from Cx. restuans in the adult stage. Herein, we provide a comprehensive review of the literature on these species of interest, and four additional, morphologically similar, Culex species, and a proposed key to adult female specimens., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.)

Introduction: A number of studies have been conducted on the relationship between the distribution of mosquito abundance and meteorological variables. However, few studies have specifically provided specific ranges of temperatures for estimating the maximum abundance of mosquitoes as an empirical basis for climatic dynamics for estimating mosquito-borne infectious disease risks., Methods: Adult mosquitoes were collected for three consecutive nights/week using Mosquito Magnet® Independence® model traps during 2018 and 2019 at US Army Garrison (USAG) Humphreys, Pyeongtaek, Gyeonggi Province, Republic of Korea (ROK). An estimate of daily mean temperatures (provided by the Korea Meteorological Administration) were distributed at the maximum abundance for selected species of mosquitoes using daily mosquito collection data after controlling for mosquito ecological cycles and environmental factors., Results: Using the Monte-Carlo simulation, the overall mosquito population abundance peaked at 22.7°C (2.5th-97.5th: 21.7°C-23.8°C). Aedes albopictus, vector of Zika, chikungunya, dengue fever and other viruses, abundance peaked at 24.6°C (2.5th-97.5th, 22.3°C-25.6°C), while Japanese encephalitis virus (JEV) vectors, e.g., Culex tritaeniorhynchus and Culex pipiens, peaked at 24.3°C (2.5th-97.5th: 21.9°C-26.3°C) and 22.6°C (2.5th-97.5th: 21.9°C-25.2°C), respectively. Members of the Anopheles Hyrcanus Group, some of which are vivax malaria vectors in the ROK, abundance peaked at 22.4°C (2.5th-97.5th: 21.5°C-23.8°C)., Conclusion: The empirical mean temperature ranges for maximum abundance were determined for each mosquito species collected at USAG Humphreys. These data contributed to the identification of relative mosquito abundance patterns for estimating mosquito-borne disease risks and developing and implementing disease prevention practices., Competing Interests: The authors have declared that no competing interests exist.

Emergent macrophytes play critical roles in water treatment processes of free-water surface constructed treatment wetlands. Management strategies for plant biomass affect wetland function and mosquito populations. Sinking of harvested macrophyte biomass is thought to provide organic carbon that enhances denitrifying bacteria important for nutrient removal while concomitantly reducing harborage for mosquitoes. The effects of sinking versus floating dried plant biomass (California bulrush [Schoenoplectus californicus]) on immature mosquito abundance and water quality (nutrient levels, oxygen demand, and physicochemical variables) were examined in mesocosms (28-m2 ponds or 1.4-m2 wading pools) under different flow regimes in 4 studies. The numbers of mosquito larvae in earthen ponds with floating vegetation were greater than in ponds with sunken vegetation on most dates but did not differ significantly between the 2 vegetation treatments in experiments using wading pools. Differences of the abundance of Anopheles larvae between the 2 vegetation management treatments were larger than for Culex larvae when naturally occurring larval mosquito predators were present. At high turnover rates (>2 pond volumes/day), water quality did not differ significantly between the vegetation management treatments and the water supply. At low turnover rates (approximately 2-6% of water volume/day), water quality differed significantly between the 2 vegetation management treatments and the water supply. Sinking vegetation can enhance the effectiveness of mosquito control but, depending on water management practices, may raise the concentrations of water quality constituents in discharges that are regulated under the Clean Water Act., (Copyright © 2020 by The American Mosquito Control Association, Inc.)

A field study was carried out on the year-long residual activity of the insect growth regulator (IGR) pyriproxyfen (Nylar 0.5G) in comparison with methoprene (Altosid® XRP Pellets) against mosquito developmental stages in catch basins in northwestern Riverside County, southern California. Pyriproxyfen was applied at 75, 100, 125, 150, 175 g per catch basin and methoprene at 3.5 g per catch basin. A total of 80 catch basins (10 per each treatment and 20 for control) were used. Posttreatment observations of catch basins were carried out at weekly intervals, with all pupal collections reared to adults. Mosquito species composition in this study, consisting mostly of Culex species (693), was predominated by Cx. quinquefasciatus (92.8%), followed by Cx. erythrothorax (5.5%), Cx. tarsalis (1.2%), Cx. stigmatosoma (0.3%), and Cx. thriambus (0.2%). Activity of both IGRs was expressed as percent inhibition of adult emergence (% IAE). Data generated on % IAE showed that, like methoprene, pyriproxyfen provided complete control of mosquitoes at 75, 125, and 175 g per catch basin up to 50 wk posttreatment at the Riverside amusement park, whereas its activity against mosquitoes in catch basins treated with 100 g and 150 g at the Eastvale site was short-lived, up to 48 wk. Water samples, bioassayed against laboratory-reared, 4th-stage larvae of Cx. quinquefasciatus 1-2 wk after the 50-wk-long study, showed evidence of significant % IAE (∼50) by pyriproxyfen at the 2 higher rates (125 g, 175 g) used at the amusement park. In conclusion, pyriproxyfen can be used to effectively control mosquitoes in catch basins for 48-50 wk, depending on the rate of application., (Copyright © 2020 by The American Mosquito Control Association, Inc.)

Introduction: Discarded vehicle tires represent a serious threat both to the environment and to public health as they have the potential to harbor important mosquito (Diptera: Culicidae) vectors., Objective: To assess the importance of used vehicle tires as larval habitats for mosquito fauna that colonize these artificial reservoirs in Jarabacoa, Dominican Republic., Materials and Methods: Used tires were sampled with pipettes at specialized tire fitting shops and scattered stockpiles of tires between June and August, 2018., Results: We sampled 396 tires; 57 (Container Index=14.4%) were positive for immature stages and contained 2,400 specimens, 11 species, and four genera (Anopheles, Aedes, Culex, and Toxorhynchites). The most abundant species was Aedes albopictus (42.3%) followed by Aedes aegypti (34.3%), and Culex quinquefasciatus (14.0%) while other species (9.4%) were less abundant. The container index varied significantly among the different tire sizes (χ2=13.4; p≤0.05). The highest infestation levels were found in the largest tires. A low positive correlation (r=0.38, n=396; p≤0.001) between the tire size and the prevalence of immature stages was recorded. The presence of organic matter had an overall positive effect on the infestation levels (U=11,430.0; p≤0.001)., Conclusions: These rubber residues, usually located nearby human populations, represent suitable breeding sites for arboviruses vectors such as dengue, chikungunya, Zika, and West Nile.

The mosquito Culex erythrothorax Dyar is a West Nile virus (WNV) vector that breeds in wetlands with emergent vegetation. Urbanization and recreational activities near wetlands place humans, birds and mosquitoes in close proximity, increasing the risk of WNV transmission. Adult Cx. erythrothorax abundance peaked in a wetland bordering the San Francisco Bay of California (USA) during the first 3 hours after sunset (5527 ± 4070 mosquitoes / trap night) while peak adult Culex tarsalis Coquillett abundance occurred during the subsequent 3 h period (83 ± 30 Cx. tarsalis). When insecticide resistance was assessed using bottle bioassay, Cx. erythrothorax was highly sensitive to permethrin, naled, and etofenprox insecticides compared to a strain of Culex pipiens that is susceptible to insecticides (LC50 = 0.35, 0.71, and 4.1 μg/bottle, respectively). The Cx. erythrothorax were 2.8-fold more resistant to resmethrin, however, the LC50 value was low (0.68 μg/bottle). Piperonyl butoxide increased the toxicity of permethrin (0.5 μg/bottle) and reduced knock down time, but a higher permethrin concentration (2.0 μg/bottle) did not have similar effects. Bulk mixed-function oxidase, alpha-esterase, or beta-esterase activities in mosquito homogenates were higher in Cx. erythrothorax relative to the Cx. pipiens susceptible strain. There was no difference in the activity of glutathione S-transferase between the two mosquito species and insensitive acetylcholine esterase was not detected. Larvicides that were applied to the site had limited impact on reducing mosquito abundance. Subsequent removal of emergent vegetation in concert with larvicide applications and reduced daily environmental temperature substantially reduced mosquito abundance. To control Cx. erythrothorax in wetlands, land managers should consider vegetation removal so that larvicide can efficiently enter the water. Vector control agencies may more successfully control adult viremic Cx. erythrothorax that enter nearby neighborhoods by applying adulticides during the 3 h that follow sunset., Competing Interests: The authors have declared that no competing interests exist.

Purpose: In southern Iran with a tropical climate, the above and underground cisterns of drinking water are the main habitats for immature stages of mosquitoes. The local people do not agree with using calcium hypochlorite granules in the water cisterns due to the change of water taste. Following the increased worldwide interest in using ultrasound technology for water disinfection, this study was conducted to assess the possible dual effects of ultrasound waves against immature mosquitoes. Materials and methods: Combinations of four power levels, two frequencies, three temperatures, and eight exposure times were applied to immature Culex pipiens . The exposure chamber of immature mosquitoes was the disposable cups placed inside a sonication bath containing tap water. Results: The mortality rate of immature mosquitos was 85-91% at 10-25 W, 30 °C and 130 kHz. The most effective ultrasound regimen for causing immature mortality included a fixed frequency of 130 kHz, a temperature of 30 °C, and a power level of 20-25 W. The lowest efficacy was observed in the egg stage. The median lethal time (LT 50 ) and ninety percent lethal time (LT 90 ) of sonication was 5.9 ± 0.6 min and 1.6 ± 0.1 min at 30 °C and a fixed frequency of 130 kHz. Conclusions: Evidence indicated effective ultrasound irradiation against immature mosquitoes in the water. This study provides a clear scientific basis for larvicidal properties induced by ultrasound emissions at laboratory conditions. The findings could be used for setting the ultrasound devices in the field investigations for achieving desired effectiveness.

The repeated usage of chemical insecticides, responsible for insecticide resistance in mosquitoes and environmental toxicity. Currently effective and environmental-safe control strategies are needed for the control disease-vector mosquitoes. Entomopathogens can be an effective alternative to chemical insecticide. Herein we isolated and tested 46 soil-borne entomopathogenic fungi belonging to six genera, namely Beauveria sp., Metarhizium sp., Fusarium sp., Aspergillus sp., Trichoderma sp., and Verticillium sp., fungi conidia were tested on Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus larvae. Bioassays results show that M. anisopliae fungal isolate causes a 100%, 98.6% and 92% mortality within six days, on Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus, respectively. M. anisopliae treated three mosquito larvae have lower lifetime with LT 50 values in A. stephensi, 2.931 days; A. aegypti, 2.676 days and C. quinquefasciatus, 3.254 days. 18 s rDNA sequence analysis confirmed that the isolated fungus are belonging to the genus of M. anisopliae-VKKH3, B. bassiana-VKBb03, and V. lecanii-VKPH1. Our results clearly show that M. anisopliae has good potential, as a low-cost, environmentally safe tool for the control of A. aegypti, A. stephensi, and C. quinquefasciatus mosquitoes., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Bacillus thuringiensis (Bt) can synthesize insecticides to efficiently control insects. In this study, Bt strain S3580-1 with mosquitocidal activity was subjected to whole genome sequencing using an Illumina HiSeq 2000 system. A novel toxin, Cry80Aa1, was identified based on the resulting data. A conserved domain analysis revealed that Cry80Aa1 includes the Ricin_B_lectin domain (located at 10-150) and the Toxin_10 domain (located at 155-353). Phylogenetic tree analysis showed that Cry80Aa1 was in a distinct clade significantly distinguished from the known Cry proteins containing the Toxin_10 domain. Bioassays demonstrated that the Cry80Aa1 protein exhibited toxicity to third instar larvae of Culex pipiens pallens (LC 50 : 71.9 μg/mL; 95% FL: 59.5-122.7 μg/mL)., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Vector-borne infectious diseases (VBDs) represent a major public health concern worldwide. Among VBDs, West Nile virus (WNV) showed an increasingly wider spread in temperate regions of Europe, including Italy. During the last decade, WNV outbreaks have been recurrently reported in mosquitoes, horses, wild birds, and humans, showing great variability in the temporal and spatial distribution pattern. Due to the complexity of the environment-host-vector-pathogen interaction and the incomplete understanding of the epidemiological pattern of the disease, WNV occurrences can be difficult to predict. The analyses of ecological drivers responsible for the earlier WNV reactivation and transmission are pivotal; in particular, variations in the vector population dynamics may represent a key point of the recent success of WNV and, more in general, of the VBDs., Methods: We investigated the variations of Culex pipiens population abundance using environmental, climatic and trapping data obtained over nine years (2010 to 2018) through the WNV entomological surveillance programme implemented in northeastern Italy. An information theoretic approach (IT-AIC c ) and model-averaging algorithms were implemented to examine the relationship between the seasonal mosquito population growth rates and both intrinsic (e.g. intraspecific competition) and extrinsic (e.g. environmental and climatic variables) predictors, to identify the most significant combinations of variables outlining the Cx. pipiens population dynamics., Results: Population abundance (proxy for intraspecific competition) and length of daylight were the predominant factors regulating the mosquito population dynamics; however, other drivers encompassing environmental and climatic variables also had a significant impact, although sometimes counterintuitive and not univocal. The analyses of the single-year datasets, and the comparison with the results obtained from the overall model (all data available from 2010 to 2018), highlighted remarkable differences in coefficients magnitude, sign and significance. These outcomes indicate that different combinations of factors might have distinctive, and sometimes divergent, effects on mosquito population dynamics., Conclusions: A more realistic acquaintance of the intrinsic and extrinsic mechanisms of mosquito population fluctuations in relation to continuous changes in environmental and climatic conditions is paramount to properly reinforce VBDs risk-based surveillance activities, to plan targeted density control measures and to implement effective early detection programmes.

On the basis of the mechanism of acylhydrazone compounds inhibiting the assembly of TMV CP and the unique structural characteristics of diketopiperazine ring, a series of optically pure indole diketopiperazine acylhydrazone were designed and synthesized. In order to systematically study the effect of the spatial configuration of the compounds on the antiviral activity, four compounds with different spatial configurations at C6 and C12 a were also prepared. The bioassay results indicated that most of these new compounds displayed moderate to good antiviral activity, among which compounds 23 , 25 , 27 , 28 , 31 , and 5d showed a significantly higher activity than that of commercial ribavirin. An in-depth structure-activity relationship investigation showed that the spatial conformation was one of the most important factors in adjusting antiviral activity; the research results provided information about the possible optimum configuration for interaction of this molecular with its target protein. At the same time, these new compounds also exhibited broad-spectrum fungicidal activities against 14 kinds of phytopathogenic fungi. What's more, some of these compounds exhibited good insecticidal activity to Plutella xylostella and Culex pipiens pallens .

Culex quinquefasciatus is a vector of many diseases that adversely impact human and animal health; however, compared to other mosquito vectors limited genome engineering technologies have been characterized for this vector. Clustered regularly interspaced short palindrome repeats-associated protein 9 (CRISPR-Cas9) based technologies are a powerful tool for genome engineering and functional genetics and consequently have transformed genetic studies in many organisms. Our objective was to improve upon the limited technologies available for genome editing in C. quinquefasciatus to create a reproducible and straightforward method for CRISPR-Cas9-targeted mutagenesis in this vector. Here we describe methods to achieve high embryo survival and mutagenesis rates and we provide details on the injection supplies and procedures, embryo handling and guide RNA (gRNA) target designs. Through these efforts, we achieved embryo survival rates and germline mutagenesis rates that greatly exceed previously reported rates in this vector. This work is also the first to characterize the white gene marker in this species, which is a valuable phenotypic marker for future transgenesis or mutagenesis of this vector. Overall, these tools provide the framework for future functional genetic studies in this important disease vector and may support the development of future gene drive and genetic technologies that can be used to control this vector., (© 2019 The Royal Entomological Society.)