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Invertebrate host–parasite associations are one of the keystones in order to understand vector-borne diseases. The study of these specific interactions provides information not only about how the vector is affected by the parasite at the gene-expression level, but might also reveal mosquito strategies for blocking the transmission of the parasites. A very well-known vector for human malaria is Anopheles gambiae. This mosquito species has been the main focus for genomics studies determining essential key genes and pathways over the course of a malaria infection. However, to-date there is an important knowledge gap concerning other non-mammophilic mosquito species, for example some species from the Culex genera which may transmit avian malaria but also zoonotic pathogens such as West Nile virus. From an evolutionary perspective, these 2 mosquito genera diverged 170 million years ago, hence allowing studies in both species determining evolutionary conserved genes essential during malaria infections, which in turn might help to find key genes for blocking malaria cycle inside the mosquito. Here, we extensively review the current knowledge on key genes and pathways expressed in Anopheles over the course of malaria infections and highlight the importance of conducting genomic investigations for detecting pathways in Culex mosquitoes linked to infection of avian malaria. By pooling this information, we underline the need to increase genomic studies in mosquito–parasite associations, such as the one in Culex–Plasmodium, that can provide a better understanding of the infection dynamics in wildlife and reduce the negative impact on ecosystems.

The vectorial capacity of mosquitoes, which influences the dynamics of vector-borne disease transmission, is intricately linked to mosquito abundance and the composition and diversity of their associated microbiomes. However, the influence of environmental factors on mosquito populations and microbiome diversity remains underexplored. Here we examined the effects of seasonality and developed land cover on Culex mosquito abundance and bacterial diversity. Biweekly field sampling of female Culex mosquitoes was conducted using gravid and CDC light traps, spanning summer to autumn across varying developed land cover levels in two urban areas in Central Illinois. Mosquito abundance was assessed by the number of mosquitoes captured per trap night and compared across seasons and developed levels. The mean mosquito abundance for gravid and light traps was 12.96 ± 2.15 and 7.67 ± 1.44, respectively. Notably, higher levels of developed land cover exhibited higher Culex abundance than the low level for light traps, but no significant difference was found between summer and early autumn. In gravid traps, no significant differences were detected across seasons or developed levels. Microbial analysis of the mosquito microbiome revealed that Proteobacteria and Wolbachia, with a mean relative abundance of 80.77 and 52.66% respectively, were identified as the most dominant bacterial phylum and genus. Their relative abundance remained consistent across seasons and developed land cover levels, with negligible variations. Alpha diversity, as measured by observed species, Chao1, Shannon, and Simpson, showed slightly higher values in early-autumn compared to late-summer. A notable pattern of bacterial diversity, as indicated by all four diversity indexes, is evident across varying levels of land development. Significantly, high or intermediate developed levels consistently showed reduced alpha diversity when compared to the lower level. This underscores the pronounced impact of anthropogenic ecological disturbances in shaping mosquito microbiomes. Beta diversity analysis revealed no significant dissimilarities in bacterial community composition across seasons and developed levels, although some separation was noted among different levels of developed land cover. These findings highlight the significant role of environmental factors in shaping mosquito abundance and their associated microbiomes, with potential implications for the vectorial capacity in the transmission of vector-borne diseases. [ABSTRACT FROM AUTHOR]

Snowshoe hare virus (SSHV) is a zoonotic arthropod-borne virus (arbovirus) circulating in colder areas of the Northern Hemisphere. SSHV is maintained in an enzootic cycle between small mammals and mosquitoes, assumably of the genera Aedes and Culiseta. Symptoms of SSHV human infection can range from asymptomatic to severe neuroinvasive disease. Studies on SSHV transmission are limited, and there is no information available on whether mosquitoes of the genus Culex are able to transmit SSHV. Therefore, we investigated six mosquito species via salivation assay for their vector competence. We demonstrated that SSHV can be transmitted by the abundant European Culex species Cx. pipiens biotype pipiens, Cx. pipiens biotype molestus, and Cx. torrentium with low transmission efficiency between 3.33% and 6.67%. Additionally, the invasive species Ae. albopictus can also transmit SSHV with a low transmission efficiency of 3.33%. Our results suggest that local transmission of SSHV after introduction to Europe seems to be possible from a vector perspective. [ABSTRACT FROM AUTHOR]

Understanding the blood-feeding patterns of mosquitoes is essential for evaluating their potential as disease vectors, especially in urban areas where mosquitoes coexist with humans, domestic animals and wildlife. This study aimed to bridge a substantial gap in regional knowledge by identifying the blood meal sources of field-collected mosquitoes in domestic and open green environments from two urbanisations of temperate Argentina, the Área Metropolitana de Buenos Aires (AMBA) and Tandil, using molecular techniques. Female mosquitoes were collected from November 2019 to March 2020 and April–May 2021. A bipartite network analysis was performed for each environment and urbanisation. A total of 103 blood meals from Aedes (2 species) and Culex (7 species) were identified. Among these, five mammal and 18 bird species were recognised as hosts. Aedes mosquitoes exclusively fed on mammals, while Culex mosquitoes exhibited a broader host range including both birds and mammals. In AMBA, the open green environments were composed by more mosquito species than the domestic environments, while both presented similar numbers of vertebrate species. In contrast, in open green environments from Tandil only blood-fed Aedes albifasciatus were collected. For open green environments of AMBA and domestic environments of Tandil, results suggested some degree of host selection. For the three main vectors of diseases in the region, Aedes aegypti , Ae. albifasciatus and Culex pipiens molestus , we present the first molecular evidence of human blood meals in South America. Epidemiological significance of the present findings is discussed. [ABSTRACT FROM AUTHOR]

Microplastics (MPs) are a growing issue because they endanger both aquatic organisms and humans. Studies have indicated that wastewater treatment plants (WWTPs) are one of the major contributors to MPs in the environment. However, studies on the abundance of MP contamination in WWTPs and its transmission into aquatic organisms are still scarce, especially in Egypt. The goal of this study was to examine the temporal fluctuations in the distribution of MPs in surface water and the dominant macroinvertebrate fauna (Culex sp. larvae) in a fixed wastewater basin in Sohag Governorate, Egypt. The average of MPs in the surface water was 3.01 ± 0.9 particles/L. The results indicated to seasonal variation of MP abundance in the wastewater basin that was significantly higher in winter than in the other seasons. The risk index for polymers (H), pollution load index (PLI), and potential ecological risk index (RI) were used to assess the degree of MP contamination. The basin has moderate H values (<1000) because of the presence of polymers with moderate hazard scores such as polyester (PES), polyethylene (PE), and polypropylene (PP). According to the PLI values, surface water is extremely contaminated with MPs (PLI: 88 to 120). The RI values of surface water showed higher ecological risk (level V). MPs in Culex sp. larvae were seasonally changed with an 85% detection rate, and an abundance average of 0.24 ± 0.65 particles/ind, MP concentration in Culex sp. larvae was influenced by the MP characters (shape, color, and polymer). The larvae of Culex sp. showed a greater preference for black and red fibrous polyester (PES) with sizes (<1000 μm) of MPs. These findings suggest that Culex sp. larvae prefer ingesting MPs that resemble their food. It is possible to overestimate Culex sp.'s preference for lower sizes because of their catabolism of MPs. To better understand the preferences of Culex sp. larvae for MPs, further controlled trials should be conducted. Practitioner Points: Wastewater is highly contaminated with microplastics (MPs) in the different seasons.First report of detection of the seasonal abundance of MP in Culex sp. larvae.Culex sp. larvae showed a stronger feeding preference for MPs with specific characteristics.Smaller size and blue polyester fibers were the dominant characteristics of MPs in wastewater. [ABSTRACT FROM AUTHOR]

Mosquitoes are of public health interest as many of them are vectors of pathogens that can cause diseases in humans. If they are not perfectly preserved, their identification can be difficult. Therefore, we tested the use of an alternative, advanced method of geometric morphometrics for the analysis of wing morphological variability in five (Anopheles, Aedes, Coquillettidia, Culex, Culiseta) genera and ten Slovenian mosquito species alone, seven autochthonous and three invasive. We analysed the intragenic and intraspecific variability in wing size and shape. The results showed that the analysed mosquito species differ in wing size and have different levels of variability, which also affects the correct identification of each taxon. Different classification approaches reach up to 80% (Culex and Coquillettidia), 98% (Aedes) and 100% (Anopheles and Culiseta) accuracy, respectively. Furthermore, additional information on wing size can help in identification when the data and wing venation are less reliable. The distances between taxa centroids in morphological space reflect genetic distances and taxonomic hierarchy: on average, they are greatest between the two subfamilies (Anophelinae, Culicinae), followed by the distances between genera within the Culicinae family and finally by the distances between species within the genera Aedes and Culex. Nevertheless, the topology of the similarity tree based on morphological data reflects the topology of the genetic phylogeny only at the subfamily level. We assessed that geometric morphometrics on wings can be used as an alternative method when other structures otherwise traditionally used for species identification are broken. [ABSTRACT FROM AUTHOR]

We aimed to demonstrate the direct physiological effects of octopamine on the auditory responses of Johnston's organ sensory neurons in Culex pipiens pipiens mosquitoes. Responses to acoustic stimulation were measured as the frequency-threshold curves and as an instantly recorded autoexcitation frequency before and after the injection of octopamine (or chlordimeform, an agonist of octopamine receptors). The autoexcitation frequency was assessed by including the auditory neurons in a positive feedback loop, when the amplified neuronal response was used to drive the stimulating loudspeaker. Our results indicate that (1) octopamine affects the properties of the auditory system in both male and female mosquitoes; (2) in female mosquitoes, octopamine significantly reduces the sensitivity of auditory neurons below 90 Hz and produces little effect in the high-frequency range (100 Hz and above); (3) in male mosquitoes, octopamine significantly increases the tuning frequency of the auditory system (ratio of tuning frequencies before and after the injection 1.32–1.55). There is a strong sexual dimorphism in the octopaminergic modulation of the mosquito auditory system. The observed physiological effects of octopamine, in both male and female mosquitoes, cannot be fully explained by the modulation of mechanical stiffness of the antenna and must include the changes in frequency tuning of the auditory neurons. [ABSTRACT FROM AUTHOR]

Mosquitoes harbor a wide diversity of microorganisms, including viruses that are human pathogens, or that are insect specific. We used metatranscriptomics, an unbiased high-throughput molecular approach, to describe the composition of viral and other microbial communities in six medically important mosquito species from across Western Australia: Aedes vigilax, Culex annulirostris, Cx. australicus, Cx. globocoxitus, Cx. pipiens biotype molestus, and Cx. quinquefasciatus. We identified 42 viral species, including 13 novel viruses, from 19 families. Culex mosquitoes exhibited a significantly higher diversity of viruses than Aedes mosquitoes, and no virus was shared between the two genera. Comparison of mosquito populations revealed a heterogenous distribution of viruses between geographical regions and between closely related species, suggesting that geography and host species may play a role in shaping virome composition. We also detected bacterial and parasitic microorganisms, among which Wolbachia bacteria were detected in three members of the Cx. pipiens complex, Cx. australicus, Cx. pipiens biotype molestus, and Cx. quinquefasciatus. In summary, our unbiased metatranscriptomics approach provides important insights into viral and other microbial diversity in Western Australian mosquitoes that vector medically important viruses. [ABSTRACT FROM AUTHOR]

Background: Hematophagous mosquitoes transmit many pathogens that cause human diseases. Pathogen acquisition and transmission occur when female mosquitoes blood feed to acquire nutrients for reproduction. The midgut epithelium of mosquitoes serves as the point of entry for transmissible viruses and parasites. Results: We studied midgut epithelial dynamics in five major mosquito vector species by quantifying PH3-positive cells (indicative of mitotic proliferation), the incorporation of nucleotide analogs (indicative of DNA synthesis accompanying proliferation and/or endoreplication), and the ploidy (by flow cytometry) of cell populations in the posterior midgut epithelium of adult females. Our results show that the epithelial dynamics of post-emergence maturation and of mature sugar-fed guts were similar in members of the Aedes, Culex, and Anopheles genera. In the first three days post-emergence, ~ 20% of cells in the posterior midgut region of interest incorporated nucleotide analogs, concurrent with both proliferative activity and a broad shift toward higher ploidy. In mature mosquitoes maintained on sugar, an average of 3.5% of cells in the posterior midgut region of interest incorporated nucleotide analogs from five to eight days post-emergence, with a consistent presence of mitotic cells indicating constant cell turnover. Oral bacterial infection triggered a sharp increase in mitosis and nucleotide analog incorporation, suggesting that the mosquito midgut undergoes accelerated cellular turnover in response to damage. Finally, blood feeding resulted in an increase in cell proliferation, but the nature and intensity of the response varied by mosquito species and by blood source (human, bovine, avian or artificial). In An. gambiae, enterocytes appeared to reenter the cell cycle to increase ploidy after consuming blood from all sources except avian. Conclusions: We saw that epithelial proliferation, differentiation, and endoreplication reshape the blood-fed gut to increase ploidy, possibly to facilitate increased metabolic activity. Our results highlight the plasticity of the midgut epithelium in mosquitoes' physiological responses to distinct challenges. [ABSTRACT FROM AUTHOR]

The grindstone process, which uses tyrosinase as a catalyst, was used to create analogues of geranylacetone. Tyrosinase was used to prepare the Mannich base under favourable reaction conditions, resulting in a high yield. All synthesized compounds were characterized using FTIR, Nuclear magnetic resonance, and mass spectral analyses. The active geranylacetone derivatives (1a-l) were investigated for larvicidal activity against Culex quinquefasciatus; compound 1b (LD5�€�:20.7 pg/mL) was noticeably more effective than geranylacetone (LD50: >100 ^g/mL) and permethrin (LD50: 24.4 pg/mL) lead compounds because of their ability to kill larvae and use them as pesticides. All compounds (1a-1l) were found to be low toxic, whereas compounds 1b, 1d, and 1k were screened for antifeedant screening of non -aquatic target for the toxicity measurement against marine fish Oreochromis mossambicus at 100 pg/mL caused 0% mortality in within 24 h. Molecular docking studies of synthesised compound 1b and permethrin docked with 3OGN, compound 1b demonstrated a greater binding affinity (-9.6 kcal/mol) compared to permethrin (-10.5 kcal/mol). According to these results, the newly synthesised geranylacetone derivatives can serve as lead molecules of larvicides agents. [ABSTRACT FROM AUTHOR]

Two members of the Culex pipiens subgroup, Culex pallens and Culex pipiens f. molestus, are known to occur in the Republic of Korea (ROK). These species exhibit morphologically similar features and are challenging to distinguish below the species level. Therefore, this study utilized wing geometric morphometrics (GM) on the right wing of the Culex pipiens subgroup, alongside sequencing of the cytochrome c oxidase subunit I (COI) region. Mosquitoes were collected from 11 locations between June and October (2020–2022) to minimize regional and seasonal variations. Additionally, Culex pipiens f. pipiens, which is not native to the ROK, was included in the analysis. Culex tritaeniorhynchus, Aedes albopictus, and Anopheles sinensis, the primary vectors in the ROK, were used as outgroups for comparison. All three taxa in the Culex pipiens subgroup could be identified with an 82.4%–97.0% accuracy using GM. However, a comparison of the COI regions of the Culex pipiens subgroup revealed no clear differences between the taxa. These data can be used for accurate identification, contributing to effective mosquito control, in addition to providing a foundation for evolutionary and ecological studies on wing shape differences. [ABSTRACT FROM AUTHOR]

Popularly known as "heartworm disease," canine dirofilariasis is a zoonosis that has a worldwide distribution, transmitted by vectors such as mosquitoes of the genera Aedes, Culex and Anopheles. In regions considered non-endemic, it is of great value to identify microfilariae with zoonotic potential such as D.immitis and D.repens, in order to minimize or prevent the occurrence of outbreaks or epidemics. It is notorious that the prevalence of D. immitis has increased in several regions of Brazil and the world in recent years. In Rio Branco, Acre, until the writing of the report, there was no notification of dogs positive for heartworm disease, despite having basic sanitation conditions, rainfall and temperature that favor the high concentration of mosquito populations, which allows the maintenance and dissemination of microfilariae between animals and humans. Therefore, this study aims to report the first occurrence of canine heartworm disease in the city of Rio Branco, Acre. [ABSTRACT FROM AUTHOR]

Wolbachia, an endosymbiotic bacterium, has a significant impact on the reproductive biology and pathogen transmission capabilities of various arthropods, including mosquitoes. This study aimed to assess the prevalence of Wolbachia in Culex molestus, a vector mosquito species, in South Korea. Mosquito samples were collected from different locations and taxonomically identified. Individual mosquito DNA was extracted, and polymerase chain reaction (PCR) was conducted targeting the Wolbachia surface protein gene (wsp) and three multi‐locus sequence typing (MLST) (gatB, hcpA, and ftsZ) genes. Gel electrophoresis was performed to determine the prevalence of Wolbachia by analyzing PCR products. Our findings revealed that Wolbachia was present in 100% of the tested Culex mosquitoes from South Korea, with one major Wolbachia strain (supergroup B) detected using the wsp gene and subgrouping determined with the gatB as well as hcpA, and ftsZ. These results underscore the widespread occurrence of Wolbachia in Cx. molestus mosquito within the studied region, thus suggesting the potential for leveraging this endosymbiont to devise novel strategies for mosquito‐borne disease control. Future research should investigate the influence of Wolbachia on mosquito biology and vector competence in South Korea. [ABSTRACT FROM AUTHOR]

Multiple studies about mosquitoes were carried out in several regions in northeastern Algeria, but the region of Guelma was not among them; therefore, the current study came to fill this void. Our inventory offered us a close knowledge of the biodiversity of Culicidae taxa in the region of Guelma that helped us define the presence of 12 species belonging to four genera (Culex, Culiseta, Aedes, and Anopheles). Culex pipiens (52.25%) was the most abundant species known in the north-east of Algeria. As well as Aedes albopictus being second (22.58%), after its new appearance in the region. Notably, results showed the first record of both Anopheles multicolour and Aedes geniculatus in the north-east of Algeria. [ABSTRACT FROM AUTHOR]

Controlling the vector by eradicating mosquito larvae is frequently done by spreading Abate powder containing 1% Temephos as a larvicide agent. However, the use of Abate powder will possibly increase pesticide residue contamination in the water. This study aimed to explore the potential use of ethanolic extract from red dragon fruit skin to kill mosquito larvae. An experimental laboratory study was performed to examine the larvicidal activity of ethanolic extract from red dragon fruit skin. Maceration method was used to extract active compounds from the fruit skin. Subsequently, the viscous extract was formulated using the wet granulation method to obtain powder form. The larvicide test was done by comparing the lethal concentration (LC50) of mosquito larvae after treatment using the extract and the reference larvicide agent, 1% Temephos Abate. The treatment groups consisted of four different extract concentrations of 1%, 2%, 3%, and 4%. The LC50 of each treatment groups was then calculated against the negative control group. Phytochemical screening showed that the ethanol extract of red dragon fruit skin contains alkaloids, flavonoids, saponins, triterpenoids, and tannins. The larvicide test on Aedes aegypti mosquito larvae showed that the extract treatment hardly killed the Aedes aegypti mosquito larvae. However, on Culex mosquito larvae, the ethanolic extract of red dragon fruit skin was able to kill mosquito larvae, with LC50 value of of 19.06%. The ethanolic extract of red dragon fruit skin was effective on killing the Culex mosquito larvae but not the Aedes aegypti mosquito larvae. [ABSTRACT FROM AUTHOR]

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Introduction: Arthropod-borne diseases are infectious diseases mediated by mosquitoes that have incurred a considerable health problem. The high prevalence of this disease (56.66%) in some rural areas, like Papua, is attributable to some environmental factors such as abundant breeding sites and favorable climatic conditions, which contribute to a high rate of disease transmission. Methods: This experimental research aims to determine the breeding site, mosquitos' diversity, and susceptibility to temephos insecticide through an entomological survey to find out the breeding site of the larvae. Mosquito larvae from positive breeding sites were captured and identified in the laboratory. Temephos susceptibility tests were conducted on each species at 0.02 ppm for Aedes and Culex, and 0.25 ppm for Anopheles. Results and Discussion: Entomology survey in Naena Muktipura village, Papua, revealed that 5 of 27 stagnant water bodies harbored mosquito breeding activity. The survey identified Aedes, Culex, and Anopheles larvae, with Culex and Anopheles preferring dirty stagnant water covered with leaves, while Aedes albopictus prefers sunlit puddles. All three species exhibited susceptibility to temephos. Conclusion: This study found that all three Diptera genera in Naena Muktipura village, despite their varied breeding site preferences, were susceptible to temephos. [ABSTRACT FROM AUTHOR]

There is burgeoning interest in how artificial light at night (ALAN) interacts with disease vectors, particularly mosquitoes. ALAN can alter mosquito behaviour and biting propensity, and so must alter disease transfer rates. However, most studies to date have been laboratory-based, and it remains unclear how ALAN modulates disease vector risk. Here, we identify five priorities to assess how artificial light can influence disease vectors in socio-ecological systems. These are to (i) clarify the mechanistic role of artificial light on mosquitoes, (ii) determine how ALAN interacts with other drivers of global change to influence vector disease dynamics across species, (iii) determine how ALAN interacts with other vector suppression strategies, (iv) measure and quantify the impact of ALAN at scales relevant for vectors, and (v) overcome the political and social barriers in implementing it as a novel vector suppression strategy. These priorities must be addressed to evaluate the costs and benefits of employing appropriate ALAN regimes in complex socio-ecological systems if it is to reduce disease burdens, especially in the developing world. This article is part of the theme issue 'Light pollution in complex ecological systems'. [ABSTRACT FROM AUTHOR]

West Nile Virus (WNV) is a mosquito vector-borne zoonosis with an increasing incidence in Europe that has become a public health concern. In Spain, although local circulation has been known for decades, until 2020, when a large outbreak occurred, West Nile Virus cases were scarce and mostly occurred in southern Spain. Since then, there have been new cases every year and the pathogen has spread to new regions. Thus, monitoring of circulating variants and lineages plays a fundamental role in understanding WNV evolution, spread and dynamics. In this study, we sequenced WNV consensus genomes from mosquito pools captured in 2022 as part of a newly implemented surveillance program in southern Spain and compared it to other European, African and Spanish sequences. Characterization of WNV genomes in mosquitoes captured in 2022 reveals the co-circulation of two WNV lineage 1 variants, the one that caused the outbreak in 2020 and another variant that is closely related to variants reported in Spain in 2012, France in 2015, Italy in 2021–2022 and Senegal in 2012–2018. The geographic distribution of these variants indicates that WNV L1 dynamics in southern Europe include an alternating dominance of variants in some territories. [ABSTRACT FROM AUTHOR]

Mosquitoes transmit many pathogens, like malaria (by Anopheles), lymphatic filariasis (by Culex), Zika virus and Dengue virus (by Aedes). Female mosquitoes have robust preferences for human odours over animal odours. A recent study showed that some olfactory glomerulus within the Aedes aegypti was significantly activated by human odours but weakly activated by animal odours. Among the odour chemicals, decanal and undecanal played dominant roles. However, most previous studies adopted lab-reared mosquitoes who were investigated in 1-way or 2-way enclosed tunnels. Here we studied wild Anopheles and Culex mosquitoes in an open environment and found that no mosquito could be caught without either CO2 or heat, indicating that both CO2 and heat gate the responses to other odorant stimuli. Only wet clothes soaked with human sweat (but not human-scented dry clothes) attracted 20–50% more mosquitoes at 37°C and showed no effect at 25°C. Thus for Anopheles and Culex, human odour may only play a significant role in the host-seeking behavior only upon sweating. And neither decanal nor undecanal attracted mosquitoes in our field study, indicating inefficacy of any single volatile substance. [ABSTRACT FROM AUTHOR]

Targeted vector surveillance informed by data on mosquito biting patterns can help limit arboviral zoonotic diseases. To characterise host‐biting networks in rural and urban equestrian facilities from temperate Argentina, adult resting mosquitoes were collected (December 2018–April 2019) with a battery‐powered aspirator. Engorged females were sorted to species, and their blood source was identified using molecular techniques. Bipartite network analysis was performed for rural and urban matrices. A total of 177 bloodmeals from 11 mosquito species of Aedes and Culex were identified, with seven mammal and 17 bird species recognised as hosts. Mammals represented 61% of the total feeds, mainly horse, dog and sheep; the best represented avian hosts were Columbiformes. Aedes species and Culex maxi fed only on mammals, while most other Culex species presented a wide range of hosts. The rural network had more nodes and interactions than its urban counterpart, both with some degree of host selection and aggregated patterns according to network indices. Culex quinquefasciatus was the strongest species in both networks, whereas Culex apicinus and Culex dolosus had a prominent role in the rural network. Bipartite network analysis will contribute to understanding the effects of urbanisation in the dynamics of vector‐borne diseases. [ABSTRACT FROM AUTHOR]

In Southeast Asia, despite the use of Japanese encephalitis vaccines and vaccination coverage, Japanese encephalitis (JE) transmission is still a major public health issue. The main vectors of this virus are mosquitoes from the genus Culex, which diversity and density are important in Southeast Asia. The main vector species of Japanese encephalitis virus (JEV) in Cambodia belong to the Vishnui subgroup. However, their morphological identification solely based on the adult stage remains challenging, making their segregation and detection difficult. In order to identify and describe the distribution of the three main JEV vector species in Cambodia, namely Culex vishnui, Cx. pseudovishnui and Cx. tritaeniorhynchus, mosquito samplings were carried out throughout the country in different environments. Phylogenetic analysis of the cytochrome c oxidase subunit I (coI) gene using maximum‐likelihood tree with ultrafast bootstrap and phylogeographic analysis were performed. The three main Culex species are phylogenetically separated, and represent two distinct clades, one with Cx. tritaeniorhynchus and the second with Cx. vishnui and Cx. pseudovishnui, the latter appearing as a subgroup of Cx. vishnui. The phylogeographic analysis shows a distribution of the Vishnui subgroup on the entire Cambodian territory with an overlapped distribution areas leading to a sympatric distribution of these species. The three JEV vector species are geographically well‐defined with a strong presence of Cx. pseudovishnui in the forest. Combined with the presence of Cx. tritaeniorhynchus and Cx. vishnui in rural, peri‐urban, and urban areas, the presence of JEV‐competent vectors is widespread in Cambodia. [ABSTRACT FROM AUTHOR]

Simple Summary: Mosquito-borne infectious diseases threaten millions of people worldwide, and it is critical that we identify new methods for controlling these insects. We recently developed a new class of mosquito insecticides that consists of yeasts which produce interfering RNA that is custom-designed to turn off mosquito genes yet does not hurt other organisms. Here, we test a newly optimized robust yeast strain which is suitable for industry-scale yeast production. Following heat inactivation and drying, the yeast was shipped to St. Augustine, Trinidad and Bangkok, Thailand, where it was evaluated in outdoor studies performed in tropical settings. The yeast effectively killed a variety of different types of mosquito larvae that died following yeast consumption. It could also be supplied to adult mosquitoes in a sugar bait solution, resulting in high levels of mosquito death. The yeast killed mosquitoes more quickly in outdoor experiments than in laboratory trials. These promising results indicate that yeast insecticides could one day be used globally as a new means of eco-friendly mosquito control and disease prevention. Eco-friendly new mosquito control innovations are critical for the ongoing success of global mosquito control programs. In this study, Sh.463_56.10R, a robust RNA interference (RNAi) yeast insecticide strain that is suitable for scaled fermentation, was evaluated under semi-field conditions. Inactivated and dried Sh.463_56.10R yeast induced significant mortality of field strain Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus larvae in semi-field larvicide trials conducted outdoors in St. Augustine, Trinidad, where 100% of the larvae were dead within 24 h. The yeast was also stably suspended in commercial bait and deployed as an active ingredient in miniature attractive targeted sugar bait (ATSB) station sachets. The yeast ATSB induced high levels of Aedes and Culex mosquito morbidity in semi-field trials conducted in Trinidad, West Indies, as well as in Bangkok, Thailand, in which the consumption of the yeast resulted in adult female mosquito death within 48 h, faster than what was observed in laboratory trials. These findings support the pursuit of large-scale field trials to further evaluate the Sh.463_56.10R insecticide, a member of a promising new class of species-specific RNAi insecticides that could help combat insecticide resistance and support effective mosquito control programs worldwide. [ABSTRACT FROM AUTHOR]

West Nile virus (WNV) is the most significant arbovirus in the United States in terms of both morbidity and mortality. West Nile exists in a complex transmission cycle between avian hosts and the arthropod vector, Culex spp. mosquitoes. Human spillover events occur when humans are bitten by an infected mosquito and predicting these rates of infection and therefore the risk to humans may be associated with fluctuations in environmental conditions. In this study, we evaluate the hydrological and meteorological drivers associated with mosquito biology and viral development to determine if these associations can be used to forecast seasonal mosquito infection rates with WNV in the Coachella Valley of California. We developed and tested a spatially resolved ensemble forecast model of the WNV mosquito infection rate in the Coachella Valley using 17 years of mosquito surveillance data and North American Land Data Assimilation System‐2 environmental data. Our multi‐model inference system indicated that the combination of a cooler and dryer winter, followed by a wetter and warmer spring, and a cooler than normal summer was most predictive of the prevalence of West Nile positive mosquitoes in the Coachella Valley. The ability to make accurate early season predictions of West Nile risk has the potential to allow local abatement districts and public health entities to implement early season interventions such as targeted adulticiding and public health messaging before human transmission occurs. Such early and targeted interventions could better mitigate the risk of WNV to humans. Plain Language Summary: West Nile virus (WNV) is the most significant arbovirus in the United States and is transmitted seasonally by mosquitoes. Humans are most at risk when they are in close proximity to infected mosquitoes. Predicting the risk to humans is not straightforward. In this study, we use deviations in climate associated with mosquito biology and viral development to forecast seasonal West Nile risk in the Coachella Valley of California. We developed a statistical model of WNV transmission in the Coachella Valley using 17 years of mosquito surveillance data and environmental data. Our model indicated that the combination of a cooler and dryer winter followed by a wetter and warmer spring and a cooler than normal summer was the combination of environmental events most associated with West Nile positive mosquitoes in the Coachella Valley. The ability to make accurate early season predictions of West Nile risk could assist local public health entities implement early season interventions to better mitigate the risk of WNV to humans in the Coachella Valley. Key Points: Environmentally informed West Nile virus (WNV) forecast modelOur forecast shows that a dry, cool winter, followed by a wet, warm spring, and a cool summer promotes WNVEarly season forecasts are a potential decision tool to inform public health and mosquito abatement intervention [ABSTRACT FROM AUTHOR]

Several emerging mosquito control technologies require mass releases of adult male mosquitoes. Previous studies resulted in the generation of a laboratory female-specific larvicidal yeast strain targeting the GGT gene, which facilitated the laboratory sex separation of male Culex quinquefasciatus mosquitoes. Global deployment of this yeast-based sex-separation technology requires engineering second generation yeast strains which can be used in industrial-scale fermentations to support global mosquito control programs. In this study, the RNA-guided Cas-CLOVER system was used in combination with piggyBac transposase to generate robust Saccharomyces cerevisiae strains with multiple integrated copies of the insecticidal GGT shRNA expression cassette. Top expressing Cas-CLOVER strains killed Culex quinquefasciatus female larvae which consumed the yeast, facilitating male sex separation. Scaled fermentation resulted in kilogram-scale production of the yeast, which can be heat-killed and dried for global deployment to mosquito mass-rearing facilities. [ABSTRACT FROM AUTHOR]

Background Culex quinquefasciatus is a notorious vector known to transmit pathogens such as Wuchereria bancrofti (causing Lymphatic filariasis) and flaviviruses such as West Nile virus in India and St. Louis Encephalitis virus in the USA. It is the vector of the Rift Valley Fever virus, also on the African continent. Mosquitoes also harbor other non-pathogenic insect-specific flaviviruses (ISFs), such as Culex flavivirus (CxFV) and Aedes flavivirus. Recent studies have implicated ISFs interfering with the vectorial efficiency of the pathogenic arbo-viruses. Methods One hundred specimens of the Cx. quinquefasciatus population in two urban areas in Kerala State, India, were screened to have an understanding of the prevalence of these flaviviruses in this vector species. Viral RNA was extracted from individual specimens and was subjected to RT-PCR towards amplification of the CxFV non-structural protein 5 (NS5) gene. Results Among the 100 specimens, 7.0% were found to be harboring CxFV infection. The phylogenetic analysis of the gene sequences showed that the virus isolates were genetically related to Kenya, with 98–99% sequence similarities. Conclusion This is the first report on the occurrence of CxFV from Cx. quinquefasciatus from India. The occurrence of these viruses in mosquitoes could play a critical role in disease vector management. [ABSTRACT FROM AUTHOR]

Purpose: Mosquitoes are important vectors that carry disease-causing agents that can affect humans and animals. DNA barcoding is a complementary identification which can be used to validate morphological characterization of mosquito species. The objectives of this study were to identify the mitochondrial sequence of the COI gene and to construct a molecular phylogeny based on the genetic divergence of the mosquito species studied. Methods: In this study, DNA extraction and the amplification of the mitochondrial cytochrome oxidase subunit I genes (COI) were performed on pooled mosquito samples collected in Nay Pyi Taw area, Myanmar. Results: Fragments of the COI gene showed 99–100% identity with sequences of Aedes aegypti, Armigeres subalbatus, Culex pipiens complex, and Cx. quinquefasciatus, respectively, deposited in GenBank. This study categorized two haplotypes from each Ar. subalbatus and Cx. pipiens complex COI gene sequence, as well as three haplotypes from Cx. quinquefasciatus COI gene sequences. The highest haplotype diversity and nucleotide diversity were observed in the Ar. subalbatus population (Hd = 1.0000; π = 0.0033), followed by the Cx. pipiens complex and Cx. quinquefasciatus populations. Conclusion: This study provides useful information on the molecular identification and genetic diversity of mosquito vectors with medical and veterinary significance, which may assist in the improvement of mosquito control programs. [ABSTRACT FROM AUTHOR]

Arboviruses, i.e., viruses transmitted by blood-sucking arthropods, trigger significant global epidemics. Over the past 20 years, the frequency of the (re-)emergence of these pathogens, particularly those transmitted by Aedes and Culex mosquitoes, has dramatically increased. Therefore, understanding how human behavior is modulating population exposure to these viruses is of particular importance. This synthesis explores human behavioral factors driving human exposure to arboviruses, focusing on household surroundings, socio-economic status, human activities, and demographic factors. Household surroundings, such as the lack of water access, greatly influence the risk of arbovirus exposure by promoting mosquito breeding in stagnant water bodies. Socio-economic status, such as low income or low education, is correlated to an increased incidence of arboviral infections and exposure. Human activities, particularly those practiced outdoors, as well as geographical proximity to livestock rearing or crop cultivation, inadvertently provide favorable breeding environments for mosquito species, escalating the risk of virus exposure. However, the effects of demographic factors like age and gender can vary widely through space and time. While climate and environmental factors crucially impact vector development and viral replication, household surroundings, socio-economic status, human activities, and demographic factors are key drivers of arbovirus exposure. This article highlights that human behavior creates a complex interplay of factors influencing the risk of mosquito-borne virus exposure, operating at different temporal and spatial scales. To increase awareness among human populations, we must improve our understanding of these complex factors. [ABSTRACT FROM AUTHOR]

Previously, we have identified an odorant receptor (OR) from the southern house mosquito Culex quinquefasciatus, CquiOR32, which responded to both odorants (agonists) and inhibitory compounds (antagonists). CquiOR32/CquiOrco-expressing oocytes responded to methyl salicylate and other odorants with inward (regular) currents but gave currents in the reverse direction when challenged with eucalyptol and other inhibitors. To determine whether hitherto unknown ORs show this intrareceptor inhibition, we have now examined two other receptors in the same cluster, CquiOR27 and CquiOR28. We cloned and tested four variants of CquiOR28, but none of the 250 compounds in our panel of odorants, including an Orco ligand candidate (OLC12), elicited inward or upward deflections of the current traces. By contrast, CquiOR27/CquiOrco-expressing oocytes gave robust, dose-dependent inward currents when challenged with γ-octalactone and other odorants. On the other hand, octylamine and other phenolic compounds elicited dose-dependent currents in the reverse direction. When stimulatory and inhibitory compounds were presented in binary mixtures, γ-octalactone-elicited inward currents were attenuated in a dose-dependent manner according to the concentration of octylamine. As part of our chemical ecology approach, we tested the repellency activity of the most potent ligands in the surface landing and feeding assay and a newly reported hand-in cage assay. Protection elicited by γ-octalactone did not differ significantly from that of DEET at the same dose. In the hand-in cage assay, a cream formulation of γ-octalactone showed 97.0 ± 1.3% protection, with 47.6 ± 8.3% and 1.4 ± 0.7% landings per trial in the hands covered with a control and γ-octalactone cream, respectively (N = 8, p = 0.0078, Wilcoxon matched-pairs signed-rank test).

Insect ommochrome biosynthesis pathways metabolize tryptophan to generate eye-color pigments and wild-type alleles of pathway genes are useful phenotypic markers in transgenesis studies. Pleiotropic effects of mutations in some genes exert a load on both survival and reproductive success in blood-feeding species. Here, we investigated the challenges imposed on mosquitoes by the increase of tryptophan metabolites resulting from blood meal digestion and the impact of disruptions of the ommochrome biosynthesis pathway. Female mosquitoes with spontaneous and induced mutations in the orthologs of the genes encoding kynurenine hydroxylase in Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus exhibited impaired survival and reproductive phenotypes that varied in type and severity among the species. A compromised midgut permeability barrier function was also observed in An. stephensi. Surprisingly, mutant mosquitoes displayed an increase in microbiota compared to controls that was not accompanied by a general induction of immune genes. Antibiotic treatment rescued some deleterious traits implicating a role for the kynurenine pathway (KP) in midgut homeostasis. Supplemental xanthurenic acid, a KP end-product, rescued lethality and limited microbiota proliferation in Ae. aegypti. These data implicate the KP in the regulation of the host/microbiota interface. These pleiotropic effects on mosquito physiology are important in the development of genetic strategies targeting vector mosquitoes.

Abstract Background Malaria vectors vary in feeding preference depending on their innate behaviour, host availability and abundance. Host preference and human biting rate in malaria vectors are key factors in establishing zooprophylaxis and zoopotentiation. This study aimed at assessing the impact of non-human hosts in close proximity to humans on the human biting rate of primary and secondary malaria vectors, with varying host preferences. Methods The effect of the presence of non-human hosts in close proximity to the human host on the mean catches per person per night, as a proxy for mosquito biting rate, was measured using mosquito-electrocuting traps (METs), in Sagamaganga, Kilombero Valley, Tanzania. Two experiments were designed: (1) a human versus a calf, each enclosed in a MET, and (2) a human surrounded by three calves versus a human alone, with each human volunteer enclosed individually in a MET spaced 10 m apart. Each experiment was conducted on alternate days and lasted for 36 nights per experiment. During each experiment, the positions of hosts were exchanged daily (except the human in experiment 2). All anopheline mosquitoes caught were assayed for Plasmodium sporozoites using enzyme-linked immunosorbent assay. Results A total of 20,574 mosquitoes were captured and identified during the study, of which 3608 were anophelines (84.4% primary and 15.6% secondary malaria vectors) and 17,146 were culicines. In experiment 1, the primary malaria vector, Anopheles arabiensis, along with Culex spp. demonstrated a preference for cattle, while the primary vectors, Anopheles funestus, preferred humans. In experiment 2, both primary vectors, An. arabiensis and An. funestus, as well as the secondary vector Anopheles rivolurum, demonstrated behaviours amenable to zooprophylaxis, whereas Culex spp. increased their attraction to humans in the presence of nearby cattle. All anopheline mosquitoes tested negative for sporozoites. Conclusions The findings of this study provide support for the zooprophylaxis model for malaria vectors present in the Kilombero Valley, and for the zoopotentiation model, as it pertains to the Culex spp. in the region. However, the factors regulating zooprophylaxis and zoopotentiation are complex, with different species-dependent mechanisms regulating these behaviours, that need to be considered when designing integrated vector management programmes.

BackgroundThe surveillance of vector mosquitoes is essential for prevention and control of mosquito-borne diseases. In this study, we developed an internet-based vector mosquito monitor, MS-300, and evaluated its efficiency for the capture of the important vector mosquitoes, Aedes albopictus and Culex quinquefasciatus, in laboratory and field trials.Methodology/principal findingsThe linear sizes of adult Ae. albopictus and Cx. quinquefasciatus were measured and an infrared window was designed based on these data. A device to specifically attract these two species and automatically transmit the number of captured mosquitoes to the internet was developed. The efficiency of the device in capturing the two species was tested in laboratory, semi-field and open field trials. The efficiency results for MS-300 for catching and identifying Ae. albopictus in laboratory mosquito-net cages were 98.5% and 99.3%, and 95.8% and 98.6%, respectively, for Cx. quinquefasciatus. In a wire-gauze screened house in semi-field trials, the efficiencies of MS-300 baited with a lure in catching Ae. albopictus and Cx. quinquefasciatus were 54.2% and 51.3%, respectively, which were significantly higher than 4% and 4.2% without the lure. The real-time monitoring data revealed two daily activity peaks for Ae. albopictus (8:00-10:00 and 17:00-19:00), and one peak for Cx. quinquefasciatus (20:00-24:00). During a 98-day surveillance trial in the field, totals of 1,118 Ae. albopictus and 2,302 Cx. quinquefasciatus were captured by MS-300. There is a close correlation between the number of captured mosquitoes and the temperature in the field, and a positive correlation in the species composition of the captured samples among the mosquitoes using MS-300, BioGents Sentinel traps and human landing catches.Conclusions/significanceThe data support the conclusion that MS-300 can specifically and efficiently capture Ae. albopictus and Cx. quinquefasciatus, and monitor their density automatically in real-time. Therefore, MS-300 has potential for use as a surveillance tool for prevention and control of vector mosquitoes.

BackgroundEndogenous circadian rhythms result from genetically-encoded molecular clocks, whose components and downstream output factors cooperate to generate cyclic changes in activity. Mating is an important activity of mosquitoes, however, the key aspects of mating rhythm patterns and their regulatory mechanisms in two vector mosquito species, Aedes albopictus and Culex quinquefasciatus, remain unclear.Methodology/principal findingsWe determined and compared the diel mating activity rhythms of these two mosquito species and discovered that Ae. albopictus had mating peaks in the light/dark transition periods (ZT0-3 and ZT9-12), while Cx. quinquefasciatus only had a mating peak at ZT12-15. Knockouts of the clock (clk) orthologous genes (Aalclk and Cxqclk) resulted in phase delay or phase reversal of the mating peaks in Ae. albopictus and Cx. quinquefasciatus, respectively. In addition, the temporal expression pattern of the desaturase orthologous genes, desat1, in both mosquito species was also different in respective wild-type strains and showed phase changes similar to the mating rhythms in clk mutant strains. Inhibition of desat1 expression resulted in decreased mating activity in male mosquitoes of both species but not females. In addition, desat1 regulated cuticular hydrocarbons' synthesis in both species. Silencing desat1 in male Ae. albopictus resulted in decreases of nonadecane and tricosane, which promoted mating, with concomitant increases of heptacosane, which inhibited mating. Silencing desat1 in male Cx. quinquefasciatus also resulted in decreases of tricosane, which promoted mating.Conclusions/significanceThese results suggest that Aalclk and Cxqclk have significant roles in the mating activity rhythms in both Ae. albopictus and Cx. quinquefasciatus by regulating the temporal expression of the desat1 gene under LD cycles, which affects sex pheromone synthesis and mating. This work provides insights into the molecular regulatory mechanism of distinct mating rhythm of Ae. albopictus and Cx. quinquefasciatus and may provide a basis for the control of these two important vector mosquitoes.

BackgroundVector control strategies typically rely on pesticides to target mosquitoes involved in enzootic and zoonotic transmission of West Nile virus (WNV). Nevertheless, increasing insecticide resistance and a desire to reduce pesticide usage provide the impetus for developing alternative strategies. Ivermectin (IVM), an antiparasitic drug which is widely used in human and veterinary medicine, is a potential alternative for targeted control because Culex mosquitoes experience increased mortality following ingestion of IVM in bloodmeals.Methodology/principal findingsWe conducted a randomized field trial to investigate the impact of treating backyard chicken flocks with IVM in urban neighborhoods across Davis, California on mosquito populations and WNV transmission dynamics. We observed a significant reduction in WNV seroconversions in treated vs. untreated chickens, suggesting a reduction in WNV transmission intensity around treated flocks. We also detected a reduction in parity rates of Cx. tarsalis near treated vs. untreated flocks and increased mortality in wild mosquitoes following a bloodmeal on treated chickens (IVM serum concentration > 5ng/mL) vs. chickens with IVM serum concentrations < 5 ng/mL. However, we did not find a significant difference in abundance or infection prevalence in mosquitoes between treatment groups associated with the reductions in seroconversions. Mosquito immigration from surrounding larval habitat, relatively low WNV activity in the study area, and variable IVM serum concentrations likely contributed to uncertainty about the impact.Conclusions/significanceTaken together, our results point to a reduction in WNV transmission due to the impact of IVM on Culex mosquito populations and support the ongoing investigation of oral administration of IVM to wild birds for local control of WNV transmission, although further work is needed to optimize dosing and understand effects on entomological endpoints.

Local vector control and public health agencies in California use the California Mosquito-Borne Virus Surveillance and Response Plan to monitor and evaluate West Nile virus (WNV) activity and guide responses to reduce the burden of WNV disease. All available data from environmental surveillance, such as the abundance and WNV infection rates in Culex tarsalis and the Culex pipiens complex mosquitoes, the numbers of dead birds, seroconversions in sentinel chickens, and ambient air temperatures, are fed into a formula to estimate the risk level and associated risk of human infections. In many other areas of the US, the vector index, based only on vector mosquito abundance and infection rates, is used by vector control programs to estimate the risk of human WNV transmission. We built models to determine the association between risk level and the number of reported symptomatic human disease cases with onset in the following three weeks to identify the essential components of the risk level and to compare California's risk estimates to vector index. Risk level calculations based on Cx. tarsalis and Cx. pipiens complex levels were significantly associated with increased human risk, particularly when accounting for vector control area and population, and were better predictors than using vector index. Including all potential environmental components created an effective tool to estimate the risk of WNV transmission to humans in California.

Metagenomic studies of mosquito viromes demonstrated a more diverse composition than just an exclusive composition of pathogenic arboviruses transmitted to humans. In our study, the virome of 866 female mosquitoes collected throughout 2020 at the São Paulo Zoo, located in the city of São Paulo/SP—Brazil, was obtained. Specifically, in this paper, we describe a new virus found by viral RNA extraction and next-generation MiSeq sequencing of a group of 23 specimens of Anopheles (Nys.) strodei. The complete genome with a length of 9709 nucleotides was characterized by a positive orientation and a single strand, with a single large ORF, which encodes a polyprotein of 2987 amino acids. The phylogenetic analysis showed an association with the viral family Iflaviridae and the Riboviria realm. We carried out comparisons with translated sequences of the capsid regions of other iflavirus, and the identities in relation to our sequence were below the minimum limit of 90%, indicating that possibly it is a new species of iflavirus. Our findings contribute to expanding knowledge of virome composition among mosquito species in Brazil and globally. Moreover, we provide a viral genome reference specific to this geographic region and Culicidae family of mosquitoes. This resource facilitates future in silico recognition and assembly of viral genomes within metagenomic datasets.

Japanese encephalitis is a disease caused by the Japanese encephalitis virus (JEV) and is a concern for U.S. military personnel stationed in the Republic of Korea (ROK). The recent literature reports a potential shift from GI to GV as the dominant genotype circulating in east Asia. In the ROK, GV has been reported in a few Culex spp., but not in the main JEV vector, Cx. tritaeniorhynchus. The goal of this surveillance was to shed light on the current knowledge of the epidemiology of JEV in the ROK by analyzing mosquito collection data from three consecutive years, 2021–2023, and molecularly detecting and genotyping JEV in all Culex spp. collected in several military locations across the ROK. In this study, we detected only JEV GI in Cx. tritaeniorhynchus in 2021 samples. In contrast, all 2022 and 2023 positive samples were GV and detected in Cx. bitaeniorhynchus, Cx. orientalis, and Cx. pipiens. Results support a shift in JEV genotype in the ROK and suggest that for GV, Culex spp. other than Cx. tritaeniorhynchus may be playing an important role.

Mosquitoes of the Culex genus are responsible for a large burden of zoonotic virus transmission globally. Collectively, they play a significant role in the transmission of medically significant diseases such as Japanese encephalitis virus and West Nile virus. Climate change, global trade, habitat transformation and increased urbanisation are leading to the establishment of Culex mosquitoes in new geographical regions. These novel mosquito incursions are intensifying concerns about the emergence of Culex-transmitted diseases and outbreaks in previously unaffected areas. New mosquito control methods are currently being developed and deployed globally. Understanding the complex interaction between pathogens and mosquitoes is essential for developing new control strategies for Culex species mosquitoes. This article reviews the role of Culex mosquitos as vectors of zoonotic disease, discussing the transmission of viruses across different species, and the potential use of Wolbachia technologies to control disease spread. By leveraging the insights gained from recent successful field trials of Wolbachia against Aedes-borne diseases, we comprehensively discuss the feasibility of using this technique to control Culex mosquitoes and the potential for the development of next generational Wolbachia-based control methods.

Abstract Species distribution modeling (SDM) has become an increasingly common approach to explore questions about ecology, geography, outbreak risk, and global change as they relate to infectious disease vectors. Here, we conducted a systematic review of the scientific literature, screening 563 abstracts and identifying 204 studies that used SDMs to produce distribution estimates for mosquito species. While the number of studies employing SDM methods has increased markedly over the past decade, the overwhelming majority used a single method (maximum entropy modeling; MaxEnt) and focused on human infectious disease vectors or their close relatives. The majority of regional models were developed for areas in Africa and Asia, while more localized modeling efforts were most common for North America and Europe. Findings from this study highlight gaps in taxonomic, geographic, and methodological foci of current SDM literature for mosquitoes that can guide future efforts to study the geography of mosquito-borne disease risk. Graphical Abstract

Abstract Background Mosquitoes in the genus Culex are primary vectors in the US for West Nile virus (WNV) and other arboviruses. Climatic drivers such as temperature have differential effects on species-specific changes in mosquito range, distribution, and abundance, posing challenges for population modeling, disease forecasting, and subsequent public health decisions. Understanding these differences in underlying biological dynamics is crucial in the face of climate change. Methods We collected empirical data on thermal response for immature development rate, egg viability, oviposition, survival to adulthood, and adult lifespan for Culex pipiens, Cx. quinquefasciatus, Cx. tarsalis, and Cx. restuans from existing literature according to the PRISMA scoping review guidelines. Results We observed linear relationships with temperature for development rate and lifespan, and nonlinear relationships for survival and egg viability, with underlying variation between species. Optimal ranges and critical minima and maxima also appeared varied. To illustrate how model output can change with experimental input data from individual Culex species, we applied a modified equation for temperature-dependent mosquito type reproduction number for endemic spread of WNV among mosquitoes and observed different effects. Conclusions Current models often input theoretical parameters estimated from a single vector species; we show the need to implement the real-world heterogeneity in thermal response between species and present a useful data resource for researchers working toward that goal. Graphical Abstract

Controlling the vector by eradicating mosquito larvae is frequently done by spreading Abate powder containing 1% Temephos as a larvicide agent. However, the use of Abate powder will possibly increase pesticide residue contamination in the water. This study aimed to explore the potential use of ethanolic extract from red dragon fruit skin to kill mosquito larvae. An experimental laboratory study was performed to examine the larvicidal activity of ethanolic extract from red dragon fruit skin. Maceration method was used to extract active compounds from the fruit skin. Subsequently, the viscous extract was formulated using the wet granulation method to obtain powder form. The larvicide test was done by comparing the lethal concentration (LC50) of mosquito larvae after treatment using the extract and the reference larvicide agent, 1% Temephos Abate. The treatment groups consisted of four different extract concentrations of 1%, 2%, 3%, and 4%. The LC50 of each treatment groups was then calculated against the negative control group. Phytochemical screening showed that the ethanol extract of red dragon fruit skin contains alkaloids, flavonoids, saponins, triterpenoids, and tannins. The larvicide test on Aedes aegypti mosquito larvae showed that the extract treatment hardly killed the Aedes aegypti mosquito larvae. However, on Culex mosquito larvae, the ethanolic extract of red dragon fruit skin was able to kill mosquito larvae, with LC50 value of of 19.06%. The ethanolic extract of red dragon fruit skin was effective on killing the Culex mosquito larvae but not the Aedes aegypti mosquito larvae.

Currently, West-Nile virus (WNV) is spreading worldwide to colder regions due to climate change. Human mortality and morbidity are prevalent and steadily increasing, associated with costs to public health systems. Therefore, the question of the impact of scientific engagement arises. What trends, barriers, and incentives for research related to global burdens are important in this context? To answer these questions, this study provides detailed insights into the publication patterns of WNV research and interprets them using several parameters, such as absolute and relative publication indices and socioeconomic and epidemiological characteristics. It is shown that national interests combined with regional outbreaks significantly influence publication intensity. Thus, a correlation between national publication volume and the number of WNV cases was observed. In contrast to most life science topics, the scientific interest in WNV significantly decreased after 2006. The USA, as the main actor in WNV research, is at the centre of international networking. Recently, European countries are also getting involved according to their new-emerging outbreaks. The results demonstrate national interest in research activities with a lack of globally focused approaches that are urgently needed to better understand and assess the distribution and characteristics of WNV.HighlightsThere is a correlation between national WNV cases and the number of publications.The declining trend in WNV research does not correspond to the growing prevalence.USA as most-publishing country is also at the centre of networking WNV research.France is the primary network partner for African and Middle Eastern countries.

Background: Mosquito-specific viruses (MSVs) comprise a variety of different virus families, some of which are known to interfere with infections of medically important arboviruses. Viruses belonging to the family Mesoniviridae or taxon Negevirus harbor several insect-specific viruses, including MSVs, which are known for their wide geographical distribution and extensive host ranges. Although these viruses are regularly identified in mosquitoes all over the world, their presence in mosquitoes in Germany had not yet been reported. Methods: A mix of three MSVs (Yichang virus [Mesoniviridae] and two negeviruses [Daeseongdong virus and Dezidougou virus]) in a sample that contained a pool of Coquillettidia richiardii mosquitoes collected in Germany was used to investigate the interaction of these viruses with different arboviruses in Culex-derived cells. In addition, small RNA sequencing and analysis of different mosquito-derived cells infected with this MSV mix were performed. Results: A strain of Yichang virus (Mesoniviridae) and two negeviruses (Daeseongdong virus and Dezidougou virus) were identified in the Cq. richiardii mosquitoes sampled in Germany, expanding current knowledge of their circulation in central Europe. Infection of mosquito-derived cells with these three viruses revealed that they are targeted by the small interfering RNA (siRNA) pathway. In Culex-derived cells, co-infection by these three viruses had varying effects on the representative arboviruses from different virus families (Togaviridae: Semliki forest virus [SFV]; Bunyavirales: Bunyamwera orthobunyavirus [BUNV]; or Flaviviridae: Usutu virus [USUV]). Specifically, persistent MSV co-infection inhibited BUNV infection, as well as USUV infection (but the latter only at specific time points). However, the impact on SFV infection was only noticeable at low multiplicity of infection (MOI 0.1) and at specific time points in combination with the infection status. Conclusions: Taken together, these results are important findings that will lead to a better understanding of the complex interactions of MSVs, mosquitoes and arboviruses. [ABSTRACT FROM AUTHOR]

Background: Mosquitoes are the deadliest organisms in the world, killing an estimated 750,000 people per year due to the pathogens they can transmit. Mosquitoes also pose a major threat to other vertebrate animals. Culex territans is a mosquito species found in temperate zones worldwide that feeds almost exclusively on amphibians and can transmit parasites; however, little is known about its ability to transmit other pathogens, including fungi. Batrachochytrium dendrobatidis (Bd) is a topical pathogenic fungus that spreads through contact. With amphibian populations around the world experiencing mass die-offs and extinctions due to this pathogen, it is critical to study all potential modes of transmission. Because Cx. territans mosquitoes are in contact with their hosts for long periods of time while blood-feeding, we hypothesize that they can transmit and pick up Bd. Methods: In this study, we first assessed Cx. territans ability to transfer the fungus from an infected surface to a clean one under laboratory conditions. We also conducted a surveillance study of Bd infections in frogs and mosquitoes in the field (Mountain Lake Biological station, VA, USA). In parallel, we determined Cx. territans host preference via blood meal analysis of field caught mosquitoes. Results: We found that this mosquito species can carry the fungus to an uninfected surface, implying that they may have the ability to transmit Bd to their amphibian hosts. We also found that Cx. territans feed primarily on green frogs (Rana clamitans) and bullfrogs (Rana catesbeiana) and that the prevalence of Bd within the frog population at our field site varied between years. Conclusions: This study provides critical insights into understanding the role of amphibian-biting mosquitoes in transmitting pathogens, which can be applied to disease ecology of susceptible amphibian populations worldwide. [ABSTRACT FROM AUTHOR]

Background Malaria vectors vary in feeding preference depending on their innate behaviour, host availability and abundance. Host preference and human biting rate in malaria vectors are key factors in establishing zooprophylaxis and zoopotentiation. This study aimed at assessing the impact of non-human hosts in close proximity to humans on the human biting rate of primary and secondary malaria vectors, with varying host preferences. Methods The efect of the presence of non-human hosts in close proximity to the human host on the mean catches per person per night, as a proxy for mosquito biting rate, was measured using mosquito-electrocuting traps (METs), in Sagamaganga, Kilombero Valley, Tanzania. Two experiments were designed: (1) a human versus a calf, each enclosed in a MET, and (2) a human surrounded by three calves versus a human alone, with each human volunteer enclosed individually in a MET spaced 10 m apart. Each experiment was conducted on alternate days and lasted for 36 nights per experiment. During each experiment, the positions of hosts were exchanged daily (except the human in experiment 2). All anopheline mosquitoes caught were assayed for Plasmodium sporozoites using enzyme-linked immunosorbent assay. Results A total of 20,574 mosquitoes were captured and identifed during the study, of which 3608 were anophelines (84.4% primary and 15.6% secondary malaria vectors) and 17,146 were culicines. In experiment 1, the primary malaria vector, Anopheles arabiensis, along with Culex spp. demonstrated a preference for cattle, while the primary vectors, Anopheles funestus, preferred humans. In experiment 2, both primary vectors, An. arabiensis and An. funestus, as well as the secondary vector Anopheles rivolurum, demonstrated behaviours amenable to zooprophylaxis, whereas Culex spp. increased their attraction to humans in the presence of nearby cattle. All anopheline mosquitoes tested negative for sporozoites. Conclusions The fndings of this study provide support for the zooprophylaxis model for malaria vectors present in the Kilombero Valley, and for the zoopotentiation model, as it pertains to the Culex spp. in the region. However, the factors regulating zooprophylaxis and zoopotentiation are complex, with diferent species-dependent mechanisms regulating these behaviours, that need to be considered when designing integrated vector management programmes. [ABSTRACT FROM AUTHOR]