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Culex species, such as Culex quinquefasciatus and Culex nigripalpus display a range of feeding habits and act as vectors for pathogens that can cause diseases in both humans and animals. Understanding their feeding habits is pivotal for enhancing disease prevention strategies. The present study introduces the application of two multiplex real‐time PCR (qPCR) followed by high‐resolution melting (HRM) as a cost‐effective and time‐efficient alternative. This investigation involved the development of two multiplex qPCR‐HRM: assay 1 detects human, dog and chicken, while assay 2 detects cat, cattle and horse in Culex sp. engorged female mosquitoes. The qPCR‐HRM reactions showed a detection limit of one copy of genomic DNA when performed as single and multiplex qPCR‐HRM. The reaction efficiencies were 97.96% for human, 100.60% for dog, 99.03% for chicken, 99.92% for feline, 99.18% for cattle and 97.68% for horse. The qPCR‐HRM method, employing multiplex 1 and 2, was applied to field‐collected mosquitoes and demonstrated the ability to detect DNA from multiple blood sources within a single sample. By analysing both multiplexes, it was possible to identify up to five distinct blood sources in Cx. quinquefasciatus and Cx. nigripalpus, and up to two sources in Culex coronatus. Sequencing corroborated the qPCR‐HRM results, confirming the presence of DNA from one to four different blood sources with 100% accuracy. The development of these molecular methods may contribute for identification of blood‐feeding patterns in mosquitoes. It contributes to studies on the dissemination and transmission of pathogens among various animals and humans, thereby bridging the gap between epidemiology and vector monitoring. [ABSTRACT FROM AUTHOR]

The diversity and abundance of vectors are essential parameters in the transmission dynamics of West Nile virus (WNV) between its avian reservoirs and clinically susceptible mammalian species. Knowing the determinants of vector abundance could be thus useful in preventing West Nile fever (WNF) cases and associated socio‐economic impact. We designed a survey at the wildlife–livestock interface to test the hypothesis that variations in environmental favourability between anthropized and wild scenarios modulate WNV vector abundance and transmission risk. In a continental Mediterranean region where WNF has recently emerged, we selected nine sampling sites and allocated three areas to every site with a decreasing gradient of wildlife–livestock interaction: A1—a horse farm where interaction is maximal; A2—a zone of intermediate interaction 500–1000 m from the farm; and A3—an entirely wild zone of low interaction 1–5 km from the farm. At a fortnightly frequency, we estimated mosquito abundance at each of the 27 study sites in May–December 2018 and April–July 2019. We estimated bird and mammal abundance, collected meteorological information and characterised mosquito habitat at the site scale. Thereafter, we studied the determinants of Culex spp., Culex pipiens sensu lato (s.l.) Linnaeus, 1758 (Diptera: Culicidae) and Culex theileri  Theobald, 1903 abundance by constructing negative binomial generalised linear mixed models. We identified 20 mosquito species, with a notable predominance of Culex spp. and, particularly, of Cx. pipiens s.l. We found differences in the spatiotemporal distribution of Culex spp. abundance and confirmed our hypothesis by finding important effects of local environmental variations in abundance. The accumulated rainfall in fortnights 4–14 and the mean temperature of the two fortnights before sampling were positively and statistically significantly associated with the abundance of Cx. pipiens s.l. (Z = 13.09, p < 0.001, and Z = 9.91, p < 0. 001, respectively) and Culex spp. (Z = 13.35, p < 0.001, and Z = 6.99, p < 0.001, respectively), while the mean temperature of the two previous fortnights was a positive statistically significant predictor (Z = 14.69, p < 0.001) of the abundance of Cx. theileri. The farm environment was the most conducive predictor to hosting Culex spp. compared with wild settings. Our results indicate that continental Mediterranean environments are favourable for WNV circulation and maintenance, especially the environment of anthropized rural settings such as farms. These results will have an impact on the spatiotemporal risk prediction of WNF emergence in continental Mediterranean environments. [ABSTRACT FROM AUTHOR]

Mosquito‐borne diseases contribute substantially to the global burden of disease, and are strongly influenced by environmental conditions. Ongoing and rapid environmental change necessitates improved understanding of the response of mosquito‐borne diseases to environmental factors like temperature, and novel approaches to mapping and monitoring risk. Recent development of trait‐based mechanistic models has improved understanding of the temperature dependence of transmission, but model predictions remain challenging to validate in the field. Using West Nile virus (WNV) as a case study, we illustrate the use of a novel remote sensing‐based approach to mapping temperature‐dependent mosquito and viral traits at high spatial resolution and across the diurnal cycle. We validate the approach using mosquito and WNV surveillance data controlling for other key factors in the ecology of WNV, finding strong agreement between temperature‐dependent traits and field‐based metrics of risk. Moreover, we find that WNV infection rate in mosquitos exhibits a unimodal relationship with temperature, peaking at ~24.6–25.2°C, in the middle of the 95% credible interval of optimal temperature for transmission of WNV predicted by trait‐based mechanistic models. This study represents one of the highest resolution validations of trait‐based model predictions, and illustrates the utility of a novel remote sensing approach to predicting mosquito‐borne disease risk. [ABSTRACT FROM AUTHOR]

The study aims to assess the larvicidal efficacy of Diospyros kaki fruit extracts (crude and solvents) against the rice field mosquito, Culex vishnui, which is the vector of Japanese encephalitis. Under simulated semi-field conditions, the larvicidal property of the most effective extract was tested against third instar Cx. vishnui larvae. The crude extract of fruits has a strong larvicidal effect. The larvicidal bioassay with chloroform: methanol extractive outperformed the other solvent extractives of fruits. After 72 h of exposure in a larvicidal bioassay, chloroform: methanol extractive had the lowest LC50 and LC90 values, which were 46 ppm and 186 ppm, respectively, against 1st instar larvae. The semi-field results showed that chloroform: methanol extract was likewise highly effective against third instar Cx. vishnui larvae, with the maximum mortality rate of 100% at 400 ppm concentration after 72 h of exposure. Three major bioactive compounds were identified namely 2,7-dibutoxy-fluoren-9-one- oxime, Pthalic acid, and Tris (2,4-ditert-butylphenyl) phosphate. Non-target organisms, such as Diplonychus annulatum nymphs, were found to be unaffected by the said extracts. Thus, we can claim that the fruits of D. kaki exhibit outstanding larvicidal efficacy against Cx. vishnui mosquito. [ABSTRACT FROM AUTHOR]

West Nile virus (WNV) and St. Louis encephalitis virus (SLEV) are closely related flaviviruses that can cause encephalitis in humans and related diseases in animals. In nature, both are transmitted by Culex, with wild birds, including jays, sparrows, and robins, serving as vertebrate hosts. WNV and SLEV circulate in the same environments and have recently caused concurrent disease outbreaks in humans. The extent that coinfection of mosquitoes or birds may alter transmission dynamics, however, is not well characterized. We therefore sought to determine if coinfection alters infection kinetics and virus levels in birds and infection rates in mosquitoes. Accordingly, American robins (Turdus migratorius), two species of mosquitoes, and vertebrate and invertebrate cells were infected with WNV and/or SLEV to assess how simultaneous exposure may alter infection outcomes. There was variable impact of coinfection in vertebrate cells, with some evidence that SLEV can suppress WNV replication. However, robins had comparable viremia and antibody responses regardless of coinfection. Conversely, in Culex cells and mosquitoes, we saw a minimal impact of simultaneous exposure to both viruses on replication, with comparable infection, dissemination, and transmission rates in singly infected and coinfected mosquitoes. Importantly, while WNV and SLEV levels in coinfected mosquito midguts were positively correlated, we saw no correlation between them in salivary glands and saliva. These results reveal that while coinfection can occur in both avian and mosquito hosts, the viruses minimally impact one another. The potential for coinfection to alter virus population structure or the likelihood of rare genotypes emerging remains unknown. [ABSTRACT FROM AUTHOR]

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. Author summary: Mosquito vectors are strongly affected by temperature, and their distributions are likely to shift under climate change. Lab studies show that mosquito abundance has a unimodal response to temperature with thermal optima, upper and lower thermal limits. However, it remains unknown how mosquito laboratory-derived thermal responses relate to the thermal responses of mosquitoes in nature. We used a global database of field-collected mosquito occurrences, geospatial environmental covariates, and species distribution models to estimate the relationship between temperature and probability of mosquito occurrence. We found that thermal minima (r = 0.87) and, to a lesser degree, thermal optima (r = 0.69) estimated from laboratory studies were correlated with those from the species distribution models. For most species, we did not detect thermal maxima. These results suggest that laboratory studies and field-based machine learning studies are complementary. Together, they can help to better understand vector response to climate change. [ABSTRACT FROM AUTHOR]

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

Nanotechnology is a trend that highly advanced over the past thirty years, addressing and resolving more challenges across different fields. In this investigation, biogenic silver nanoparticles (Ag-NPs) were formed using Brevibacillus agri as an endophytic bacterial strain and utilized as a safe approach to control the spread of medical insect, Culex pipens. The formed Ag-NPs were characterized by UV-vis analysis that showed maximum surface plasmon resonance at a wavenumber of 420 nm. Fourier transforms infrared analysis exhibits the role of various biomolecules in bacterial filtrate reduction and capping as-formed NPs. The sizes of biogenic NPs were 20 - 50 nm with spherical shapes with weight and atomic percentages of Ag ions of 46.20% and 13.96% respectively as shown by SEM-EDX analysis. The crystalline structure was confirmed by XRD. The mortality of various instar larvae and pupae of C. pipens were concentration-dependent and showed maximum percentages of 88.4±0.95, 76.3±0.8, 69.6±1.1, 52.1±1.4, and 47.3±1.01% for first to fourth instar larvae and pupae respectively at a concentration of 30 ppm. The concentrations of proteins, carbohydrates, and lipids of III instar larvae were decreased from (172.1 ± 4.31, 118.1±4.6, and 463.1±1.32 µg/g) for the control to (54.2±1.72, 91.3±2.17, 204.6±9.31 µg/g) respectively upon treatment with 30 ppm of Ag-NPs. [ABSTRACT FROM AUTHOR]

Simple Summary: Mosquitoes transmit pathogens through cycles involving animals and humans. Understanding which animals mosquitoes feed on helps us to monitor these diseases. In our study, we examined blood-fed female mosquitoes from various sites in French Guiana to determine their feeding preferences. We used DNA (genetic material) present in their blood meals to identify the animals they had fed on. We discovered that most mosquitoes we studied, especially those from the Culex group, prefer feeding on mammals like humans and other animals, followed by birds, amphibians, and reptiles. We identified 46 different animal species in the blood meals. Our findings provide important information on mosquito feeding habits and help improve the tracking of diseases spread by mosquitoes, which is crucial for public health. Pathogens transmitted by mosquitoes (Diptera, Culicidae) in sylvatic or urban cycles involve wild or domestic animals and humans, driven by various mosquito species with distinct host preferences. Understanding mosquito–host associations is crucial for ecological insights and pathogen surveillance. In this study, we analyzed mosquito blood meals from coastal French Guiana by amplifying and sequencing host DNA from blood-fed females. Using the 12S ribosomal RNA gene and Sanger sequencing, we identified blood meals from 26 mosquito species across six genera, with 59% belonging to the Culex genus. Nanopore sequencing of selected samples showed 12 mosquito species with one to three mixed blood-meal sources. Mammals were the primary hosts (88%), followed by birds (7%), squamates (3%), and amphibians (2%), indicating a strong preference for mammalian hosts. A total of 46 vertebrate host species were identified, demonstrating high host diversity. This research provides insights into mosquito host usage and highlights the complexities of monitoring arboviruses of public health concern. [ABSTRACT FROM AUTHOR]

Advances in diagnostic techniques coupled with ongoing environmental changes have resulted in intensified surveillance and monitoring of arbovirus circulation in the Amazon. This increased effort has resulted in increased detection of insect-specific viruses among hematophagous arthropods collected in the field. This study aimed to document the first isolation of Agua Salud alphavirus in mosquitoes collected within the Brazilian Amazon. Arthropods belonging to the family Culicidae were collected within a forest fragment located in the Environmental Protection Area of the metropolitan region of Belem. Subsequently, these specimens were meticulously identified to the species level. Afterward, the collected batches were macerated, and the resulting supernatant was then inoculated into C6/36 and Vero cell cultures to facilitate viral isolation. The presence of arboviruses within the inoculated cell cultures was determined through indirect immunofluorescence analysis. Furthermore, positive supernatant samples underwent nucleotide sequencing to precisely identify the viral strains present. Notably, a batch containing Culex (Melanoconion) mosquitoes was identified to be positive for the genus Alphavirus via indirect immunofluorescence. This study is the first report on insect-specific alphavirus isolation in Brazil and the first-ever description of Agua Salud alphavirus isolation within Amazon Forest remnants. [ABSTRACT FROM AUTHOR]

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An assessment of the phytotelmata as mosquito larval habitats in the Darjeeling hill region, India was carried out from July to September 2021 coinciding with the rainy season. Applying random sampling of the selected sites along the different altitudes from the Darjeeling hills, revealed that at least six different varieties of phytotelmata are available with differences in the habitat features and the mosquito immature productivity. The plants like Musa balbisiana (Banana), Colocasia esculenta (Cocoyam), Arundo donax (Narkat), Pandanus furcatus (Tarika), Neoregelia regia (Bromeliad) were observed to be positive for the mosquitoes along with the bamboo stumps. The groves of four bamboo plants namely, Bambusa nutans (Malabas), Dendrocalamus giganteus, (Bhalubas), Dendrocalamus hamiltonii (Choyabas), and Yushania maling (Malingo) were found to be positive for the mosquito larvae. In almost all instances, the dominance of the Aedes mosquitoes was observed with the occasional presence of the chironomid larvae as the dominant insects in the phytotelmata. The phytotelmata types varied with the water content, mosquito larval abundance, and water quality justifying the differences in the mosquito larval habitats. Apparently, the mosquito availability in the Darjeeling Hill region appears to be facilitated by the existence of these diverse phytotelmata as mosquito larval habitats. Interestingly, we also observed altitudinal variations in the pupal productivity irrespective of the phytotelmata types and the differences in the pupal productivity of each of the phytotelmata types. [ABSTRACT FROM AUTHOR]

The aim of this review was to delve into the extent of mosquito virome coverage (proportion of viral reads) via meta-viromic sequencing and uncover potential factors of heterogeneity that could impact this coverage. Data sources were PubMed, Web of Science, Embase, Scopus, Science-Direct, Google Scholar, and the China National Knowledge Infrastructure. Pooled coverage was estimated using random-effects modeling, and subgroup analyses further reveal potential heterogeneous factors. Within the three mosquito genera studied, Culex exhibited the highest pooled viral coverage of mosquito viromes at 7.09% (95% CI: 3.44–11.91%), followed by Anopheles at 5.28% (95% CI: 0.45–14.93%), and Aedes at 2.11% (95% CI: 0.58–7.66%). Subgroup analyses showed that multiple processing methods significantly affected the viral coverage of mosquito viromes, especially pre-treatment of mosquito samples with saline buffer/medium and antibiotics prior to DNase/RNase treatment and removal of the host genome prior to RNA library construction. In conclusion, the results of this study demonstrate that the viral coverage of mosquito viromes varies between mosquito genera and that pre-treatment of mosquito samples with saline buffer/medium and antibiotics before DNase/RNase treatment and removing host genomes prior to RNA library construction are critical for the detection of RNA viruses in mosquito vectors using meta-viromic sequencing. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract Background Over the last few decades, river ecosystem is highly modified through various anthropogenic activities which are resulted to alter ecosystem functions and services. This modified ecosystem rendering conducive environment to mosquitoes through various ecological links for the self-sustaining populations. However, deciphering the community assemblage of immature mosquitoes with reference to water quality at modified ecosystem is very essential to make suitable control measure to curtail mosquito populations. In order to understand how the water quality influences the larval density, habitat specificity and community assemblage of immature mosquito populations, a study was conducted at different ecosystems (urban, semi-urban and rural) along the Vaigai river. The physicochemical parameters such as pH, TDS, salinity, conductivity, turbidity, DO, were analyzed at each study site. Results Our results clearly revealed that Anopheline species were highly preferred to breed less polluted habitat than Culicine species. Community assemblage by Anopheline and Culicine mosquitoes were found to be higher at all the studies whilst community assemblage by Anopheline were maximum at rural and semi-urban sites. Among the Anopheline species, Anopheles subpictus able to breed at high polluted habitat, particularly higher turbid level (28.49 ± 2.18 NTU) than other Anopheles species. Cx. gelidus mostly breed at sewage disposal habitats with high salinity level (1.01 ± 0.08) whilst Cx. bitaeniorhynchus bred in only fresh water bodies particularly low turbid habitats (3.97 ± 0.40 NTU). Grouping of immature mosquitoes based on the habitat similarity, An. subpictus, Cx. vishnui, An. vagus, Cx. tritaeniorhynchus, Cx. gelidus and Cx. quinquefasciatus were able to breed in highly polluted habitats which are resulted fell in group A than group B mosquitoes. Cx. vishnui and An. subpictus have strong habitat similarity (0.96) and can able to share their habitats with more number of Anopheline and Culicine mosquitoes. Conclusions From the study we concluded that, Cx. vishnui and An. subpictus were most prevalent species and strong habitats similarity along the Vaigai river basin. An. subpictus and An. vagus can adapt to breed in polluted habitats and this may be adequate to extend the vectorial capacity and disease outbreak along the Vaigai river basin.

In the current study, the effect of lethal concentrations of drug solutions and chemicals against pupae of Culex pipiens molestus mosquitoes was investigated after 24 hours and up to 3 days of the experiment using different concentrations. Regarding the drug solutions, the concentration of 100 ppm of the niclosamide drug solution showed a 100% lethal effect on the second day of the experiment, and the value of LC50 was 113.7 ppm. The concentration of 800 ppm of the sulfasalazine drug solution caused the death of all the pupa, with a death rate of 100% on the third day of the experiment, and the value of LC50 was 854.7 ppm. The concentration of 2000 ppm for both piperazine and isoniazid medicinal solutions caused a death rate of 100% on the second and third day of the experiment, respectively, and the LC50 value for both piperazine and isoniazid was 2220.7 and 2273.0 ppm, respectively. The concentration of 14,000 ppm of the drug solution 4-aminoantipyrine caused the death of all pupal mosquitoes on the second day of the experiment, and the value of LC50 was 14,360.0 ppm. As for solutions of chemicals, the concentration of 150 ppm of the chemical solution ethyl salicylate showed a lethal effect on all pupal mosquitoes, with a 100% death rate on the second day of the experiment, and the value of LC50 was 176.2 ppm. The death rate of mosquito pups reached 100% when using chemical solutions of thymol, sulfamic acid, sulfanilamide, and 2,6-dihydroxytoluene at concentrations of 240, 2000, 2500 and 10,000 ppm, respectively. The value of LC50 for each of thymol and sulfamic acid was, Sulfanilamide and 2,6-dihydroxytoluene 352.1, 2940.8, 2582.5 and 10750.0 ppm, respectively.

Ivermectin (IVM)-treated birds provide the potential for targeted control of Culex mosquitoes to reduce West Nile virus (WNV) transmission. Ingestion of IVM increases mosquito mortality, which could reduce WNV transmission from birds to humans and in enzootic maintenance cycles affecting predominantly bird-feeding mosquitoes and from birds to humans. This strategy might also provide an alternative method for WNV control that is less hampered by insecticide resistance and the logistics of large-scale pesticide applications. Through a combination of field studies and modeling, we assessed the feasibility and impact of deploying IVM-treated birdfeed in residential neighborhoods to reduce WNV transmission. We first tracked 105 birds using radio telemetry and radio frequency identification to monitor their feeder usage and locations of nocturnal roosts in relation to five feeder sites in a neighborhood in Fort Collins, Colorado. Using these results, we then modified a compartmental model of WNV transmission to account for the impact of IVM on mosquito mortality and spatial movement of birds and mosquitoes on the neighborhood level. We found that, while the number of treated lots in a neighborhood strongly influenced the total transmission potential, the arrangement of treated lots in a neighborhood had little effect. Increasing the proportion of treated birds, regardless of the WNV competency status, resulted in a larger reduction in infection dynamics than only treating competent birds. Taken together, model results indicate that deployment of IVM-treated feeders could reduce local transmission throughout the WNV season, including reducing the enzootic transmission prior to the onset of human infections, with high spatial coverage and rates of IVM-induced mortality in mosquitoes. To improve predictions, more work is needed to refine estimates of daily mosquito movement in urban areas and rates of IVM-induced mortality. Our results can guide future field trials of this control strategy.

The assembly of host‐associated microbial communities is influenced by multiple factors, but the effect of microbiomes on host phenotypes is often not well understood. To address questions of food‐web effects on host microbiome assembly, we manipulated the resource environment (grass only [G] vs. grass + nutrients [GN]), competition type (intra‐ vs. inter‐specific) and density (high vs. low) for Culex restuans mosquito larvae. We predicted the microbial communities in fourth‐instar larvae would differ between these environmental treatments and that these treatments would translate into differences in the adult phenotype.Resource environment and density influenced the larval microbiome. In addition, the larval microbiome exhibited notable differences compared to the free‐living microbial communities.Resource‐driven differences in the larval samples can be attributed to Arcobacteraceae being more abundant in larvae reared in the GN treatments relative to those reared in the G treatments and Comamonadaceae being more abundant in the G treatment. Although significant, the difference in community structure between density treatments was difficult to discern. This appears to be driven by Weeksellaceae only being abundant in the high‐density, interspecific, GN treatment.Rearing larvae to adulthood under severe food limitation resulted in low survival (<25%) in both resource environments. Approximately 60% of survivors to adulthood were male. Larvae reared in the intraspecific, G treatment had the shortest development time to adulthood and emerged as the smallest adults.These results demonstrate how environmental variation can significantly alter the alpha and beta diversity of free‐living microbes, which in turn can significantly affect host phenotype and critical life history traits, such as development time, size at adulthood, and survival. These findings highlight the importance of considering environmental influences on microbiome diversity to understand and predict host outcomes, offering valuable insights for diverse applications in fields such as ecology, public health, and agriculture. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Mosquitoes transmit quite a lot of deadly diseases, including chikungunya, brain fever, dengue fever, Japanese encephalitis, hemorrhagic fever, malaria, filariasis, yellow fever, and Zika fever. Every year, mosquito-borne diseases affect millions of people. For that reason, mosquito control is vital for public health, especially in tropical countries. To manage mosquitoes, various synthetic chemicals are used, each with its advantages and disadvantages. Chemical management has a satisfactory effect on reducing mosquito populations, even though mosquito resistance and the presence of pesticide residues in the environment and the development of resistance in mosquito control tactics are crucial concerns. It is difficult to control whether an insecticide-resistant mosquito population causes to the reappearance of vector-borne diseases. In this emergency circumstance, we need to look for alternatives. Alternatively, plant-based phytochemicals are still more widely used for mosquito control programs, in terms of cost since they are efficient and economical, eco-friendly, safe for humans and the environment, and may be used as an effective alternative to chemical larvicides. The current state of data on phytochemical sources has variation in larvicidal activity according to mosquito species, instar specificity, the polarity of solvents used during extraction, the nature of the active ingredient, and promising advances made in biological control of mosquitoes by plant-derived reviewed in this article. [ABSTRACT FROM AUTHOR]

BACKGROUND: The incompatible insect technique (IIT) has been used for Aedes mosquito population suppression to curb the transmission of dengue. However, its wide application is limited owing to the low output of male mosquitoes and the risk of population replacement from the release of fertile Wolbachia‐infected females. This study aims to improve IIT efficiency for broader adoption. RESULTS: We assessed the impact of 10% pyriproxyfen (PPF) sticky powder exposure on Wolbachia (from Culex molestus)‐transinfected Aedes albopictus Guangzhou line (GUA line) (GC) mosquitoes. We found that the exposure caused chronic toxicity in adult mosquitoes without affecting the cytoplasmic incompatibility (CI)‐inducing capability of males. The PPF‐contaminated GC females exhibited significant sterilization and the ability to disseminate lethal doses of PPF to breeding sites. Subsequently, we conducted a field trial combining PPF with IIT aiming to suppress the Ae. albopictus population. This combined approach, termed boosted IIT (BIIT), showed a notable enhancement in population suppression efficiency. The improved efficacy of BIIT was attributed to the dispersion of PPF particles in the field via the released PPF‐contaminated male mosquitoes. During the BIIT field trial, no Wolbachia wPip‐positive Ae. albopictus larvae were detected, indicating the effective elimination of the risk of Wolbachia‐induced population replacement. Additionally, the field trial of BIIT against Ae. albopictus resulted in the suppression of the nontarget mosquito species Culex quinquefasciatus. CONCLUSION: Our results highlight the remarkable efficiency and feasibility of combining IIT with PPF in suppressing mosquito populations, facilitating the widespread implementation of IIT‐based management of mosquito‐borne diseases. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background and purpose: Culicidae plays a crucial role in the spread of diseases like Malaria and arboviral diseases such as Japanese Encephalitis, Zika virus, Chikungunya, Dengue fever, Yellow fever, and West Nile fever. Different mosquito species have different abundance and distribution due to a variety of factors, such as physicochemical factors, weather conditions, vegetation, and human activities. The characteristics of larval habitats are among the factors affecting the population, and diversity, and determining the preferred breeding places of the species. For this reason, a study was conducted to examine the species diversity of Culicidae and their larval habitats in Shushtar County. Materials and methods: This cross-sectional study was conducted in Shushtar County from the central, Mian Ab and Shuaibiya districts, 2021. Larvae were collected from the breeding places in the sampling areas including Noormohammadi, Konarpir, Saheli, Abu Gerva, and Pirdalu by using the dipping technique in spring and summer and kept in lactophenol solution. Then, a microscopic slide was prepared from the samples using licidophor, which were identified with valid diagnostic keys. Characteristics of larval habitat such as habitat type, water condition, vegetation, substrate type, water quality, and light condition were also recorded. To analyze the data and determine the relationship between the frequency and characteristics of larval habitats, SPSS version 16 software and a chi-square statistical test were used. Qualitative variables were also reported as numbers (percentages). Abundance and distribution maps of samples were prepared using ArcGIS software version 10.5. Results: In this study, a total of 527 larvae were identified, including five species of Anopheles stephensi, Culex pusillus, and Cx. sinaiticus, Cx. theileri and, Cx. Perexiguus. Anopheles stephensi with the lowest frequency (8.48%) was reported only from the Saheli area. This species was collected from temporary, stationary, without vegetation, mud substrate, cloudy water, sunny, and natural habitats. Culex theileri, one of the important vectors of arboviral diseases, was observed in temporary and natural habitats, stagnant waters without vegetation, and cloudy and sunny waters habitates. Culex pusillus was the only species observed in Pirdalo and Abu Gerva. By examining the larval habitats of Culicidae in Shushtar County, Cx. pusillus was the most abundant collected species, from temporary habitat (82.7%), stagnant waters (73.6%), mud substrate (100%), turbid waters (78.6%), natural (95%), and sunny (82.7%). Also, the abundance of collected larvae in temporary habitats, stagnant waters, in areas with vegetation, and mud substrate was high. Besides, it was more abundant in turbid waters and sunny areas, and these differences were statistically significant (P

Background and Aim: Some Culex mosquitoes are competent vectors for Japanese encephalitis virus (JEV) and Zika virus (ZIKV), which cause public health problems worldwide, especially in South-east Asia. Xenosurveillance of Culex mosquitoes remains limited compared with other common mosquito-borne diseases. This study aimed to identify JEV and ZIKV in field-caught Culex mosquitoes collected from Ubon Ratchathani province. Materials and Methods: We investigated the presence of JEV and ZIKV in Culex mosquitoes from two districts in Ubon Ratchathani province, Thailand, and examined their role in viral interepidemic circulation. Female Culex mosquitoes (5,587) were collected using a mechanical aspirator from indoors and outdoors. The consensus sequences of the E and NS1 genes of JEV and the E gene of ZIKV were identified using real-time reverse transcription-polymerase chain reaction. Results: From 335 sample pools that contain a total of 5587 adult female Culex mosquitoes collected from Don Yung, Mueang district (4,406) and Phon Duan, Det Udom district (1,181), none of the collected mosquitoes tested positive for either JEV or ZIKV. Conclusion: This study did not find JEV and ZIKV in Culex mosquitoes collected from the area of collection, which may be due to the low circulating amount of the virus in the vectors in the area, making it undetectable, or it may be because Culex mosquitoes are not suitable vector for the virus being tested. However, further xenosurveillance study of JEV and ZIKV in mosquito is suggested to prepare for the next outbreak.

Abstract Background Mosquitoes are important vectors of pathogens. They are usually collected with CO2-baited traps and subsequently identified by morphology. This procedure is very time-consuming. Automatic counting traps could facilitate timely evaluation of the local risk for mosquito-borne pathogen transmission or decision-making on vector control measures, but the counting accuracy of such devices has rarely been validated in the field. Methods The Biogents (BG)-Counter 2 automatically counts mosquitoes by discriminating the size of captured objects directly in the field and transmits the data to a cloud server. To assess the accuracy of this counting device, 27 traps were placed at 19 sampling sites across Germany and used in daily, weekly or bimonthly intervals from April until October 2021. The BG-Counter 2 was attached to a CO2-trap (BG-Pro trap = CO2-Pro) and the same trap was converted to also attract gravid mosquitoes (upside-down BG-Pro trap with a water container beneath = CO2-Pro-gravid). All captured mosquitoes were identified by morphology. The number of females (unfed and gravid), mosquito diversity and the number of identified specimens in relation to the counting data of the BG-Counter were compared between the two trapping devices to evaluate sampling success and counting accuracy. Results In total 26,714 mosquitoes were collected during 854 trap days. The CO2-Pro-gravid trap captured significantly more mosquitoes per trap day for all specimens, gravid females and non-gravid females, while there was no difference in the mosquito diversity. The linear model with the captured mosquitoes as a response and the counted specimens as a predictor explained only a small degree of the variation within the data (R 2 = 0.16), but per individual trap the value could reach up to 0.62 (mean R 2 = 0.23). The counting accuracy for the daily samples had a significant positive correlation with sample size, resulting in higher accuracy for the CO2-Pro-gravid trap and higher accuracy for sites and sampling months with high mosquito abundance. Conclusions While the accuracy of the BG-Counter 2 is quite low, the device is able to depict mosquito phenology and provide information about local population dynamics. Graphical Abstract

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

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

Background and purpose: Culicidae plays a crucial role in the spread of diseases like Malaria and arboviral diseases such as Japanese Encephalitis, Zika virus, Chikungunya, Dengue fever, Yellow fever, and West Nile fever. Different mosquito species have different abundance and distribution due to a variety of factors, such as physicochemical factors, weather conditions, vegetation, and human activities. The characteristics of larval habitats are among the factors affecting the population, and diversity, and determining the preferred breeding places of the species. For this reason, a study was conducted to examine the species diversity of Culicidae and their larval habitats in Shushtar County. Materials and methods: This cross-sectional study was conducted in Shushtar County from the central, Mian Ab and Shuaibiya districts, 2021. Larvae were collected from the breeding places in the sampling areas including Noormohammadi, Konarpir, Saheli, Abu Gerva, and Pirdalu by using the dipping technique in spring and summer and kept in lactophenol solution. Then, a microscopic slide was prepared from the samples using licidophor, which were identified with valid diagnostic keys. Characteristics of larval habitat such as habitat type, water condition, vegetation, substrate type, water quality, and light condition were also recorded. To analyze the data and determine the relationship between the frequency and characteristics of larval habitats, SPSS version 16 software and a chi-square statistical test were used. Qualitative variables were also reported as numbers (percentages). Abundance and distribution maps of samples were prepared using ArcGIS software version 10.5. Results: In this study, a total of 527 larvae were identified, including five species of Anopheles stephensi, Culex pusillus, and Cx. sinaiticus, Cx. theileri and, Cx. Perexiguus. Anopheles stephensi with the lowest frequency (8.48%) was reported only from the Saheli area. This species was collected from temporary, stationary, without vegetation, mud substrate, cloudy water, sunny, and natural habitats. Culex theileri, one of the important vectors of arboviral diseases, was observed in temporary and natural habitats, stagnant waters without vegetation, and cloudy and sunny waters habitates. Culex pusillus was the only species observed in Pirdalo and Abu Gerva. By examining the larval habitats of Culicidae in Shushtar County, Cx. pusillus was the most abundant collected species, from temporary habitat (82.7%), stagnant waters (73.6%), mud substrate (100%), turbid waters (78.6%), natural (95%), and sunny (82.7%). Also, the abundance of collected larvae in temporary habitats, stagnant waters, in areas with vegetation, and mud substrate was high. Besides, it was more abundant in turbid waters and sunny areas, and these differences were statistically significant (P<0.0001). Conclusion: In this study, Cx.pusillus was the most common species, while An. stephensi was the least common abundance among collected species. It was observed significant difference between this species and the type of habitat, water flow, vegetation, water quality, and light conditions. Conducting more comprehensive studies in this field, especially regarding the possible role of different species of Culicidae, helped to solve the health and medical problems of humans and veterinary medicine in the future. [ABSTRACT FROM AUTHOR]

The present study investigated the relative abundance of mosquitoes larvae in three different breeding habitats in seven districts of Beni-Suef Governorate, Egypt during two years (2019–2020). The results showed that the species composition of mosquitoes was represented by four genera and eight species; four of Culex (Cx. pipiens Linnaeus,1758, Cx. theileri Theobald,1903, Cx. perexiguus Theobald, 1903 and Cx. antennatus (Becker,1903)), two of Anopheles (An. coustani Laveran, 1900 and An. multicolor Cambouliu, 1902) and one each of Aedes and Culiseta (Ae. caspius (Pallas, 1771) and Cs. longiareolata (Macquart, 1838)). The results were demonstrated also, that Cx. pipiens (4934 larvae) was the most common species, followed by Cx. perexiguus (2991 larvae) in all breeding habitats. Particularly, this species distributed in drains more than irrigation tanks and agriculture canals, the other species of Anopheles and Aedes were very little or absence in all test breeding habitats. The densities of almost mosquitoes species with higher collections were found in Saft, Elfant and El-Maimun villages and rate of collection decreased in Tizmant and Bedahl. These information on the species composition and their abundance of mosquitoes in Beni-Suef may help better planning for the integrated pest management for these vectors and transmitted diseases. [ABSTRACT FROM AUTHOR]

Culex tarsalis Coquillett (Diptera: Culicidae) mosquitoes are capable of vectoring numerous pathogens affecting public and animal health. Unfortunately, the probing behaviors of mosquitoes are poorly understood because they occur in opaque tissues. Electropenetrography (EPG) has the potential to elucidate these behaviors by recording the electrical signals generated during probing. We used an AC–DC EPG with variable input resistors (Ri levels) to construct a waveform library for Cx. tarsalis feeding on human hands. Biological events associated with mosquito probing were used to characterize waveforms at four Ri levels and with two electrical current types. The optimal settings for EPG recordings of Cx. tarsalis probing on human hands was an Ri level of 107 Ohms using an applied signal of 150 millivolts alternating current. Waveforms for Cx. tarsalis included those previously observed and associated with probing behaviors in Aedes aegypti L. (Diptera: Culicidae): waveform families J (surface salivation), K (stylet penetration through the skin), L (types 1 and 2, search for a blood vessel/ingestion site), M (types 1 and 2, ingestion), N (type 1, an unknown behavior which may be a resting and digestion phase), and W (withdrawal). However, we also observed variations in the waveforms not described in Ae. aegypti, which we named types L3, M3, M4, and N2. This investigation enhances our understanding of mosquito probing behaviors. It also provides a new tool for the automated calculation of peak frequency. This work will facilitate future pathogen acquisition and transmission studies and help identify new pest and disease management targets. [ABSTRACT FROM AUTHOR]

Simple Summary: The main mosquito species capable of transmitting arboviruses belong to the genera Aedes spp., Psorophora spp., Anopheles spp., Culex spp., Mansonia spp., Coquillettidia spp., Haemagogus spp., Sabethes spp., Culiseta spp., and Wyeomyia spp. Some neglected mosquito species have the potential to become significant disease vectors due to parameters such as global distribution, rapid adaptation to urban areas, and anthropophilic habits. This review discusses the epidemiological importance and biology of three neglected mosquitoes, Aedes scapularis, Aedes vittatus, and Aedes japonicus japonicus, in the context of vectorial capacity and how urbanization, climate change, and globalization alter disease transmission dynamics and may increase the participation of neglected species in propagating diseases. More than 3550 species of mosquitoes are known worldwide, and only a fraction is involved in the transmission of arboviruses. Mosquitoes in sylvatic and semi-sylvatic habitats may rapidly adapt to urban parks and metropolitan environments, increasing human contact. Many of these mosquitoes have been found naturally infected with arboviruses from the Alphaviridae, Flaviviridae, and Bunyaviridae families, with many being the cause of medically important diseases. However, there is a gap in knowledge about the vector status of newly invasive species and their potential threat to human and domestic animal populations. Due to their rapid distribution, adaptation to urban environments, and anthropophilic habits, some neglected mosquito species may deserve more attention regarding their role as secondary vectors. Taking these factors into account, we focus here on Aedes (Ochlerotatus) scapularis (Rondani), Aedes japonicus japonicus (Theobald), and Aedes (Fredwardsius) vittatus (Bigot) as species that have the potential to become important disease vectors. We further discuss the importance of these neglected mosquitoes and how factors such as urbanization, climate change, and globalization profoundly alter the dynamics of disease transmission and may increase the participation of neglected species in propagating diseases. [ABSTRACT FROM AUTHOR]

Background: West Nile virus (WNV), Everglades virus (EVEV), and five species of Orthobunyavirus were isolated from mosquitoes collected in the Everglades in 2016–2017. Prior studies of blood meals of mosquitoes in southern Florida have related findings to acquisition and transmission of EVEV, St. Louis encephalitis virus, and WNV, but not the Orthobunyavirus viruses associated with the subgenus Melanoconion of the genus Culex. Materials and Methods: In the present study, blood-fed mosquitoes were collected in the Everglades in 2016, 2017, 2021, and 2022, and from an industrial site in Naples, FL in 2017. Blood meals were identified to host species by PCR assays using mitochondrial cytochrome b gene. Results: Blood meals were identified from Anopheles crucians complex and 11 mosquito species captured in the Florida Everglades and from 3 species collected from an industrial site. The largest numbers of blood-fed specimens were from Culex nigripalpus, Culex erraticus, Culex cedecei, and Aedes taeniorhynchus. Cx. erraticus fed on mammals, birds, and reptiles, particularly American alligator. This mosquito species could transmit WNV to American alligator in the wild. Cx. nigripalpus acquired blood meals primarily from birds and mammals and frequently fed on medium-sized mammals and white-tailed deer. Water and wading birds were the primary avian hosts for Cx. nigripalpus and Cx. erraticus in the Everglades. Wading birds are susceptible to WNV and could serve as reservoir hosts. Cx. cedecei fed on five species of rodents, particularly black and hispid cotton rats. EVEV and three different species of Orthobunyavirus have been isolated from the hispid cotton rat and Cx. cedecei in the Everglades. Cx. cedecei is likely acquiring and transmitting these viruses among hispid cotton rats and other rodents. The marsh rabbit was a frequent host for An. crucians complex. An. crucians complex, and other species could acquire Tensaw virus from rabbits. Conclusions: Our study contributes to a better understanding of the host and viral associations of mosquito species in southwestern Florida. [ABSTRACT FROM AUTHOR]

The spatial pattern of many pathogens that are transmitted by mosquitoes (Diptera: Culicidae) is changing due to globalization and climate change. Thus, surveillance of mosquito vectors is becoming increasingly widespread as basis for risk assessment and control. Species identification by loop‐mediated isothermal amplification (LAMP) assays is simple, fast and reliable. Specific primers for several mosquito species are available (Aedes aegypti, Aedes albopictus, Aedes geniculatus, Aedes japonicus, Aedes koreicus, species of the Anopheles funestus group and An. gambiae complex). In the present work, LAMP assays targeting the rDNA internal transcribed spacer region were developed for Ae. cretinus, a minor sister taxon of Ae. albopictus, and for Culex pipiens/Culex quinquefasciatus and Culex torrentium. The specificities of the primers designed in silico were confirmed by in vitro tests with DNA form non‐target species. Further, the release of DNA from mosquito stages (eggs, larvae, pupae, adults) was investigated, revealing that an incubation for 5 min at 80°C in water is suitable for LAMP. With this method, one specimen (egg, larva, pupa, adult) of a target species could be detected among 49 non‐targets. Thus, the assays are suitable for fast and reliable identification of mosquito species of all life stages by colour change visible to the naked eye, and they are operable under field conditions. [ABSTRACT FROM AUTHOR]

A study was carried out in 345 water ecosystems (rivers, streams, ditches, lagoons, ponds, puddles, rockpools, and artificial containers) throughout the autonomous community of Galicia (NW Spain) during different seasons between 2020 and 2023. The results revealed the first detections of Anopheles (Anopheles) petragnani Del Vechio, 1939 (n  = 185) and Culex (Culex) mimeticus Noè, 1899 (n  = 12) in the territory, allowing to update and expand the overall knowledge about their distribution and ecology. The breeding preferences of the species were analyzed considering habitat characteristics (land use, water body type, and degree of insolation), geographical variables (latitude, longitude, and altitude), and physical–chemical water parameters (temperature, pH, dissolved oxygen, electrical conductivity, salinity, total dissolved solids, and turbidity). In addition, the relationship between the occurrence of these species and other mosquitoes present in the study area was discussed. Anopheles petragnani is widely distributed in the region and shows breeding preferences for water bodies of fluvial origin, forest environments, and shaded situations. Culex mimeticus was detected sporadically breeding in a pond and in a river in the south of the region, both in industrial and agricultural land uses, and always exposed to the sun. Although the presence of these species may not currently pose a health risk in the region, it is important not neglecting their study since the adequate characterization of their larval biotopes is relevant regarding vector surveillance and control measures. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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

Lymphatic filariasis (LF) is a crippling and disfiguring parasitic condition. India accounts for 55% of the world's LF burden. The filarial parasite Wuchereria bancrofti is known to cause 99.4% of the cases while, Brugia malayi accounts for 0.6% of the issue occurring mainly in some pockets of Odisha and Kerala states. The Balasore (Baleswar) district of Odisha has been a known focus of B. malayi transmission. We employed molecular xenomonitoring to detect filarial parasite DNA in vectors. In six selected villages, Gravid traps were used to collect Culex mosquitoes and hand catch method using aspirators was followed for collection of mansonioides. A total of 2903 mosquitoes comprising of Cx. quinquefasciatus (n = 2611; 89.94%), Cx. tritaeniorhynchus (n = 100; 3.44%), Mansonia annuliferea (n = 139; 4.78%) and Mansonia uniformis (n = 53; 1.82%) were collected from six endemic villages. The species wise mosquitoes were made into 118 pools, each with a maximum of 25 mosquitoes, dried and transported to the laboratory at VCRC, Puducherry. The mosquito pools were subjected to parasite DNA extraction, followed by Real-time PCR using LDR and HhaI probes to detect W. bancrofti and B. malayi infections, respectively. Seven pools (6.66%) of Cx. quinquefasciatus, showed infection with only W. bancrofti while none of the pools of other mosquito species showed infection with either W. bancrofti or B. malayi. Although the study area is endemic to B. malayi, none of the vectors of B. malayi was found with parasite infection. This study highlights the ongoing transmission of bancroftian filariasis in the study villages of Balasore district of Odisha and its implications for evaluating LF elimination programme. [ABSTRACT FROM AUTHOR]

Usutu virus (USUV), an arbovirus from the Flaviviridae family, genus Flavivirus, has recently gained increasing attention because of its potential for emergence. After his discovery in South Africa, USUV spread to other African countries, then emerged in Europe where it was responsible for epizootics. The virus has recently been found in Asia. USUV infection in humans is considered to be most often asymptomatic or to cause mild clinical signs. However, a few cases of neurological complications such as encephalitis or meningo-encephalitis have been reported in both immunocompromised and immunocompetent patients. USUV natural life cycle involves Culex mosquitoes as its main vector, and multiple bird species as natural viral reservoirs or amplifying hosts, humans and horses can be incidental hosts. Phylogenetic studies carried out showed eight lineages, showing an increasing genetic diversity for USUV. This work describes the development and validation of a novel whole-genome amplicon-based sequencing approach to Usutu virus. This study was carried out on different strains from Senegal and Italy. The new approach showed good coverage using samples derived from several vertebrate hosts and may be valuable for Usutu virus genomic surveillance to better understand the dynamics of evolution and transmission of the virus. [ABSTRACT FROM AUTHOR]

Arboviruses transmitted by mosquitoes, including Japanese encephalitis virus (JEV), present a substantial global health threat. JEV is transmitted by mosquitoes in the genus Culex, which are common in both urban and rural areas in Vietnam. In 2020, we conducted a 1-year survey of Culex mosquito abundance in urban, suburban, and peri-urban areas of Hanoi using CDC-light traps. Mosquitoes were identified to species and sorted into pools based on species, sex, and trap location. The mosquito pools were also investigated by RT-qPCR for detection of JEV. In total, 4829 mosquitoes were collected over a total of 455 trap-nights, across 13 months. Collected mosquitoes included Culex, Aedes, Anopheles, and Mansonia species. Culex mosquitoes, primarily Cx. quinquefasciatus, predominated, especially in peri-urban areas. Most Culex mosquitoes were caught in the early months of the year. The distribution and abundance of mosquitoes exhibited variations across urban, suburban, and peri-urban sites, emphasizing the influence of environmental factors such as degree of urbanization, temperature and humidity on Culex abundance. No JEV was detected in the mosquito pools. This study establishes baseline knowledge of Culex abundance and temporal variation, which is crucial for understanding the potential for JEV transmission in Hanoi. [ABSTRACT FROM AUTHOR]

Background: A treated fabric device for emanating the volatile pyrethroid transfluthrin was recently developed in Tanzania that protected against night-biting Anopheles and Culex mosquitoes for several months. Here perceptions of community end users provided with such transfluthrin emanators, primarily intended to protect them against day-active Aedes vectors of human arboviruses that often attack people outdoors, were assessed in Port-au-Prince, Haiti. Methods: Following the distribution of transfluthrin emanators to participating households in poor-to-middle class urban neighbourhoods, questionnaire surveys and in-depth interviews of end-user households were supplemented with conventional and Photovoice-based focus group discussions. Observations were assessed synthetically to evaluate user perceptions of protection and acceptability, and to solicit advice for improving and promoting them in the future. Results: Many participants viewed emanators positively and several outlined various advantages over current alternatives, although some expressed concerns about smell, health hazards, bulkiness, unattractiveness and future cost. Most participants expressed moderate to high satisfaction with protection against mosquitoes, especially indoors. Protection against other arthropod pests was also commonly reported, although satisfaction levels were highly variable. Diverse use practices were reported, some of which probably targeted nocturnal Culex resting indoors, rather than Aedes attacking them outdoors during daylight hours. Perceived durability of protection varied: While many participants noted some slow loss over months, others noted rapid decline within days. A few participants specifically attributed efficacy loss to outdoor use and exposure to wind or moisture. Many expressed stringent expectations of satisfactory protection levels, with even a single mosquito bite considered unsatisfactory. Some participants considered emanators superior to fans, bedsheets, sprays and coils, but it is concerning that several preferred them to bed nets and consequently stopped using the latter. Conclusions: The perspectives shared by Haitian end-users are consistent with those from similar studies in Brazil and recent epidemiological evidence from Peru that other transfluthrin emanator products can protect against arbovirus infection. While these encouraging sociological observations contrast starkly with evidence of essentially negligible effects upon Aedes landing rates from parallel entomological assessments across Haiti, Tanzania, Brazil and Peru, no other reason to doubt the generally encouraging views expressed herein by Haitian end users could be identified. [ABSTRACT FROM AUTHOR]

Background: Culex tritaeniorhynchus is widely distributed in China, from Hainan Island in the south to Heilongjiang in the north, covering tropical, subtropical, and temperate climate zones. Culex tritaeniorhynchus carries 19 types of arboviruses. It is the main vector of the Japanese encephalitis virus (JEV), posing a serious threat to human health. Understanding the effects of environmental factors on Culex tritaeniorhynchus can provide important insights into its population structure or isolation patterns, which is currently unclear. Results: In total, 138 COI haplotypes were detected in the 552 amplified sequences, and the haplotype diversity (Hd) value increased from temperate (0.534) to tropical (0.979) regions. The haplotype phylogeny analysis revealed that the haplotypes were divided into two high-support evolutionary branches. Temperate populations were predominantly distributed in evolutionary branch II, showing some genetic isolation from tropical/subtropical populations and less gene flow between groups. The neutral test results of HNQH (Qionghai) and HNHK(Haikou) populations were negative (P < 0.05), indicating many low-frequency mutations in the populations and that the populations might be in the process of expansion. Moreover, Wolbachia infection was detected only in SDJN (Jining) (2.24%), and all Wolbachia genotypes belonged to supergroup B. To understand the influence of environmental factors on mosquito-borne viruses, we examined the prevalence of Culex tritaeniorhynchus infection in three ecological environments in Shandong Province. We discovered that the incidence of JEV infection was notably greater in Culex tritaeniorhynchus from lotus ponds compared to those from irrigation canal regions. In this study, the overall JEV infection rate was 15.27 per 1000, suggesting the current risk of Japanese encephalitis outbreaks in Shandong Province. Conclusions: Tropical and subtropical populations of Culex tritaeniorhynchus showed higher genetic diversity and those climatic conditions provide great advantages for the establishment and expansion of Culex tritaeniorhynchus. There are differences in JEV infection rates in wild populations of Culex tritaeniorhynchus under different ecological conditions. Our results suggest a complex interplay of genetic differentiation, population structure, and environmental factors in shaping the dynamics of Culex tritaeniorhynchus. The low prevalence of Wolbachia in wild populations may reflect the recent presence of Wolbachia invasion in Culex tritaeniorhynchus. [ABSTRACT FROM AUTHOR]

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 FROM AUTHOR]

Mosquitoes, belonging to the family Culicidae are small blood-feeding, two-winged dipteran insects consisting of 3,600 species. The larvae of mosquitoes were collected from the different locations at regular intervals (January to December 2022) from some of the selected districts of the National Capital Regions: Sonipat, Meerut, Muradnagar, Hapur, Baghpat, Ghaziabad, Delhi and Pilkhuwa. Sampling was done from different water containers using the dipping or pipetting method. Larvae of the mosquitoes were identified by applying morphological keys. The study revealed that the mosquito breeding was high in the summer in comparison to the winter season. Population of the mosquito larvae reached a peak during July to September (monsoon/rainy season). Although, Aedes, Culex and Anopheles mosquitoes shared the common breeding habitats but Aedes preferred household water and smaller containers in comparison to Culex which breed in every aquatic habitat. The highest house indices were found in Sonipat (76.47%) followed by Meerut (75.49%), Muradnagar (66.67%), Hapur (62.5%), Baghpat (62.5%), Ghaziabad (53.7%), Delhi (47.82%), and Pilkhuwa (44.44%). Total house index for all locations was 64.33, whereas Breteau index for all locations was 86.94. Aedes (62.31%) was at its highest prevalence in humid and warm climatic conditions as compared to Culex (34.20%) and Anopheles (3.48%). The high prevalence of Aedes mosquito larvae (62.31%) in comparison to Culex (34.20%) and Anopheles (3.48%) shows high risks of Dengue, Chikungunya, Yellow fever and Zika virus transmission in National Capital Region, India. [ABSTRACT FROM AUTHOR]

Background: Mosquitoes are important vectors of pathogens. They are usually collected with CO2-baited traps and subsequently identified by morphology. This procedure is very time-consuming. Automatic counting traps could facilitate timely evaluation of the local risk for mosquito-borne pathogen transmission or decision-making on vector control measures, but the counting accuracy of such devices has rarely been validated in the field. Methods: The Biogents (BG)-Counter 2 automatically counts mosquitoes by discriminating the size of captured objects directly in the field and transmits the data to a cloud server. To assess the accuracy of this counting device, 27 traps were placed at 19 sampling sites across Germany and used in daily, weekly or bimonthly intervals from April until October 2021. The BG-Counter 2 was attached to a CO2-trap (BG-Pro trap = CO2-Pro) and the same trap was converted to also attract gravid mosquitoes (upside-down BG-Pro trap with a water container beneath = CO2-Pro-gravid). All captured mosquitoes were identified by morphology. The number of females (unfed and gravid), mosquito diversity and the number of identified specimens in relation to the counting data of the BG-Counter were compared between the two trapping devices to evaluate sampling success and counting accuracy. Results: In total 26,714 mosquitoes were collected during 854 trap days. The CO2-Pro-gravid trap captured significantly more mosquitoes per trap day for all specimens, gravid females and non-gravid females, while there was no difference in the mosquito diversity. The linear model with the captured mosquitoes as a response and the counted specimens as a predictor explained only a small degree of the variation within the data (R2 = 0.16), but per individual trap the value could reach up to 0.62 (mean R2 = 0.23). The counting accuracy for the daily samples had a significant positive correlation with sample size, resulting in higher accuracy for the CO2-Pro-gravid trap and higher accuracy for sites and sampling months with high mosquito abundance. Conclusions: While the accuracy of the BG-Counter 2 is quite low, the device is able to depict mosquito phenology and provide information about local population dynamics. [ABSTRACT FROM AUTHOR]

Mosquitoes (Diptera: Culicidae) are one of the most impactful pests to human society, both as a nuisance and a potential vector of human and animal pathogens. Mosquito larvae develop in still aquatic environments. Eliminating these habitats near high human density or managing them to reduce the suitability for mosquitoes will reduce mosquito populations in these human environments and decrease the overall negative impact of mosquitoes on humans. One common source of standing water in urban and suburban environments is the water that pools in stormwater control measures. Previous studies have shown that some stormwater control measures generate large numbers of mosquitoes while others harbor none, and the reason for this difference remains unclear. Our study focuses on elucidating the factors that cause a stormwater control measure to be more or less suitable for mosquitoes. During the summers of 2021 and 2022, we collected and identified mosquito larvae from thirty stormwater control measures across central Ohio to assess variation in mosquito abundance and diversity among sites. Our goal was to determine if specific types of stormwater control measures (retention ponds, detention ponds, or constructed wetlands) harbored different abundances of mosquitoes or different community structures. We also assessed environmental parameters of these sites to elucidate their effects on mosquito abundance and diversity. Overall, we recorded the highest number of mosquito larvae and species in constructed wetlands. However, these sites were dominated by the innocuous species, Culex territans. Conversely, detention ponds held fewer mosquitoes but a higher proportion of known vector species, including Culex pipiens and Aedes vexans. The total number of mosquitoes across all sites was correlated with higher vegetation, more shade, lower water temperatures, and lower pH, suggesting stormwater control measures with these features may also be hotspots for mosquito proliferation. [ABSTRACT FROM AUTHOR]

The Culicidae mosquito family breeds in various natural and artificial water bodies. Phytotelmata are plant structures that hold water and are used by many mosquito species during their life cycle. Mosquitoes are disease vectors, and phytotelmata play an important role in harbouring them. Investigating these phytotelmata will facilitate to develop appropriate site specific vector control strategy. Approximately 750 phytotelmata in 16 locations on Car Nicobar Island was examined. Mosquito larvae were collected from different habitats such as tree holes, root holes, leaf axils, and bamboo stumps. On average, the number of larvae per 10 ml of water ranged from 0.003 to 3.833. Among the 16 mosquito species identified, there were vectors of malaria (Anopheles sundaicus), dengue (Aedes aegypti and Aedes albopictus) and filariasis (Culex quinquefasciatus), belonging to seven genera. The most common phytotelmata were coconut trees (17.5%) followed by Java apple trees (10.3%) and bamboo stumps (7.1%). The most widespread mosquito species was Aedes edwardsi , followed by Aedes malayensis and Ae. albopictus , and the per cent of phytotelmata that contained these species were 42.6, 22.0, and 16.8% respectively. Anopheles mosquitoes had the highest container index (6.1%) in root holes, while Aedes and Culex mosquitoes were more frequently found in bamboo stumps. The alpha biodiversity indices indicated that the mosquito population in Car Nicobar Island was highly diverse (S = 16; DMg = 36.9). Tree holes showed the highest diversity, as all 16 species were collected from them. Assessment of phytotelmata, there is a significant relationship observed between mosquito genus, village, habitat, and tree families, on the remote island of Car Nicobar. Considering the presence of disease-transmitting mosquito vectors in the phytotelmata, these habitats should be considered when implementing control measures to eliminate mosquito-borne infections. [ABSTRACT FROM AUTHOR]

Information obtained about the blood feeding behavior of mosquito populations in nature is important for understanding their vectorial capacity and evaluating their potential role in the transmission of vector-borne diseases. This study investigated the feeding patterns of Culex pipiens, Anopheles sacharovi, and Cx. tritaeniorhynchus by using the ELISA method. A total of 435 blood-fed females were collected from the Aegean and Mediterranean regions of Türkiye between May 2021 and September 2023. The types and numbers of potential domestic animal hosts were recorded in each village. Of the 372 blood meals tested, 358 gave a single host positive result mainly on cow and human, chicken, respectively, in both villages. On the other hand, the host tendency for chicken was found in a very low degree in both villages. The human and other hosts were underselected, highlighting a marked zoophily for the vectors. Overall, these hosts were mainly selected in proportion to their abundance. These results highlight the zoophilic and exophagic behavior of these vectors. In conclusion, understanding the blood feeding behavior of mosquito populations is important for vector biology of these vectors. More research is needed to study the feeding behavior of outdoor mosquito populations and the influence of domestic animals on disease epidemiology. [ABSTRACT FROM AUTHOR]