West Nile Virus Vector Research Articles

Environmental determinants of West Nile virus vector abundance at the wildlife-livestock interface.

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.

Monitoring of mosquito breeding sites in the municipality of Cascais, Portugal

Abstract Culex pipiens is the primary West Nile Virus vector in Portugal. Additionally, in 2017, Aedes albopictus, vector of various arboviruses, was detected in Northern Portugal. To mitigate viral transmission risks, effective surveillance is crucial, including monitoring egg, larva, and pupa stages. Located on the west of Lisbon, Cascais implemented a surveillance based on monitoring invasive species at Points of Entry and, complementarily, on detecting and intervening in breeding sites. Active searches for new breeding sites and monitoring those that were already identified are conducted weekly during the peak mosquito season (May to October) and biweekly during the colder season (November to April), following the methodology outlined below: • Upon detection, samples of immature stages are collected for submission to the National Institute of Health (INSA); • Corrective measures are requested to the managing entities of the locations where the sites are found; • Date of action, location, collection point, date of report, and status (pending or intervened) are recorded on a database; • Locations with pending status undergo continuous monitoring. In 2023, 16 artificial breeding sites were identified. These findings resulted from 23 actions conducted across 8 locations. 35 samples of immatures were collected, followed by their submission and notification to the respective managing entities. Recurrent detections were noted in 9 breeding sites. The managing entities implemented measures at 13 of the identified sites, leaving 3 with pending intervention. The identification of breeding sites demands continuous monitoring efforts and requires an effective coordination between the Public Health services and the management entities. Based on the achieved outcomes, it is evident that the implementation of corrective measures has been successful, making a significant contribution to reducing mosquito density. Key messages • Effective surveillance and intervention strategies are crucial in mitigating West Nile Virus and arbovirus transmission risks in Portugal. • Continuous monitoring and coordination between public health services and management entities are key for maintaining effective vector control measures.

Mosquito dynamics and their drivers in peri-urban Antananarivo, Madagascar: insights from a longitudinal multi-host single-site survey

BackgroundAntananarivo, the capital city of Madagascar, is experiencing a steady increase in population growth. Due to the abundance of mosquito vectors in this locality, the population exposed to mosquito-borne diseases is therefore also increasing, as is the risk of epidemic episodes. The aim of the present study was to assess, in a resource-limited setting, the information on mosquito population dynamics and disease transmission risk that can be provided through a longitudinal entomological study carried out in a multi-host single site.MethodsMosquitoes were collected every 15 days over 16 months (from January 2017 to April 2018) using six CDC-light traps in a peri-urban area of Antananarivo. Multivariable generalised linear models were developed using indoor and outdoor densities of the predominant mosquito species as response variables and moon illumination, environmental data and climatic data as the explanatory variables.ResultsOverall, 46,737 mosquitoes belonging to at least 20 species were collected, of which Culex antennatus (68.9%), Culex quinquefasciatus (19.8%), Culex poicilipes (3.7%) and Anopheles gambiae sensu lato (2.3%) were the most abundant species. Mosquito densities were observed to be driven by moon illumination and climatic factors interacting at different lag periods. The outdoor models demonstrated biweekly and seasonal patterns of mosquito densities, while the indoor models demonstrated only a seasonal pattern.ConclusionsAn important diversity of mosquitoes exists in the peri-urban area of Antananarivo. Some well-known vector species, such as Cx. antennatus, a major vector of West Nile virus (WNV) and Rift-Valley fever virus (RVFV), Cx. quinquefasciatus, a major vector of WNV, Cx. poicilipes, a candidate vector of RVFV and An. gambiae sensu lato, a major vector of Plasmodium spp., are abundant. Importantly, these four mosquito species are present all year round, even though their abundance declines during the cold dry season, with the exception of Cx. quinquefasciatus. The main drivers of their abundance were found to be temperature, relative humidity and precipitation, as well as—for outdoor abundance only—moon illumination. Identifying these drivers is a first step towards the development of pathogen transmission models (R0 models), which are key to inform public health stakeholders on the periods of most risk for vector-borne diseases.Graphical

Eilat virus (EILV) causes superinfection exclusion against West Nile virus (WNV) in a strain-specific manner in Culex tarsalis mosquitoes.

West Nile virus (WNV) is the leading cause of mosquito-borne illness in the USA. There are currently no human vaccines or therapies available for WNV, and vector control is the primary strategy used to control WNV transmission. The WNV vector Culex tarsalis is also a competent host for the insect-specific virus (ISV) Eilat virus (EILV). ISVs such as EILV can interact with and cause superinfection exclusion (SIE) against human pathogenic viruses in their shared mosquito host, altering vector competence for these pathogenic viruses. The ability to cause SIE and their host restriction make ISVs a potentially safe tool to target mosquito-borne pathogenic viruses. In the present study, we tested whether EILV causes SIE against WNV in mosquito C6/36 cells and C. tarsalis mosquitoes. The titres of both WNV strains - WN02-1956 and NY99 - were suppressed by EILV in C6/36 cells as early as 48-72 h post-superinfection at both m.o.i. values tested in our study. The titres of WN02-1956 at both m.o.i. values remained suppressed in C6/36 cells, whereas those of NY99 showed some recovery towards the final timepoint. The mechanism of SIE remains unknown, but EILV was found to interfere with NY99 attachment in C6/36 cells, potentially contributing to the suppression of NY99 titres. However, EILV had no effect on the attachment of WN02-1956 or internalization of either WNV strain under superinfection conditions. In C. tarsalis, EILV did not affect the infection rate of either WNV strain at either timepoint. However, in mosquitoes, EILV enhanced NY99 infection titres at 3 days post-superinfection, but this effect disappeared at 7 days post-superinfection. In contrast, WN02-1956 infection titres were suppressed by EILV at 7 days post-superinfection. The dissemination and transmission of both WNV strains were not affected by superinfection with EILV at either timepoint. Overall, EILV caused SIE against both WNV strains in C6/36 cells; however, in C. tarsalis, SIE caused by EILV was strain specific potentially owing to differences in the rate of depletion of shared resources by the individual WNV strains.

Evaluating larvicidal, ovicidal and growth inhibiting activity of five medicinal plant extracts on Culex pipiens (Diptera: Culicidae), the West Nile virus vector

Mosquitoes, one of the deadliest animals on the planet, cause millions of fatalities each year by transmitting several human illnesses. Synthetic pesticides were previously used to prevent the spread of diseases by mosquitoes, which was effective in protecting humans but caused serious human health problems, environmental damage, and developed mosquito pesticide resistance. This research focuses on exploring new, more effective, safer, and environmentally friendly compounds to improve mosquito vector management. Phytochemicals are possible biological agents for controlling pests and many are target-specific, rapidly biodegradable, and eco-friendly. The potential of extracts of Lantana camara, Melia azedarach, Nerium oleander, Ricinus communis, and Withania somnifera against 3rd instar Culex pipiens (Common house mosquito) larvae was evaluated. Methanol extracts had more toxic effects against Cx. pipiens larvae (95–100%, 24 h post-treatment) than aqueous extracts (63–91%, 24 h post-treatment). The methanol extracts of Nerium oleander (LC50 = 158.92 ppm) and Ricinus communis (LC50 = 175.04 ppm) were very effective at killing mosquito larvae, 24 h after treatment. N. oleander (LC50 = 373.29 ppm) showed high efficacy in aqueous plant extracts. Among the different extracts of the five plants screened, the methanol extract of R. communis recorded the highest ovicidal activity of 5% at 800 ppm concentration. Total developmental duration and growth index were highly affected by R. communis and M. azedarach methanol extracts. In field tests it was clear that plant extracts decreased mosquito larval density, especially when mixed with mosquito Bti briquette, with stability up to seven days for N. oleander. GC–MS results showed that the methanol extract had a higher number of chemical compounds, particularly with more terpene compounds. A high-performance liquid chromatography (HPLC) technique was used to detect the existence of non-volatile polyphenols and flavonoids. All five methanol extracts showed high concentrations of active ingredients such as gallic acid, chlorogenic acid (more than 100 μg/ml) and the rosmarinic acid was also found in all the five extracts in addition to 17 active polyphenols and flavonoids presented at moderate to low concentrations. Molecular modeling of 18 active ingredients detected by the HPLC were performed to the vicinity of one of the fatty acid binding proteins of lm-FABP (PDB code: 2FLJ). Rutin, Caffeic acid, coumaric acid and rosmarinic acid which presented densely in R. communis and N. oleander showed multiple and stable intermolecular hydrogen bonding and π–π stacking interactions. The inhibition ability of the fatty acid binding protein, FABP4, was evaluated with remarkable receptor inhibition evident, especially with R. communis and N. oleander having inhibitory concentrations of IC50 = 0.425 and 0.599 µg/mL, respectively. The active phytochemical compounds in the plants suggest promising larvicidal and ovicidal activity, and have potential as a safe and effective alternative to synthetic insecticides.

Biological effects of Bougainvillea glabra, Delonix regia, Lantana camara, and Platycladus orientalis extracts and their possible metabolomics therapeutics against the West Nile virus vector, Culex pipiens (Diptera: Culicidae)

Plants are a treasure trove of biological materials containing a wide range of potential phytochemicals that are target-specific, rapidly biodegradable, and environmentally friendly, with multiple medicinal effects. Unfortunately, the development of resistance to synthetic pesticides and antibiotics led to the discovery of new antibiotics, antioxidants, and biopesticides. This has also led to the creation of new medications that work very well. The current study aimed to prove that ornamental plants contain specialized active substances that are used in several biological processes. Mosquitoes, one of the deadliest animals on the planet, cause millions of fatalities each year by transmitting several human illnesses. Phytochemicals are possible biological agents for controlling pests that are harmful. The potential of leaf extracts of Bougainvillea glabra, Delonix regia, Lantana camara, and Platycladus orientalis against Culex pipiens and microbial agents was evaluated. Acetone extracts had more toxic effects against Cx. pipiens larvae (99.0–100 %, 72 h post-treatment), and the LC50 values were 142.8, 189.5, 95.4, and 71.1 ppm for B. glabra, D. regia, L. camara, and P. orientalis, respectively. Plant extracts tested in this study showed high insecticidal, antimicrobial, and antioxidant potential. GC-MS and HPLC analyses showed a higher number of terpenes, flavonoids, and phenolic compounds. The ADME analysis of element, caryophyllene oxide, caryophyllene, and copaene showed that they were similar to drugs and that they were better absorbed by the body and able to pass through the blood-brain barrier. Our results confirm the ability of ornamental plants to have promising larvicidal and antimicrobial activity and biotechnology.

A single-cell atlas of the Culex tarsalis midgut during West Nile virus infection.

The mosquito midgut functions as a key interface between pathogen and vector. However, studies of midgut physiology and associated virus infection dynamics are scarce, and in Culex tarsalis - an extremely efficient vector of West Nile virus (WNV) - nonexistent. We performed single-cell RNA sequencing on Cx. tarsalis midguts, defined multiple cell types, and determined whether specific cell types are more permissive to WNV infection. We identified 20 cell states comprised of 8 distinct cell types, consistent with existing descriptions of Drosophila and Aedes aegypti midgut physiology. Most midgut cell populations were permissive to WNV infection. However, there were higher levels of WNV RNA (vRNA) in enteroendocrine cells and cells enriched for mitochondrial genes, suggesting enhanced replication in these populations. In contrast, proliferating intestinal stem cell (ISC) populations had the lowest levels of vRNA, a finding consistent with studies suggesting ISC proliferation in the midgut is involved in viral control. Notably, we did not detect significant WNV-infection induced upregulation of canonical mosquito antiviral immune genes (e.g., AGO2, R2D2, etc.) at the whole-midgut level. Rather, we observed a significant positive correlation between immune gene expression levels and vRNA in individual cells, suggesting that within midgut cells, high levels of vRNA may trigger antiviral responses. Our findings establish a Cx. tarsalis midgut cell atlas, and provide insight into midgut infection dynamics of WNV by characterizing cell-type specific enhancement/restriction of, and immune response to, infection at the single-cell level.

Temperature and time of host-seeking activity impact the efficacy of chemical control interventions targeting the West Nile virus vector, Culex tarsalis.

West Nile virus (WNV) is the leading mosquito-borne disease causing-pathogen in the United States. Concerningly, there are no prophylactics or drug treatments for WNV and public health programs rely heavily on vector control efforts to lessen disease incidence. Insecticides can be effective in reducing vector numbers if implemented strategically, but can diminish in efficacy and promote insecticide resistance otherwise. Vector control programs which employ mass-fogging applications of insecticides, often conduct these methods during the late-night hours, when diel temperatures are coldest, and without a-priori knowledge on daily mosquito activity patterns. This study's aims were to 1) quantify the effect of temperature on the toxicity of two conventional insecticides used in fogging applications (malathion and deltamethrin) to Culex tarsalis, an important WNV vector, and 2) quantify the time of host-seeking of Cx. tarsalis and other local mosquito species in Maricopa County, Arizona. The temperature-toxicity relationship of insecticides was assessed using the WHO tube bioassay, and adult Cx. tarsalis, collected as larvae, were exposed to three different insecticide doses at three temperature regimes (15, 25, and 35°C; 80% RH). Time of host-seeking was assessed using collection bottle rotators with encephalitis vector survey traps baited with dry ice, first at 3h intervals during a full day, followed by 1h intervals during the night-time. Malathion became less toxic at cooler temperatures at all doses, while deltamethrin was less toxic at cooler temperatures at the low dose. Regarding time of host-seeking, Cx. tarsalis, Aedes vexans, and Culex quinquefasciatus were the most abundant vectors captured. During the 3-hour interval surveillance over a full day, Cx. tarsalis were most-active during post-midnight biting (00:00-06:00), accounting for 69.0% of all Cx. tarsalis, while pre-midnight biting (18:00-24:00) accounted for 30.0% of Cx. tarsalis. During the 1-hour interval surveillance overnight, Cx. tarsalis were most-active during pre-midnight hours (18:00-24:00), accounting for 50.2% of Cx. tarsalis captures, while post-midnight biting (00:00-06:00) accounted for 49.8% of Cx. tarsalis. Our results suggest that programs employing large-scale applications of insecticidal fogging should consider temperature-toxicity relationships coupled with time of host-seeking data to maximize the efficacy of vector control interventions in reducing mosquito-borne disease burden.

Larval Competition between Aedes and Culex Mosquitoes Carries over to Higher Arboviral Infection during Their Adult Stage.

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

Larvicidal activity of six Moroccan Artemisia essential oils and a correlation study of their major components against Culex pipiens

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

The potential role of the Asian bush mosquito Aedes japonicus as spillover vector for West Nile virus in the Netherlands

BackgroundIn recent years the Asian bush mosquito Aedes japonicus has invaded Europe, including the Netherlands. This species is a known vector for a range of arboviruses, possibly including West Nile virus (WNV). As WNV emerged in the Netherlands in 2020, it is important to investigate the vectorial capacity of mosquito species present in the Netherlands to estimate the risk of future outbreaks and further spread of the virus. Therefore, this study evaluates the potential role of Ae. japonicus in WNV transmission and spillover from birds to dead-end hosts in the Netherlands.MethodsWe conducted human landing collections in allotment gardens (Lelystad, the Netherlands) in June, August and September 2021 to study the diurnal and seasonal host-seeking behaviour of Ae. japonicus. Furthermore, their host preference in relation to birds using live chicken-baited traps was investigated. Vector competence of field-collected Ae. japonicus mosquitoes for two isolates of WNV at two different temperatures was determined. Based on the data generated from these studies, we developed a Susceptible-Exposed-Infectious-Recovered (SEIR) model to calculate the risk of WNV spillover from birds to humans via Ae. japonicus, under the condition that the virus is introduced and circulates in an enzootic cycle in a given area.ResultsOur results show that Ae. japonicus mosquitoes are actively host seeking throughout the day, with peaks in activity in the morning and evening. Their abundance in August was higher than in June and September. For the host-preference experiment, we documented a small number of mosquitoes feeding on birds: only six blood-fed females were caught over 4 full days of sampling. Finally, our vector competence experiments with Ae. japonicus compared to its natural vector Culex pipiens showed a higher infection and transmission rate when infected with a local, Dutch, WNV isolate compared to a Greek isolate of the virus. Interestingly, we also found a small number of infected Cx. pipiens males with virus-positive leg and saliva samples.ConclusionsCombining the field and laboratory derived data, our model predicts that Ae. japonicus could act as a spillover vector for WNV and could be responsible for a high initial invasion risk of WNV when present in large numbers.Graphical

Broadscale spatial synchrony in a West Nile virus mosquito vector across multiple timescales

Insects often exhibit irruptive population dynamics determined by environmental conditions. We examine if populations of the Culex tarsalis mosquito, a West Nile virus (WNV) vector, fluctuate synchronously over broad spatial extents and multiple timescales and whether climate drives synchrony in Cx. tarsalis, especially at annual timescales, due to the synchronous influence of temperature, precipitation, and/or humidity. We leveraged mosquito collections across 9 National Ecological Observatory Network (NEON) sites distributed in the interior West and Great Plains region USA over a 45-month period, and associated gridMET climate data. We utilized wavelet phasor mean fields and wavelet linear models to quantify spatial synchrony for mosquitoes and climate and to calculate the importance of climate in explaining Cx. tarsalis synchrony. We also tested whether the strength of spatial synchrony may vary directionally across years. We found significant annual synchrony in Cx. tarsalis, and short-term synchrony during a single period in 2018. Mean minimum temperature was a significant predictor of annual Cx. tarsalis spatial synchrony, and we found a marginally significant decrease in annual Cx. tarsalis synchrony. Significant Cx. tarsalis synchrony during 2018 coincided with an anomalous increase in precipitation. This work provides a valuable step toward understanding broadscale synchrony in a WNV vector.

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

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

An update on the ecology, seasonality and distribution of Culex modestus in England

Abstract Culex (Barraudius) modestus is an important vector of West Nile virus (WNV) in Europe and it has the potential to play a bridge vector role in future WNV transmission in the UK. Here we provide an update on the known distribution of the species in England based on adult and larval data, characterise the preferred Cx. modestus larval habitats, and present adult and larval data from sites where the species is known to occur. Culex modestus is primarily found in the south-east of England, particularly in North Kent, the Thames Estuary, and along the Essex coast, and as far east as Orford Ness, Suffolk, and in Rainham Marshes, Essex, in the West. Adult numbers peak in mid-late July, with larval numbers highest in late August / early September. Preferred habitats in North Kent are warm, shallow, narrow ditches, with an abundance of marginal, submerged, and floating vegetation. Such environmental data on the distribution, seasonality and habitat preference of Cx. modestus are critical for informing WNV surveillance programmes, identifying at risk areas (associated with this species) and providing information for a targeted control strategy in the event of WNV transmission.

Impacts of ground ultra-low volume adulticide applications on Culex pipiens and Culex restuans (Diptera: Culicidae) abundance, age structure, and West Nile virus infection in Cook County, Illinois.

Since the introduction of West Nile virus (WNV) to the United States over 20 years ago, thousands of cases of human disease and death have been reported. Yearly seasonal outbreaks continue to persist, and the city and suburbs of Chicago, Illinois, is considered a "hot spot" for WNV activity. To interrupt WNV transmission, ground ultra-low volume (ULV) adulticide applications are regularly used to reduce Culex pipiens L. and Culex restuans Theobold (Diptera: Culicidae) abundance and infection. The real-world effectiveness of adulticide applications has not been comprehensively assessed, and prior studies, including our own investigation, have yielded inconclusive or conflicting results. Therefore, we expanded our prior work and evaluated the effects of 5 sequential weekly truck-mounted ULV adulticide applications in large residential areas in the northern suburbs of Chicago, Illinois, in 2019 and 2020. Each day, Cx. pipiens and Cx. restuans host-seeking and gravid mosquitoes were collected to assess abundance, age structure, and WNV infection rates. Adulticide applications resulted in significant reductions of both host-seeking and gravid abundance on the night of treatment. The reduction in host-seeking mosquitoes was followed by a reduction in gravid mosquitoes trapped 3 and 4 days after adulticide application and an increase in the proportion of nulliparous mosquitoes. WNV infection rates were significantly reduced in treatment sites as compared to untreated sites when infection rates were higher in 2020. This large-scale study provides comprehensive evidence that ground ULV adulticide applications are an effective tool in an integrated mosquito management program for combating WNV vectors and infection risk.

Blood-feeding patterns of Culex pipiens biotype pipiens and pipiens/molestus hybrids in relation to avian community composition in urban habitats

BackgroundCulex pipiens sensu stricto (s.s.) is considered the primary vector of Usutu virus and West Nile virus, and consists of two morphologically identical but behaviourally distinct biotypes (Cx. pipiens biotype pipiens and Cx. pipiens biotype molestus) and their hybrids. Both biotypes are expected to differ in their feeding behaviour, and pipiens/molestus hybrids are presumed to display intermediate feeding behaviour. However, the evidence for distinct feeding patterns is scarce, and to date no studies have related differences in feeding patterns to differences in host abundance.MethodsMosquitoes were collected using CO2-baited traps. We collected blood-engorged Cx. pipiens/torrentium specimens from 12 contrasting urban sites, namely six city parks and six residential areas. Blood engorged Cx. pipiens/torrentium mosquitoes were identified to the species and biotype/hybrid level via real-time polymerase chain reaction (PCR). We performed blood meal analysis via PCR and Sanger sequencing. Additionally, avian host communities were surveyed via vocal sounds and/or visual observation.ResultsWe selected 64 blood-engorged Cx. pipiens/torrentium mosquitoes of which we successfully determined the host origin of 55 specimens. Of these, 38 belonged to biotype pipiens, 14 were pipiens/molestus hybrids and the identity of three specimens could not be determined. No blood-engorged biotype molestus or Cx. torrentium specimens were collected. We observed no differences in feeding patterns between biotype pipiens and pipiens/molestus hybrids across different habitats. Avian community composition differed between city parks and residential areas, whereas overall avian abundance did not differ between the two habitat types.ConclusionsOur results show the following: (1) Cx. pipiens s.s. feeding patterns did not differ between city parks and residential areas, regardless of whether individuals were identified as biotype pipiens or pipiens/molestus hybrids. (2) We detected differences in host availability between city parks and residential areas. (3) We show that in both urban habitat types, biotype pipiens and pipiens/molestus hybrids fed on both mammalian and avian hosts. This underscores the potential role in arbovirus transmission of biotype pipiens and pipiens/molestus hybrids.Graphical

Culex pipiens and Culex restuans larval interactions shape the bacterial communities in container aquatic habitats.

Container aquatic habitats host a community of aquatic insects, primarily mosquito larvae that browse on container surface microbial biofilm and filter-feed on microorganisms in the water column. We examined how the bacterial communities in these habitats respond to feeding by larvae of two container-dwelling mosquito species, Culex pipiens and Cx. restuans. We also investigated how the microbiota of these larvae is impacted by intra- and interspecific interactions. Microbial diversity and richness were significantly higher in water samples when mosquito larvae were present, and in Cx. restuans compared to Cx. pipiens larvae. Microbial communities of water samples clustered based on the presence or absence of mosquito larvae and were distinct from those of mosquito larvae. Culex pipiens and Cx. restuans larvae harbored distinct microbial communities when reared under intraspecific conditions and similar microbial communities when reared under interspecific conditions. These findings demonstrate that mosquito larvae play a major role in structuring the microbial communities in container habitats and that intra- and interspecific interactions in mosquito larvae may shape their microbiota. This has important ecological and public health implications since larvae of the two mosquito species are major occupants of container habitats while the adults are vectors of West Nile virus.

Implications of migratory and exotic birds and the mosquito community on West Nile virus transmission

Background Vector-borne diseases like West Nile virus (WNV) pose a global health challenge, with rising incidence and distribution. Culex mosquitoes are crucial WNV vectors. Avian species composition and bird community diversity, along with vector communities, influence WNV transmission patterns. However, limited knowledge exists on their impact in southwestern Spain, an area with active WNV circulation in wild birds, mosquitoes, and humans. Methods To address this, we conducted a comprehensive study investigating the contributions of migratory and exotic bird species to WNV transmission and the influence of mosquito community composition. Results Analysing 1194 serum samples from 44 avian species, we detected WNV antibodies in 32 samples from 11 species, four for the first time in Europe. Migratory birds had higher WNV exposure likelihood than native and exotic species, and higher phylogenetic diversity in bird communities correlated with lower exposure rates. Moreover, in 5859 female mosquitoes belonging to 12 species, we identified WNV competent vectors like Cx. pipiens s.l. and the Univittatus subgroup. Birds with WNV antibodies were positively associated with competent vector abundance, but negatively with overall mosquito species richness. Conclusions These findings highlight the complex interactions between bird species, their phylogenetics, and mosquito vectors in WNV transmission. Understanding these dynamics will help to implement effective disease control strategies in southwestern Spain.

Do It Yourself: A Modernized Gravid Trap Design for Mosquito Surveillance.

Gravid traps have become a common and frequently essential surveillance tool for parous Culex spp. vectors of West Nile virus and other encephalitis-causing pathogens. The recent closing of BioQuip Products Inc., an entomological supply company, has jeopardized the commercial availability of gravid traps. The Salt Lake City Mosquito Abatement District presents herein a template for making your own gravid trap, but with some modernizations for quieter fans and longer lasting, light weight, lithium battery packs. At the time of writing, the materials cost for the fan ($14 USD), toolbox ($13), cables ($9), ABS pipe ($2.50), aluminum brackets ($10), catch container with lid ($9), trap net ($10), USB battery pack ($35) and the negligible amount of 3D-printed filament ($2), is approximately half the cost (not including labor) of the formerly available commercial model. Additionally, performance validation in the laboratory (t4,9 = 0.1191, P < 0.9109) and within two field sites (χ2 = 0.107, P < 0.744) demonstrated no significant differences in collections of gravid Culex pipiens. We do not present an overhaul of the previous gravid trap blueprint, but the quality-of-life updates to the trap design, the feasibility of in-house manufacture, and the mirrored collection efficacy to the commercial model can allow improved maintenance of gravid trap surveillance networks without a commercial supplier.

Interactions between West Nile Virus and the Microbiota of Culex pipiens Vectors: A Literature Review.

The flavivirus West Nile virus (WNV) naturally circulates between mosquitoes and birds, potentially affecting humans and horses. Different species of mosquitoes play a role as vectors of WNV, with those of the Culex pipiens complex being particularly crucial for its circulation. Different biotic and abiotic factors determine the capacity of mosquitoes for pathogen transmission, with the mosquito gut microbiota being recognized as an important one. Here, we review the published studies on the interactions between the microbiota of the Culex pipiens complex and WNV infections in mosquitoes. Most articles published so far studied the interactions between bacteria of the genus Wolbachia and WNV infections, obtaining variable results regarding the directionality of this relationship. In contrast, only a few studies investigate the role of the whole microbiome or other bacterial taxa in WNV infections. These studies suggest that bacteria of the genera Serratia and Enterobacter may enhance WNV development. Thus, due to the relevance of WNV in human and animal health and the important role of mosquitoes of the Cx. pipiens complex in its transmission, more research is needed to unravel the role of mosquito microbiota and those factors affecting this microbiota on pathogen epidemiology. In this respect, we finally propose future lines of research lines on this topic.