Arbovirus surveillance: first dengue virus detection in local Aedes albopictus mosquitoes in Europe, Catalonia, Spain, 2015.

Dengue has emerged as the most important viral mosquito-borne disease globally. The current risk of dengue outbreaks in Europe appeared with the introduction of the vector Aedes albopictus mosquito in Mediterranean countries. Considering the increasing frequency of dengue epidemics worldwide and the movement of viraemic hosts, it is expected that new autochthonous cases will occur in the future in Europe. Arbovirus surveillance started in Catalonia in 2015 to monitor imported cases and detect possible local arboviral transmission. During 2015, 131 patients with a recent travel history to endemic countries were tested for dengue virus (DENV) and 65 dengue cases were detected. Twenty-eight patients with a febrile illness were viraemic, as demonstrated by a positive real-time RT-PCR test for DENV in serum samples. Entomological investigations around the viraemic cases led to the detection of DENV in a pool of local Ae. albopictus captured in the residency of one case. The sequence of the DENV envelope gene detected in the mosquito pool was identical to that detected in the patient. Our results show how entomological surveillance conducted around viraemic travellers can be effective for early detection of DENV in mosquitoes and thus might help to prevent possible autochthonous transmission.

First Data on Resistance to Pyrethroids in Wild Populations of Aedes albopictus from Spain.

The invasive mosquito, Aedes albopictus, found in Spain since 2004, is a competent vector of yellow fever, Zika, dengue, and chikungunya viruses among other diseases. Although controversial, the use of adulticiding is a relevant tool for vector control and could be crucial for the management of any possible outbreak of imported diseases. We present the 1st study in Spain on the susceptibility of field populations from Barcelona, Peñíscola, Castellón, and Mallorca of Ae. albopictus to several pyrethroids using either bioassays under the World Health Organization methodology and biochemical tests. In the bioassays, the discriminating concentrations were calculated using a local, susceptible laboratory strain. Different susceptibility levels were found for some combinations of populations and products. The biochemical tests carried out by enzymatic analysis supported these results, showing an overexpression of glutathione S-transferase activity in 1 population.

Simple Indices Provide Insight to Climate Attributes Delineating the Geographic Range of Aedes albopictus (Diptera: Culicidae) Prior to Worldwide Invasion.

Aedes albopictus (Skuse) has expanded its distribution worldwide during the past decades. Despite attempts to explain and predict its geographic occurrence, analyses of the distribution of Ae. albopictus in the context of broad climatic regions (biomes) has not been performed. We analyzed climate conditions at its distribution sites in the range before the worldwide invasions (from the easternmost Hawaii through westernmost Madagascar) by using thermal and aridity-humidity indices descriptive of major biomes. A significant advantage of this approach is that it uses simple indices clearly related to the population dynamics of Ae. albopictus. Although Ae. albopictus has been regarded as a forest species preferring humid climate, in areas with significant human habitation, the distribution sites extended from the perhumid, rain forest zone to the semiarid, steppe zone. This pattern was common from the tropics through the temperate zone. Across the distribution range, there was no seasonal discordance between temperature and precipitation; at sites where winter prevents Ae. albopictus reproduction (monthly means<10°C), precipitation was concentrated in warm months (>10°C) under the Asian summer monsoon. Absence of the species in northern and eastern coastal Australia and eastern coastal Africa was not attributable solely to climate conditions. However, Asia west of the summer monsoon range was climatically unsuitable because of low precipitation throughout the year or in warm months favorable to reproduction (concentration of precipitation in winter). We hypothesized that Ae. albopictus originated in continental Asia under the monsoon climate with distinct dry seasons and hot, wet summer, enabling rapid population growth.

Ground ultra-low volume adulticiding field trials using pyrethroids against Aedes albopictus in the Baix Llobregat region, Spain.

Ground ultra-low volume (ULV) application against Aedes albopictus is a challenging methodology as its efficacy is strongly related to environmental conditions, insect behavior, and the pesticide formulation. Such operations are very relevant in many control programs, although their real efficacy is often unknown. Trials were carried out as part of real-world applications in urban scenarios in the Baix Llobregat region to evaluate the efficacy of 2 active ingredients, delatamethrin and alpha-cypermethrin. Both pyrethroids were tested against caged mosquitoes placed at a range of distances and under varying environmental conditions. Additionally, persistence on plants placed at the same distances was evaluated after 5 h, as well as droplet distribution using water-sensitive papers. The deltamethrin formulation used yielded significantly higher 24-h mortality than alpha-cypermethrin (100% direct mortality up to 25 m versus 10 m; and 73.9% residual 5-h mortality at 20 m compared to <50% at 5 m). The influence of environmental factors, a comparison with other field tests, and the real effect on uncaged mosquitoes is discussed.

[First record of Aedes albopictus in Galicia, obtained by citizen science through Mosquito Alert]. / Primera cita de Aedes albopictus para Galicia, obtenida mediante ciencia ciudadana por Mosquito Alert.

The first detection of the tiger mosquito, Aedes (Stegomyia) albopictus (Skuse, 1894), in the autonomous community of Galicia (Spain) is reported. The finding has been possible thanks to the collaboration between citizens, the citizen science application Mosquito Alert and the Rede Galega de Vixilancia de Vectores (ReGaViVec). At the beginning of August 2023, a same person submitted through the app several reports consistent with the tiger mosquito in the municipality of Moaña, in Pontevedra. The ReGaViVec entomological team confirmed the species and conducted vector surveillance in the area by placing traps (11 ovitraps and 3 BG-Sentinel 2 with BG-Lure attractant) with a weekly collection frequency. This finding represents the most northwestern detection of the tiger mosquito in the Iberian Peninsula and shows the crucial role of citizen science in vector surveillance.

A genotyping array for the globally invasive vector mosquito, Aedes albopictus.

BACKGROUND: Although whole-genome sequencing (WGS) is the preferred genotyping method for most genomic analyses, limitations are often experienced when studying genomes characterized by a high percentage of repetitive elements, high linkage, and recombination deserts. The Asian tiger mosquito (Aedes albopictus), for example, has a genome comprising up to 72% repetitive elements, and therefore we set out to develop a single-nucleotide polymorphism (SNP) chip to be more cost-effective. Aedes albopictus is an invasive species originating from Southeast Asia that has recently spread around the world and is a vector for many human diseases. Developing an accessible genotyping platform is essential in advancing biological control methods and understanding the population dynamics of this pest species, with significant implications for public health. METHODS: We designed a SNP chip for Ae. albopictus (Aealbo chip) based on approximately 2.7 million SNPs identified using WGS data from 819 worldwide samples. We validated the chip using laboratory single-pair crosses, comparing technical replicates, and comparing genotypes of samples genotyped by WGS and the SNP chip. We then used the chip for a population genomic analysis of 237 samples from 28 sites in the native range to evaluate its usefulness in describing patterns of genomic variation and tracing the origins of invasions. RESULTS: Probes on the Aealbo chip targeted 175,396 SNPs in coding and non-coding regions across all three chromosomes, with a density of 102 SNPs per 1 Mb window, and at least one SNP in each of the 17,461 protein-coding genes. Overall, 70% of the probes captured the genetic variation. Segregation analysis found that 98% of the SNPs followed expectations of single-copy Mendelian genes. Comparisons with WGS indicated that sites with genotype disagreements were mostly heterozygotes at loci with WGS read depth < 20, while there was near complete agreement with WGS read depths > 20, indicating that the chip more accurately detects heterozygotes than low-coverage WGS. Sample sizes did not affect the accuracy of the SNP chip genotype calls. Ancestry analyses identified four to five genetic clusters in the native range with various levels of admixture. CONCLUSIONS: The Aealbo chip is highly accurate, is concordant with genotypes from WGS with high sequence coverage, and may be more accurate than low-coverage WGS.

Laboratory tests on the efficacy of VBC60035, a combined larvicidal formulation of Bacillus thuringiensis israelensis (strain AM65-52) and Bacillus sphaericus (strain 2362) against Aedes albopictus in simulated catch basins.

Catch basins are a common breeding site for mosquito larvae in urban areas. The arrival in Spain of the Asian tiger mosquito, Aedes albopictus, has turned catch basins into a challenging public health issue in some areas such as the Baix Llobregat region, where more than 135,000 units are treated per year in a large-scale application program. Although biorational larvicides are used, modern formulations providing longer efficacy periods could lead to manpower savings. Laboratory indoor trials are presented on the formulation VBC60035 (Vectomax FG), which is under registration in Spain. Indoor microcosms simulating real catch basins were treated and recolonized weekly using insectary-reared Ae. albopictus larvae. The trial took place over 2 seasons in the same water at dosages of 10, 50, and 577 kg/ha. At all 3 concentrations, the efficiency was close to 100% for up to 345 days posttreatment with 14.1% mean mortality in the control. Field factors including flushing, bacterial degradation, and toxic effluents in the drainage pools are discussed as limitations of indoor tests.

Laboratory tests of the residual effect of deltamethrin on vegetation against Aedes albopictus.

Aedes albopictus, the Asian tiger mosquito, was 1st detected in Spain in 2004 and since then has spread along the Mediterranean coast. New adulticiding strategies are being tested to enhance control of the species. The residual effect of applications of 2 formulations containing deltamethrin over vegetation was evaluated in bioassay tests for 34 days. Hedera helix plants were treated with K-othrine SC25 (deltamethrin 2.56% SC) and Aqua K-othrine (deltamethrin 2.03% EW). Bioassays consisted of exposing batches of Ae. albopictus females to treated plants. Significant mortality (> 80%) for up to 12 days was obtained with Aqua K-othrine, and for up to 5 days using K-othrine SC25. Results show that the type of formulation has an impact on the residual effect of deltamethrin against Ae. albopictus. This validates the strategy of applying pyrethroid insecticides to vegetal barriers for the control of this species, as carried out in several countries.

Drivers of longevity of wild-caught Aedes albopictus populations.

BACKGROUND: Age structure and longevity constitute fundamental determinants of mosquito populations' capacity to transmit pathogens. However, investigations on mosquito-borne diseases primarily focus on aspects such as abundance or dispersal rather than survival and demography. Here, we examine the post-capture longevity of wild-caught populations of the Asian tiger mosquito Aedes albopictus to investigate the influence of environmental factors and individual frailty on longevity. METHODS: We captured females of Ae. albopictus from June to November 2021 in a vegetated and an urban area by two methods of capture (BG traps and Human Landing catch). They were kept in semi-controlled conditions in the field, and survival was monitored daily across the 859 individuals captured. We studied the differences in longevity per capture method and location and the influence on longevity of seasonal, climatic and individual factors. RESULTS: Photoperiod, GDD, minimum and maximum temperature and relative humidity showed an effect on the risk of death of females in the field. Females captured in urban area with Human Landing catch methods had greater longevity than females captured in non-urban areas with BG traps. Individual variance, reflecting individual frailties, had an important effect on the risk of death: the greater the frailty, the shorter the post-capture longevity. Overall, longevity is affected not only by climate and seasonal drivers like temperature and photoperiod but also by the individual frailty of mosquitoes. CONCLUSION: This work unravels environmental drivers of key demographic parameters such as longevity, as modulated by individual frailty, in disease vectors with strong seasonal dynamics. Further demographic understanding of disease vectors in the wild is needed to adopt new surveillance and control strategies and improve our understanding of disease risk and spread.

A Survey on Native and Invasive Mosquitoes and Other Biting Dipterans in Northern Spain.

PURPOSE: Haematophagous Diptera, such as mosquitoes (Culicidae), biting midges (Ceratopogonidae), and black flies (Simuliidae), are important insects for public and animal health due to their capacity to bite and transmit pathogens. Outdoor recreation areas are usually affected by biting species and provide suitable habitats to both adult and immature stages. This study aimed to determine the species diversity and larval sites of these Diptera groups in two golf courses. METHODS: A multi-method collection approach using ultraviolet-CDC traps, human landing catches, collection in breeding sites, and ovitraps was implemented during summer 2020 in northern Spain. Insects were determined by morphological features accompanied by DNA barcoding. RESULTS: A total of ten native mosquito species were recorded either as adults or as larval stages. The invasive species Aedes japonicus was collected only at egg or pupa stage in ovitraps. Culex pipiens s.l. and Culex torrentium were both common mosquito species accounting for 47.9% of the total larval site collections and their larvae might be found in a wide range of natural and artificial sites. Culiseta longiareolata specimens were also prominent (30.1% of the total) and occurred exclusively in man-made water-filled containers. A total of 13 Culicoides species were identified, 10 of which were captured by ultraviolet-CDC traps, particularly members of the Obsoletus complex (Culicoides obsoletus/Culicoides scoticus, 74.9%) and seven species by emergence traps, being the two most abundant C. kibunensis (44.8%) and C. festivipennis (34.9%). Simulium cryophilum was also collected hovering around the operator under field sampling. CONCLUSION: A comprehensive representation of the blood-sucking Diptera fauna and their larval sites was obtained by the multi-method approach in two Spanish golf courses.

Mosquito alert: leveraging citizen science to create a GBIF mosquito occurrence dataset.

The Mosquito Alert dataset includes occurrence records of adult mosquitoes collected worldwide in 2014-2020 through Mosquito Alert, a citizen science system for investigating and managing disease-carrying mosquitoes. Records are linked to citizen science-submitted photographs and validated by entomologists to determine the presence of five targeted European mosquito vectors: Aedes albopictus, Ae. aegypti, Ae. japonicus, Ae. koreicus, and Culex pipiens. Most records are from Spain, reflecting Spanish national and regional funding, but since autumn 2020 substantial records from other European countries are included, thanks to volunteer entomologists coordinated by the AIM-COST Action, and to technological developments to increase scalability. Among other applications, the Mosquito Alert dataset will help develop citizen science-based early warning systems for mosquito-borne disease risk. It can also be reused for modelling vector exposure risk, or to train machine-learning detection and classification routines on the linked images, to assist with data validation and establishing automated alert systems.

Multiple invasions, Wolbachia and human-aided transport drive the genetic variability of Aedes albopictus in the Iberian Peninsula.

The Asian tiger mosquito, Aedes albopictus, is one of the most invasive species in the world. Native to the tropical forests of Southeast Asia, over the past 30 years it has rapidly spread throughout tropical and temperate regions of the world. Its dramatic expansion has resulted in public health concerns as a consequence of its vector competence for at least 16 viruses. Previous studies showed that Ae. albopictus spread has been facilitated by human-mediated transportation, but much remains unknown about how this has affected its genetic attributes. Here we examined the factors that contributed to shaping the current genetic constitution of Ae. albopictus in the Iberian Peninsula, where the species was first found in 2004, by combining population genetics and Bayesian modelling. We found that both mitochondrial and nuclear DNA markers showed a lack of genetic structure and the presence of worldwide dominant haplotypes, suggesting regular introductions from abroad. Mitochondrial DNA showed little genetic diversity compared to nuclear DNA, likely explained by infection with maternally transmitted bacteria of the genus Wolbachia. Multilevel models revealed that greater mosquito fluxes (estimated from commuting patterns and tiger mosquito population distribution) and spatial proximity between sampling sites were associated with lower nuclear genetic distance, suggesting that rapid short- and medium-distance dispersal is facilitated by humans through vehicular traffic. This study highlights the significant role of human transportation in shaping the genetic attributes of Ae. albopictus and promoting regional gene flow, and underscores the need for a territorially integrated surveillance across scales of this disease-carrying mosquito.

Publishing data to support the fight against human vector-borne diseases.

Vector-borne diseases are responsible for more than 17% of human cases of infectious diseases. In most situations, effective control of debilitating and deadly vector-bone diseases (VBDs), such as malaria, dengue, chikungunya, yellow fever, Zika and Chagas requires up-to-date, robust and comprehensive information on the presence, diversity, ecology, bionomics and geographic spread of the organisms that carry and transmit the infectious agents. Huge gaps exist in the information related to these vectors, creating an essential need for campaigns to mobilise and share data. The publication of data papers is an effective tool for overcoming this challenge. These peer-reviewed articles provide scholarly credit for researchers whose vital work of assembling and publishing well-described, properly-formatted datasets often fails to receive appropriate recognition. To address this, GigaScience's sister journal GigaByte partnered with the Global Biodiversity Information Facility (GBIF) to publish a series of data papers, with support from the Special Programme for Research and Training in Tropical Diseases (TDR), hosted by the World Health Organisation (WHO). Here we outline the initial results of this targeted approach to sharing data and describe its importance for controlling VBDs and improving public health.

AIMSurv: First pan-European harmonized surveillance of Aedes invasive mosquito species of relevance for human vector-borne diseases.

Human and animal vector-borne diseases, particularly mosquito-borne diseases, are emerging or re-emerging worldwide. Six Aedes invasive mosquito (AIM) species were introduced to Europe since the 1970s: Aedes aegypti, Ae. albopictus, Ae. japonicus, Ae. koreicus, Ae. atropalpus and Ae. triseriatus. Here, we report the results of AIMSurv2020, the first pan-European surveillance effort for AIMs. Implemented by 42 volunteer teams from 24 countries. And presented in the form of a dataset named "AIMSurv Aedes Invasive Mosquito species harmonized surveillance in Europe. AIM-COST Action. Project ID: CA17108". AIMSurv2020 harmonizes field surveillance methodologies for sampling different AIMs life stages, frequency and minimum length of sampling period, and data reporting. Data include minimum requirements for sample types and recommended requirements for those teams with more resources. Data are published as a Darwin Core archive in the Global Biodiversity Information Facility- Spain, comprising a core file with 19,130 records (EventID) and an occurrences file with 19,743 records (OccurrenceID). AIM species recorded in AIMSurv2020 were Ae. albopictus, Ae. japonicus and Ae. koreicus, as well as native mosquito species.

Host-feeding patterns of native Culex pipiens and invasive Aedes albopictus mosquitoes (Diptera: Culicidae) in urban zones from Barcelona, Spain.

The feeding patterns of haematophagous arthropods are of major importance in the amplification and transmission of infectious disease agents to vertebrate hosts, including humans. The establishment of new vector populations in nonnative range might alter transmission networks. The Asian tiger mosquito Aedes albopictus (Skuse) represents an example of how an invasive species can alter the risk of viral transmission to humans. Blood meal molecular identification from two sympatric mosquito species (the invasive Ae. albopictus and the native Culex pipiens) was carried out by polymerase chain reaction-based methods. Samples were collected in Barcelona metropolitan area, Spain, from June to October 2009 as part of a monitoring-control program. Blood meals were identified to the species level in 30 Ae. albopictus and 43 Cx. pipiens. Ae. albopictus acquired blood exclusively from human hosts (100%), whereas Cx. pipiens fed on a diversity of avian and mammalian hosts, including 35.7% of blood meals from humans. Based on mosquito diet, our results suggest that the Ae. albopictus invasion in Spain might increase the risk of virus transmission to humans and could support local outbreaks of imported tropical viruses such as dengue and chikungunya. However, in the studied area, the presence of this invasive species would have a negligible effect on the transmission of zoonotic agents such as West Nile virus. However, Cx. pipiens could amplify and transmit West Nile virus, but avian contribution to its diet was lower than that reported in North America. Feeding patterns of these mosquito species may help to understand the flavivirus outbreaks recently reported in southwestern Europe.

Deep learning identification for citizen science surveillance of tiger mosquitoes.

Global monitoring of disease vectors is undoubtedly becoming an urgent need as the human population rises and becomes increasingly mobile, international commercial exchanges increase, and climate change expands the habitats of many vector species. Traditional surveillance of mosquitoes, vectors of many diseases, relies on catches, which requires regular manual inspection and reporting, and dedicated personnel, making large-scale monitoring difficult and expensive. New approaches are solving the problem of scalability by relying on smartphones and the Internet to enable novel community-based and digital observatories, where people can upload pictures of mosquitoes whenever they encounter them. An example is the Mosquito Alert citizen science system, which includes a dedicated mobile phone app through which geotagged images are collected. This system provides a viable option for monitoring the spread of various mosquito species across the globe, although it is partly limited by the quality of the citizen scientists' photos. To make the system useful for public health agencies, and to give feedback to the volunteering citizens, the submitted images are inspected and labeled by entomology experts. Although citizen-based data collection can greatly broaden disease-vector monitoring scales, manual inspection of each image is not an easily scalable option in the long run, and the system could be improved through automation. Based on Mosquito Alert's curated database of expert-validated mosquito photos, we trained a deep learning model to find tiger mosquitoes (Aedes albopictus), a species that is responsible for spreading chikungunya, dengue, and Zika among other diseases. The highly accurate 0.96 area under the receiver operating characteristic curve score promises not only a helpful pre-selector for the expert validation process but also an automated classifier giving quick feedback to the app participants, which may help to keep them motivated. In the paper, we also explored the possibilities of using the model to improve future data collection quality as a feedback loop.

At the tip of an iceberg: citizen science and active surveillance collaborating to broaden the known distribution of Aedes japonicus in Spain.

BACKGROUND: Active surveillance aimed at the early detection of invasive mosquito species is usually focused on seaports and airports as points of entry, and along road networks as dispersion paths. In a number of cases, however, the first detections of colonizing populations are made by citizens, either because the species has already moved beyond the implemented active surveillance sites or because there is no surveillance in place. This was the case of the first detection in 2018 of the Asian bush mosquito, Aedes japonicus, in Asturias (northern Spain) by the citizen science platform Mosquito Alert. METHODS: The collaboration between Mosquito Alert, the Ministry of Health, local authorities and academic researchers resulted in a multi-source surveillance combining active field sampling with broader temporal and spatial citizen-sourced data, resulting in a more flexible and efficient surveillance strategy. RESULTS: Between 2018 and 2020, the joint efforts of administrative bodies, academic teams and citizen-sourced data led to the discovery of this species in northern regions of Spain such as Cantabria and the Basque Country. This raised the estimated area of occurrence of Ae. japonicus from < 900 km2 in 2018 to > 7000 km2 in 2020. CONCLUSIONS: This population cluster is geographically isolated from any other population in Europe, which raises questions about its origin, path of introduction and dispersal means, while also highlighting the need to enhance surveillance systems by closely combining crowd-sourced surveillance with public health and mosquito control agencies' efforts, from local to continental scales. This multi-actor approach for surveillance (either passive and active) shows high potential efficiency in the surveillance of other invasive mosquito species, and specifically the major vector Aedes aegypti which is already present in some parts of Europe.

Correction: Phenotypic insecticide resistance in arbovirus mosquito vectors in Catalonia and its capital Barcelona (Spain).

[This corrects the article DOI: 10.1371/journal.pone.0217860.].

Phenotypic insecticide resistance in arbovirus mosquito vectors in Catalonia and its capital Barcelona (Spain).

A range of mosquito species that belong to the Culicidae family are responsible for the worldwide transmission of infectious arboviral diseases such as dengue fever, Zika, West Nile fever and Chikungunya fever. Spain is at risk of arbovirus outbreaks, as various arboviral diseases are frequently introduced and it has established competent vector populations. Autochthonous human cases of West Nile virus have been reported infrequently since 2004, and since October 2018 three autochthonous human case of dengue fever have been confirmed. In response to an outbreak of any arboviral disease, space spraying or fogging will be implemented to control adult mosquito populations. To ensure adulticiding is cost-effective, the insecticide susceptibility status of vectors throughout Catalonia, an autonomous region in north-eastern Spain, was assessed through standardized WHO tube and CDC bottle bioassays. All Culex pipiens populations tested were resistant to at least one of the pyrethroids tested, whereas Aedes albopictus populations were susceptible to all pyrethroids tested. More detailed studies on the Cx. pipiens populations from the Barcelona area (the capital and largest city of Catalonia) revealed resistance to all four classes of public health insecticides available (pyrethroids, carbamates, organophosphates and organochlorides). All Ae. albopictus populations were susceptible to those classes, except for one of the tests performed with pirimiphos-methyl (an organophosphate). Pyrethroids are currently the first line chemical class to be used in space spray operations in response to an outbreak of an arboviral disease. While pyrethroids can be effective in reducing Ae. albopictus populations, this class may not be effective to control Cx. pipiens populations.

Description of a Culex pipiens Gynandromorph Captured in La Rioja (Spain).

A gynandromorph is an imago combining well-defined male and female structures, caused by different processes, and it is infrequent in wild-caught mosquitoes. A Culex pipiens gynandromorph was caught in La Grajera wetland (La Rioja, Spain) in 2016. The phenotypic sex of wings was determined using linear discriminating analysis and the training error for the model on a previous database of sex-classified wing biometry data. It is suggested that the individual had an uncommon layout categorized as a top quarter male gynandromorph.