Automated age grading of female Culex pipiens by an optical sensor system coupled to a mosquito trap.

Background: The age distribution of a mosquito population is a major determinant of its vectorial capacity. To contribute to disease transmission, a competent mosquito vector, carrying a pathogen, must live longer than the extrinsic incubation period of that pathogen to enable transmission to a new host. As such, determining the age of female mosquitoes is of significant interest for vector-borne diseases surveillance and control programs.

Vector Competence Assays for RVFV in Mosquitoes.

Rift Valley fever virus (RVFV) is the causative agent of Rift Valley fever, a mosquito-borne zoonotic febrile illness that primarily affects ruminants, camels, and humans. It is endemic in sub-Saharan Africa and the Arabian Peninsula, albeit with the potential to spread to and establish in non-endemic countries where mosquito vectors are present. Assessing the vector competence (VC) of local mosquito populations is essential for risk evaluation and to develop more accurate control and surveillance strategies.

Tissue distribution and transmission of Rift Valley fever phlebovirus in European and mosquitoes following intrathoracic inoculation.

Rift Valley fever virus (, RVFV) poses significant economic challenges, particularly in African nations, causing substantial livestock losses and severe haemorrhagic disease in humans. In Europe, the risk of RVFV transmission is deemed moderate due to the presence of competent vectors like and , along with susceptible animal vertebrate hosts across member states. This study investigates RVFV infection dynamics in European mosquito populations, aiming to enhance our understanding of their vectorial capacity and virus transmission, which can be useful for future investigations to improve RVFV surveillance, control programmes, and preventive treatments.

West Nile Virus Seroprevalence in Wild Birds and Equines in Madrid Province, Spain.

West Nile virus (WNV) is a re-emerging flavivirus, primarily circulating among avian hosts and mosquito vectors, causing periodic outbreaks in humans and horses, often leading to neuroinvasive disease and mortality. Spain has reported several outbreaks, most notably in 2020 with seventy-seven human cases and eight fatalities. WNV has been serologically detected in horses in the Community of Madrid, but to our knowledge, it has never been reported from wild birds in this region.

Spatial scale influences the distribution of viral diversity in the eukaryotic virome of the mosquito .

Our knowledge of the diversity of eukaryotic viruses has recently undergone a massive expansion. This diversity could influence host physiology through yet unknown phenomena of potential interest to the fields of health and food production. However, the assembly processes of this diversity remain elusive in the eukaryotic viromes of terrestrial animals.

Infravec2 guidelines for the design and operation of containment level 2 and 3 insectaries in Europe.

With the current expansion of vector-based research and an increasing number of facilities rearing arthropod vectors and infecting them with pathogens, common measures for containment of arthropods as well as manipulation of pathogens are becoming essential for the design and running of such research facilities to ensure safe work and reproducibility, without compromising experimental feasibility. These guidelines and comments were written by experts of the Infravec2 consortium, a Horizon 2020-funded consortium integrating the most sophisticated European infrastructures for research on arthropod vectors of human and animal diseases. They reflect current good practice across European laboratories with experience of safely handling different mosquito species and the pathogens they transmit.

Zika vector competence data reveals risks of outbreaks: the contribution of the European ZIKAlliance project.

First identified in 1947, Zika virus took roughly 70 years to cause a pandemic unusually associated with virus-induced brain damage in newborns. Zika virus is transmitted by mosquitoes, mainly Aedes aegypti, and secondarily, Aedes albopictus, both colonizing a large strip encompassing tropical and temperate regions. As part of the international project ZIKAlliance initiated in 2016, 50 mosquito populations from six species collected in 12 countries were experimentally infected with different Zika viruses.

A novel optical sensor system for the automatic classification of mosquitoes by genus and sex with high levels of accuracy.

Background: Every year, more than 700,000 people die from vector-borne diseases, mainly transmitted by mosquitoes. Vector surveillance plays a major role in the control of these diseases and requires accurate and rapid taxonomical identification. New approaches to mosquito surveillance include the use of acoustic and optical sensors in combination with machine learning techniques to provide an automatic classification of mosquitoes based on their flight characteristics, including wingbeat frequency.

Widespread Circulation of Flaviviruses in Horses and Birds in Northeastern Spain (Catalonia) between 2010 and 2019.

The surveillance for West Nile virus (WNV) in Catalonia (northeastern Spain) has consistently detected flaviviruses not identified as WNV. With the aim of characterizing the flaviviruses circulating in Catalonia, serum samples from birds and horses collected between 2010 and 2019 and positive by panflavivirus competition ELISA (cELISA) were analyzed by microneutralization test (MNT) against different flaviviruses. A third of the samples tested were inconclusive by MNT, highlighting the limitations of current diagnostic techniques.

Microbiota Variation Across Life Stages of European Field-Caught and During Laboratory Colonization: New Insights for Malaria Research.

The potential use of bacteria for developing novel vector control approaches has awakened new interests in the study of the microbiota associated with vector species. To set a baseline for future malaria research, a high-throughput sequencing of the bacterial 16S ribosomal gene V3-V4 region was used to profile the microbiota associated with late-instar larvae, newly emerged females, and wild-caught females of a sylvan population from a former malaria transmission area of Spain. Field-acquired microbiota was then assessed in non-blood-fed laboratory-reared females from the second, sixth, and 10th generations.

Alteration in the Culex pipiens transcriptome reveals diverse mechanisms of the mosquito immune system implicated upon Rift Valley fever phlebovirus exposure.

Rift Valley fever phlebovirus (RVFV) causes an emerging zoonotic disease and is mainly transmitted by Culex and Aedes mosquitoes. While Aedes aegypti-dengue virus (DENV) is the most studied model, less is known about the genes involved in infection-responses in other mosquito-arboviruses pairing. The main objective was to investigate the molecular responses of Cx.

Evidence of Zika virus horizontal and vertical transmission in from Spain but not infectious virus in saliva of the progeny.

mosquitoes have been experimentally demonstrated to be a competent vector for Zika virus (ZIKV) in different countries, but there are still some gaps related to the importance of in ZIKV transmission. Recent studies on Spanish populations showed controversial results for ZIKV transmission and no studies have been performed yet to detect infectious ZIKV in saliva of progeny of infected female mosquitoes. Herein, the horizontal transmission (HT) and vertical transmission (VT) of ZIKV in field-collected mosquitoes from Spain were evaluated for ZIKV strains (African I and Asian lineages) to better estimate the risk of ZIKV transmission by .

The Role of Birds of Prey in West Nile Virus Epidemiology.

Reported human cases of West Nile virus (WNV) in Europe increased dramatically in 2018. Lineage 1 strains had been circulating in Euro-Mediterranean countries since the early 1990s. The subsequent introduction of WNV lineage 2 has been responsible for the remarkable upsurge of European WNV outbreaks since 2004, including the dramatic increase in human cases observed since 2018.

High sensitivity of one-step real-time reverse transcription quantitative PCR to detect low virus titers in large mosquito pools.

Background: Mosquitoes are the deadliest animals in the world. Their ability to carry and spread diseases to humans causes millions of deaths every year. Due to the lack of efficient vaccines, the control of mosquito-borne diseases primarily relies on the management of the vector.

Laboratory colonization and maintenance of Anopheles atroparvus from the Ebro Delta, Spain.

Background: Historically, Anopheles atroparvus has been considered one of the most important malaria vectors in Europe. Since malaria was eradicated from the European continent, the interest in studying its vectors reduced significantly. Currently, to better assess the potential risk of malaria resurgence on the continent, there is a growing need to update the data on susceptibility of indigenous Anopheles populations to imported Plasmodium species.

Viromics on Honey-Baited FTA Cards as a New Tool for the Detection of Circulating Viruses in Mosquitoes.

Worldwide, emerging and re-emerging infectious diseases (EIDs) are a major burden on public and animal health. Arthropod vectors, with mosquitoes being the main contributors of global disease, transmit more than 70% of the recognized EIDs. To assess new alternatives for arthropod-borne viral diseases surveillance, and for the detection of new viruses, honey-baited Flinders Technology Associates (FTA) cards were used as sugar bait in mosquito traps during entomological surveys at the Llobregat River Delta (Catalonia, Spain).

Field-captured Aedes vexans (Meigen, 1830) is a competent vector for Rift Valley fever phlebovirus in Europe.

Background: Aedes vexans (Meigen) is considered a nuisance species in central Europe and the Mediterranean region. It is an anthropophilic and mammalophilic floodwater mosquito involved in the transmission of several arboviruses. Rift Valley fever (RVF) is a relevant mosquito-borne zoonosis, affecting mainly humans and ruminants, that causes severe impact in public health and economic loses.

Usefulness of Eurasian Magpies () for West Nile virus Surveillance in Non-Endemic and Endemic Situations.

In September 2017, passive surveillance allowed the detection of West Nile virus (WNV) lineage 2 for the first time in northern Spain in a northern goshawk (). However, a cross sectional study carried out in Eurasian magpies () in a nearby area evidenced that WNV had been circulating two months earlier. Therefore, active surveillance in Eurasian magpies proved its effectiveness for the early detection of WNV in a non-endemic area.

European Aedes caspius mosquitoes are experimentally unable to transmit Zika virus.

Background: Aedes caspius (Pallas, 1771) is a floodwater mosquito species widely distributed in the Western Palaearctic. As an anthropophilic species, its role as an arbovirus vector may be the key for understanding the transmission cycle of certain diseases in Europe such as Zika virus (ZIKV). Concerning vector competence for ZIKV, studies related to Ae.

Limited risk of Zika virus transmission by five Aedes albopictus populations from Spain.

Background: Aedes albopictus, the Asian tiger mosquito, is an exotic invasive species in Europe. It has substantial public health relevance due to its potential role in transmitting several human pathogens. Out of the European countries, Spain has one of the highest risk levels of autochthonous arbovirus transmission due to both the high density of Ae.

Detection of West Nile virus lineage 2 in North-Eastern Spain (Catalonia).

In September 2017, West Nile virus (WNV) lineage 2 was detected in Catalonia (Northern Spain) in northern goshawks by passive surveillance. The phylogenetic analyses showed that it was related to the Central/Southern European strains, evidencing WNV lineage 2 spread to Western Europe. WNV local transmission was later detected in bearded vultures housed at the Wildlife Recovery center where the goshawk was transferred to.

Arbovirus surveillance: first dengue virus detection in local 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 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.

West Nile virus and other mosquito-borne viruses present in Eastern Europe.

Eastern Europe (EE) has been severely affected by mosquito-borne viruses (moboviruses). In this review, we summarize the epidemiology of moboviruses, with particular attention to West Nile virus (WNV). The study of WNV human cases in EE between 2010 and 2016, revealed that the epidemiology of WNV in EE is complex with the combination of introduction of different WNV strains from lineages 1 and 2, and the establishment of endemic cycles.