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The efficacy of polymers (paints) enriched with microencapsulated insecticides against Aedes albopictus was evaluated under laboratory conditions using the forced contact technique (WHO). Three types of paints were evaluated: pyrethroids (PY), organophosphates (OP) and pyriproxyfen (PPF). These paints were applied on two types of surfaces and at two concentrations. Adult nulliparous females without blood ingestion were used to initially determine the knock-down (KD) effect in the first 30 minutes followed by the mortality rate and the time to 50% mortality (TD50) up to 72 hours. Ten bioassays were performed from a single initial treatment over the course of 27 months. A greater effect was confirmed when the surface was more porous and at a double concentration. Each formulation exhibited differences in effect intensities on KD or TD50 but showed greater effectiveness when the formulation contained PY. Implications for vector control are discussed based on studies on different formulations of microencapsulated insecticides on different surfaces and concentrations in laboratory and field bioassays.

En 2016 se estableció un sistema anual de vigilancia entomológica para la detección de Aedes albopictus en la comunidad autónoma de La Rioja. En el verano de 2020, se detectó por primera vez la presencia de huevos en una zona urbana, lo que constituye el primer registro de esta especie en la región. La identificación de este mosquito invasor supone un cambio en el riesgo de aparición de casos de arbovirosis en La Rioja. La vigilancia entomológica es crucial para el diseño de estrategias de control que, junto con la vigilancia epidemiológica, puedan evitar la aparición de brotes autóctonos de arbovirosis. En este trabajo se detalla el proceso de la primera detección de mosquito tigre en La Rioja, así como las intervenciones llevadas a cabo y las perspectivas de futuro. In 2016, an annual entomological surveillance program for the detection of Aedes albopictus in the autonomous community of La Rioja was established. In the summer of 2020, the presence of eggs was detected for the first time in an urban area, which constitutes the first record of this species in the region. The identification of this invasive mosquito implies a change in the risk of occurrence of arbovirus cases in La Rioja. Entomological surveillance is crucial for the design of control strategies that, together with epidemiological surveillance, can prevent the appearance of autochthonous arbovirus outbreaks. This work is a detailed description of the first detection of tiger mosquito in La Rioja, as well as the completed interventions, and the perspectives for the future.

Este artículo contiene 12 páginas, 2 tablas, 2 figuras., 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 frst 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 frst 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 research‑ ers resulted in a multi-source surveillance combining active feld sampling with broader temporal and spatial citizensourced data, resulting in a more fexible and efcient surveillance strategy. Results: Between 2018 and 2020, the joint eforts 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 sur‑ veillance systems by closely combining crowd-sourced surveillance with public health and mosquito control agencies’ eforts, from local to continental scales. This multi-actor approach for surveillance (either passive and active) shows high potential efciency in the surveillance of other invasive mosquito species, and specifcally the major vector Aedes aegypti which is already present in some parts of Europe., RE and FB from Mosquito Alert received funding from “la Caixa” Foundation (ID 100010434), under agreement HR18-00336 (Community-Driven Big Data Intel‑ ligence to Fight Mosquito-Borne Disease). This project has also been funded by the European Union’s Horizon 2020 research and innovation program, under the grant agreement 874735 (VEO). Samplings in the Basque Country were funded by the Department of Public Health and the Department of Economic Development and Infrastructures of the Basque Government, and co-funded by the project EU-LIFE 18 IPC/ES/000001 (Urban Klima 2050).

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.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.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.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.

An important element of vector control and surveillance of mosquito-borne diseases is updated information on vector species distribution. The aim of this study was to collect available information about mosquito species reported in Extremadura between 1920 and 2020 and create a catalog that would combine both published data and our recent field identifications. An exhaustive list is hereby presented, including species status and detailed distribution maps at a municipal level as well as their importance for public health. A total of 33 species, classified into five genera: Anopheles (five species), Aedes (14), Culex (nine), Culiseta (four), and Orthopodomyia (one) has been recorded, including 31 autochthonous, one invasive, Aedes (Stegomyia) albopictus, and one disappeared since 1953, Aedes (Stegomyia) aegypti. For the first time in Extremadura, we report the presence of important vectors such as Aedes (Aedimorphus) vexans vexans and Culex (Culex) perexiguus, and the new record of six species in the province of Badajoz, namely: Aedes (Dahliana) echinus, Aedes (Fredwardsius) vittatus, Aedes (Ochlerotatus) berlandi, Aedes (Ochlerotatus) pulcritarsis, Culex (Culex) mimeticus, and Culiseta (Culiseta) subochrea. Nineteen of these species are potential vectors of medical and veterinary relevance.

Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 (Diptera: Psychodidae) has been found in several European countries. In Spain, sporadic records were reported in the early ’80s in Catalonia (Northeast Spain), and it was never detected again. Recent entomological surveys carried out between 2004 and 2020 revealed the presence of several specimens of P. mascittii in Spain. The species identification was confirmed by both morphological and molecular analyses. The analyzed specimens belonged to the haplotype (COI_2) defined by one polymorphic site compared to other European specimens. Phlebotomus mascittii was found in low population densities in rural areas associated with livestock farms and in an urban cemetery during the summer season. This study provides the first records of this species in various localities along the Cantabrian cornice (Northern Spain) and represents its westernmost observation in the Palearctic region. The implications of the finding of this uncommon species are discussed at different levels, with emphasis on its suspected role in the transmission of leishmaniosis.

Changes in environmental conditions, whether related or not to human activities, are continuously modifying the geographic distribution of vectors, which in turn affects the dynamics and distribution of vector-borne infectious diseases. Determining the main ecological drivers of vector distribution and how predicted changes in these drivers may alter their future distributions is therefore of major importance. However, the drivers of vector populations are largely specific to each vector species and region. Here, we identify the most important human-activity-related and bioclimatic predictors affecting the current distribution and habitat suitability of the mosquito Culex pipiens and potential future changes in its distribution in Spain. We determined the niche of occurrence (NOO) of the species, which considers only those areas lying within the range of suitable environmental conditions using presence data. Although almost ubiquitous, the distribution of Cx. pipiens is mostly explained by elevation and the degree of urbanization but also, to a lesser extent, by mean temperatures during the wettest season and temperature seasonality. The combination of these predictors highlights the existence of a heterogeneous pattern of habitat suitability, with most suitable areas located in the southern and northeastern coastal areas of Spain, and unsuitable areas located at higher altitude and in colder regions. Future climatic predictions indicate a net decrease in distribution of up to 29.55%, probably due to warming and greater temperature oscillations. Despite these predicted changes in vector distribution, their effects on the incidence of infectious diseases are, however, difficult to forecast since different processes such as local adaptation to temperature, vector-pathogen interactions, and human-derived changes in landscape may play important roles in shaping the future dynamics of pathogen transmission. info:eu-repo/semantics/acceptedVersion

1) Background: Aedes albopictus has rapidly expanded throughout Europe, becoming a public health concern in the Mediterranean Basin. 2) Methods: Following the detection of Ae. albopictus in the southwestern French region of Aquitaine in 2012, an entomological surveillance programme was implemented in the Basque Country (Northern Spain) in 2013. 3) Results: Ae. albopictus eggs were first detected in 2014 in a transited parking area in the northeastern sampling point, 22 km away from the nearest French site with recorded presence of tiger mosquito. At this site, eggs were found throughout the study (2014&ndash, 2018). Other western and southern municipalities became positive in 2017 and 2018. Ae. albopictus adults were first captured in 2018 by aspiration of the vegetation in an area where eggs had been detected since 2015, suggesting a progressive establishment of a self-sustained population. Incidence of insect bites in humans was roughly constant over the study period except for a significant increase in 2018 in the Health County where eggs had been detected since 2014. Densities of Ae. albopictus eggs in positive areas remained at similar levels over the years. 4) Conclusion: Multiple approaches and standardized methods are necessary to successfully control this vector.

(1) Background: Aedes albopictus has rapidly expanded throughout Europe, becoming a public health concern in the Mediterranean Basin. (2) Methods: Following the detection of Ae. albopictus in the southwestern French region of Aquitaine in 2012, an entomological surveillance programme was implemented in the Basque Country (Northern Spain) in 2013. (3) Results: Ae. albopictus eggs were first detected in 2014 in a transited parking area in the northeastern sampling point, 22 km away from the nearest French site with recorded presence of tiger mosquito. At this site, eggs were found throughout the study (2014–2018). Other western and southern municipalities became positive in 2017 and 2018. Ae. albopictus adults were first captured in 2018 by aspiration of the vegetation in an area where eggs had been detected since 2015, suggesting a progressive establishment of a self-sustained population. Incidence of insect bites in humans was roughly constant over the study period except for a significant increase in 2018 in the Health County where eggs had been detected since 2014. Densities of Ae. albopictus eggs in positive areas remained at similar levels over the years. (4) Conclusion: Multiple approaches and standardized methods are necessary to successfully control this vector.

Se expone la primera cita de Aedes (Fredwardsius) vittatus (Bigot, 1861) en la comunidad autónoma de Galicia, conseguida gracias a la plataforma de ciencia ciudadana Mosquito Alert. Aun cuando este proyecto está enfocado hacia la detección y seguimiento de dos especies de culícidos exóticos invasores: Aedes (Stegomyia) albopictus (Skuse 1894) y Aedes (Stegomyia) aegypti (L.), se evidencia la gran capacidad existente para el estudio de la biodiversidad así como la detección de otras especies por parte de plataformas que se apoyan en miles de ciudadanos motivados y un equipo experto. We present here the first record of Aedes (Fredwardsius) vittatus (Bigot, 1861) in the autonomous region of Galicia, obtained by the means of the citizen science project Mosquito Alert. Whereas this project only aims to the monitoring and early detection of the two invasive mosquito species Aedes (Stegomyia) albopictus (Skuse 1894) and Aedes (Stegomyia) aegypti (L.), it is worth noting its capacities in regards to the general biodiversity study and the detection of mosquito species if a large pool of users combines with the expert knowledge.

On October 3rd 2018, dengue virus (DENV) infection was confirmed in three family members (symptoms onset between August 18th and 27th) without travel history outside of Spain. They had been together in the Autonomous Communities (AC) of Murcia and Andalusia. By the end of October, a second cluster of two dengue cases (symptoms onset on September 27th and 30th) was confirmed in the AC of Murcia. DENV type 1 sequence was identical to the first cluster, and the epidemiological link was a visit from a case of the first cluster to a fruit-farm neighboring the small village of residence of the second cluster. The entomological investigation found Aedes albopictus activity in this area although all mosquitoes were PCR-negative for DENV. This is the first autochthonous dengue outbreak identified in Spain. This outbreak highlights challenges to timely detect and respond to DENV transmission and opens questions on dengue dynamics in a non-endemic context.

Background: A number of mosquito-borne viruses such as dengue virus (DENV), Usutu virus (USUV), West Nile virus (WNV) are autochthonously transmitted in Europe and six invasive mosquito species have been detected in this temperate region. This has increased the risk for the emergence of further mosquito-borne diseases. However, there is a paucity of information on whether European populations of invasive mosquito species are competent to transmit arboviruses. In this study, the susceptibility of Aedes albopictus originating from Spain and a laboratory-adapted colony of Aedes aegypti, was assessed for infection with, and transmission of Zika virus (ZIKV). Vertical transmission in both species was also assessed. Methods: Aedes albopictus colonised from eggs collected in Spain and an existing colony of Ae. aegypti were fed infectious blood meals containing ZIKV (Polynesian strain) at 1.6 × 10 7 PFU/ml. Blood-fed mosquitoes were separated and maintained at 20 °C or 25 °C. Legs, saliva and bodies were sampled from specimens at 7, 14 and 21 days post-infection (dpi) in order to determine infection, dissemination and transmission rates. All samples were analysed by real-time RT-PCR using primers targeting the ZIKV NS1 gene. Results: At 14 dpi and 21 dpi, ZIKV RNA was detected in the bodies of both species at both temperatures. However, live virus only was detected in the saliva of Ae. aegypti at 25 °C with a transmission rate of 44%. No evidence for virus expectoration was obtained for Ae. albopictus under any condition. Notably, ZIKV RNA was not detectable in the saliva of Ae. aegypti at 20 °C after 21 days. No vertical transmission of ZIKV was detected in this study. Conclusions: Experimental infection of Ae. albopictus colonized from Spain with ZIKV did not result in expectoration of virus in saliva in contrast to results for Ae. aegypti. No evidence of vertical transmission of virus was observed in this study. This suggests that this strain of Ae. albopictus is not competent for ZIKV transmission under the conditions tested. © 2019 The Author(s).

[Background] Aedes japonicus is an invasive vector mosquito from Southeast Asia which has been spreading across central Europe since the year 2000. Unlike the Asian Tiger mosquito (Aedes albopictus) present in Spain since 2004, there has been no record of Ae. japonicus in the country until now., [Results] Here, we report the first detection of Ae. japonicus in Spain, at its southernmost location in Europe. This finding was triggered by the citizen science platform Mosquito Alert. In June 2018, a citizen sent a report via the Mosquito Alert app from the municipality of Siero in the Asturias region (NW Spain) containing pictures of a female mosquito compatible with Ae. japonicus. Further information was requested from the participant, who subsequently provided several larvae and adults that could be classified as Ae. japonicus. In July, a field mission confirmed its presence at the original site and in several locations up to 9 km away, suggesting a long-time establishment. The strong media impact in Asturias derived from the discovery raised local participation in the Mosquito Alert project, resulting in further evidence from surrounding areas., [Conclusions] Whilst in the laboratory Ae. japonicus is a competent vector for several mosquito-borne pathogens, to date only West Nile virus is a concern based on field evidence. Nonetheless, this virus has yet not been detected in Asturias so the vectorial risk is currently considered low. The opportunity and effectiveness of combining citizen-sourced data to traditional surveillance methods are discussed., Mosquito Alert is currently promoted by the ‘la Caixa’ Banking Foundation and Dipsalut from the Diputació de Girona, and sponsored by the Company Lokimica S.A.

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Background Leishmania infantum is the parasite responsible for the disease in humans known as zoonotic visceral leishmaniasis (ZVL). Dogs are considered the main domestic reservoir of ZVL and sand flies are the proven vectors. The use of systemic insecticides in dogs has been studied as an alternative strategy to control ZVL in endemic areas. One systemic insecticide in dogs, fluralaner, has a proven anti-sand fly effect in membrane-fed studies. However, the efficacy and duration on sand flies directly feeding from dogs treated with fluralaner remains unknown. Methods Direct feeding bioassays were performed on 10 beagle dogs that had been randomly assigned to two groups: one with five dogs orally treated with Bravecto® (fluralaner) and other five as a control. About 30 females of Phlebotomus papatasi were allowed to directly feed from dogs at seven days before the administration of the treatment and Days 3, 17, 31, 45 and 73 post-treatment. Sand fly mortality after feeding was observed every 24 h for 5 days. The Kaplan-Meyer method, Henderson-Tilton formula and a negative binomial mixed model were used to respectively calculate: (i) mortality and its 95% confidence interval (CI); (ii) efficacy of the insecticide at killing sand flies in 24 h; and (iii) differences in the risk of sand fly death at 24 h after feeding. Results Control sand fly mortality 24 h after feeding was always ≤ 20% and mortality in the fluralaner group ranged from 2% (95% CI: 0–4%) 7 days before treatment to 100% at 3 days post-treatment. Fluralaner efficacy was 100, 93, 94 and 75% at Days 3, 17, 31 and 45, respectively (P < 0.0001). The increase in the risk of sand fly death was 32.9 (95% CI: 4–263), 76 (95% CI: 8–705), 95.8 (95% CI: 9–1029) and 10.6 times (95% CI: 1.43–79) on Days 3, 17, 31 and 45, respectively Conclusions The efficacy of fluralaner, orally administered to dogs, against sand-flies was above 90% for 31 days. Fluralaner administered to dogs should be further evaluated as a control strategy in ZVL endemic areas. Electronic supplementary material The online version of this article (10.1186/s13071-018-3231-8) contains supplementary material, which is available to authorized users.

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, Peniscola, Castellon, 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.

Este artículo contiene 13 páginas, 8 figuras, 2 tablas., Recent outbreaks of Zika, chikungunya and dengue highlight the importance of better understanding the spread of disease-carrying mosquitoes across multiple spatio-temporal scales. Traditional surveillance tools are limited by jurisdictional boundaries and cost constraints. Here we show how a scalable citizen science system can solve this problem by combining citizen scientists’ observations with expert validation and correcting for sampling effort. Our system provides accurate early warning information about the Asian tiger mosquito (Aedes albopictus) invasion in Spain, well beyond that available from traditional methods, and vital for public health services. It also provides estimates of tiger mosquito risk comparable to those from traditional methods but more directly related to the human–mosquito encounters that are relevant for epidemiological modelling and scalable enough to cover the entire country. These results illustrate how powerful public participation in science can be and suggest citizen science is positioned to revolutionize mosquito-borne disease surveillance worldwide., A. Torrell (Departament de Territori i Sostenibilitat, Generalitat de Catalunya) and the support of the Spanish Ministry of Public Health, CCAES (Centro de Coordinación de Alertas y Emergencias Sanitarias) and the Health Department of the Generalitat de Catalunya. J.R.B.P. is funded by the European Union’s Horizon 2020 research and innovation programme under Marie Skłodowska-Curie grant agreement no. 657956. The research leading to these results has received funding from the Spanish Ministry of Economy and Competitiveness (MINECO, Plan Estatal I+D+I CGL2013-43139-R) and ‘la Caixa’ Banking Foundation. Mosquito Alert is currently promoted by ‘la Caixa’ Banking Foundation. Other contributors during 2014-15 are the Fundación Española de Ciencia y Tecnología (FCT-13-7019), the program RecerCaixa-2013, and the company Lokimica S.A.

After the recent detection of Aedes albopictus (Skuse, 1894) in Southwestern France, the entomological surveillance in the Basque Country, since 2013, has found two positive traps in Irun (Guipuzkoa). The differences in environmental conditions, which could influence its establishment, between this area and the Spa- nish Levante populations are discussed. Also, this is the first record of Culex hortensis Ficalbi, 1889 and Culiseta longiareolata (Macquart, 1838) for the Basque Country and the Guipuzkoa pro- vince. Also it is the first record of Cx. pipiens Linnaeus, 1758 for the Guipuzkoa province.

In 2004, Aedes (Stegomyia) albopictus (Skuse, 1894) was observed for the first time in Catalonia, northeastern Spain. A decade later, it has spread throughout the eastern Mediterranean region of the country and the Balearic Islands. Framed within a national surveillance project, we present the results of monitoring in 2013 in the autonomous communities of the mainland Levante. The current study reveals a remarkable increase in the spread of the invasive mosquito in relation to results from 2012; the species was present and well-established in 48 municipalities, most of which were along the Mediterranean coastline from the Valencian Community to the Region of Murcia. Financial support: Directorate General of Public Health, Quality and Innovation, Ministry of Health, Social Services and Equity in Spain (Entomological surveillance at airports and ports against imported vectors of exotic infectious diseases and surveillance of potential indigenous vectors of such diseases project, OTRI 2013/0215) Sí

First findings of Aedes (Stegomyia) albopictus (Skuse, 1894) in the province of Valencia, Spain Framed within the project "Entomological surveillance at airports and ports against vectors of imported infectious diseases, and sur- veillance of potential native vectors of these diseases", on Septem- ber 2011 Aedes albopictus (Diptera, Culicidae) is reported for the first time in the province of Valencia in the coastal municipalities of Sueca, Cullera and Gandia, using oviposition traps.

Two gynandromorphs of Culex theileri are described. The individuals were caught in the Centers for Disease Control and Prevention miniature light traps in the municipality of Alcollarin, Caceres, Spain, in 2011. One of the specimens presented polar gynandromorphism, whereas the other showed a combination of polar and transverse gynandromorphic features. Details are given of the main characteristics of both mosquitoes, with particular emphasis that this gynandromorphic finding is only the second reported for this mosquito species worldwide.

espanolSe reporta la estrecha relacion entre uno de los principales vectores del virus del Nilo Occidental (VNO), Culex pipiens (Diptera, Culicidae), y la planta invasora acuatica Ludwigia grandiflora (Spermatophyta, Onagraceae) en el marjal de Xeraco-Xeresa, un humedal al sur de la provincia de Valencia. Los hidrofitos aloctonos desempenan en ocasiones un papel primordial en la proliferacion descontrolada de poblaciones de mosquitos y, en consecuencia, tambien en la transmision y propagacion de muchas enfermedades emergentes y reemergentes transmitidas por estos dipteros. Teniendo en cuenta la situacion geografica de este humedal y los diferentes factores climaticos y biologicos, la aparicion de una epidemia del VNO con incidencia humana es un hecho mas que probable en esta vertiente mediterranea del pais EnglishThe close relationship between one of the main vectors of West Nile virus (WNV), Culex pipiens (Diptera, Culicidae), and invasive aquatic plant Ludwigia grandiflora (Spermatophyta, Onagraceae) is reported on Xeraco-Xeresa marsh, a wetland in the south of the province of Valencia. Allochthonous hydrophytes sometimes play a major role in the uncontrolled proliferation of culicid populations and, consequently, also in the transmission and spread of many emerging and re-emerging mosquito-borne diseases. Considering the location of the wetland and the different climatic and biological factors, the emergence of West Nile virus infection outbreak in humans is a fairly likely at this Mediterranean side of the country.

The sandfly Phlebotomus perniciosus is the most widespread vector of Leishmania infantum in Spain. Laboratory colonisation represents the most feasible source of information on the biology of these insects, but in conducting any study, the density of individuals in the colony may drop to such an extent that it is sometimes difficult to recover the initial population levels. A new technique was tested for the recovery of sandfly eggs in three different colonies; the recovery rate was studied by comparing the standard method of mass rearing with this new method of colony management. The results demonstrate a mean increase of 18.4% in adult production, a growth in colony productivity that justifies the inclusion of this process in the routine maintenance of any colony of sandflies.

The control of arthropod vectors of pathogens that affect human and animal health is important for the eradication of vector-borne diseases. Recent evidences showed a reduction in the survival and/or fertility of mosquitoes, sand flies and poultry red mites fed in vitro with antibodies against the recombinant Aedes albopictus akirin. These experiments were the first step toward the development of a multi-target arthropod vaccine. In this study, we showed that the oviposition of A. albopictus and Phlebotomus perniciosus fed on mice vaccinated with recombinant A. albopictus akirin was reduced by 17% and 31%, respectively when compared to controls. However, Aedes aegypti mosquitoes were not affected after feeding on vaccinated mice. These results showed that recombinant A. albopictus akirin could be used to vaccinate hosts for the control of mosquito and sand fly infestations and suggested new experiments to develop improved vaccine formulations., This research was supported by the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Spain (project FAU2008-00014-00-00) and the Consejería de Educación y Ciencia, JCCM, Spain (project PEII09-0118-8907). J.A. Moreno-Cid is a recipient of a JCCM fellowship. M. Canales was funded by the Wellcome Trust under the “Animal Health in the Developing World” initiative (project 0757990). M. Villar was funded by the JAE-DOC program (CSIC-FSE), Spain.

Background Japanese encephalitis virus (JEV) is the principal cause of mosquito-borne encephalitis in human populations within Asia. If introduced into new geographic areas, it could have further implications for public and animal health. However, potential mosquito vectors for virus transmission have not been fully investigated. The Asian tiger mosquito, Aedes albopictus, has emerged in Europe and is now expanding its geographical range into more northerly latitudes. Culex quinquefasciatus, although absent from Europe, has been detected in Turkey, a country with territory in Europe, and could act as a vector for JEV in other regions. To assess the risk of these invasive species acting as vectors for JEV and therefore potentially contributing to its geographical expansion, we have investigated the vector competence of Ae. albopictus and Cx. quinquefasciatus. Methods Two colonised lines of Ae. albopictus (Italy and Spain) and a line of Cx. quinquefasciatus (Tanzania) were compared for susceptibility to infection by oral feeding with JEV strain SA-14, genotype III at 106 PFU/ml and maintained at 25 °C. Specimens were processed at 7 and 14 days post-inoculation (dpi). Rates of infection, dissemination and transmission were assessed through detection of viral RNA by real-time polymerase chain reaction (RT-PCR) in mosquito body, legs and saliva, respectively, at each time point. Where possible, infection and dissemination were confirmed by immunohistochemical (IHC) detection of the JEV envelope protein. Results Aedes albopictus from Italy showed no susceptibility to infection with JEV strain SA-14. Conversely, Ae. albopictus colonised in Spain was susceptible and 100% of infected mosquitoes that were subjected to saliva screening expressed viral RNA at 14 dpi. Culex quinquefasciatus was highly susceptible to infection as early as 7 dpi and 50% of infected mosquitoes that were subjected to saliva screening expressed viral RNA at 14 dpi. Infection and dissemination were confirmed in Cx. quinquefasciatus by IHC detection of JEV envelope protein in both the mid-gut and salivary glands. Conclusions Aedes albopictus from two different locations in Europe range from being susceptible to JEV and capable of transmission through to being resistant. Culex quinquefasciatus also appears highly susceptible; therefore, both species could potentially act as vectors for JEV and facilitate the emergence of JEV into new regions. Graphical Abstract

Ten years have gone by since the Asian tiger mosquito Aedes albopictus was recorded for the first time in Spain. In this paper, all relevant published information about this vector in Spain for the period 2004-2014 is reviewed. The known distribution for 2014 is provided, including all historical records (published and unpublished data) and the results from samplings of the last year. The consequences on public health about the presence of the Asian tiger mosquito in Spain are also highlighted. Further, legal aspects and control plans related to the management and diseases transmitted by this invasive vector species are also discussed. Part of the samplings are supported by the contract “Entomological surveillance at airports and ports against imported vectors of exotic infectious diseases and surveillance of potential indigenous vectors of such diseases”, funding by the Spanish Ministery of Health, Social Services and Equality, and the project “The tiger mosquito invasion in Spain: A public health risk caused by global change”, funding by the Spanish Ministry of Economy and Competitiveness. The Generalitat de Catalunya has provided their unpublished sampling data from 2011 to 2014, what was indispensable to complete the known distribution. The Instituto Universitario de Enfermedades Tropicales y Salud Pública de Canarias is involved in the entomological surveillance of airports and harbours of Canary Islands and the regional administration (Consejería de Sanidad) supports, in part, this work. The companies Compañía de Tratamientos de Levante S.L., Rafaela Belmonte Nortes and Campos Serrano Biólogos have collaborated in the samplings of 2014 in the Spanish Mediterranean side, exclusive Catalonia. The data of the project Atrapaeltigre.com has significantly helped to find some possible new localities for Ae. albopictus located far way from the known range. Sí