BACKGROUND: Invasive mosquitoes of the genus Aedes are quickly spreading around the world. The presence of these alien species is concerning for both their impact on the native biodiversity and their high vector competence. The surveillance of Aedes invasive mosquito (AIM) species is one of the most important steps in vector-borne disease control and prevention. METHODS: In 2020, the monitoring of AIM species was conducted in five areas (Bratislava, Zvolen, Banská Bystrica, Prešov, Košice) of Slovakia. The sites were located at points of entry (border crossings with Austria and Hungary) and in the urban and rural zones of cities and their surroundings. Ovitraps were used at the majority of sites as a standard method of monitoring. The collected specimens were identified morphologically, with subsequent molecular identification by conventional PCR (cox1) and Sanger sequencing. The phylogenetic relatedness of the obtained sequences was inferred by the maximum likelihood (ML) method. The nucleotide heterogeneity of the Slovak sequences was analysed by the index of disparity. RESULTS: A bush mosquito, Aedes japonicus japonicus, was found and confirmed by molecular methods in three geographically distant areas of Slovakia-Bratislava, Zvolen and Prešov. The presence of AIM species is also likely in Košice; however, the material was not subjected to molecular identification. The nucleotide sequences of some Slovak strains confirm their significant heterogeneity. They were placed in several clusters on the ML phylogenetic tree. Moreover, Ae. j. japonicus was discovered in regions of Slovakia that are not close to a point of entry, where the mosquitoes could find favourable habitats in dendrothelms in city parks or forests. CONCLUSION: Despite being a first record of the Ae. j. japonicus in Slovakia, our study indicates that the established populations already exist across the country, underlining the urgent need for intensified surveillance of AIM species as well as mosquito-borne pathogens.

Department of Biology and General Ecology Technical University in Zvolen T G Masaryka 24 960 01 Zvolen Slovakia

Department of Breeding and Diseases of Game Fish and Bees Ecology and Cynology University of Veterinary Medicine and Pharmacy in Košice Komenského 73 041 81 Košice Slovakia

Department of Ecology Faculty of Humanities and Natural Sciences 17 Novembra č 1 081 16 Prešov Slovakia

Department of Ecology Faculty of Natural Sciences Comenius University in Bratislava Iľkovičova 6 842 15 Bratislava Slovakia

Department of Ecosystem Biology Faculty of Science University of South Bohemia Branišovská 1760 370 05 České Budějovice Czech Republic

Department of Microbiology and Virology Faculty of Natural Sciences Comenius University in Bratislava Ilkovičova 6 842 15 Bratislava Slovakia

Institute of Botany Plant Science and Biodiversity Center Slovak Academy of Sciences Dúbravská cesta 9 845 23 Bratislava Slovakia

Institute of Virology Biomedical Research Center Slovak Academy of Sciences Dúbravská cesta 9 845 05 Bratislava Slovakia

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