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Background: The expansion of invasive mosquitoes throughout Europe has increased in recent decades. In northern Spain, Aedes albopictus was detected for the first time in 2014, and Aedes japonicus was detected in the three Basque provinces in 2020. This study aimed to evaluate the distribution of these mosquito species and their association with factors related to urbanization., Methods: In 2021, a total of 568 ovitraps were deployed in 113 sampling sites from 45 municipalities with > 10,000 inhabitants. Oviposition substrate sticks were replaced each fortnight and examined for Aedes eggs from June to November. Aedes eggs were counted, and the eggs from a selection of positive oviposition sticks, encompassing at least one stick from each positive ovitrap, were hatched following their life cycle until the adult stage. When egg hatching was not successful, PCR targeting the COI gene and sequencing of amplicons were carried out., Results: Eggs were detected in 66.4% of the sampling sites and in 32.4% of the ovitraps distributed in the three provinces of the Basque Country. Aedes albopictus and Ae. japonicus were widespread in the studied area, confirming their presence in 23 and 26 municipalities, respectively. Co-occurrence of both species was observed in 11 municipalities. The analysis of the presence of Aedes invasive mosquitoes and the degree of urbanization (urban, suburban, peri-urban) revealed that Ae. albopictus showed a 4.39 times higher probability of being found in suburban areas than in peri-urban areas, whereas Ae. japonicus had a higher probability of being found in peri-urban areas. Moreover, the presence of Ae. albopictus was significantly associated with municipalities with a higher population density (mean = 2983 inh/km 2 ), whereas Ae. japonicus was associated with lower population density (mean = 1590 inh/km 2 )., Conclusions: The wide distribution of Ae. albopictus and Ae. japonicus observed confirmed the spread and establishment of these species in northern Spain. A new colonization area of Ae. japonicus in Europe was confirmed. Due to the potential impact of Aedes invasive mosquitoes on public health and according to our results, surveillance programs and control plans should be designed considering different urbanization gradients, types of environments, and population density., (© 2023. The Author(s).)

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., Competing Interests: The authors declare no conflict of interests. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.