Indigo-dyed cellulose fibers and synthetic polymers in surface-feeding seabird chick regurgitates from the Gulf of Alaska.

We provide evidence of anthropogenic materials ingestion in seabirds from a remote oceanic area, using regurgitates obtained from black-legged kittiwake (Rissa tridactyla) chicks from Middleton Island (Gulf of Alaska, USA). By means of GPS tracking of breeding adults, we identified foraging grounds where anthropogenic materials were most likely ingested. They were mainly located within the continental shelf of the Gulf of Alaska and near the Alaskan coastline. Anthropogenic cellulose fibers showed a high prevalence (85 % occurrence), whereas synthetic polymers (in the micro- and mesoplastics dimensional range) were less frequent (20 %). Most fibers (60 %) were blue and we confirmed the presence of indigo-dyed cellulosic fibers, characteristic of denim fabrics. In terms of mass, contamination levels were 0.077 µg g-1 wet weight and 0.009 µg g-1 wet weight for anthropogenic microfibers and synthetic polymers, respectively. These results represent the only recent report of contamination by anthropogenic fibers in seabirds from the Gulf of Alaska.

The great tit HapMap project: A continental-scale analysis of genomic variation in a songbird.

A major aim of evolutionary biology is to understand why patterns of genomic diversity vary within taxa and space. Large-scale genomic studies of widespread species are useful for studying how environment and demography shape patterns of genomic divergence. Here, we describe one of the most geographically comprehensive surveys of genomic variation in a wild vertebrate to date; the great tit (Parus major) HapMap project. We screened ca 500,000 SNP markers across 647 individuals from 29 populations, spanning ~30 degrees of latitude and 40 degrees of longitude - almost the entire geographical range of the European subspecies. Genome-wide variation was consistent with a recent colonisation across Europe from a South-East European refugium, with bottlenecks and reduced genetic diversity in island populations. Differentiation across the genome was highly heterogeneous, with clear 'islands of differentiation', even among populations with very low levels of genome-wide differentiation. Low local recombination rates were a strong predictor of high local genomic differentiation (FST), especially in island and peripheral mainland populations, suggesting that the interplay between genetic drift and recombination causes highly heterogeneous differentiation landscapes. We also detected genomic outlier regions that were confined to one or more peripheral great tit populations, probably as a result of recent directional selection at the species' range edges. Haplotype-based measures of selection were related to recombination rate, albeit less strongly, and highlighted population-specific sweeps that likely resulted from positive selection. Our study highlights how comprehensive screens of genomic variation in wild organisms can provide unique insights into spatio-temporal evolutionary dynamics.

Birds of a feather flock together: a dataset for Clock and Adcyap1 genes from migration genetics studies.

Birds in seasonal habitats rely on intricate strategies for optimal timing of migrations. This is governed by environmental cues, including photoperiod. Genetic factors affecting intrinsic timekeeping mechanisms, such as circadian clock genes, have been explored, yielding inconsistent findings with potential lineage-dependency. To clarify this evidence, a systematic review and phylogenetic reanalysis was done. This descriptor outlines the methodology for sourcing, screening, and processing relevant literature and data. PRISMA guidelines were followed, ultimately including 66 studies, with 34 focusing on candidate genes at the genotype-phenotype interface. Studies were clustered using bibliographic coupling and citation network analysis, alongside scientometric analyses by publication year and location. Data was retrieved for allele data from databases, article supplements, and direct author communications. The dataset, version 1.0.2, encompasses data from 52 species, with 46 species for the Clock gene and 43 for the Adcyap1 gene. This dataset, featuring data from over 8000 birds, constitutes the most extensive cross-species collection for these candidate genes, used in studies investigating gene polymorphisms and seasonal bird migration.

A Chromosome-Level Reference Genome for the Black-Legged Kittiwake (Rissa tridactyla), a Declining Circumpolar Seabird.

Amidst the current biodiversity crisis, the availability of genomic resources for declining species can provide important insights into the factors driving population decline. In the early 1990s, the black-legged kittiwake (Rissa tridactyla), a pelagic gull widely distributed across the arctic, subarctic, and temperate zones, suffered a steep population decline following an abrupt warming of sea surface temperature across its distribution range and is currently listed as Vulnerable by the International Union for the Conservation of Nature. Kittiwakes have long been the focus for field studies of physiology, ecology, and ecotoxicology and are primary indicators of fluctuating ecological conditions in arctic and subarctic marine ecosystems. We present a high-quality chromosome-level reference genome and annotation for the black-legged kittiwake using a combination of Pacific Biosciences HiFi sequencing, Bionano optical maps, Hi-C reads, and RNA-Seq data. The final assembly spans 1.35 Gb across 32 chromosomes, with a scaffold N50 of 88.21 Mb and a BUSCO completeness of 97.4%. This genome assembly substantially improves the quality of a previous draft genome, showing an approximately 5× increase in contiguity and a more complete annotation. Using this new chromosome-level reference genome and three more chromosome-level assemblies of Charadriiformes, we uncover several lineage-specific chromosome fusions and fissions, but find no shared rearrangements, suggesting that interchromosomal rearrangements have been commonplace throughout the diversification of Charadriiformes. This new high-quality genome assembly will enable population genomic, transcriptomic, and phenotype-genotype association studies in a widely studied sentinel species, which may provide important insights into the impacts of global change on marine systems.

Rectal Administration of Leishmania Cells Elicits a Specific, Th1-Associated IgG2a Response in Mice: New Perspectives for Mucosal Vaccination against Leishmaniasis, after the Repurposing of a Study on an Anti-Viral Vaccine Candidate.

The mucosal immune system plays a pivotal role in the control of infections, as it represents the first line of defense against most pathogens, from respiratory viruses to intestinal parasites. Mucosal vaccination is thus regarded as a promising strategy to protect animals, including humans, from infections that are acquired by ingestion, inhalation or through the urogenital system. In addition, antigens delivered at the mucosal level can also elicit systemic immune responses. Therefore, mucosal vaccination is potentially effective also against systemic infections acquired through non-mucosal routes, for example, through the bite of hematophagous insects, as in the case of leishmaniasis, a widespread disease that affects humans and dogs. Here, we explored the potential of antigen rectal administration for the generation of anti-Leishmania immunity. Mice were immunized through rectal administration of whole cells of the model parasite Leishmania tarentolae (using a clone engineered to express the spike protein of the SARS-CoV-2 virus generated in a previous study). A specific anti-Leishmania IgG antibody response was detected. In addition, the recorded IgG2a/IgG1 ratio was higher than that of animals injected subcutaneously; therefore, suggesting a shift to a Th1-biased immune response. Considering the importance of a Th1 polarization as a protective response against Leishmania infections, we suggest that further investigation should be focused on the development of novel types of vaccines against these parasites based on rectal immunization.

Experimental nest cooling reveals dramatic effects of heatwaves on reproduction in a Mediterranean bird of prey.

Future climatic scenarios forecast increases in average temperatures as well as in the frequency, duration, and intensity of extreme events, such as heatwaves. Whereas behavioral adjustments can buffer direct physiological and fitness costs of exposure to excessive temperature in wild animals, these may prove more difficult during specific life stages when vagility is reduced (e.g., early developmental stages). By means of a nest cooling experiment, we tested the effects of extreme temperatures on different stages of reproduction in a cavity-nesting Mediterranean bird of prey, the lesser kestrel (Falco naumanni), facing a recent increase in the frequency of heatwaves during its breeding season. Nest temperature in a group of nest boxes placed on roof terraces was reduced by shading them from direct sunlight in 2 consecutive years (2021 and 2022). We then compared hatching failure, mortality, and nestling morphology between shaded and non-shaded (control) nest boxes. Nest temperature in control nest boxes was on average 3.9°C higher than in shaded ones during heatwaves, that is, spells of extreme air temperature (>37°C for ≥2 consecutive days) which hit the study area during the nestling-rearing phase in both years. Hatching failure markedly increased with increasing nest temperature, rising above 50% when maximum nest temperatures exceeded 44°C. Nestlings from control nest boxes showed higher mortality during heatwaves (55% vs. 10% in shaded nest boxes) and those that survived further showed impaired morphological growth (body mass and skeletal size). Hence, heatwaves occurring during the breeding period can have both strong lethal and sublethal impacts on different components of avian reproduction, from egg hatching to nestling growth. More broadly, these findings suggest that the projected future increases of summer temperatures and heatwave frequency in the Mediterranean basin and elsewhere in temperate areas may threaten the local persistence of even relatively warm-adapted species.

Wing morphology covaries with migration distance in a highly aerial insectivorous songbird.

According to classical prediction of aerodynamic theory, birds and other powered fliers that migrate over long distances should have longer and more pointed wings than those that migrate less. However, the association between wing morphology and migratory behavior can be masked by contrasting selective pressures related to foraging behavior, habitat selection and predator avoidance, possibly at the cost of lower flight energetic efficiency. We studied the handwing morphology of Eurasian barn swallows Hirundo rustica from four populations representing a migration distance gradient. This species is an aerial insectivore, so it flies extensively while foraging, and may migrate during the day using a 'fly-and-forage' migration strategy. Prolonged foraging flights may reinforce the effects of migration distance on flight morphology. We found that two wings' aerodynamic properties-isometric handwing length and pointedness, both favoring energetically efficient flight, were more pronounced in barn swallows from populations undertaking longer seasonal migrations compared to less migratory populations. Our result contrast with two recent interspecific comparative studies that either reported no relationship or reported a negative relationship between pointedness and the degree of migratory behavior in hirundines. Our results may thus contribute to confirming the universality of the rule that longer migrations are associated with more pointed wings.

Eco-evolutionary drivers of avian migratory connectivity.

Migratory connectivity, reflecting the extent by which migrants tend to maintain their reciprocal positions in seasonal ranges, can assist in the conservation and management of mobile species, yet relevant drivers remain unclear. Taking advantage of an exceptionally large (~150,000 individuals, 83 species) and more-than-a-century-long dataset of bird ringing encounters, we investigated eco-evolutionary drivers of migratory connectivity in both short- and long-distance Afro-Palearctic migratory birds. Connectivity was strongly associated with geographical proxies of migration costs and was weakly influenced by biological traits and phylogeny, suggesting the evolutionary lability of migratory behaviour. The large intraspecific variability in avian migration strategies, through which most species geographically split into distinct migratory populations, explained why most of them were significantly connected. By unravelling key determinants of migratory connectivity, our study improves knowledge about the resilience of avian migrants to ecological perturbations, providing a critical tool to inform transboundary conservation and management strategies at the population level.

Time trees and clock genes: a systematic review and comparative analysis of contemporary avian migration genetics.

Timing is a crucial aspect for survival and reproduction in seasonal environments leading to carefully scheduled annual programs of migration in many species. But what are the exact mechanisms through which birds (class: Aves) can keep track of time, anticipate seasonal changes, and adapt their behaviour? One proposed mechanism regulating annual behaviour is the circadian clock, controlled by a highly conserved set of genes, collectively called 'clock genes' which are well established in controlling the daily rhythmicity of physiology and behaviour. Due to diverse migration patterns observed within and among species, in a seemingly endogenously programmed manner, the field of migration genetics has sought and tested several candidate genes within the clock circuitry that may underlie the observed differences in breeding and migration behaviour. Among others, length polymorphisms within genes such as Clock and Adcyap1 have been hypothesised to play a putative role, although association and fitness studies in various species have yielded mixed results. To contextualise the existing body of data, here we conducted a systematic review of all published studies relating polymorphisms in clock genes to seasonality in a phylogenetically and taxonomically informed manner. This was complemented by a standardised comparative re-analysis of candidate gene polymorphisms of 76 bird species, of which 58 are migrants and 18 are residents, along with population genetics analyses for 40 species with available allele data. We tested genetic diversity estimates, used Mantel tests for spatial genetic analyses, and evaluated relationships between candidate gene allele length and population averages for geographic range (breeding- and non-breeding latitude), migration distance, timing of migration, taxonomic relationships, and divergence times. Our combined analysis provided evidence (i) of a putative association between Clock gene variation and autumn migration as well as a putative association between Adcyap1 gene variation and spring migration in migratory species; (ii) that these candidate genes are not diagnostic markers to distinguish migratory from sedentary birds; and (iii) of correlated variability in both genes with divergence time, potentially reflecting ancestrally inherited genotypes rather than contemporary changes driven by selection. These findings highlight a tentative association between these candidate genes and migration attributes as well as genetic constraints on evolutionary adaptation.

Extra food provisioning does not affect behavioral lateralization in nestling lesser kestrels.

Costs and benefits of brain lateralization may depend on environmental conditions. Growing evidence indicates that the development of brain functional asymmetries is adaptively shaped by the environmental conditions experienced during early life. Food availability early in life could act as a proxy of the environmental conditions encountered during adulthood, but its potential modulatory effect on lateralization has received little attention. We increased food supply from egg laying to early nestling rearing in a wild population of lesser kestrels Falco naumanni, a sexually dimorphic raptor, and quantified the lateralization of preening behavior (head turning direction). As more lateralized individuals may perform better in highly competitive contexts, we expected that extra food provisioning, by reducing the level of intra-brood competition for food, would reduce the strength of lateralization. We found that extra food provisioning improved nestling growth, but it did not significantly affect the strength or direction of nestling lateralization. In addition, maternal body condition did not explain variation in nestling lateralization. Independently of extra food provisioning, the direction of lateralization differed between the sexes, with female nestlings turning more often toward their right. Our findings indicate that early food availability does not modulate behavioral lateralization in a motor task, suggesting limited phenotypic plasticity in this trait.

A chromosome-level reference genome and pangenome for barn swallow population genomics.

Insights into the evolution of non-model organisms are limited by the lack of reference genomes of high accuracy, completeness, and contiguity. Here, we present a chromosome-level, karyotype-validated reference genome and pangenome for the barn swallow (Hirundo rustica). We complement these resources with a reference-free multialignment of the reference genome with other bird genomes and with the most comprehensive catalog of genetic markers for the barn swallow. We identify potentially conserved and accelerated genes using the multialignment and estimate genome-wide linkage disequilibrium using the catalog. We use the pangenome to infer core and accessory genes and to detect variants using it as a reference. Overall, these resources will foster population genomics studies in the barn swallow, enable detection of candidate genes in comparative genomics studies, and help reduce bias toward a single reference genome.

Efficacy of mucosal vaccination using a protozoan parasite as a vehicle for antigen delivery: IgG and neutralizing response after rectal administration of LeCoVax-2, a candidate vaccine against COVID-19.

Mucosal vaccination is regarded as a promising alternative to classical, intramuscular vaccine delivery. However, only a limited number of vaccines have been licensed for mucosal administration in humans. Here we propose Leishmania tarentolae, a protozoan parasite, as a potential antigen vehicle for mucosal vaccination, for administration via the rectal or oral routes. To test this hypothesis, we exploited L. tarentolae for the production and delivery of SARS-CoV-2 antigens. Two antigens were assayed in BALB/c mice: Lt-spike, a L. tarentolae clone engineered for the surface expression of the SARS-CoV-2 spike protein; RBD-SD1, a purified portion of the spike protein, produced by another engineered clone of the protozoon. Immune response parameters were then determined at different time points. Both antigens, administered either separately or in combination (Lt-spike + RBD-SD1, hereafter LeCoVax-2), determined significant IgG seroconversion and production of neutralizing antibodies after subcutaneous administration, but only in the presence of adjuvants. After rectal administration, the purified RBD-SD1 antigen did not induce any detectable immune response, in comparison with the intense response observed after administration of LeCoVax-2 or Lt-spike alone. In rectal administration, LeCoVax-2 was also effective when administered without adjuvant. Our results show that L. tarentolae is an efficient and safe scaffold for production and delivery of viral antigens, to be used as vaccines. In addition, rectal vaccination experiments prove that L. tarentolae is suitable as a vaccine vehicle and adjuvant for enteral vaccination. Finally, the combined preparation LeCoVax-2 can be considered as a promising candidate vaccine against SARS-CoV-2, worthy of further investigation.

Across the deserts and sea: inter-individual variation in migration routes of south-central European barn swallows (Hirundo rustica).

BACKGROUND: The spatiotemporal organization of migratory routes of long-distance migrants results from trade-offs between minimizing the journey length and en route risk of migration-related mortality, which may be reduced by avoiding crossing inhospitable ecological barriers. Despite flourishing avian migration research in recent decades, little is still known about inter-individual variability in migratory routes, as well as the carry-over effects of spatial and temporal features of migration on subsequent migration stages. METHODS: We reconstructed post- and pre-breeding migration routes, barrier crossing behaviour and non-breeding movements of the largest sample (N = 85) analysed to date of individual barn swallows breeding in south-central Europe, which were tracked using light-level geolocators. RESULTS: Most birds spent their non-breeding period in the Congo basin in a single stationary area, but a small fraction of itinerant individuals reaching South Africa was also observed. Birds generally followed a 'clockwise loop migration pattern', moving through the central Mediterranean and the Sahara Desert during post-breeding (north to south) migration yet switching to a more western route, along the Atlantic coast of Africa, Iberia and western Mediterranean during the pre-breeding (south to north) migration. Southward migration was straighter and less variable, while northward migration was significantly faster despite the broader detour along the Atlantic coast and Iberia. These patterns showed limited sex-related variability. The timing of different circannual events was tightly linked with previous migration stages, considerably affecting migration route and speed of subsequent movements. Indeed, individuals departing late from Africa performed straighter and faster pre-breeding migrations, partly compensating for the initial departure delays, but likely at the cost of performing riskier movements across ecological barriers. CONCLUSIONS: Different spatiotemporal migration strategies during post- and pre-breeding migration suggest that conditions en route may differ seasonally and allow for more efficient travelling along different migration corridors in either season. While highlighting patterns of inter-individual variability, our results support increasing evidence for widespread loop migration patterns among Afro-Palearctic avian migrants. Also, they suggest that carry-over effects acting across different phases of the annual cycle of migratory species can have major impacts on evolutionary processes.

The cloacal microbiome of a cavity-nesting raptor, the lesser kestrel (Falco naumanni).

Background: Microbial communities are found on any part of animal bodies exposed to the environment, and are particularly prominent in the gut, where they play such a major role in the host metabolism and physiology to be considered a "second genome". These communities, collectively known as "microbiome", are well studied in humans and model species, while studies on wild animals have lagged behind. This is unfortunate, as different studies suggested the central role of the gut microbiome in shaping the evolutionary trajectories of species and their population dynamics. Among bird species, only few descriptions of raptor gut microbiomes are available, and mainly carried out on captive individuals. Objectives: In this study, we aimed at improving the knowledge of raptor microbiomes by providing the first description of the gut microbiome of the lesser kestrel (Falco naumanni), a cavity-nesting raptor. Results: The gut microbiome of the lesser kestrel was dominated by Actinobacteria (83.9%), Proteobacteria (8.6%) and Firmicutes (4.3%). We detected no differences in microbiome composition between males and females. Furthermore, the general composition of the microbiome appears similar to that of phylogenetically distant cavity-nesting species. Conclusions: Our results broaden the knowledge of raptor gut microbial communities and let us hypothesize that the distinct nest environment in terms of microclimate and presence of organic material from previous breeding attempts, to which cavity-nesting species that reuse the nest are exposed, might be an important driver shaping microbiomes.

Vegetation height and structure drive foraging habitat selection of the lesser kestrel (Falco naumanni) in intensive agricultural landscapes.

Habitat selection in animals is a fundamental ecological process with key conservation implications. Assessing habitat selection in endangered species and populations occupying the extreme edges of their distribution range, or living in highly anthropized landscapes, may be of particular interest as it may provide hints to mechanisms promoting potential range expansions. We assessed second- and third-order foraging habitat selection in the northernmost European breeding population of the lesser kestrel (Falco naumanni), a migratory falcon of European conservation interest, by integrating results obtained from 411 direct observations with those gathered from nine GPS-tracked individuals. The study population breeds in the intensively cultivated Po Plain (northern Italy). Direct observations and GPS data coincide in showing that foraging lesser kestrels shifted their habitat preferences through the breeding cycle. They positively selected alfalfa and other non-irrigated crops during the early breeding season, while winter cereals were selected during the nestling-rearing phase. Maize was selected during the early breeding season, after sowing, but significantly avoided later. Overall, vegetation height emerged as the main predictor of foraging habitat selection, with birds preferring short vegetation, which is likely to maximise prey accessibility. Such a flexibility in foraging habitat selection according to spatio-temporal variation in the agricultural landscape determined by local crop management practices may have allowed the species to successfully thrive in one of the most intensively cultivated areas of Europe. In the southeastern Po Plain, the broad extent of hay and non-irrigated crops is possibly functioning as a surrogate habitat for the pseudo-steppe environment where most of the European breeding population is settled, fostering the northward expansion of the species in Europe. In intensive agricultural landscapes, the maintenance of alfalfa and winter cereals crops and an overall high crop heterogeneity (deriving from crop rotation) is fundamental to accommodate the ecological requirements of the species in different phases of its breeding cycle.

Annual egg productivity predicts female-biased mortality in avian species.

Among avian species, the differential cost entailed by either sex in competition for mates has been regarded as the main evolutionary influence on sex differences in mortality rates. However, empirical evidence suggests that sex-biased adult mortality is mainly related to differential energy investment in gamete production, with a greater annual mass devoted to egg production leading to higher female mortality. We explicitly tested the generality of this pattern in a comparative framework. Annual egg production can be relatively large in some species (up to 200% of female body mass) and annual mortality is generally biased toward females. We showed that greater annual egg productivity resulted in higher mortality rates of females relative to males. Mating system was secondarily important, with species in which males were more involved in mating competition having more equal mortality rates between the sexes. However, both traits explained only a limited fraction of the interspecific variation in female-biased mortality. Other traits, such as sexual size dimorphism and parental care, had much weaker influences on female-biased mortality. Our results suggest that both annual mass devoted to gamete production by females and mating system contribute to the evolution of the fundamental life-history trade-off between reproduction and survival in avian taxa.

The hidden cost of following currents: Microplastic ingestion in a planktivorous seabird.

Microplastics are increasingly pervasive pollutants, particularly abundant in the neuston where they drift with currents. We assessed dietary microplastic ingestion in the Mediterranean storm petrel (Hydrobates pelagicus melitensis), a small pelagic seabird that forages on plankton and inhabit the Mediterranean sea, one of the most polluted seas worldwide. We collected spontaneous regurgitates from 30 chick-rearing individuals and used GPS tracking data from 7 additional individuals to locate foraging areas. Birds foraged in pelagic areas characterized by water stirring and mixing, and regurgitates from 14 individuals (i.e. 45 %) contained microplastics. Fibers were the dominant shape (56 %), with polyester, polyethylene and nylon being the most frequent polymers. Our findings highlight the potential sensitivity of this species of conservation interest to plastic pollution and suggest that storm petrel regurgitates can be a valuable matrix to investigate microplastic ingestion in planktonic foragers, providing a characterization of spatio-temporal patterns of microplastic exposure in pelagic environments.

Year-round variation in the isotopic niche of Scopoli's shearwater (Calonectris diomedea) breeding in contrasting sea regions of the Mediterranean Sea.

Top marine predators are key components of marine food webs. Among them, long-distance migratory seabirds, which travel across different marine ecosystems over the year, may experience important year-round changes in terms of oceanographic conditions and availability of trophic resources. We tested whether this was the case in the Scopoli's shearwater (Calonectris diomedea), a trans-equatorial migrant and top predator, by sampling birds breeding in three environmentally different regions of the Mediterranean Sea. The analysis of positional data and stable isotopes (δ1³C and δ15N) of target feathers revealed that birds from the three regions were spatially segregated during the breeding period while they shared non-breeding areas in the Atlantic Ocean. Isotopic baseline levels of N and C (meso-zooplankton) were significantly different among marine regions during breeding. Such variation was reflected at the higher trophic levels of pelagic and demersal fish muscles as well as in shearwater feathers grown in the Mediterranean. δ15N- and δ13C-adjusted values of shearwaters were significantly different among populations suggesting that birds from different breeding areas relied on prey species from different trophic levels. Conversely, the non-breeding spatial and isotopic niches overlapped greatly among the three populations. Shearwater trophic niches during breeding were narrower and segregated compared to the non-breeding period, revealing a high plasticity in trophic resource use. Overall, this study highlights seasonal and region-specific use of trophic resources by Scopoli's shearwater, suggesting a broad trophic plasticity and possibly a high adaptability to environmental changes.

Assessing the distribution of invasive Asian mosquitoes in Northern Italy and modelling the potential spread of Aedes koreicus in Europe.

In the last decade, Aedes koreicus and Aedes japonicus japonicus mosquitoes, which are competent vectors for various arboviruses of public health relevance, colonised Italy and other European countries. Nevertheless, information about their current and potential distribution is partial. Accordingly, in this study four regions of Northern Italy (Lombardy, Liguria, Piedmont and Aosta Valley) were surveyed during 2021 for the presence of eggs, larvae and pupae of these two invasive species. We found evidence for a widespread presence of Ae. koreicus in pre-Alpine territories of Lombardy and Piedmont. Larvae from the invasive subspecies of Ae. j. japonicus were also collected in the same geographic areas, though they were less frequent. Occurrence data from this study and results from previous monitoring campaigns were used to generate a Maxent model for the prediction of habitat suitability for Ae. koreicus mosquitoes in Northern Italy and the rest of Europe. Peri-urban areas located in proximity to forests, pastures and vineyards were revealed as highly suitable environments for colonisation by this invasive species. Maps of the potential distribution also suggest the presence of further suitable areas in currently uncolonized countries. We conclude that this invasive mosquito species has the potential for a broad expansion at the European level in the coming decades.