Detection, identification and genotyping of Borrelia spp. in ticks of Coastal-Karst and Littoral-Inner Carniola regions in Slovenia.

The density and spread of tick vector species have increased throughout Europe in the last 30 years, leading to an increase of Lyme borreliosis cases, including in Slovenia. The aim of this study was to isolate Borrelia strains and determine the prevalence of B. burgdorferi sensu lato and B. miyamotoi in adults of Ixodes ricinus (Linnaeus) collected in 2019 in the two regions of the country (Coastal-Karst and Littoral-Inner Carniola) by cultivation and PCR. We isolated B. burgdorferi s.l. by culture method in 28/559 (5%) ticks from both regions. Culture-negative samples (531/559, i.e., 95%) were additionally tested by real-time PCR. In 155/531 (29.2%) PCR-positive samples, a fragment of flaB or glpQ was amplified and further sequenced to identify species of the Borrelia. Using both methods, cultivation and PCR, Borrelia spp. prevalence was 32.7% in the Coastal-Karst region and 33.0% in the Littoral-Inner Carniola region. Genotyping of the Borrelia spp. isolates revealed that 17/28 (60%) were B. garinii subtype Mlg2. Of all tick samples tested for B. miyamotoi 8/398 (2%) were PCR positive. Based on previous studies in these regions, we had expected more ticks to be infected with B. afzelii, but genotyping revealed that B. garinii was the most abundant.

Aedes koreicus, a vector on the rise: Pan-European genetic patterns, mitochondrial and draft genome sequencing.

BACKGROUND: The mosquito Aedes koreicus (Edwards, 1917) is a recent invader on the European continent that was introduced to several new places since its first detection in 2008. Compared to other exotic Aedes mosquitoes with public health significance that invaded Europe during the last decades, this species' biology, behavior, and dispersal patterns were poorly investigated to date. METHODOLOGY/PRINCIPAL FINDINGS: To understand the species' population relationships and dispersal patterns within Europe, a fragment of the cytochrome oxidase I (COI or COX1) gene was sequenced from 130 mosquitoes, collected from five countries where the species has been introduced and/or established. Oxford Nanopore and Illumina sequencing techniques were combined to generate the first complete nuclear and mitochondrial genomic sequences of Ae. koreicus from the European region. The complete genome of Ae. koreicus is 879 Mb. COI haplotype analyses identified five major groups (altogether 31 different haplotypes) and revealed a large-scale dispersal pattern between European Ae. koreicus populations. Continuous admixture of populations from Belgium, Italy, and Hungary was highlighted, additionally, haplotype diversity and clustering indicate a separation of German sequences from other populations, pointing to an independent introduction of Ae. koreicus to Europe. Finally, a genetic expansion signal was identified, suggesting the species might be present in more locations than currently detected. CONCLUSIONS/SIGNIFICANCE: Our results highlight the importance of genetic research of invasive mosquitoes to understand general dispersal patterns, reveal main dispersal routes and form the baseline of future mitigation actions. The first complete genomic sequence also provides a significant leap in the general understanding of this species, opening the possibility for future genome-related studies, such as the detection of 'Single Nucleotide Polymorphism' markers. Considering its public health importance, it is crucial to further investigate the species' population genetic dynamic, including a larger sampling and additional genomic markers.

First record of Aedes koreicus (Diptera, Culicidae) in Slovenia.

The first record of Aedes koreicus was made in the village of Lovrenc na Dravskem Polju, north-eastern part of the country. The discovery of Ae. koreicus in various continental European countries motivated us to revise samples of the collected Aedes japonicus japonicus. We found Ae. koreicus in samples from 2013, where the larvae were misidentified as Ae. j. japonicus. The species was identified morphologically and molecularly. The first discovery of Ae. koreicus advocates an urgent need for a nationwide mosquito surveillance programme.