Giovanni Naro

Mosquitoes are vectors of numerous pathogens that pose significant threats to human and animal health, making their monitoring crucial for effective disease control. Environmental DNA (eDNA)–based approaches enable the detection of multiple organisms in their habitats without the need for morphological identification. However, they require strict and extensive protocol optimization. In this study, we focused on the need for novel monitoring strategies, particularly ones capable of overcoming the limitations of traditional methods. Given the central role of water in the life cycle of mosquitoes, we explored whether eDNA collected from aquatic habitats could provide an effective tool for mosquito detection and taxonomic identification. Specifically, we aimed to evaluate whether an eDNA‐based approach can reliably detect multiple mosquito species and accurately discriminate among them under different community compositions. To this end, we developed and validated a protocol for an eDNA‐based detection and identification method for mosquito monitoring. First, we assessed its performance by conducting laboratory experiments on samples containing nine different combinations of larvae from five mosquito species. eDNA occupancy analysis revealed varying species‐specific detection probabilities, with Aedes albopictus showing the highest detection rate and Aedes koreicus the lowest. Overall detection probability was high at moderate to high larval densities (5 to 50 ind/L), whereas it decreased at 0.5 ind/L, indicating reduced sensitivity at very low larval concentrations. Then, we collected 165 water samples from three types of field locations and assessed the influence of ecological variables, including land use and site type, on mosquito detection probability. Our results indicate that eDNA represents a promising tool for mosquito monitoring, particularly in contexts where larval densities are not extremely low, and may support its future integration into vector surveillance and control program.

The Ceratopogonidae (Diptera, Nematocera) is a highly diverse family of insects inhabiting a wide range of habitat types that is only absent from polar regions and remote islands. Despite this broad diversity, scientific knowledge of this family remains incomplete, including the genus Leptoconops Skuse, 1889. Although several Leptoconops species have been reported from Europe, no detailed study has yet been conducted to resolve the taxonomy of this genus and clarify species occurrence. Here, the first comprehensive study of Leptoconops diversity in southwest Spain is provided based on extensive sampling conducted in 2023–2024. In total, eight Leptoconops species were collected using carbon-dioxide suction traps: Leptoconops (Leptoconops) noei Clastrier & Coluzzi, 1973, Leptoconops (Leptoconops) irritans Noé, 1905, and four new species: Leptoconops (Leptoconops) nigrithorax González & Tanase, sp. nov ., Leptoconops (Leptoconops) triangularis González & Tanase, sp. nov., Leptoconops (Leptoconops) pseudoirritans González & Tanase, sp. nov ., and Leptoconops (Leptoconops) communis González & Tanase, sp. nov . Additionally, Leptoconops (Leptoconops) bidentatus Gutsevich, 1960, was recorded for the first time from the Iberian Peninsula. One species of the subgenus Holoconops : Leptoconops (Holoconops) cf. kerteszi Kieffer, 1908, was also identified. Molecular support for the morphological identification is provided using COX1 barcode sequences. A brief review of Leptoconops species occurrence in Europe is also included, with detailed morphological descriptions supported by high-quality images of the newly described species, and a complete identification key for dry-preserved and slide-mounted adult females of the subgenus L. ( Leptoconops ).

The Ceratopogonidae family, comprising over 6000 described species, includes the genus Leptoconops , which has been understudied despite its ecological significance and biting nuisance to humans. Here, we document the presence of inland Leptoconops noei populations for the first time in Italy in an area previously considered environmentally atypical for this species. Our findings expand the known range of L. noei , traditionally confined to coastal areas, and highlight its potential to colonise diverse habitats. Interestingly, despite the thorough morphological and molecular identification of sampled individuals and their clear assignment to the species L. noei , a notable morphological variability was observed in the sensory structures of the maxillary palps, suggesting the possibility of a new morphotype or environment‐driven adaptations. Phylogenetic analyses of COX1 sequences revealed negligible genetic differentiation between the newly discovered inland population and coastal populations, suggesting recent separation or gene‐flow connection. These findings underscore the ecological adaptability of L. noei , which poses risks of human‐biting nuisance, allergic reactions and even possible economic losses in recreative areas, in a range of locations wider than previously assumed. Additionally, this study reports the first complete mitochondrial genome for the genus Leptoconops , offering valuable genetic insights for taxonomic clarification, evolutionary studies and future research on the biology of Ceratopogonidae.