Aedes japonicus and Aedes koreicus are two invasive mosquitoes native to East Asia that are quickly establishing in temperate regions of Europe. Both species are vectors of arboviruses, but we currently lack a clear understanding of their evolution. Here, we present new short-read, shallow genome sequencing of A. japonicus and A. koreicus individuals from northern Italy, which we used for downstream phylogenetic and barcode analyses. We explored associated microbial DNA and found high occurrences of Delftia bacteria in both samples, but neither Asaia nor Wolbachia. We then assembled complete mitogenomes and used these data to infer divergence times estimating the split of A. japonicus from A. koreicus in the Oligocene, which was more recent than that previously reported using mitochondrial markers. We recover a younger age for most other nodes within Aedini and other Culicidae. COI barcoding and phylogenetic analyses indicate that A. japonicus yaeyamensis, A. japonicus amamiensis, and the two A. koreicus sampled from Europe should be considered as separate species within a monophyletic species complex. Our studies further clarify the evolution of A. japonicus and A. koreicus, and indicate the need to obtain whole-genome data from putative species in order to disentangle their complex patterns of evolution.

As a first line of defence for the central nervous system (CNS), microglia play a critical role in maintaining homeostasis within the brain. Upon detection of damage or threats, activated cells release factors to communicate and potentiate immune responses. This activation also increases activity of the kynurenine pathway (KP) and alters expression of key KP enzymes such as indoleamine 2,3-dioxygenase (IDO-1) and kynurenine 3-monooxygenase (KMO), both major contributors in the pathology of several neurodegenerative and psychiatric disorders. This study investigated the impact of pro-inflammatory stimuli on the C20 human microglial cell line, focussing on the regulation of KMO and IDO-1 expression, and the production of cytokines. Additionally, we explored whether the anti-inflammatory effects of dexamethasone (DEXA) influenced these outcomes. This additional characterisation of a physiologically relevant human microglial cell line offers a novel and reliable platform for investigating human-specific microglial biology and function. C20 were challenged for 24 hours with cytokines or lipopolysaccharide (LPS). Gene expression was measured by RT-qPCR and excreted cytokines were quantified using a multiplex array. Our results showed up-regulation of IDO-1 and KMO transcripts, and increased release of pro- and anti-inflammatory cytokines. Notably, these effects were significantly dampened by pre-incubation with DEXA. Furthermore, transcriptomic analyses supported these data by highlighting TNF-α-activated enriched pathways, as well as those down-regulated in samples co-treated with DEXA. This study contributes to the understanding of key mechanisms regulated in human microglia by immune challenges and supports the crucial role of synthetic glucocorticoids (GCs) in moderating the microglial immune response induced by pro-inflammatory signals. These data support the use of GCs as possible therapeutic interventions for diseases associated with neuroinflammation, particularly those with altered KP metabolism.

Blood feeding (hematophagy) is widespread across Diptera (true flies), yet the underlying genetic basis remains poorly understood. Using phylogenomics, we show that four gene families associated with neuromodulation, immune responses, embryonic development, and iron metabolism have undergone independent expansions within mosquitoes and sandflies. Our findings illuminate the underlying genetic basis for blood-feeding adaptations in these important disease vectors.

Multiple selective pressures can shape the evolution of color vision in animals, by acting on the co- evolution of the opsin genes. How do adaptive processes shape the duplications of opsins, the evolution of their amino acids and the modification of their patterns of expression? At large phylogenetic scales, natural selection due to the contrasted light environments has been found to have a profound impact on the evolution of the opsin gene family. However, in closely-related species, species interactions due to sexual selection or competition may also influence opsin evolution. Here, we investigate the diversification of opsin sequences and their expression in closely-related blue Morpho butterfly species, living in different microhabitats, to shed light on the effect of biotic and abiotic selective pressures shaping the evolution of their opsin gene family. First, we combined genomics, transcriptomics and immunochemistry to precisely characterize the expression and the spatial distribution of the opsin proteins found in the eyes of Morpho helenor . We found unique ommatidial types compared to other butterfly species. We then investigated the evolution of opsin genes among 18 Morpho species, found signature of positive selection on two opsin genes, and identified key co-evolving amino-acids shaping the diversification of the Morpho visual system. We showed that such opsin evolution was correlated to both light environment and wing coloration, highlighting the joint effect of several selective pressures in the evolution of those proteins. Overall, our study underlines the peculiar evolution of visual systems in closely-related species specialized in divergent microhabitats.

Mosquitoes are the deadliest vectors of disease. They impose a huge health burden on human populations spreading parasites as disparate as protozoans (malaria), viruses (yellow fever and more), and nematodes (filariasis), which cause life-threatening conditions. In recent years, mating has been proposed as a putative target for population control. Mosquitoes mate mid-air in swarms initiated by males and triggered by a combination of internal and external stimuli. As the number of females in a swarm is limited, there is intense competition among males. Indeed, males ‘retune’ their physiology for this demanding behaviour but the underlying genetic changes are largely unknown. Interestingly, recent evidence indicates that the upregulation of circadian clock genes may be involved in the swarming of malaria mosquitoes of the genus Anopheles . Here, we use whole-head RNA-seq to identify gene expression changes in Aedes aegypti males who are engaged in swarming in a laboratory setting. Our results suggest that swarming males tend to lower some housekeeping functions while increasing the remodelling of the cytoskeleton and neuronal connectivity; the transcription of circadian clock genes is unaffected.