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Although West Nile virus (WNV) and other arthropod-borne viruses are a major public health problem, the mechanisms of antiviral immunity in mosquitoes are poorly understood. Dicer-2, responsible for the RNAi-mediated response through the C-terminal RNase-III domain, also contains an N-terminal DExD/H-box helicase domain similar to mammalian RIG-I/MDA5 which, in Drosophila , was found to be required for activation of an antiviral gene, Vago . Here we show that the Culex orthologue of Vago (Cx Vago) is up-regulated in response to WNV infection in a Dicer-2–dependent manner. Further, our data show that Cx Vago is a secreted peptide that restricts WNV infection by activation of the Jak-STAT pathway. Thus, Vago appears to function as an IFN-like antiviral cytokine in mosquitoes.

The taxonomically intricate genus Culex Linnaeus includes numerous known vector species of parasites and viruses to humans. The aim of this article is to comprehensively review the Culex species which occur in French Guiana to provide a stronger taxonomic foundation for future studies on this genus in South America. The occurrence of Culex species was investigated in light of current taxonomic knowledge through an extensive examination of voucher specimens deposited in the entomological collections of four French depositories and additional specimens recently collected at various localities in French Guiana. Based on this review, 104 Culex species classified in eight subgenera are confirmed to occur in French Guiana. Compared to the most recent checklist, 18 species are added, and 10 species excluded, resulting in a total number of 242 valid mosquito species known to occur in French Guiana. Three nominal species are synonymized, three others are newly described, and a last one is transferred to another informal infrasubgeneric group. Overall, this review also highlights the limits of the use of only bibliographic data when dealing with taxonomically complex groups of insects.

We conducted a retrospective phylogenetic analysis of Mayaro virus (MAYV) detected in French Guiana during 1996-2024. Analysis revealed circulation of MAYV genotype D sublineage 2 and suggested introduction from Brazil and spread to Haiti and Venezuela. Phylogenetic findings support endemic circulation and reinforce the need for MAYV surveillance in the region.

Oropouche fever is a zoonotic dengue-like syndrome caused by Oropouche virus. In August-September 2020, dengue-like syndrome developed in 41 patients in a remote rainforest village in French Guiana. By PCR or microneutralization, 23 (82.1%) of 28 tested patients were positive for Oropouche virus, documenting its emergence in French Guiana.

With dengue virus (DENV) becoming endemic in tropical and subtropical regions worldwide, there is a pressing global demand for effective strategies to control the mosquitoes that spread this disease. Recent advances in genetic engineering technologies have made it possible to create mosquitoes with reduced vector competence, limiting their ability to acquire and transmit pathogens. Here we describe the development of Aedes aegypti mosquitoes synthetically engineered to impede vector competence to DENV. These mosquitoes express a gene encoding an engineered single-chain variable fragment derived from a broadly neutralizing DENV human monoclonal antibody and have significantly reduced viral infection, dissemination, and transmission rates for all four major antigenically distinct DENV serotypes. Importantly, this is the first engineered approach that targets all DENV serotypes, which is crucial for effective disease suppression. These results provide a compelling route for developing effective genetic-based DENV control strategies, which could be extended to curtail other arboviruses.

Among mosquitoes (Diptera: Culicidae), the genus Culex Linnaeus is one of the most diverse in the world and includes numerous known vector species of parasites and viruses to humans. Morphological identification of Culex species is notoriously difficult and relies mostly on the examination of properly dissected male genitalia which largely prevents female and immature identification during entomological, ecological or arboviral surveys. The aims of this study were (i) to establish a DNA barcode library for Culex mosquitoes of French Guiana based on the mitochondrial gene cytochrome c oxidase I (COI) marker, (ii) to compare three approaches of molecular delimitation of species to morphological identification, (iii) to test the effectiveness of the COI marker at a broader geographical scale across South America, and (iv) to discuss the internal classification of the genus Culex as regard to our phylogenetic analysis. Mosquitoes used in this study were sampled in French Guiana between 2013 and 2023. We provide 246 COI sequences for 90 morphologically identified species of Culex , including five new country records and two newly described species. Overall, congruence between morphological identification and molecular delimitations using the COI barcode was high. The Barcode of Life Data clustering approach into Barcode Index Numbers gives the best result in terms of species delimitation. Inconsistencies between morphological identification and molecular delimitation can be explained by introgression, incomplete lineage sorting, imperfect taxonomy or the effect of geographical gap in sampling. This increases by almost two-fold the number of mosquito species for which a DNA barcode is available in French Guiana, including 75% of the Culex species currently known in the territory. Finally, this study confirms the usefulness of the COI barcode in identifying Culex of South America, but also points the limits of this marker for some groups of species within the subgenera Culex and Melanoconion .

Madagascar has an important diversity of fleas (Siphonaptera), which almost species do not exist elsewhere. Scientists have provided huge efforts to improve knowledge on Malagasy fleas since the middle of 1900s particularly by investigating topics such as taxonomy, systematics, biogeography, and flea vector role. Since then, new species discovery has increased and currently, 48 species are recorded which the majority is endemic.Therefore, it becomes necessary to have updated keys to identify species membership.This paper presents morphological-based keys to identify at genus and species levels adult fleas reported as occurring in Madagascar. Illustrations are proposed to make easier the observation of morphological criteria, which may be tricky for sibling species.

Aedes aegypti mosquitoes, main vectors for numerous flaviviruses, have olfactory preferences and are capable of olfactory learning especially when seeking their required environmental conditions to lay their eggs. In this study, we showed that semiochemical conditions during Aedes aegypti larval rearing affected future female choice for oviposition: water-reared mosquitoes preferred to lay eggs in water or p-cresol containers, while skatole reared mosquitoes preferred skatole sites. Using two independent behavioural assays, we showed that this skatole preference was lost in mosquitoes infected with dengue virus. Viral RNA was extracted from infected female mosquito heads, and an increase of virus load was detected from 3 to 10 days post infection, indicating replication in the insect head and possibly in the central nervous system. Expression of selected genes, potentially implied in olfactory learning processes, were also altered during dengue infection. Based on these results, we hypothesise that dengue virus infection alters gene expression in the mosquito’s head and is associated with a loss of olfactory preferences, possibly modifying oviposition site choice of female mosquitoes.

Despite their importance as vectors of human and livestock diseases, relatively little is known about innate antiviral immune pathways in mosquitoes and other insects. Previous work has shown that Culex Vago (CxVago), which is induced and secreted from West Nile virus (WNV)-infected mosquito cells, acts as a functional homolog of interferon, by activating Jak-STAT pathway and limiting virus replication in neighbouring cells. Here we describe the Dicer-2-dependent pathway leading to WNV-induced CxVago activation. Using a luciferase reporter assay, we show that a NF-κB-like binding site in CxVago promoter region is conserved in mosquito species and is responsible for induction of CxVago expression following WNV infection. Using dsRNA-based gene knockdown, we show that the NF-κB ortholog, Rel2, plays significant role in the signaling pathway that activates CxVago in mosquito cells in vitro and in vivo. Using similar approaches, we also show that TRAF, but not TRAF-3, is involved in activation of Rel2 after viral infection. Overall the study shows that a conserved signaling pathway, which is similar to mammalian interferon activation pathway, is responsible for the induction and antiviral activity of CxVago.

Background Mosquitoes are responsible for transmission of viruses, including dengue, West Nile and chikungunya viruses. Female mosquitoes are infected when they blood-feed on vertebrates, a required step for oogenesis. During this process, mosquitoes encounter high iron loads. Since iron is an essential nutrient for most organisms, including pathogens, one of the defense mechanisms for the host includes sequestration of iron away from the invading pathogen. Here, we determine whether iron availability affects viral replication in mosquitoes. Methods To elucidate effect of iron availability on mosquito cells during infection, Culex cells were treated with either ferric ammonium citrate (FAC) or the iron chelator, deferoxamine (DFX). Real time RT-PCR was performed using ferritin (heavy chain) and NRAMP as a measure of iron homeostasis in cells. To determine iron requirement for viral replication, Culex cells were knocked down for NRAMP using dsRNA. Finally, the results were validated in Culex mosquito-infection model, by treating infected mosquitoes with DFX to reduce iron levels. Results Our results show that infection of Culex cells led to induction in levels of ferritin (heavy chain) and NRAMP mRNAs in time-dependent manner. Results also showed that treatment of cells with FAC, reduced expression of NRAMP (iron transporter) and increase levels of ferritin (heavy chain). Interestingly, increasing iron levels increased viral titers; while reducing intracellular iron levels, either by NRAMP knock-down or using DFX, reduced viral titers. The results from Culex mosquito infection showed that mosquitoes treated with DFX had reduced viral titers compared with untreated controls in midgut as well as carcass 8 days pi. Saliva from mosquitoes treated with DFX also showed reduced viral titers compared with untreated controls, indicating low viral transmission capacity. Conclusions Our results indicate that iron is required for viral replication in mosquito cells. Mosquitoes respond to viral infection, by inducing expression of heavy chain ferritin, which sequesters available iron, reducing its availability to virus infected cells. The data indicates that heavy chain ferritin may be part of an immune mechanism of mosquitoes in response to viral infections.