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Abstract Background Although whole-genome sequencing (WGS) is the preferred genotyping method for most genomic analyses, limitations are often experienced when studying genomes characterized by a high percentage of repetitive elements, high linkage, and recombination deserts. The Asian tiger mosquito (Aedes albopictus), for example, has a genome comprising up to 72% repetitive elements, and therefore we set out to develop a single-nucleotide polymorphism (SNP) chip to be more cost-effective. Aedes albopictus is an invasive species originating from Southeast Asia that has recently spread around the world and is a vector for many human diseases. Developing an accessible genotyping platform is essential in advancing biological control methods and understanding the population dynamics of this pest species, with significant implications for public health. Methods We designed a SNP chip for Ae. albopictus (Aealbo chip) based on approximately 2.7 million SNPs identified using WGS data from 819 worldwide samples. We validated the chip using laboratory single-pair crosses, comparing technical replicates, and comparing genotypes of samples genotyped by WGS and the SNP chip. We then used the chip for a population genomic analysis of 237 samples from 28 sites in the native range to evaluate its usefulness in describing patterns of genomic variation and tracing the origins of invasions. Results Probes on the Aealbo chip targeted 175,396 SNPs in coding and non-coding regions across all three chromosomes, with a density of 102 SNPs per 1 Mb window, and at least one SNP in each of the 17,461 protein-coding genes. Overall, 70% of the probes captured the genetic variation. Segregation analysis found that 98% of the SNPs followed expectations of single-copy Mendelian genes. Comparisons with WGS indicated that sites with genotype disagreements were mostly heterozygotes at loci with WGS read depth 20, indicating that the chip more accurately detects heterozygotes than low-coverage WGS. Sample sizes did not affect the accuracy of the SNP chip genotype calls. Ancestry analyses identified four to five genetic clusters in the native range with various levels of admixture. Conclusions The Aealbo chip is highly accurate, is concordant with genotypes from WGS with high sequence coverage, and may be more accurate than low-coverage WGS. Graphical Abstract

Abstract Mosquitoes have profoundly affected human history and continue to threaten human health through the transmission of a diverse array of pathogens. The phylogeny of mosquitoes has remained poorly characterized due to difficulty in taxonomic sampling and limited availability of genomic data beyond the most important vector species. Here, we used phylogenomic analysis of 709 single copy ortholog groups from 256 mosquito species to produce a strongly supported phylogeny that resolves the position of the major disease vector species and the major mosquito lineages. Our analyses support an origin of mosquitoes in the early Triassic (217 MYA [highest posterior density region: 188–250 MYA]), considerably older than previous estimates. Moreover, we utilize an extensive database of host associations for mosquitoes to show that mosquitoes have shifted to feeding upon the blood of mammals numerous times, and that mosquito diversification and host-use patterns within major lineages appear to coincide in earth history both with major continental drift events and with the diversification of vertebrate classes.

Mixed dispersal strategies are a form of bet hedging in which a species or population utilizes different dispersal strategies dependent upon biotic or abiotic conditions. Here we provide an example of a mixed dispersal strategy in the Aedes albopictus / Ascogregarina taiwanensis host/parasite system, wherein upon host emergence, the gregarine parasite is either carried with an adult mosquito leaving the larval habitat, or released back into the larval habitat. We show that the parasite invests a larger proportion of its dispersing (oocyst) life stage into adult female mosquitoes as opposed to adult male mosquitoes at low parasite exposure levels. However, as the exposure level of parasite increases, so does the parasite investment in adult males, whereas there is no change in the proportion of oocysts in the adult female, regardless of dose. Thus, A. taiwanensis is utilizing several dispersal strategies, depending upon host sex and intraspecific density. Furthermore, we demonstrate that this parasite reduces body size, increases time to emergence in females, and leads to a reduction in estimates of per capita growth rate of the host.

Two invasive mosquitoes, Aedes aegypti and Aedes albopictus , have been interacting during the course of a rapid range expansion by A. albopictus. We investigated the potential for interspecific feeding interference by male mosquitoes interacting with females within and between these species. A. aegypti feeding on both sugar and blood was suppressed when females of this species were exposed to A. albopictus males, but no change was observed when exposed to conspecifics. A. albopictus feeding was not affected by males of either species. The potential consequences of these behaviors are discussed within the context of other known interspecific effects, all of which appear to favor the displacement of A. aegypti by A. albopictus .