A genotyping array for the globally invasive vector mosquito, Aedes albopictus.

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, while there was near complete agreement with WGS read depths > 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.

Population genetics of an invasive mosquito vector, Aedes albopictus in the Northeastern USA.

The Asian tiger mosquito (Aedes albopictus) arrived in the USA in the 1980's and rapidly spread throughout eastern USA within a decade. The predicted northern edge of its overwintering distribution on the East Coast of the USA roughly falls across New York, Connecticut, and Massachusetts, where the species has been recorded as early as 2000. It is unclear whether Ae. albopictus populations have become established and survive the cold winters in these areas or are recolonized every year. We genotyped and analyzed populations of Ae. albopictus from the northeast USA using 15 microsatellite markers and compared them with other populations across the country and to representatives of the major global genetic clades to investigate their connectivity and stability. Founder effects or bottlenecks were rare at the northern range of the Ae. albopictus distribution in the northeastern USA, with populations displaying high levels of genetic diversity and connectivity along the East Coast. There is no evidence of population turnover in Connecticut during the course of three consecutive years, with consistent genetic structure throughout this period. Overall, these results support the presence of established populations of Ae. albopictus in New York, Connecticut, and Massachusetts, successfully overwintering and migrating in large numbers. Given the stability and interconnectedness of these populations, Ae. albopictus has the potential to continue to proliferate and expand its range northward under mean warming conditions of climate change. Efforts to control Ae. albopictus in these areas should thus focus on vector suppression rather than eradication strategies, as local populations have become firmly established and are expected to reemerge every summer.

Sex-specific ornament evolution is a consistent feature of climatic adaptation across space and time in dragonflies.

Adaptation to different climates fuels the origins and maintenance of biodiversity. Detailing how organisms optimize fitness for their local climates is therefore an essential goal in biology. Although we increasingly understand how survival-related traits evolve as organisms adapt to climatic conditions, it is unclear whether organisms also optimize traits that coordinate mating between the sexes. Here, we show that dragonflies consistently adapt to warmer climates across space and time by evolving less male melanin ornamentation-a mating-related trait that also absorbs solar radiation and heats individuals above ambient temperatures. Continent-wide macroevolutionary analyses reveal that species inhabiting warmer climates evolve less male ornamentation. Community-science observations across 10 species indicate that populations adapt to warmer parts of species' ranges through microevolution of smaller male ornaments. Observations from 2005 to 2019 detail that contemporary selective pressures oppose male ornaments in warmer years; and our climate-warming projections predict further decreases by 2070. Conversely, our analyses show that female ornamentation responds idiosyncratically to temperature across space and time, indicating the sexes evolve in different ways to meet the demands of the local climate. Overall, these macro- and microevolutionary findings demonstrate that organisms predictably optimize their mating-related traits for the climate just as they do their survival-related traits.

Aedes albopictus Populations and Larval Habitat Characteristics across the Landscape: Significant Differences Exist between Urban and Rural Land Use Types.

One of the most profound recent global changes has been the proliferation of urban metropolitan areas. A consequence of urbanization is a reduction in abundance, or diversity, of wildlife. One exception, is the proliferation of vectors of disease; recent years have seen the emergence and resurgence of diseases vectored by species closely associated with humans. Aedes albopictus, a mosquito with a near global range and broad ecological niche, has been described as an urban, suburban, or rural vector, or a forest edge species depending on local conditions. We tested the hypothesis that abundance and phenological patterns of this species vary among different land use types in a temperate city because of the variation in the biotic and abiotic conditions characteristic of those habitat types. A. albopictus populations in urban and suburban areas were an order of magnitude larger than in rural areas and were detected several weeks earlier in the season. Additionally, we found fewer overall mosquito species, higher temperatures, lower nitrogen, higher pH, and faster water evaporation in larval habitats in urban vs. rural areas. By understanding the ecological differences that facilitate a species in one habitat and not another, we can potentially exploit those differences for targeted control.

Aedes albopictus (Diptera: Culicidae) Has Not Become the Dominant Species in Artificial Container Habitats in a Temperate Forest More Than a Decade After Establishment.

Aedes albopictus (Skuse) (Diptera: Culicidae) is one of the most invasive species globally, and has led to rapid declines and local extirpations of resident mosquitoes where it becomes established. A potential mechanism behind these displacements is the superior competitive ability of Ae. albopictus in larval habitats. Research on the context-dependent nature of competitive displacement predicts that Ae. albopictus will not replace native Aedes triseriatus (Say) (Diptera: Culicidae) in treeholes but could do so in artificial container habitats. Aedes albopictus remains rare in temperate treeholes but less is known about how Ae. albopictus fares in artificial containers in forests. Tyson Research Center (TRC) is a field station composed of mostly oak-hickory forest located outside Saint Louis, MO. The container community has been studied regularly at TRC since 2007 with permanently established artificial containers on the property since 2013. Aedes albopictus was detected each year when these communities were sampled; however, its abundance remains low and it fails to numerically dominate other species in these communities. We present data that show Ae. albopictus numbers have not increased in the last decade. We compare egg counts from 2007 to 2016 and combine larval sample data from 2012 to 2017.We present average larval densities and prevalence of Ae. albopictus and two competitors, Ae. triseriatus and Aedes japonicus (Theobald) (Diptera: Culicidae), as well as monthly averages by year. These data highlight a circumstance in which Ae. albopictus fails to dominate the Aedes community despite it doing so in more human-impacted habitats. We present hypotheses for these patterns based upon abiotic and biotic environmental conditions.

Cold Nights, City Lights: Artificial Light at Night Reduces Photoperiodically Induced Diapause in Urban and Rural Populations of Aedes albopictus (Diptera: Culicidae).

As the planet becomes increasingly urbanized, it is imperative that we understand the ecological and evolutionary consequences of urbanization on species. One common attribute of urbanization that differs from rural areas is the prevalence of artificial light at night (ALAN). For many species, light is one of the most important and reliable environmental cues, largely governing the timing of daily and seasonal activity patterns. Recently, it has been shown that ALAN can alter behavioral, phenological, and physiological traits in diverse taxa. For temperate insects, diapause is an essential trait for winter survival and commences in response to declining daylight hours in the fall. Diapause is under strong selection pressure in the mosquito, Aedes albopictus (Skuse); local adaptation and rapid evolution has been observed along a latitudinal cline. It is unknown how ALAN affects this photosensitive trait or if local adaptation has occurred along an urbanization gradient. Using a common garden experiment, we experimentally demonstrated that simulated ALAN reduces diapause incidence in this species by as much as 40%. There was no difference, however, between urban and rural demes. We also calculated diapause incidence from wild demes in urban areas to determine whether wild populations exhibited lower than predicted incidence compared to estimates from total nocturnal darkness. In early fall, lower than predicted diapause incidence was recorded, but all demes reached nearly 100% diapause before terminating egg laying. It is possible that nocturnal resting behavior in vegetation limits the amount of ALAN exposure this species experiences potentially limiting local adaptation.

Host food quality and quantity differentially affect Ascogregarina barretti parasite burden, development and within-host competition in the mosquito Aedes triseriatus.

Host condition depends in large part on the quality and quantity of available food and heavily influences the outcome of parasite infection. Although parasite fitness traits such as growth rate and size may depend on host condition, whether host food quality or quantity is more important to parasite fitness and within-host interactions is poorly understood. We provided individual mosquito hosts with a standard dose of a gregarine parasite and reared mosquitoes on two food types of different quality and two quantities. We measured host size, total parasite count and area, and average size of parasites within each treatment. Food quality significantly influenced the number of parasites in a host; hosts fed a low-quality diet were infected with more parasites than those provided a high-quality diet. In addition, we found evidence of within-host competition; there was a negative relationship between parasite size and count though this relationship was dependent on host food quality. Host food quantity significantly affected total parasite area and parasite size; lower food quantity resulted in smaller parasites and reduced overall parasite area inside the host. Thus both food quality and quantity have the potential to influence parasite fitness and population dynamics.

Invasive species reduces parasite prevalence and neutralizes negative environmental effects on parasitism in a native mosquito.

Invasive species research often focuses on direct effects of invasion on native ecosystems and less so on complex effects such as those influencing host-parasite interactions. However, invaders could have important effects on native host-parasite dynamics. Where infectious stages are ubiquitous and native host-pathogen specificity is strong, invasive less-competent hosts may reduce the pool of infectious stages, effectively reducing native host-parasite encounter rate. Alternatively, invasive species could alter transmission via changes in native species abundance. Biotic and abiotic environmental factors can also impact disease dynamics by altering host or parasite condition. However, little is known about potential interactive effects of invasion and environmental context on native species disease dynamics. Moreover, experimental examinations of the mechanisms driving dilution effects are limited, but serve to provide tests of predictions leading to diversity-disease relationships. Using field and laboratory experiments, we tested competing hypotheses that an invasive species reduces the prevalence of a native parasite in its host by removing infectious propagules from the environment or by reducing native host abundance. In addition, we evaluated the role of detritus quantity as a resource base in mediating effects of the invasive species. Native parasite prevalence was reduced when the invasive species was present. Prevalence was also higher in high detritus habitats, although this effect was lost when the invasive species was present. The invasive species significantly reduced infectious propagules from the aquatic habitats. Presence of the invasive species had no effect on the native species abundance; thus, the reduction in parasitism was not due to changes in host density but through a reduction in infectious propagule encounters. We conclude that an invasive species can facilitate a native species by reducing parasite prevalence via a dilution effect and that these effects can be modified by resource level. Reductions in parasitism may have ripple effects throughout the community, altering the strength of competitive interactions, predation rates or coinfection with other pathogens. We advocate considering potential positive effects of invasive species on recipient communities, in addition to effects of invasions on host-parasite interactions to gain a broader understanding of the complex consequences of invasion.

Genetic and environmental influences on the size-fecundity relationship in Aedes albopictus (Diptera: Culicidae): Impacts on population growth estimates?

Population growth models are integral to ecological studies by providing estimates of population performance across space and time. Several models have been developed that estimate population growth through correlates of demographic traits, as measuring each parameter of the model can be prohibitive in experimental studies. Since differences in female size can accurately reflect changes in fecundity for many taxa, Livdahl and Sugihara developed a population growth index that incorporates size-fecundity relationships as a proxy for fecundity. To investigate the extent to which this model is robust to variation of this proxy, we tested if genetic (source population), temperature and resource treatments affect the size-fecundity relationship in Aedes albopictus (Skuse), the Asian tiger mosquito. We then determined if variation in the size-fecundity relationship alters the population growth estimates, lambda (λ'), when applied to Livdahl and Sugihara's model. We performed 2 laboratory experiments in which we reared cohorts of four different geographic populations of A. albopictus across 5 temperature treatments (18, 21, 25, 18, 31°C) and three resource treatments (low, medium, high larval resources). We determined if the slope of the size-fecundity relationship varied by source population, temperature, or resource; and if variation in this relationship affects lambda (λ') estimates in a competition study between A. albopictus and Culex pipiens (Linnaeus), the northern house mosquito. Temperature treatments significantly affected the size-fecundity relationship, resource level marginally affected the relationship, while source population had no effect. We found positive relationships between size and fecundity when mosquito larvae were reared at high temperatures and low resource levels but the relationship disappeared when mosquitoes were reared at a low temperature or with high levels of resources. The variation in the size-fecundity relationship produced from different temperatures resulted in statistically different lambda (λ') estimates. However, these changes in lambda (λ') did not alter the trends in the population performance across treatments or conclusions of the competition study. This study provides evidence that the population growth model is sensitive to variation in size-fecundity relationships and we recommend biologists apply the most compatible size-fecundity relationship to the models to obtain the most accurate estimates of population performance.

Landscape Physiognomy Influences Abundance of the Lone Star Tick, Amblyomma americanum (Ixodida: Ixodidae), in Ozark Forests.

The lone star tick, Amblyomma americanum Linnaeus (Ixodida: Ixodidae), is emerging as an important human disease vector in the United States. While some recent studies have modeled broad-scale (regional or county-level) distribution patterns of A. americanum, less is known about how local-scale habitat characteristics drive A. americanum abundance. Such local-scale information is vital to identify targets for tick population control measures within land management units. We investigated how habitat features predict host-seeking A. americanum adult and nymph abundance within a 12-ha oak-hickory forest plot in the Missouri Ozarks. We trapped ticks using CO2-baited traps at 40 evenly spaced locations for three 24-h periods during the summer of 2015, and we measured biotic and abiotic variables surrounding each location. Of 2,008 A. americanum captured, 1,009 were nymphs, and 999 were adults. We observed spatial heterogeneity in local tick abundance (min = 0 ticks, max = 112 ticks, mean = 16.7 ticks per trap night). Using generalized linear mixed models, we found that both nymphs and adults had greater abundance in valleys as well as on northern-facing aspects. Moreover, nymph abundance was negatively related to temperature variance, while adult abundance had a negative relationship with elevation. These results demonstrate that managers in this region may be able to predict local tick abundance through simple physiognomic factors and use these parameters for targeted management action.

Human-aided and natural dispersal drive gene flow across the range of an invasive mosquito.

Human-aided transport is responsible for many contemporary species introductions, yet the contribution of human-aided transport to dispersal within non-native regions is less clear. Understanding dispersal dynamics for invasive species can streamline mitigation efforts by targeting routes that contribute disproportionally to spread. Because of its limited natural dispersal ability, rapid spread of the Asian tiger mosquito (Aedes albopictus) has been attributed to human-aided transport, but until now, the relative roles of human-aided and natural movement have not been rigorously evaluated. Here, we use landscape genetics and information-theoretic model selection to evaluate 52 models representing 9240 pairwise dispersal paths among sites across the US range for Ae. albopictus and show that recent gene flow reflects a combination of natural and human-aided dispersal. Highways and water availability facilitate dispersal at a broad spatial scale, but gene flow is hindered by forests at the current distributional limit (range edge) and by agriculture among sites within the mosquito's native climatic niche (range core). Our results show that highways are important to genetic structure between range-edge and range-core pairs, suggesting a role for human-aided mosquito transport to the range edge. In contrast, natural dispersal is dominant at smaller spatial scales, reflecting a shifting dominance to natural movement two decades after introduction. These conclusions highlight the importance of (i) early intervention for species introductions, particularly those with readily dispersed dormant stages and short generation times, and (ii) strict monitoring of commercial shipments for transported immature stages of Ae. albopictus, particularly towards the northern edge of the US range.

Biological Invasions Across Spatial Scales: Intercontinental, Regional, and Local Dispersal of Cladoceran Zooplankton.

The frequency of dispersal of invertebrates among lakes depends upon perspective and spatial scale. Effective passive dispersal requires both the transport of propagules and the establishment of populations large enough to be detected. At a global scale, biogeographic patterns of cladoceran zooplankton species suggest that effective dispersal among continents was originally rare, but greatly increased in the past century with expanded commerce. Genetic analysis allows some reconstruction of past dispersal events. Allozyme and mitochondrial DNA comparisons among New World and Old-World populations of several exotic cladocerans have provided estimates for likely source populations of colonists, their dispersal corridors, and timing of earlier dispersal events. Detecting the Old-World tropical exotic Daphnia lumholtzi early in its invasion of North America has allowed detailed analysis of its spatial spread. Twelve years of collection records indicate a rapid invasion of reservoirs in the United States, by both regional spread and long-distance jumps to new regions. Combining landscape features with zooplankton surveys from south-central US reservoirs revealed higher colonization rates of D. lumholtzi at lower landscape positions, a result which can be explained by either greater propagule load or by higher susceptibility of these downstream reservoirs. Because invaded reservoirs provide a source of propagules for nearby floodplain ponds, the rarity of this species in ponds suggests limitation by local environments. Such analyses of invading species over multiple spatial scales allow a better understanding of ecological processes governing invasion dynamics.