Light pollution disrupts circadian clock gene expression in two mosquito vectors during their overwintering dormancy.

Artificial light at night (ALAN) is an increasingly important form of environmental disturbance as it alters Light:Dark cycles that regulate daily and seasonal changes in physiology and phenology. The Northern house mosquito (Culex pipiens) and the tiger mosquito (Aedes albopictus) enter an overwintering dormancy known as diapause that is cued by short days. These two species differ in diapause strategy: Cx. pipiens diapause as adult females while Ae. albopictus enter a maternally-programmed, egg diapause. Previous studies found that ALAN inhibits diapause in both species, but the mechanism is unknown. As the circadian clock is implicated in the regulation of diapause in many insects, we examined whether exposure to ALAN altered the daily expression of core circadian cloc genes (cycle, Clock, period, timeless, cryptochrome 1, cryptochrome 2, and Par domain protein 1) in these two species when reared under short-day, diapause-inducing conditions. We found that exposure to ALAN altered the abundance of several clock genes in adult females of both species, but that clock gene rhythmicity was maintained for most genes. ALAN also had little effect on clock gene abundance in mature oocytes that were dissected from female Ae. albopictus that were reared under short day conditions. Our findings indicate that ALAN may inhibit diapause initiation through the circadian clock in two medically-important mosquitoes.

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.

Know Your Enemy: Effects of a Predator on Native and Invasive Container Mosquitoes.

We tested the effect of the native container-dwelling predator Toxorhynchites rutilus on the codominant container-dwelling mosquitoes: native Aedes triseriatus and invasive Aedes japonicus. We established two predator treatments (predator, no predator) by removing T. rutilus from all containers, and stocking T. rutilus larvae (1/3.5 liters) in the predator treatment. Removal and stocking was repeated every 3 d and established significantly different predator abundances in both large and small containers. Repeated-measures analysis of variance (ANOVA) on standard samples showed larvae+pupae/liter of A. japonicus was greater without versus with predation, and this difference increased across samples. In contrast, repeated-measures ANOVA showed larvae+pupae/liter of A. triseriatus was statistically indistinguishable for predation treatments and was greater in small versus large containers. Thus, predation reduced invasive A. japonicus while having no detectable effect on A. triseriatus larvae and pupae. A final destructive census of pupae showed that predation reduced pupae/liter of both species, but this effect was greater and more consistent across container sizes for A. japonicus. Predator effects on abundances were not products of the nonlethal effect of predator avoidance by ovipositing females, as T. rutilus presence did not lead to reduced egg inputs by either Aedes, nor by Aedes spp. as a group. Effects of predation thus are best explained by differential success of developing larvae due to the greater lethal effect of T. rutilus on A. japonicus than on A. triseriatus. Thus, this system is consistent with the hypothesis that native predators can limit success and potential impacts of invasive mosquitoes.

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.

No detectable role for predators mediating effects of aquatic habitat size and permanence on populations and communities of container­dwelling mosquitoes.

General theory from aquatic ecology predicts that smaller aquatic habitats have shorter hydroperiods favouring species that are better resource competitors and complete development quickly. Larger habitats are predicted to have longer hydroperiods enabling longer-lived predators to persist. Habitats with long hydroperiods and predators are predicted to favour slower-developing, predator resistant species, rather than competitive species.In a field experiment, we manipulated independently habitat size and hydroperiod in water-filled containers, to test these hypotheses about processes structuring aquatic communities. We used human-made containers that are dominated by mosquitoes that vary in desiccation resistance, competitive ability, and predation resistance.Habitat size and drying had significant effects on abundances of larvae of the common species in these communities. There was sorting of species by habitat size and by drying, with species that are better competitors relatively more abundant in smaller, more ephemeral habitats, and predator resistant, slower-developing species relatively more abundant in larger or permanently flooded habitats. There were no detectable effects of habitat size or drying on the dominant predator.Habitat size and its interaction with drying affected inputs of eggs to containers. Habitat size also affected relative abundances of the two dominant species in the egg population.Although habitat size and hydroperiod significantly affected composition of these communities, these impacts did not appear to be mediated through effects on predator abundance. Species specific differences in habitat size and drying regime preferences, and habitat-dependent larval performance appear to be the main forces shaping these communities.

The roles of history: age and prior exploitation in aquatic container habitats have immediate and carry-over effects on mosquito life history.

Per-capita resource availability in aquatic habitats is influenced directly by consumer density via resource competition and indirectly via delayed resource competition when temporally non-overlapping cohorts of larvae exploit the same resources. In detritus-based systems, resources are likely to be influenced by the age of the aquatic habitat, as detritus changes in quality over time and may be replenished by new inputs.For aquatic insects that exploit detritus-based habitats, feeding conditions experienced during immature stages can influence fitness directly via effects on development and survivorship, but also indirectly by influencing adult traits such as fecundity and longevity.Larval habitat age and prior resource exploitation were manipulated in a field experiment using the container mosquito Aedes triseriatus.It was found that A. triseriatus from older habitats had greater larval survival, faster development and greater adult longevity. Exploitation of larval habitats by a prior cohort of larvae had a significant negative effect on subsequent cohorts of larvae by delaying development.It is suggested that extended conditioning of detritus probably resulted in conversion of recalcitrant resources to more available forms which improved the quality of the habitat.In a parallel study, evidence was found of carry-over effects of habitat age and prior exploitation on adult longevity for A. triseriatus and Aedes japonicus collected from unmanipulated aquatic habitats.These results indicate the importance of detritus dynamics and the discontinuous nature of resource competition in these mosquito-dominated aquatic systems.

How do Nutritional Stress and La Crosse Virus Infection Interact? Tests for Effects on Willingness to Blood Feed and Fecundity in Aedes albopictus (Diptera: Culicidae).

Evolutionary theory predicts that vector-borne pathogens should have low virulence for their vector because of selection against pathogens that harm the vector sufficiently to reduce transmission. Environmental factors such as nutritional stress can alter vector-pathogen associations by making the vectors more susceptible to pathogens (condition-dependent competence) and vulnerable to the harm caused by pathogen replication (condition-dependent virulence). We tested the hypotheses of condition-dependent competence and condition-dependent virulence by examining the interactive effects of short-term sugar deprivation and exposure to La Crosse virus (LACV) in female Aedes albopictus (Skuse). We predicted that infection status interacts with sugar deprivation to alter willingness to blood feed and fecundity in the second gonotrophic cycle (condition-dependent virulence). Sugar deprivation had no effect on body infection or disseminated infection rates. Infection status, sugar treatment, and their interaction had no effect on fecundity. Mosquitoes that had intermittent access to sugar were significantly more willing to take a second bloodmeal compared with those that had continuous access to sugar. Infection status and the interaction with sugar treatment had no effect on blood-feeding behavior. Thus, we found no evidence of short-term sugar deprivation leading to condition-dependent competence for, or condition-dependent virulence of, LACV in Ae. albopictus.

La Crosse Encephalitis Virus Infection in Field-Collected Aedes albopictus, Aedes japonicus, and Aedes triseriatus in Tennessee.

La Crosse virus (LACV) is a mosquito-borne virus and a major cause of pediatric encephalitis in the USA. La Crosse virus emerged in Tennessee and other states in the Appalachian region in 1997. We investigated LACV infection rates and seasonal abundances of the native mosquito vector, Aedes triseriatus, and 2 recently introduced mosquito species, Ae. albopictus and Ae. japonicus, in an emerging disease focus in Tennessee. Mosquitoes were collected using multiple trapping methods specific for Aedes mosquitoes at recent human case sites. Mosquito pools were tested via reverse transcriptase-polymerase chain reaction (RT-PCR) of the S segment to detect multiple Bunyamwera and California serogroup viruses, including LACV, as well as real-time RT-PCR of the M segment. A total of 54 mosquito pools were positive, including wild-caught adult females and laboratory-reared adults, demonstrating transovarial transmission in all 3 species. Maximum likelihood estimates (per 1,000 mosquitoes) were 2.72 for Ae. triseriatus, 3.01 for Ae. albopictus, and 0.63 for Ae. japonicus. We conclude that Ae. triseriatus and Ae. albopictus are important LACV vectors and that Ae. japonicus also may be involved in virus maintenance and transmission.