uafR: An R package that automates mass spectrometry data processing.

Chemical information has become increasingly ubiquitous and has outstripped the pace of analysis and interpretation. We have developed an R package, uafR, that automates a grueling retrieval process for gas -chromatography coupled mass spectrometry (GC -MS) data and allows anyone interested in chemical comparisons to quickly perform advanced structural similarity matches. Our streamlined cheminformatics workflows allow anyone with basic experience in R to pull out component areas for tentative compound identifications using the best published understanding of molecules across samples (pubchem.gov). Interpretations can now be done at a fraction of the time, cost, and effort it would typically take using a standard chemical ecology data analysis pipeline. The package was tested in two experimental contexts: (1) A dataset of purified internal standards, which showed our algorithms correctly identified the known compounds with R2 values ranging from 0.827-0.999 along concentrations ranging from 1 × 10-5 to 1 × 103 ng/µl, (2) A large, previously published dataset, where the number and types of compounds identified were comparable (or identical) to those identified with the traditional manual peak annotation process, and NMDS analysis of the compounds produced the same pattern of significance as in the original study. Both the speed and accuracy of GC -MS data processing are drastically improved with uafR because it allows users to fluidly interact with their experiment following tentative library identifications [i.e. after the m/z spectra have been matched against an installed chemical fragmentation database (e.g. NIST)]. Use of uafR will allow larger datasets to be collected and systematically interpreted quickly. Furthermore, the functions of uafR could allow backlogs of previously collected and annotated data to be processed by new personnel or students as they are being trained. This is critical as we enter the era of exposomics, metabolomics, volatilomes, and landscape level, high-throughput chemotyping. This package was developed to advance collective understanding of chemical data and is applicable to any research that benefits from GC -MS analysis. It can be downloaded for free along with sample datasets from Github at github.org/castratton/uafR or installed directly from R or RStudio using the developer tools: 'devtools::install_github("castratton/uafR")'.

Assessing effective mechanical and chemical strategies for managing Eucosma giganteana (Lepidoptera: Tortricidae) in the perennial oilseed crop, Silphium integrifolium (Asteraceae: Heliantheae).

Eucosma giganteana (Riley) is a native specialist pest of silflower, Silphium integrifolium Michx., which is currently being domesticated as a perennial oilseeds crop. The larvae of this moth attack silflower capitula and root crowns, causing both seed damage and long-term degradation of plants. To determine methods to manage E. giganteana in silflower crop fields, we conducted a laboratory bioassay and 3 field experiments to assess the effects of a suite of organic, conventional, and mechanical treatments on E. giganteana mortality and colonization of flower heads. Pyrethroids (permethrin, cyfluthrin), chlorantraniliprole, and methoxyfenozide each had significant insecticidal effects on E. giganteana in at least 2 of the experiments conducted. Nematodes marginally increased larva mortality in the laboratory bioassay and could be further investigated as a soil-applied biological control. In 2 separate field experiments, trimming the top 15% of silflower plants to delay flowering did not alone reduce E. giganteana colonization of flower heads throughout the growing season. However, when trimming was paired with a single chlorantraniliprole application, colonization of capitula was reduced by 83% over untreated control plants. Collectively, these experiments provide evidence for several treatments that could be further tested and incorporated into an integrated pest management strategy for E. giganteana.

Nutrition vs association: plant defenses are altered by arbuscular mycorrhizal fungi association not by nutritional provisioning alone.

BACKGROUND: While it is known that arbuscular mycorrhizal fungi (AMF) can improve nutrient acquisition and herbivore resistance in crops, the mechanisms by which AMF influence plant defense remain unknown. Plants respond to herbivory with a cascade of gene expression and phytochemical biosynthesis. Given that the production of defensive phytochemicals requires nutrients, a commonly invoked hypothesis is that the improvement to plant defense when grown with AMF is simply due to an increased availability of nutrients. An alternative hypothesis is that the AMF effect on herbivory is due to changes in plant defense gene expression that are not simply due to nutrient availability. In this study, we tested whether changes in plant defenses are regulated by nutritional provisioning alone or the response of plant to AMF associations. Maize plants grown with or without AMF and with one of three fertilizer treatments (standard, 2 × nitrogen, or 2 × phosphorous) were infested with fall armyworm (Spodoptera frugiperda; FAW) for 72 h. We measured general plant characteristics (e.g. height, number of leaves), relative gene expression (rtPCR) of three defensive genes (lox3, mpi, and pr5), total plant N and P nutrient content, and change in FAW mass per plant. RESULTS: We found that AMF drove the defense response of maize by increasing the expression of mpi and pr5. Furthermore, while AMF increased the total phosphorous content of maize it had no impact on maize nitrogen. Fertilization alone did not alter upregulation of any of the 3 induced defense genes tested, suggesting the mechanism through which AMF upregulate defenses is not solely via increased N or P plant nutrition. CONCLUSION: This work supports that maize defense may be optimized by AMF associations alone, reducing the need for artificial inputs when managing FAW.

Field Trapping and Flight Capacity of Eucosma giganteana (Riley) (Lepidoptera: Tortricidae) in Response to Behaviorally Active Congeneric Semiochemicals in Novel Silflower Agroecosystems.

Silphium integrifolium is a novel perennial crop being developed for oilseed and biofuel in the midwestern US. One of the primary pests in this system is Eucosma giganteana (Lepidoptera: Tortricidae). Little is known about the chemical ecology or flight behavior of E. giganteana, but many semiochemicals have been identified from other closely related Eucosma species. Some of these compounds include: (Z)- and (E)-8-dodecenyl acetate, (E)-9-dodecenyl acetate, (Z)-8-dodecenol, (E,E)-8,10-dodecadienyl acetate, and (Z,E)-9,12-tetradecadienyl acetate. The goals of this study were to evaluate whether any of these compounds could improve capture of E. giganteana on clear sticky cards in the field, and the most attractive volatiles might affect flight behavior on a computer-automated flight mill assay. We found that there was significant attraction to (E)-8-dodecenyl acetate in two years in the field, which may possibly be a component in the pheromone blend for E. giganteana. On flight mills, E. giganteana flew an average of 23 km in a 24 h period. The presence of attractive stimuli (e.g., (E)-8-dodecenyl acetate) had arresting properties and decreasing flight distance on the mill by 78 to 80%. The longest flight distances were registered in the morning (4:00-12:00) and were 1.8-fold greater than flight distances and durations at night (20:00-4:00). (E)-8-dodecenyl acetate may be useful in behaviorally based monitoring and management strategies for E. giganteana. Overall, our research expands the knowledge on the chemical ecology of adult E. giganteana.

Native Flowering Border Crops Attract High Pollinator Abundance and Diversity, Providing Growers the Opportunity to Enhance Pollination Services.

Over the past century, habitat loss from agricultural intensification has contributed to pollinator decline. One way to mitigate the harmful effects of agricultural intensification is through the re-introduction of native flowering plants as border strips that provide supplemental floral and nesting resources to pollinators. However, border crop species vary in bloom period and flower densities, and are thus likely to attract different suites of pollinator species. Resulting differences in pollinator community composition are likely to affect their ability to provide pollination services to adjacent crop habitat. To address these issues, we implemented a two-year study on the impact of different flowering border crops on pollinator abundance, richness, and community composition. We also examined which crop features (bloom duration, number of flowers, floral area) were most likely to affect pollinator densities. We found that native flowering plant border crops of diverse prairie mix and monocultures of silflower (Silphium integrifolium Michx.) and cup plant (Silphium perfoliatum L.) attracted the highest abundance and species richness of bees and pollinator groups combined, while alfalfa (Medicago sativa L.) attracted the highest lepidopteran abundance and species richness. We also found a significant, positive relationship between pollinator abundance and floral resource amount and bloom duration. These findings offer valuable insight into the impacts of different land management strategies on different pollinator groups, and thus provide landowners with management options for attracting specific pollinator groups and species.

Weight Gain of Spodoptera frugiperda Larvae (Lepidoptera: Noctuidae) on Leaf and Floral Tissues of Silphium integrifolium (Asterales: Asteraceae) Differs by Plant Genotype.

Silflower (Silphium integrifolium (Michaux)) is a native North American relative of sunflower that is undergoing domestication as a perennial oilseed crop. As silflower incurs pest damage from multiple insect species, it is necessary to screen genotypes for their effect on insect performance such that more pest tolerant/resistant accessions can be incorporated into future silflower breeding programs. We present a bioassay protocol for silflower using the generalist herbivore fall armyworm (Spodoptera frugiperda (J. E. Smith)). In this study, fall armyworm larvae were placed on leaf and flower tissue from eleven silflower genotypes, one cup plant (Silphium perfoliatum (L.) (Asterales: Asteraceae)) genotype, and an inbred sunflower line (Helianthus annuus (L.) (Asterales: Asteraceae), HA89). Caterpillar weight gained during a 4-d feeding period significantly differed on leaf and floral tissue from different silflower genotypes, between the Silphium species (silflower and cup plant), and between Silphium genotypes and annual sunflower. Two wild silflower genotypes produced lower larval weight gain on both the floral and leaf tissue than all other genotypes, suggesting these genotypes have either lower nutrition or greater resistance to fall armyworm. However, nonsignificant correlations between larval growth on floral versus leaf tissue across all plant species tested and among all silflower accessions suggest insect performances on these tissue types in silflower are independent. Along with identifying germplasm of interest for silflower breeding programs, we established an easily replicable bioassay protocol using fall armyworm on silflower floral and leaf tissues.

Cover Crop Soil Legacies Alter Phytochemistry and Resistance to Fall Armyworm (Lepidoptera: Noctuidae) in Maize.

Plant-soil feedbacks can mediate aboveground plant-herbivore interactions by impacting plant chemistry. Given that soil legacies and agricultural practices are closely tied, a better understanding of soil legacy cascades and their application in pest management are needed. We tested how cover crop legacies alter resistance to fall armyworm (Spodoptera frugiperda Smith, Lepidoptera: Noctuidae) in maize (Zea mays L., Poales: Poaceae). We compared herbivore performance and behavior of fall armyworm larvae on maize grown after four cover crop treatments: a leguminous mycorrhizal cover crop (pea: Pisum sativum L., Fabales: Fabaceae), a nonleguminous mycorrhizal cover crop (triticale: x Triticosecale Wittm. Ex A. Camus, Poales: Poaceae), a nonleguminous nonmycorrhizal cover crop (radish: Raphanus sativus L., Brassicales: Brassicaceae), and no cover crops (fallow). Soil inorganic N was highest in pea treatments and lowest in triticale treatments, while maize AMF colonization was greatest when grown after mycorrhizal cover crops compared to nonmycorrhizal or no cover crops. Cover crop legacies altered the emission of maize volatiles and fall armyworm larvae oriented toward odors emitted by maize grown after radish more frequently than triticale in olfactometer assays. Additionally, larvae performed better and consumed more leaf tissue when feeding on maize grown after radish and poorest on plants grown after triticale. When damaged by fall armyworm, maize grown after triticale expressed higher levels of lipoxygenase-3 (lox3), while plants grown after radish upregulated maize proteinase inhibitor (mpi) gene expression. Our results highlight the importance of appropriate cover crop selection and suggest that triticale could strengthen maize resistance to fall armyworm.

Cover crop mixture expression is influenced by nitrogen availability and growing degree days.

Cover crop mixtures can provide multiple ecosystem services but provisioning of these services is contingent upon the expression of component species in the mixture. From the same seed mixture, cover crop mixture expression varied greatly across farms and we hypothesized that this variation was correlated with soil inorganic nitrogen (N) concentrations and growing degree days. We measured fall and spring biomass of a standard five-species mixture of canola (Brassica napus L.), Austrian winter pea (Pisum sativum L), triticale (x Triticosecale Wittm.), red clover (Trifolium pratense L.) and crimson clover (Trifolium incarnatum L.) seeded at a research station and on 8 farms across Pennsylvania and New York in two consecutive years. At the research station, soil inorganic N (soil iN) availablity and cumulative fall growing degree days (GDD) were experimentally manipulated through fertilizer additions and planting date. Farmers seeded the standard mixture and a "farm-tuned" mixture of the same five species with component seeding rates adjusted to achieve farmer-desired services. We used Structural Equation Modeling to parse out the effects of soil iN and GDD on cover crop mixture expression. When soil iN and fall GDD were high, canola dominated the mixture, especially in the fall. Low soil iN favored legume species while a shorter growing season favored triticale. Changes in seeding rates influenced mixture composition in fall and spring but interacted with GDD to determine the final expression of the mixture. Our results show that when soil iN availability is high at the time of cover crop planting, highly competitive species can dominate mixtures which could potentially decrease services provided by other species, especially legumes. Early planting dates can exacerbate the dominance of aggressive species. Managers should choose cover crop species and seeding rates according to their soil iN and GDD to ensure the provision of desired services.

Fungal community shifts in soils with varied cover crop treatments and edaphic properties.

Cover cropping is proposed to enhance soil microbial diversity and activity, with cover crop type affecting microbial groups in different ways. We compared fungal community compositions of bulk soils differing by cover crop treatment, season, and edaphic properties in the third year of an organic, conventionally tilled rotation of corn-soybean-wheat planted with winter cover crops. We used Illumina amplicon sequencing fungal assemblages to evaluate effects of nine treatments, each replicated four times, consisting of six single winter cover crop species, a three-species mixture, a six-species mixture, and fallow. Alpha-diversity of fungal communities was not affected by cover crop species identity, function, or diversity. Sampling season influenced community composition as well as genus-level abundances of arbuscular mycorrhizal (AM) fungi. Cover crop mixtures, specifically the three-species mixture, had distinct AM fungal community compositions, while cereal rye and forage radish monocultures had unique Core OTU compositions. Soil texture, pH, permanganate oxidizable carbon, and chemical properties including Cu, and P were important variables in models of fungal OTU distributions across groupings. These results showed how fungal composition and potential functions were shaped by cover crop treatment as well as soil heterogeneity.

Can agricultural practices that mitigate or improve crop resilience to climate change also manage crop pests?

Sustainable agricultural practices that are promoted for mitigating climate change have the potential to also improve pest management. The author highlights recent studies that demonstrate effects of climate-mitigating agricultural practices on arthropod pests and predators in agronomic cropping systems. Promising practices for suppressing pests and/or improving biological control include: plant species diversification, especially via the addition of perennial species; cover cropping; tillage practices that retain crop residue; application of organic fertilizers such as compost and manure; and water management practices such as irrigation and sustainable rice intensification. More research is needed that explicitly tests pest and predator responses to agricultural practices under climate change conditions, if these practices are to be effectively promoted and implemented as agricultural pest management strategies.

Warming Alters Prey Density and Biological Control in Conventional and Organic Agricultural Systems.

SYNOPSIS: Studies have shown that organically farmed fields promote natural predator populations and often have lower pest populations than conventional fields, due to a combination of increased predation pressure and greater plant resistance to pest damage. It is unknown how pest populations and predator efficacy may respond in these farming systems as global temperatures increase. To test these questions, we placed enclosures in eight alfalfa fields farmed using conventional (n = 4) or organic (n = 4) practices for 25 years. We stocked enclosures with pea aphids and 0, 2, or 4 predaceous ladybeetles. Half of the enclosures per field were then either left at ambient temperature or plastic-wrapped to warm them by 2 °C. Aphid abundances were similar in conventional and organic fields under ambient conditions, but were significantly more abundant in conventional than in organic fields when enclosures were warmed. Predator efficacy was reduced under low predator abundance (Hippodamia convergens = 2) in conventional fields under warming conditions; predation strength in organic fields was unaffected by warming. Alfalfa biomass increased with increased predators in all farming and temperature treatments. Our study suggests that biological control may be more easily maintained in organic than in conventional systems as global temperature increases.

Effect of two commercial herbicides on life history traits of a human disease vector, Aedes aegypti, in the laboratory setting.

Some mosquito species utilize the small niches of water that are abundant in farmland habitats. These niches are susceptible to effects from agricultural pesticides, many of which are applied aerially over large tracts of land. One principal form of weed control in agricultural systems involves the development of herbicide-tolerant crops. The impact of sub-agricultural levels of these herbicides on mosquito survival and life-history traits of resulting adults have not been determined. The aim of this study was to test the effect of two commercial herbicides (Beyond and Roundup) on the survivorship, eclosion time, and body mass of Aedes aegypti. First instar A. aegypti larvae were exposed to varying concentrations (270, 550 and 820 µg/m(2) of glyphosate and 0.74, 1.49, 2.24 µL imazamox/m(2)), all treatments being below recommended application rates, of commercial herbicides in a controlled environment and resulting adult mosquitoes were collected and weighed. Exposure to Roundup had a significant negative effect on A. aegypti survivorship at medium and high sub-agricultural application concentrations, and negatively affected adult eclosion time at the highest concentration. However, exposure to low concentrations of Beyond significantly increased A. aegypti survivorship, although adult female mass was decreased at medium sub-agricultural concentrations. These results demonstrate that low concentrations of two different herbicides, which can occur in rural larval habitats as a result of spray drift, can affect the same species of mosquito in both positive and negative ways depending on the herbicide applied. The effects of commercial herbicides on mosquito populations could have an important effect on disease transmission within agricultural settings, where these and other herbicides are extensively applied to reduce weed growth.

Contributions of temporal segregation, oviposition choice, and non-additive effects of competitors to invasion success of Aedes japonicus (Diptera: Culicidae) in North America.

The mosquito Aedes japonicus (Diptera: Culicidae) has spread rapidly through North America since its introduction in the 1990s. The mechanisms underlying its establishment in container communities occupied by competitors Aedes triseriatus and Aedes albopictus are unclear. Possibilities include (A) temporal separation of A. japonicus from other Aedes, (B) oviposition avoidance by A. japonicus of sites containing heterospecific Aedes larvae, and (C) non-additive competitive effects in assemblages of multiple Aedes. Containers sampled throughout the summer in an oak-hickory forest near Eureka, MO showed peak abundance for A. japonicus occurring significantly earlier in the season than either of the other Aedes species. Despite this, A. japonicus co-occurred with one other Aedes species in 53 % of samples when present, and co-occurred with both other Aedes in 18 % of samples. In a field oviposition experiment, A. japonicus laid significantly more eggs in forest edge containers than in forest interior containers, but did not avoid containers with low or high densities of larvae of A. triseriatus, A. albopictus, or both, compared to containers without larvae. Interspecific competitive effects (measured as decrease in the index of performance, λ') of A. triseriatus or A. albopictus alone on A. japonicus larvae were not evident at the densities used, but the effect of both Aedes combined was significantly negative and super-additive of effects of individual interspecific competitors. Thus, neither oviposition avoidance of competitors nor non-additive competitive effects contribute to the invasion success of A. japonicus in North America. Distinct seasonal phenology may reduce competitive interactions with resident Aedes.

A multifaceted trophic cascade in a detritus-based system: density-, trait-, or processing-chain-mediated effects?

We investigated three pathways by which predators on an intermediate trophic level may produce a trophic cascade in detritus-based systems. Predators may increase lower trophic levels (bacteria) by reducing density of bacteriovores, by altering behavior of bacteriovores, and by processing living bacteriovores into carcasses, feces, and dissolved nutrients that are substrates for bacteria. We tested these pathways in laboratory experiments with mosquitoes in water-filled containers. Larval Toxorhynchites rutilus prey on larval Aedes triseriatus, which feed on bacteria. Using containers stocked with oak leaf infusion as a bacterial substrate, we compared bacterial productivity at 7 and 14 days for: prey alone; prey with a predator; and prey with predation cues but no predator. Controls contained no larvae, either with predation cues or without cues. Predation cues in the control treatment increased bacterial abundance at 7 days, but this effect waned by 14 days. Aedes triseriatus larvae reduced bacterial abundance significantly at 14 days. Predator cues and real predation both eliminated the negative effect of A. triseriatus on bacterial abundance. Predation cues reduced survivorship of A. triseriatus larvae at 14 days, however this effect was smaller than the effect of real predation. We further tested effects of residues from predation as cues or as detritus in a second experiment in which A. triseriatus were killed at similar rates by: real predators; mechanical damage without the predator and carcasses left as detritus; or mechanical damage and carcasses removed. No prey larvae were killed in controls. Bacterial productivity was greater with real predation than in all other treatments and greater when prey larvae were killed or killed and removed, than in controls. Thus we find evidence that all three pathways contribute to the trophic cascade from T. rutilus to bacteria in tree hole systems.

Conventional and organic soil fertility management practices affect corn plant nutrition and Ostrinia nubilalis (Lepidoptera: Crambidae) larval performance.

Few studies compare how different soil fertilization practices affect plant mineral content and insect performance in organic systems. This study examined: 1) The European corn borer, Ostrinia nubilalis (Hübner), larval response on corn (Zea mays L.) grown in field soils with different soil management histories; and 2) resilience of these plants to O. nubilalis herbivory. Treatments included: 1) standard organic--organically managed soil fertilized with dairy manure and 2 yr of alfalfa (Medicago sativa L.) in the rotation; 2) basic cation saturation ratio--organically managed soil fertilized with dairy manure and alfalfa nitrogen credits, plus addition of gypsum (CaSO4·2H2O) according to the soil balance hypothesis; and 3) conventional--conventionally managed soil fertilized with synthetic fertilizers. Corn plants were reared to maturity in a greenhouse, and then infested with 0-40 O. nubilalis larvae for 17 d. O. nubilalis exhibited negative competitive response to increasing larval densities. Mean development time was significantly faster for larvae consuming basic cation saturation ratio plants than those on standard organic plants, with intermediate development time on conventional plants. Neither total yield (number of kernels) nor proportion kernels damaged differed among soil fertility treatments. Soil nutrients differed significantly in S and in Ca:Mg and Ca:K ratios, but principal components analysis of plant tissue samples taken before O. nubilalis infestation showed that S, Fe, and Cu contributed most to differences in plant nutrient profiles among soil fertility treatments. Results demonstrate that different fertilization regimens can significantly affect insect performance within the context of organic systems, but the effects in this study were relatively minor compared with effects of intraspecific competition.

Intrinsic and extrinsic drivers of succession: Effects of habitat age and season on an aquatic insect community.

1. Classical studies of succession, largely dominated by plant community studies, focus on intrinsic drivers of change in community composition, such as interspecific competition and changes to the abiotic environment. They often do not consider extrinsic drivers of colonization, such as seasonal phenology, that can affect community change. 2. We investigated both intrinsic and extrinsic drivers of succession for dipteran communities that occupy ephemeral pools, such as those in artificial containers. By initiating communities at different times in the season and following them over time, we compared the relative importance of intrinsic (i.e., habitat age) vs. extrinsic (i.e., seasonal phenology) drivers of succession. 3. We placed water-filled artificial containers in a deciduous forest with 20 containers initiated in each of three months. Containers were sampled weekly to assess community composition. Repeated-measures mixed-effects analysis of community correspondence analysis (CA) scores enabled us to partition intrinsic and extrinsic effects on succession. Covariates of temperature and precipitation were also tested. 4. Community trajectories (as defined by CA) differed significantly with habitat age and season, indicating that both intrinsic and extrinsic effects influence succession patterns. Comparisons of AICcs showed that habitat age was more important than season for species composition. Temperature and precipitation did not explain composition changes beyond those explained by habitat age and season. 5. Quantification of relative strengths of intrinsic and extrinsic effects on succession in dipteran and other ephemeral communities enables us to disentangle processes that must be understood for predicting changes in community composition.

Attracted to the enemy: Aedes aegypti prefers oviposition sites with predator-killed conspecifics.

Oviposition habitat choices of species with aquatic larvae are expected to be influenced by both offspring risk of mortality due to predation, and offspring growth potential. Aquatic predators may indirectly influence growth potential for prey by reducing prey density and, for filter-feeding prey, by increasing bacterial food for prey via added organic matter (feces, partially eaten victims), creating the potential for interactive effects on oviposition choices. We tested the hypothesis that the mosquito Aedes aegypti preferentially oviposits in habitats with predatory Toxorhynchites larvae because of indirect effects of predation on chemical cues indicating bacterial abundance. We predicted that A. aegypti would avoid oviposition in sites with Toxorhynchites, but prefer to oviposit where bacterial food for larvae is abundant, and that predation by Toxorhynchites would increase bacterial abundances. Gravid A. aegypti were offered paired oviposition sites representing choices among: predator presence; the act of predation; conspecific density; dead conspecific larvae; and bacterial activity. A. aegypti preferentially oviposited in sites with Toxorhynchites theobaldi predation, and with killed conspecific larvae, but failed to detect preferences for other treatments. The antibiotic tetracycline eliminated the strongest oviposition preference. Both predation by Toxorhynchites and killed larvae increased bacterial abundances, suggesting that oviposition attraction is cued by bacteria. Our results show the potential for indirect effects, like trophic cascades, to influence oviposition choices and community composition in aquatic systems. Our results suggest that predators like Toxorhynchites may be doubly beneficial as biocontrol agents because of the attraction of ovipositing mosquitoes to bacterial by-products of Toxorhynchites feeding.

Phylogeography of Aedes aegypti (yellow fever mosquito) in South Florida: mtDNA evidence for human-aided dispersal.

The invasive dengue vector Aedes aegypti has persisted for > 200 years in South Florida in the United States. We tested the hypotheses that Florida's landscape creates dispersal barriers and corridors and that long-distance human-aided dispersal structures populations of Ae. aegypti. We evaluated the phylogeography of 362 individuals from Florida's East and West Coasts with a 760-bp (418- and 342-bp fragments of ND5 and ND4, respectively) mitochondrial sequence. Populations from these two coasts were not significantly differentiated, suggesting that limited urbanization in central Florida is not a strong barrier to gene flow. Evidence for long-distance dispersal between Ft. Lauderdale and the West and Ft. Myers and the East indicates the importance of human-aided dispersal. West Coast populations showed no genetic differentiation, indicating that West Coast rivers and bays did not significantly impede gene flow. Phylogeographic analysis of haplotypes showed two distinct matrilines with no geographic patterns, suggesting multiple introductions or balancing selection.

Predation resistance does not trade off with competitive ability in early-colonizing mosquitoes.

The tradeoff between colonization and competitive ability has been proposed as a mechanism for ecological succession, and this tradeoff has been demonstrated in multiple successional communities. The tradeoff between competitive ability and predation resistance is also a widely-described phenomenon; however, this tradeoff is not usually postulated as a cause of ecological succession. Early successional species that arrive before predator colonization could be either (1) less vulnerable to predation than their successors, by virtue of being poor competitors (direct competition-predation tradeoff); or (2) equally or more vulnerable to predation, because they normally colonize ahead of predators in succession and therefore are not evolutionarily adapted to avoid predators that they rarely encounter (no competition-predation tradeoff). To test these alternative hypotheses, we established water-filled containers in an oak-hickory forest. We allowed half of the containers to be naturally colonized by early-successional Culex mosquitoes, mid-successional Aedes mosquitoes, and the mosquito predator Toxorhynchites rutilus. In the other half of the containers, we prevented Aedes colonization via systematic removal of Aedes eggs, but allowed Culex and T. rutilus to colonize. The numbers of mature Culex larvae and pupae, and later the total number of Culex, were significantly greater in containers where Aedes had been removed, which suggests that Culex are competitively suppressed by Aedes. Toxorhynchites rutilus abundance and colonization rate were unaffected by the removal of Aedes, and densities of both Culex and Aedes decreased significantly with T. rutilus abundance in both treatments. In-laboratory bioassays showed that Culex were significantly more vulnerable to predation by T. rutilus than were Aedes. These data are consistent with the hypothesis that Culex and Aedes demonstrate a direct colonization-competition tradeoff, and are inconsistent with the hypothesis of a direct competition-predation tradeoff.

Competitive abilities in experimental microcosms are accurately predicted by a demographic index for R*.

Resource competition theory predicts that R*, the equilibrium resource amount yielding zero growth of a consumer population, should predict species' competitive abilities for that resource. This concept has been supported for unicellular organisms, but has not been well-tested for metazoans, probably due to the difficulty of raising experimental populations to equilibrium and measuring population growth rates for species with long or complex life cycles. We developed an index (R(index)) of R* based on demography of one insect cohort, growing from egg to adult in a non-equilibrium setting, and tested whether R(index) yielded accurate predictions of competitive abilities using mosquitoes as a model system. We estimated finite rate of increase (λ') from demographic data for cohorts of three mosquito species raised with different detritus amounts, and estimated each species' R(index) using nonlinear regressions of λ' vs. initial detritus amount. All three species' R(index) differed significantly, and accurately predicted competitive hierarchy of the species determined in simultaneous pairwise competition experiments. Our R(index) could provide estimates and rigorous statistical comparisons of competitive ability for organisms for which typical chemostat methods and equilibrium population conditions are impractical.