Comparing modern identification methods for wild bees: Metabarcoding and image-based morphological taxonomic assignment.

With the decline of bee populations worldwide, studies determining current wild bee distributions and diversity are increasingly important. Wild bee identification is often completed by experienced taxonomists or by genetic analysis. The current study was designed to compare two methods of identification including: (1) morphological identification by experienced taxonomists using images of field-collected wild bees and (2) genetic analysis of composite bee legs (multiple taxa) using metabarcoding. Bees were collected from conservation grasslands in eastern Iowa in summer 2019 and identified to the lowest taxonomic unit using both methods. Sanger sequencing of individual wild bee legs was used as a positive control for metabarcoding. Morphological identification of bees using images resulted in 36 unique taxa among 22 genera, and >80% of Bombus specimens were identified to species. Metabarcoding was limited to genus-level assignments among 18 genera but resolved some morphologically similar genera. Metabarcoding did not consistently detect all genera in the composite samples, including kleptoparasitic bees. Sanger sequencing showed similar presence or absence detection results as metabarcoding but provided species-level identifications for cryptic species (i.e., Lasioglossum). Genus-specific detections were more frequent with morphological identification than metabarcoding, but certain genera such as Ceratina and Halictus were identified equally well with metabarcoding and morphology. Genera with proportionately less tissue in a composite sample were less likely to be detected using metabarcoding. Image-based methods were limited by image quality and visible morphological features, while genetic methods were limited by databases, primers, and amplification at target loci. This study shows how an image-based identification method compares with genetic techniques, and how in combination, the methods provide valuable genus- and species-level information for wild bees while preserving tissue for other analyses. These methods could be improved and transferred to a field setting to advance our understanding of wild bee distributions and to expedite conservation research.

Conserved grasslands support similar pollinator diversity as pollinator-specific practice regardless of proximal cropland and pesticide exposure.

Pollinator diversity and abundance are declining globally. Cropland agriculture and the corresponding use of agricultural pesticides may contribute to these declines, while increased pollinator habitat (flowering plants) can help mitigate them. Here we tested whether the relative effect of wildflower plantings on pollinator diversity and counts were modified by proportion of nearby agricultural land cover and pesticide exposure in 24 conserved grasslands in Iowa, USA. Compared with general grassland conservation practices, wildflower plantings led to only a 5% increase in pollinator diversity and no change in counts regardless of the proportion of cropland agriculture within a 1 km radius. Pollinator diversity increased earlier in the growing season and with per cent flower cover. Unexpectedly, neither insecticide nor total pesticide concentrations on above-ground passive samplers were related to pollinator diversity. However, pollinator community composition was most strongly related to date of sampling, total pesticide concentration, and forb or flower cover. Our results indicate very little difference in pollinator diversity between grassland conservation practices with and without wildflower plantings. Given the relatively high economic costs of wildflower plantings, our research provides initial evidence that investment in general grassland conservation may efficiently conserve pollinator diversity in temperate regions of intensive cropland agriculture.

Ecological Harm and Economic Damages of Chemical Contamination to Linked Aquatic-Terrestrial Food Webs: A Study-Design Tool for Practitioners.

Contamination of aquatic ecosystems can have cascading effects on terrestrial consumers by altering the availability and quality of aquatic insect prey. Comprehensive assessment of these indirect food-web effects of contaminants on natural resources and their associated services necessitates using both ecological and economic tools. In the present study we present an aquatic-terrestrial assessment tool (AT2), including ecological and economic decision trees, to aid practitioners and researchers in designing contaminant effect studies for linked aquatic-terrestrial insect-based food webs. The tool is tailored to address the development of legal claims by the US Department of the Interior's Natural Resource Damage Assessment and Restoration Program, which aims to restore natural resources injured by oil spills and hazardous substance releases into the environment. Such cases require establishing, through scientific inquiry, the existence of natural resource injury as well as the determination of the monetary or in-kind project-based damages required to restore this injury. However, this tool is also useful to researchers interested in questions involving the effects of contaminants on linked aquatic-terrestrial food webs. Stylized cases exemplify how application of AT2 can help practitioners and researchers design studies when the contaminants present at a site are likely to lead to injury of terrestrial aerial insectivores through loss of aquatic insect prey and/or dietary contaminant exposure. Designing such studies with ecological endpoints and economic modeling inputs in mind will increase the relevance and cost-effectiveness of studies, which can in turn improve the outcomes of cases and studies involving the ecological effects of contaminants on food webs. Environ Toxicol Chem 2023;42:2029-2039. Published 2023. This article is a U.S. Government work and is in the public domain in the USA. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Complex Life Histories Alter Patterns of Mercury Exposure and Accumulation in a Pond-Breeding Amphibian.

Quantifying how contaminants change across life cycles of species that undergo metamorphosis is critical to assessing organismal risk, particularly for consumers. Pond-breeding amphibians can dominate aquatic animal biomass as larvae and are terrestrial prey as juveniles and adults. Thus, amphibians can be vectors of mercury exposure in both aquatic and terrestrial food webs. However, it is still unclear how mercury concentrations are affected by exogenous (e.g., habitat or diet) vs endogenous factors (e.g., catabolism during hibernation) as amphibians undergo large diet shifts and periods of fasting during ontogeny. We measured total mercury (THg), methylmercury (MeHg), and isotopic compositions (δ 13C, δ15N) in boreal chorus frogs (Pseudacris maculata) across five life stages in two Colorado (USA) metapopulations. We found large differences in concentrations and percent MeHg (of THg) among life stages. Frog MeHg concentrations peaked during metamorphosis and hibernation coinciding with the most energetically demanding life cycle stages. Indeed, life history transitions involving periods of fasting coupled with high metabolic demands led to large increases in mercury concentrations. The endogenous processes of metamorphosis and hibernation resulted in MeHg bioamplification, thus decoupling it from the light isotopic proxies of diet and trophic position. These step changes are not often considered in conventional expectations of how MeHg concentrations within organisms are assessed.

Wild Bee Exposure to Pesticides in Conservation Grasslands Increases along an Agricultural Gradient: A Tale of Two Sample Types.

Conservation efforts have been implemented in agroecosystems to enhance pollinator diversity by creating grassland habitat, but little is known about the exposure of bees to pesticides while foraging in these grassland fields. Pesticide exposure was assessed in 24 conservation grassland fields along an agricultural gradient at two time points (July and August) using silicone band passive samplers (nonlethal) and bee tissues (lethal). Overall, 46 pesticides were detected including 9 herbicides, 19 insecticides, 17 fungicides, and a plant growth regulator. For the bands, there were more frequent/higher concentrations of herbicides in July (maximum: 1600 ng/band in July; 570 ng/band in August), while insecticides and fungicides had more frequent/higher concentrations in August (maximum: 110 and 65 ng/band in July; 1500 and 1700 ng/band in August). Pesticide concentrations in bands increased 16% with every 10% increase in cultivated crops. The bee tissues showed no difference in detection frequency, and concentrations were similar among months; maximum concentrations of herbicides, insecticides, and fungicides in July and August were 17, 27, and 180 and 19, 120, and 170 ng/g, respectively. Pesticide residues in bands and bee tissues did not always show the same patterns; of the 20 compounds observed in both media, six (primarily fungicides) showed a detection-concentration relationship between the two media. Together, the band and bee residue data can provide a more complete understanding of pesticide exposure and accumulation in conserved grasslands.

Increased Mercury and Reduced Insect Diversity in Linked Stream-Riparian Food Webs Downstream of a Historical Mercury Mine.

Historical mining left a legacy of abandoned mines and waste rock in remote headwaters of major river systems in the western United States. Understanding the influence of these legacy mines on culturally and ecologically important downstream ecosystems is not always straightforward because of elevated natural levels of mineralization in mining-impacted watersheds. To test the ecological effects of historic mining in the headwaters of the upper Salmon River watershed in Idaho (USA), we measured multiple community and chemical endpoints in downstream linked aquatic-terrestrial food webs. Mining inputs impacted downstream food webs through increased mercury accumulation and decreased insect biodiversity. Total mercury (THg) in seston, aquatic insect larvae, adult aquatic insects, riparian spiders, and fish at sites up to 7.6 km downstream of mining was found at much higher concentrations (1.3-11.3-fold) and was isotopically distinct compared with sites immediately upstream of mining inputs. Methylmercury concentrations in bull trout and riparian spiders were sufficiently high (732-918 and 347-1140 ng MeHg g-1 dry wt, respectively) to affect humans, birds, and piscivorous fish. Furthermore, the alpha-diversity of benthic insects was locally depressed by 12%-20% within 4.3-5.7 km downstream from the mine. However, because total insect biomass was not affected by mine inputs, the mass of mercury in benthic insects at a site (i.e., ng Hg m-2 ) was extremely elevated downstream (10-1778-fold) compared with directly upstream of mining inputs. Downstream adult aquatic insect-mediated fluxes of THg were also high (~16 ng THg m-2 day-1 ). Abandoned mines can have ecologically important effects on downstream communities, including reduced biodiversity and increased mercury flux to higher order consumers, including fish, birds, and humans. Environ Toxicol Chem 2022;41:1696-1710. Published 2022. This article is a U.S. Government work and is in the public domain in the USA.

Contaminant fluxes across ecosystems mediated by aquatic insects.

Metals and organic contaminants in aquatic systems affect the coupling of aquatic and terrestrial ecosystems through two pathways: contaminant-induced effects on insect emergence and emergence-induced contaminant transfer. Consequently, the impact of aquatic contaminants on terrestrial ecosystems can be driven by modifications in the quantity and quality of adult aquatic insects serving as prey or contaminants entering terrestrial food webs as part of the diet of terrestrial predators. Here, we provide an overview of recent advances in the field, separating metals from organic contaminants due to their differential propensity to bioaccumulate and thus their potential contribution to either of the two pathways. Finally, this review highlights the knowledge gap in the relative impact of these pathways on terrestrial insectivores.

Cross-Ecosystem Fluxes of Pesticides from Prairie Wetlands Mediated by Aquatic Insect Emergence: Implications for Terrestrial Insectivores.

Contaminants alter the quantity and quality of insect prey available to terrestrial insectivores. In agricultural regions, the quantity of aquatic insects emerging from freshwaters can be impacted by insecticides originating from surrounding croplands. We hypothesized that, in such regions, adult aquatic insects could also act as vectors of pesticide transfer to terrestrial food webs. To estimate insect-mediated pesticide flux from wetlands embedded in an important agricultural landscape, semipermanetly and temporarily ponded wetlands were surveyed in cropland and grassland landscapes across a natural salinity gradient in the Prairie Pothole Region of North Dakota (USA) during the bird breeding season in 2015 and 2016 (n = 14 and 15 wetlands, respectively). Current-use pesticides, including the herbicide atrazine and the insecticides bifenthrin and imidacloprid, were detected in newly emerged insects. Pesticide detections were similar in insects emerging from agricultural and grassland wetlands. Biomass of emerging aquatic insects decreased 43%, and insect-mediated pesticide flux increased 50% along the observed gradient in concentrations of insecticides in emerging aquatic insects (from 3 to 577 ng total insecticide g-1 insect). Overall, adult aquatic insects were estimated to transfer between 2 and 180 µg total pesticide wetland-1 d-1 to the terrestrial ecosystem. In one of the 2 study years, biomass of emerging adult aquatic insects was also 73% lower from agricultural than grassland wetlands and was dependent on salinity. Our results suggest that accumulated insecticides reduce the availability of adult aquatic insect prey for insectivores and potentially increase insectivore exposure to insect-borne pesticides. Adult aquatic insects retain pesticides across metamorphosis and may expose insectivores living near both agricultural and grassland wetlands to dietary sources of toxic chemicals. Environ Toxicol Chem 2021;40:2282-2296. Published 2021. This article is a U.S. Government work and is in the public domain in the USA.

Variation in metal concentrations across a large contamination gradient is reflected in stream but not linked riparian food webs.

Aquatic insects link food web dynamics across freshwater-terrestrial boundaries and subsidize terrestrial consumer populations. Contaminants that accumulate in larval aquatic insects and are retained across metamorphosis can increase dietary exposure for riparian insectivores. To better understand potential exposure of terrestrial insectivores to aquatically-derived trace metals, metal concentrations in water and tissues were analyzed from different components of streams and riparian food webs across a large (2-3 orders of magnitude) metal gradient (e.g., Zn, Cu, Cd, Pb) in the Rocky Mountains (USA). Our research indicates that the trace metal concentration gradient present among streams was lost during metamorphosis of aquatic larval insects into terrestrially flying adults, decoupling terrestrial exposures from aquatic concentrations. This pattern was caused by declines in 1) among-stream variation in trace metal concentrations, 2) relationships between metal concentrations in paired water and food web components, and 3) mean metal concentrations within aquatic food webs and across the aquatic-terrestrial boundary. Specifically, among-stream variation in trace metal concentrations was highest for water and aquatic vegetation, intermediate for aquatic insect larvae (~30% lower than water) and lowest for adult aquatic insects and riparian spiders (~65% lower). Metal concentrations in paired water and food web components ranged from highly related across the stream-metal gradient (slopes ~1) for water and aquatic vegetation, to less related (slopes closer to 0) for aquatic vegetation and aquatic insect larvae, to unrelated (slopes ~0) for aquatic larval and adult insects. Finally, mean metal concentrations were highest in aquatic vegetation and lowest in adult aquatic insects emerging from streams (~50% lower than aquatic vegetation). Our results indicate less efficient trophic transfer and higher metamorphic loss of trace metals from high metal streams (i.e., exposure-dependent transfer). For many trace metals, aquatic-terrestrial dietary transfer is unlikely to be an important source of exposure for terrestrial insectivores of adult aquatic insects.

Riparian spiders indicate the magnitude and sources of polychlorinated biphenyl contamination at a large contaminated sediment site.

We investigated polychlorinated biphenyl (PCB) contamination at the Ashtabula River (northeast OH, USA) area of concern following remedial dredging using araneid and tetragnathid spiders. The PCB concentrations remain elevated in the area of concern compared with reference conditions. Patterns of contamination were strikingly similar between taxa, but were higher in tetragnathids at the most contaminated sites. Spider PCB homolog distributions identified 2 PCB sources to the area of concern. Based on these findings, we recommend situations where these taxa can be used alone, in concert, or combined into a composite "spider" sample to assess environmental contamination. Environ Toxicol Chem 2018;37:2467-2474. Published 2018 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

In vivo isotopic fractionation of zinc and biodynamic modeling yield insights into detoxification mechanisms in the mayfly Neocloeon triangulifer.

Diversity and biomass of aquatic insects decline in metal-rich aquatic environments, but the mechanisms by which insects from such environments cope with potentially toxic metal concentrations to survive through adulthood are less well understood. In this study, we measured Zn concentrations and isotopes in laboratory-reared diatoms and mayflies (Neocloeon triangulifer) from larval through adult stages. The larvae were fed Zn-enriched diatoms, and bio-concentrated Zn by a factor of 2.5-5 relative to the diatoms but maintained the same Zn-isotopic ratio. These results reflect the importance of dietary uptake and the greater rate of uptake relative to excretion or growth. Upon metamorphosis to subimago, Zn concentrations declined by >70%, but isotopically heavy Zn remained in the subimago bodies. We surmised that the loss of isotopically light Zn during metamorphosis was due to the loss of detoxified Zn and retention of metabolically useful Zn. Through the transition from subimago to imago, Zn concentrations and isotope ratios were virtually unchanged. Because the decrease in Zn body concentration and increase in heavier Zn are seen in the subimagos relative to the larvae, the compartmentalization of Zn must be occurring within the larvae. A biodynamic model was constructed, allowing for isotopic fractionation and partitioning of Zn between metabolically essential and detoxified Zn reservoirs within larvae. The model provides a consistent set of rate and fractionation constants that successfully describe the experimental observations. Specifically, metabolically essential Zn is isotopically heavier and is tightly held once assimilated, and excess, isotopically light Zn is sequestered, detoxified, and ultimately lost during the metamorphosis of larvae to subimagos. To our knowledge, this is the first documentation of in vivo isotopic fractionation in insects, offering an improved understanding of the mechanisms and rates by which the N. triangulifer larvae regulate excess Zn in their bodies.

Metamorphosis Affects Metal Concentrations and Isotopic Signatures in a Mayfly (Baetis tricaudatus): Implications for the Aquatic-Terrestrial Transfer of Metals.

Insect metamorphosis often results in substantial chemical changes that can alter contaminant concentrations and fractionate isotopes. We exposed larval mayflies (Baetis tricaudatus) and their food (periphyton) to an aqueous zinc gradient (3-340 µg Zn/l) and measured zinc concentrations at different stages of metamorphosis: larval, subimago, and imago. We also measured changes in stable isotopes (δ15N and δ13C) in unexposed mayflies. Larval zinc concentrations were positively related to aqueous zinc, increasing 9-fold across the exposure gradient. Adult zinc concentrations were also positively related to aqueous zinc, but were 7-fold lower than larvae. This relationship varied according to adult substage and sex. Tissue concentrations in female imagoes were not related to exposure concentrations, but the converse was true for all other stage-by-sex combinations. Metamorphosis also increased δ15N by ∼0.8‰, but not δ13C. Thus, the main effects of metamorphosis on insect chemistry were large declines in zinc concentrations coupled with increased δ15N signatures. For zinc, this change was largely consistent across the aqueous exposure gradient. However, differences among sexes and stages suggest that caution is warranted when using nitrogen isotopes or metal concentrations measured in one insect stage (e.g., larvae) to assess risk to wildlife that feed on subsequent life stages (e.g., adults).

A modeling approach to compare ΣPCB concentrations between congener-specific analyses.

Changes in analytical methods over time pose problems for assessing long-term trends in environmental contamination by PCBs. Congener-specific analyses vary widely in the number and identity of the 209 distinct PCB chemical configurations (congeners) that are quantified, leading to inconsistencies among summed PCB concentrations (ΣPCB) reported by different studies. Here, we present a modeling approach using linear regression to compare ΣPCB concentrations derived from different congener-specific analyses measuring different co-eluting groups. The approach can be used to develop a specific conversion model between any 2 sets of congener-specific analytical data from similar samples (similar matrix and geographic origin). We demonstrate the method by developing a conversion model for an example data set that includes data from 2 different analytical methods, a low resolution method quantifying 119 congeners and a high resolution method quantifying all 209 congeners. We used the model to show that the 119-congener set captured most (93%) of the total PCB concentration (i.e., Σ209 PCB) in sediment and biological samples. ΣPCB concentrations estimated using the model closely matched measured values (mean relative percent difference = 9.6). General applications of the modeling approach include 1) generating comparable ΣPCB concentrations for samples that were analyzed for different congener sets; and 2) estimating the proportional contribution of different congener sets to ΣPCB. This approach may be especially valuable for enabling comparison of long-term remediation monitoring results even as analytical methods change over time. Integr Environ Assess Manag 2017;13:227-232. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

Riparian spiders as sentinels of polychlorinated biphenyl contamination across heterogeneous aquatic ecosystems.

Riparian spiders are being used increasingly to track spatial patterns of contaminants in and fluxing from aquatic ecosystems. However, our understanding of the circumstances under which spiders are effective sentinels of aquatic pollution is limited. The present study tests the hypothesis that riparian spiders may be effectively used to track spatial patterns of sediment pollution by polychlorinated biphenyls (PCBs) in aquatic ecosystems with high habitat heterogeneity. The spatial pattern of ΣPCB concentrations in 2 common families of riparian spiders sampled in 2011 to 2013 generally tracked spatial variation in sediment ΣPCBs across all sites within the Manistique River Great Lakes Area of Concern (AOC), a rivermouth ecosystem located on the south shore of the Upper Peninsula, Manistique (MI, USA) that includes harbor, river, backwater, and lake habitats. Sediment ΣPCB concentrations normalized for total organic carbon explained 41% of the variation in lipid-normalized spider ΣPCB concentrations across 11 sites. Furthermore, 2 common riparian spider taxa (Araneidae and Tetragnathidae) were highly correlated (r2 > 0.78) and had similar mean ΣPCB concentrations when averaged across all years. The results indicate that riparian spiders may be useful sentinels of relative PCB availability to aquatic and riparian food webs in heterogeneous aquatic ecosystems like rivermouths where habitat and contaminant variability may make the use of aquatic taxa less effective. Furthermore, the present approach appears robust to heterogeneity in shoreline development and riparian vegetation that support different families of large web-building spiders. Environ Toxicol Chem 2017;36:1278-1286. Published 2016 Wiley Periodicals, Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Metamorphosis alters contaminants and chemical tracers in insects: implications for food webs.

Insects are integral to most freshwater and terrestrial food webs, but due to their accumulation of environmental pollutants they are also contaminant vectors that threaten reproduction, development, and survival of consumers. Metamorphosis from larvae to adult can cause large chemical changes in insects, altering contaminant concentrations and fractionation of chemical tracers used to establish contaminant biomagnification in food webs, but no framework exists for predicting and managing these effects. We analyzed data from 39 studies of 68 analytes (stable isotopes and contaminants), and found that metamorphosis effects varied greatly. δ(15)N, widely used to estimate relative trophic position in biomagnification studies, was enriched by ∼ 1‰ during metamorphosis, while δ(13)C used to estimate diet, was similar in larvae and adults. Metals and polycyclic aromatic hydrocarbons (PAHs) were predominantly lost during metamorphosis leading to ∼ 2 to 125-fold higher larval concentrations and higher exposure risks for predators of larvae compared to predators of adults. In contrast, manufactured organic contaminants (such as polychlorinated biphenyls) were retained and concentrated in adults, causing up to ∼ 3-fold higher adult concentrations and higher exposure risks to predators of adult insects. Both food web studies and contaminant management and mitigation strategies need to consider how metamorphosis affects the movement of materials between habitats and ecosystems, with special regard for aquatic-terrestrial linkages.

Cross-ecosystem impacts of stream pollution reduce resource and contaminant flux to riparian food webs.

The effects of aquatic contaminants are propagated across ecosystem boundaries by aquatic insects that export resources and contaminants to terrestrial food webs; however, the mechanisms driving these effects are poorly understood. We examined how emergence, contaminant concentration, and total contaminant flux by adult aquatic insects changed over a gradient of bioavailable metals in streams and how these changes affected riparian web-building spiders. Insect emergence decreased 97% over the metal gradient, whereas metal concentrations in adult insects changed relatively little. As a result, total metal exported by insects (flux) was lowest at the most contaminated streams, declining 96% among sites. Spiders were affected by the decrease in prey biomass, but not by metal exposure or metal flux to land in aquatic prey. Aquatic insects are increasingly thought to increase exposure of terrestrial consumers to aquatic contaminants, but stream metals reduce contaminant flux to riparian consumers by strongly impacting the resource linkage. Our results demonstrate the importance of understanding the contaminant-specific effects of aquatic pollutants on adult insect emergence and contaminant accumulation in adults to predict impacts on terrestrial food webs.

Emergence flux declines disproportionately to larval density along a stream metals gradient.

Effects of contaminants on adult aquatic insect emergence are less well understood than effects on insect larvae. We compared responses of larval density and adult emergence along a metal contamination gradient. Nonlinear threshold responses were generally observed for larvae and emergers. Larval densities decreased significantly at low metal concentrations but precipitously at concentrations of metal mixtures above aquatic life criteria (cumulative criterion accumulation ratio (CCAR) ≥ 1). In contrast, adult emergence declined precipitously at low metal concentrations (CCAR ≤ 1), followed by a modest decline above this threshold. Adult emergence was a more sensitive indicator of the effect of low metals concentrations on aquatic insect communities compared to larvae, presumably because emergence is limited by a combination of larval survival and other factors limiting successful emergence. Thus effects of exposure to larvae are not manifest until later in life (during metamorphosis and emergence). This loss in emergence reduces prey subsidies to riparian communities at concentrations considered safe for aquatic life. Our results also challenge the widely held assumption that adult emergence is a constant proportion of larval densities in all streams.

Fluxes of terrestrial and aquatic carbon by emergent mosquitoes: a test of controls and implications for cross-ecosystem linkages.

Adult aquatic insects are a common resource for many terrestrial predators, often considered to subsidize terrestrial food webs. However, larval aquatic insects themselves consume both aquatic primary producers and allochthonous terrestrial detritus, suggesting that adults could provide aquatic subsidy and/or recycled terrestrial energy to terrestrial consumers. Understanding the source of carbon (aquatic vs. terrestrial) driving aquatic insect emergence is important for predicting magnitude of emergence and effects on recipient food web dynamics; yet direct experimental tests of factors determining source are lacking. Here, we use Culex mosquitoes in experimental pools as an exemplar to test how variation in general factors common to aquatic systems (terrestrial plant inputs and light) may alter the source and amount of energy exported to terrestrial ecosystems in adult aquatic insects that rely on terrestrial resources as larvae. We found strong sequential effects of terrestrial plant inputs and light on aquatic insect oviposition, diet, and emergence of Culex mosquitoes. Ovipositing mosquitoes laid ~3 times more egg masses in high terrestrial input pools under low light conditions. This behavior increased adult emergence from pools under low light conditions; however, high input pools (which had the highest mosquito densities) showed low emergence rates due to density-dependent mortality. Mosquito diets consisted mainly of terrestrial resources (~70-90 %). As a result, the amount of aquatic carbon exported from pools by mosquitoes during the experiment was ~18 times higher from low versus high light pools, while exports of terrestrial carbon peaked from pools receiving intermediate levels of inputs (3-6 times higher) and low light (~6 times higher). Our results suggest that understanding the interplay among terrestrial plant inputs, light availability and biotic responses of aquatic insects may be key in predicting source and magnitude of emergence, and thus the strength and effects of aquatic-terrestrial linkages in freshwater systems.