Environmental DNA (eDNA) metabarcoding is an increasingly popular method for rapid biodiversity assessment. As with any ecological survey, false negatives can arise during sampling and, if unaccounted for, lead to biased results and potentially misdiagnosed environmental assessments. We developed a multi-scale, multi-species occupancy model for the analysis of community biodiversity data resulting from eDNA metabarcoding; this model accounts for imperfect detection and additional sources of environmental and experimental variation. We present methods for model assessment and model comparison and demonstrate how these tools improve the inferential power of eDNA metabarcoding data using a case study in a coastal, marine environment. Using occupancy models to account for factors often overlooked in the analysis of eDNA metabarcoding data will dramatically improve ecological inference, sampling design, and methodologies, empowering practitioners with an approach to wield the high-resolution biodiversity data of next-generation sequencing platforms.

Threatened freshwater ecosystems urgently require improved tools for effective management. Food web analysis is currently under-utilized, yet can be used to generate metrics to support biomonitoring assessments by measuring the stability and robustness of ecosystems. Using a previously developed analysis pipeline, we combined taxonomic outputs from DNA metabarcoding with a text-mining routine to extract trait information directly from the literature. This pipeline allowed us to generate heuristic food webs for sites within the lower Saint John/Wolastoq River and the Grand Lake Meadows (hereafter called the “GLM complex”), Atlantic Canada's largest freshwater wetland. While these food webs are derived from empirical traits and their structure has been shown to discriminate sites both spatially and temporally, the accuracy of their properties have not been assessed against other methods of trophic analysis. We explored two approaches to validate the utility of heuristic food webs. First, we qualitatively compared how well-trophic position derived from heuristic food webs recovered spatial and temporal differences across the GLM complex in comparison to traditional stable isotope approaches. Second, we explored how the trophic position of invertebrates, derived from heuristic food webs, predicted trophic position measured from δ¹⁵N values. In general, both heuristic food webs and stable isotopes were able to detect seasonal changes in maximum trophic position in the GLM complex. Samples from the entire GLM complex demonstrated that prey-averaged trophic position measured from heuristic food webs strongly predicted trophic position inferred from stable isotopes (R² = 0.60), and even stronger relationships were observed for some individual models (R² = 0.78 for best model). Beyond their areas of congruence, heuristic food web and stable isotope analyses also appear to complement one another, suggesting a surprising degree of independence between community trophic niche width (assessed from stable isotopes) and food web size and complexity (assessed from heuristic food webs). Collectively, these analyses indicate that trait-based networks have properties that correspond to those of actual food webs, supporting the routine adoption of food web metrics for ecosystem biomonitoring.

Leaf litter provides an important nutrient subsidy to headwater streams, but little is known about how tree genetics influence energy pathways from litter to higher trophic levels. Despite the charge to quantify carbon (C) and nitrogen (N) pathways from decomposing litter, the relationship between litter decomposition and aquatic consumers remains unresolved. We measured litter preference (attachments to litter), C and N assimilation rates, and growth rates of a shredding caddisfly (Hesperophylax magnus, Limnephilidae) in response to leaf litter of different chemical and physical phenotypes using Populus cross types (P. fremontii, P. angustifolia, and F₁ hybrids) and genotypes within P. angustifolia. We combined laboratory mesocosm studies using litter from a common garden with a field study using doubly labeled litter (¹³C and ¹⁵N) grown in a greenhouse and incubated in Oak Creek, Arizona, USA. We found that, in the lab, shredders initially chose relatively labile (low lignin and condensed tannin concentrations, rapidly decomposing) cross type litter, but preference changed within 4 d to relatively recalcitrant (high lignin and condensed tannin concentrations, slowly decomposing) litter types. Additionally, in the lab, shredder growth rates were higher on relatively recalcitrant compared to labile cross type litter. Over the course of a three‐week field experiment, shredders also assimilated more C and N from relatively recalcitrant compared to labile cross type litter. Finally, among P. angustifolia genotypes, N assimilation by shredders was positively related to litter lignin and C:N, but negatively related to condensed tannins and decomposition rate. C assimilation was likewise positively related to litter C:N, and also to litter %N. C assimilation was not associated with condensed tannins or lignin. Collectively, these findings suggest that relatively recalcitrant litter of Populus cross types provides more nutritional benefit, in terms of N fluxes and growth, than labile litter, but among P. angustifolia genotypes the specific trait of litter recalcitrance (lignin or tannins) determines effects on C or N assimilation. As shredders provide nutrients and energy to higher trophic levels, the influence of these genetically based plant decomposition pathways on shredder preference and performance may affect community and food web structure.

Ecologists often equate litter quality with decomposition rate. In soil and sediments, litter that is rapidly decomposed by microbes often has low concentrations of tannin and lignin and low C:N ratios. Do these same traits also favor element transfer to higher trophic levels in streams, where many insects depend on litter as their primary food source? We test the hypothesis that slow decomposition rates promote element transfer from litter to insects, whereas rapid decomposition favors microbes. We measured carbon and nitrogen fluxes from four plant species to a leaf‐shredding caddisfly using isotopically labeled litter. Caddisflies assimilated a higher percentage of litter carbon and nitrogen lost from slowly decomposing litters (Platanus wrightii and Quercus gambelii). In contrast, rapidly decomposing litters (Fraxinus velutina and Populus fremontii) supported higher microbial biomass. These results challenge the view that rapidly decomposing litter is higher quality by demonstrating that slowly decomposing litters provide a critical resource for insects.