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  1. The use of environmental DNA (eDNA) to assess aquatic biodiversity is a growing field with great potential for monitoring and managing threatened species, like freshwater mussel (Unionidae) populations. Freshwater mussels are globally imperiled and serve essential roles in aquatic systems as a food source and as a natural water filter making their management essential for ecosystem health. Unfortunately, mussel populations are often understudied, and challenges exist to accurately and efficiently describe the full suite of species present. Multispecies eDNA approaches may also be more challenging where freshwater mussel populations are most diverse due to ongoing and significant taxonomic restructuring that has been further complicated by molecular phylogenies using mitochondrial genes. For this study, we developed a microfluidic metabarcoding array that targets a wide range of species, from invertebrates to fishes, with an emphasis on detecting unionid mussels known to be present in the Sipsey River, Alabama. We compared mussel species diversity across six sites with well-studied mussel assemblages using eDNA surveys and traditional quadrat surveys in 2016. We examined how factors such as mussel population density, biomass and location in the river substrate impacted our ability to detect certain species; and investigated unexpected eDNA detections through phylogenetic analysis. Our eDNA results for fish and mussel species were broadly consistent with the data from traditional electrofishing and quadrat-based field surveys, although both community eDNA and conventional sampling detected species unique to that method. Our phylogenetic analysis agreed with other studies that treat Pleurobema decisum and P. chattanoogaense as synonymous species; however, they are still listed as unique species in molecular databases which complicates their identity in a metabarcoding assay. We also found that Fusconaia flava and F. cerina are indistinguishable from one another using a portion of the NADH dehydrogenase Subunit 1 (ND1) marker, which may warrant further investigation into whether or not they are synonymous. Our results show that many factors impacted our ability to detect and correctly identify Unionidae mussel species. Here we describe the obstacles we faced, including the murky phylogeny of Unionidae mussels and turbid river conditions, and our development of a potentially impactful freshwater mussel monitoring eDNA assay. 
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  2. Abstract

    Models of ecosystem development and response to environmental variation must incorporate change in vertical soil space as well as over time. Insufficient measurement of subsurface soil properties represents a major observational bias in ecosystem studies.

    We address these changes in horizontal (time) and vertical (soil profile) space along a three‐million‐year, semi‐arid, piñon‐juniper woodland substrate age gradient with characteristic progressive and retrogressive ecosystem development phases and a shift from nitrogen (N) and water to phosphorus (P) limitation. We present a novel pedological approach using isotopic tracers and biogeochemical analyses to address fine root distribution, depth of plant uptake and relative nutrient availabilities.

    We show that (a) the quantity of fine roots remains constant with ecosystem development but their distribution in the soil profile becomes increasingly deeper and less concentrated in the surface soil; (b) mean depth of tree uptake becomes deeper with substrate age and follows the relative availability of P as P‐limitation develops and (c) soil P transformations in the oldest soil profiles resemble the theoretical changes with age to produce a depth gradient of relative N and P availability.

    Synthesis. The expanding role of deep roots in this model system is tightly linked to phases of ecosystem development and relative nutrient availability. The inclusion of whole soil profiles is vital to investigating the intersections of biota, soil and geologic substrate and developing a more complete understanding of ecosystem structure and function.

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  3. Abstract

    Dissolved organic carbon (DOC) and nitrogen (DON) are important energy and nutrient sources for aquatic ecosystems. In many northern temperate, freshwater systems DOC has increased in the past 50 years. Less is known about how changes in DOC may vary across latitudes, and whether changes in DON track those of DOC. Here, we present long‐term DOC and DON data from 74 streams distributed across seven sites in biomes ranging from the tropics to northern boreal forests with varying histories of atmospheric acid deposition. For each stream, we examined the temporal trends of DOC and DON concentrations and DOC:DON molar ratios. While some sites displayed consistent positive or negative trends in stream DOC and DON concentrations, changes in direction or magnitude were inconsistent at regional or local scales. DON trends did not always track those of DOC, though DOC:DON ratios increased over time for ~30% of streams. Our results indicate that the dissolved organic matter (DOM) pool is experiencing fundamental changes due to the recovery from atmospheric acid deposition. Changes in DOC:DON stoichiometry point to a shifting energy‐nutrient balance in many aquatic ecosystems. Sustained changes in the character of DOM can have major implications for stream metabolism, biogeochemical processes, food webs, and drinking water quality (including disinfection by‐products). Understanding regional and global variation in DOC and DON concentrations is important for developing realistic models and watershed management protocols to effectively target mitigation efforts aimed at bringing DOM flux and nutrient enrichment under control.

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  4. Abstract

    Lake Superior receives inputs from approximately 2,800 tributaries that provide nutrients and dissolved organic matter (DOM) to the nearshore zone of this oligotrophic lake. Here, we review the magnitude and timing of tributary export and plume formation in Lake Superior, how these patterns and interactions may shift with global change, and how emerging technologies can be used to better characterize tributary–lake linkages. Peak tributary export occurs during snowmelt‐driven spring freshets, with additional pulses during rain‐driven storms. Instream processing and transformation of nitrogen, phosphorus, and dissolved organic carbon (DOC) can be rapid but varies seasonally in magnitude. Tributary plumes with elevated DOC concentration, higher turbidity, and distinct DOM character can be detected in the nearshore during times of high runoff, but plumes can be quickly transported and diluted by in‐lake currents and mixing. Understanding the variability in size and load of these tributary plumes, how they are transported within the lake, and how long they persist may be best addressed with environmental sensors and remote sensing using autonomous and unmanned vehicles. The connections between Lake Superior and its tributaries are vulnerable to climate change, and understanding and predicting future changes to these valuable freshwater resources will require a nuanced and detailed consideration of tributary inputs and interactions in time and space.

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  5. Abstract

    A comprehensive cross‐biome assessment of major nitrogen (N) species that includes dissolved organic N (DON) is central to understanding interactions between inorganic nutrients and organic matter in running waters. Here, we synthesize stream water N chemistry across biomes and find that the composition of the dissolved N pool shifts from highly heterogeneous to primarily comprised of inorganic N, in tandem with dissolved organic matter (DOM) becoming more N‐rich, in response to nutrient enrichment from human disturbances. We identify two critical thresholds of total dissolved N (TDN) concentrations where the proportions of organic and inorganic N shift. With low TDN concentrations (0–1.3 mg/L N), the dominant form of N is highly variable, and DON ranges from 0% to 100% of TDN. At TDN concentrations above 2.8 mg/L, inorganic N dominates the N pool and DON rarely exceeds 25% of TDN. This transition to inorganic N dominance coincides with a shift in the stoichiometry of the DOM pool, where DOM becomes progressively enriched in N and DON concentrations are less tightly associated with concentrations of dissolved organic carbon (DOC). This shift in DOM stoichiometry (defined as DOC:DON ratios) suggests that fundamental changes in the biogeochemical cycles of C and N in freshwater ecosystems are occurring across the globe as human activity alters inorganic N and DOM sources and availability. Alterations to DOM stoichiometry are likely to have important implications for both the fate of DOM and its role as a source of N as it is transported downstream to the coastal ocean.

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