Linking Water Quality to Drinking Water Treatment Costs Using Time Series Analysis: Examining the Effect of a Treatment Plant Upgrade in Ohio.

We estimate a cost function for a water treatment plant in Ohio to assess the avoided-treatment costs resulting from improved source water quality. Regulations and source water concerns motivated the treatment plant to upgrade its treatment process by adding a granular activated carbon building in 2012. The cost function uses daily observations from 2013 to 2016; this allows us to compare the results to a cost function estimated for 2007-2011 for the same plant. Both models focus on understanding the relationship between treatment costs per 1,000 gallons (per 3.79 m3) of produced drinking water and predictor variables such as turbidity, pH, total organic carbon, deviations from target pool elevation, final production, and seasonal variables. Different from the 2007-2011 model, the 2013-2016 model includes a harmful algal bloom toxin variable. We find that the new treatment process leads to a different cost model than the one that covers 2007-2011. Both total organic carbon and algal toxin are important drivers for the 2013-2016 treatment costs. This reflects a significant increase in cyanobacteria cell densities capable of producing toxins in the source water between time periods. The 2013-2016 model also reveals that positive and negative shocks to treatment costs affect volatility, the changes in the variance of costs through time, differently. Positive shocks, or increased costs, lead to higher volatility compared to negative shocks, or decreased costs, of similar magnitude. After quantifying the changes in treatment costs due to changes in source water quality, we discuss how the study results inform policy-relevant decisions.

Increasingly severe cyanobacterial blooms and deep water hypoxia coincide with warming water temperatures in reservoirs.

Cyanobacterial blooms are expected to intensify and become more widespread with climate change and sustained nutrient pollution, subsequently increasing threats to lentic ecosystems, water quality, and human health. However, little is known about their rates of change because long-term monitoring data are rare, except for some well-studied individual lakes, which typically are large and broadly dispersed geographically. Using monitoring data spanning 1987-2018 for 20 temperate reservoirs located in the USA, we found that cyanobacteria cell densities mostly posed low-to-moderate human health risks until 2003-2005, after which cell densities rapidly increased. Increases were greatest in reservoirs with extensive agriculture in their watersheds, but even those with mostly forested watersheds experienced increases. Since 2009, cell densities posing high human health risks have become frequent with 75% of yearly observations exceeding 100,000 cells ml-1 , including 53% of observations from reservoirs with mostly forested watersheds. These increases coincided with progressively earlier and longer summer warming of surface waters, evidence of earlier onset of stratification, lengthening durations of deep-water hypoxia, and warming deep waters in non-stratifying reservoirs. Among years, higher cell densities in stratifying reservoirs were associated with greater summer precipitation, warmer June surface water temperatures, and higher total Kjeldahl nitrogen concentrations. These trends are evidence that expected increases in cyanobacterial blooms already are occurring as changing climate conditions in some regions increasingly favor their proliferation. Consequently, their negative effects on ecosystems, human health, and socioeconomic wellbeing could increase and expand if warming trends and nutrient pollution continue.

DNA metabarcoding effectively quantifies diatom responses to nutrients in streams.

Nutrient pollution from human activities remains a common problem facing stream ecosystems. Identifying ecological responses to phosphorus and nitrogen can inform decisions affecting the protection and management of streams and their watersheds. Diatoms are particularly useful because they are a highly diverse group of unicellular algae found in nearly all aquatic environments and are sensitive responders to increased nutrient concentrations. Here, we used DNA metabarcoding of stream diatoms as an approach to quantifying effects of total phosphorus (TP) and total nitrogen (TN). Threshold indicator taxa analysis (TITAN) identified operational taxonomic units (OTUs) that increased or decreased along TP and TN gradients along with nutrient concentrations at which assemblages had substantial changes in the occurrences and relative abundances of OTUs. Boosted regression trees showed that relative abundances of gene sequence reads for OTUs identified by TITAN as low P, high P, low N, or high N diatoms had strong relationships with nutrient concentrations, which provided support for potentially using these groups of diatoms as metrics in monitoring programs. Gradient forest analysis provided complementary information by characterizing multi-taxa assemblage change using multiple predictors and results from random forest models for each OTU. Collectively, these analyses showed that notable changes in diatom assemblage structure and OTUs began around 20 µg TP/L, low P diatoms decreased substantially and community change points occurred from 75 to 150 µg/L, and high P diatoms became increasingly dominant from 150 to 300 µg/L. Diatoms also responded to TN with large decreases in low N diatoms occurring from 280 to 525 µg TN/L and a transition to dominance by high N diatoms from 525-850 µg/L. These diatom responses to TP and TN could be used to inform protection efforts (i.e., anti-degradation) and management goals (i.e., nutrient reduction) in streams and watersheds. Our results add to the growing support for using diatom metabarcoding in monitoring programs.

Stable isotopes of algae and macroinvertebrates in streams respond to watershed urbanization, inform management goals, and indicate food web relationships.

Watershed development and anthropogenic sources of nitrogen are among leading causes of negative impacts to aquatic ecosystems around the world. The δ15N of aquatic biota can be used as indicators of anthropogenic sources of nitrogen enriched in 15N, but this mostly has been done at small spatial extents or to document effects of point sources. In this study, we sampled 77 sites along a forest to urban land cover gradient to examine food webs and the use of δ15N of periphyton and macroinvertebrate functional feeding groups (FFGs) as indicators of watershed development and nitrogen effects on streams. Functional feeding groups had low δ15N variability among taxa within sites. Mean absolute differences between individual taxa and their respective site FFG means were < 0.55‰, whereas site means of δ15N of FFGs had ranges of approximately 7-12‰ among sites. The δ15N of periphyton and macroinvertebrate FFGs distinguished least disturbed streams from those with greater watershed urbanization, and they were strongly correlated with increasing nitrogen concentrations and watershed impervious cover. Nonmetric multidimensional scaling, using δ15N of taxa, showed that changes in macroinvertebrate assemblages as a whole were associated with forest-to-urban and increasing nitrogen gradients. Assuming an average +3.4‰ per trophic level increase, δ15N of biota indicated that detrital pathways likely were important to food web structure, even in streams with highly developed watersheds. We used periphyton and macroinvertebrate FFG δ15N to identify possible management goals that can inform decisions affecting nutrients and watershed land use. Overall, the δ15N of periphyton and macroinvertebrates were strong indicators of watershed urban development effects on stream ecosystems, and thus, also could make them useful for quantifying the effectiveness of nitrogen, stream, and watershed management efforts.

Quantifying Urban Watershed Stressor Gradients and Evaluating How Different Land Cover Datasets Affect Stream Management.

Watershed management and policies affecting downstream ecosystems benefit from identifying relationships between land cover and water quality. However, different data sources can create dissimilarities in land cover estimates and models that characterize ecosystem responses. We used a spatially balanced stream study (1) to effectively sample development and urban stressor gradients while representing the extent of a large coastal watershed (>4400 km(2)), (2) to document differences between estimates of watershed land cover using 30-m resolution national land cover database (NLCD) and <1-m resolution land cover data, and (3) to determine if predictive models and relationships between water quality and land cover differed when using these two land cover datasets. Increased concentrations of nutrients, anions, and cations had similarly significant correlations with increased watershed percent impervious cover (IC), regardless of data resolution. The NLCD underestimated percent forest for 71/76 sites by a mean of 11 % and overestimated percent wetlands for 71/76 sites by a mean of 8 %. The NLCD almost always underestimated IC at low development intensities and overestimated IC at high development intensities. As a result of underestimated IC, regression models using NLCD data predicted mean background concentrations of NO3 (-) and Cl(-) that were 475 and 177 %, respectively, of those predicted when using finer resolution land cover data. Our sampling design could help states and other agencies seeking to create monitoring programs and indicators responsive to anthropogenic impacts. Differences between land cover datasets could affect resource protection due to misguided management targets, watershed development and conservation practices, or water quality criteria.

Roles of benthic algae in the structure, function, and assessment of stream ecosystems affected by acid mine drainage.

Tens of thousands of stream kilometers worldwide are degraded by a legacy of acid loads, high metal concentrations, and altered habitat caused by acid mine drainage (AMD) from abandoned underground and surface mines. As the primary production base in streams, the condition of algal-dominated periphyton communities is particularly important to nutrient cycling, energy flow, and higher trophic levels. Here, we synthesize current knowledge regarding how AMD-associated stressors affect (i) algal communities and their use as ecological indicators, (ii) their functional roles in stream ecosystems, and (iii) how these findings inform management decisions and evaluation of restoration effectiveness. A growing body of research has found ecosystem simplification caused by AMD stressors. Species diversity declines, productivity decreases, and less efficient nutrient uptake and retention occur as AMD severity increases. New monitoring approaches, indices of biological condition, and attributes of algal community structure and function effectively assess AMD severity and effectiveness of management practices. Measures of ecosystem processes, such as nutrient uptake rates, extracellular enzyme activities, and metabolism, are increasingly being used as assessment tools, but remain in their infancy relative to traditional community structure-based approaches. The continued development, testing, and implementation of functional measures and their use alongside community structure metrics will further advance assessments, inform management decisions, and foster progress toward restoration goals. Algal assessments will have important roles in making progress toward improving and sustaining the water quality, ecological condition, and ecosystem services of streams in regions affected by the legacy of unregulated coal mining.

Inferences based on diatom compositions improve estimates of nutrient concentrations in streams.

Nutrient concentrations in streams vary strongly with flow conditions, and routinely gathered field measurements of nutrients reflect this variability. Diatom assemblage composition has been used in previous studies to infer nutrient concentrations, and because diatoms integrate nutrient concentrations over longer periods of time, diatom inferences may be less susceptible to fluctuations in streamflow. We tested this hypothesis by leveraging differences in the flashiness of streams across a large continental data set. More specifically, we tested whether the variabilities of direct measurements and diatom inferences of dissolved phosphorus and nitrate were greater in flashy versus non-flashy streams. We further considered whether models linking landscape predictor variables to nutrient concentrations yielded consistent results across flashy and non-flashy streams. Our analysis indicated that measured nutrient concentrations were more variable in flashy compared to non-flashy streams and that landscape models identified different important predictors of nutrient concentrations when fit using data from flashy vs. non-flashy streams. In contrast, variabilities of diatom-inferred nutrient concentrations were similar among stream types, as were the important predictor variables (e.g., manure application rates for nitrate and number of wet days for dissolved phosphorus). These analyses indicate that use of diatom-inferred nutrient concentrations can potentially improve efforts to quantify stream nutrient concentrations.

Using DNA metabarcoding to characterize national scale diatom-environment relationships and to develop indicators in streams and rivers of the United States.

Recent advancements in DNA techniques, metabarcoding, and bioinformatics could help expand the use of benthic diatoms in monitoring and assessment programs by providing relatively quick and increasingly cost-effective ways to quantify diatom diversity in environmental samples. However, such applications of DNA-based approaches are relatively new, and in the United States, unknowns regarding their applications at large scales exist because only a few small-scale studies have been done. Here, we present results from the first nationwide survey to use DNA metabarcoding (rbcL) of benthic diatoms, which were collected from 1788 streams and rivers across nine ecoregions spanning the conterminous USA. At the national scale, we found that diatom assemblage structure (1) was strongly associated with total phosphorus and total nitrogen concentrations, conductivity, and pH and (2) had clear patterns that corresponded with differences in these variables among the nine ecoregions. These four variables were strong predictors of diatom assemblage structure in ecoregion-specific analyses, but our results also showed that diatom-environment relationships, the importance of environmental variables, and the ranges of these variables within which assemblage changes occurred differed among ecoregions. To further examine how assemblage data could be used for biomonitoring purposes, we used indicator species analysis to identify ecoregion-specific taxa that decreased or increased along each environmental gradient, and we used their relative abundances of gene reads in samples as metrics. These metrics were strongly correlated with their corresponding variable of interest (e.g., low phosphorus diatoms with total phosphorus concentrations), and generalized additive models showed how their relationships compared among ecoregions. These large-scale national patterns and nine sets of ecoregional results demonstrated that diatom DNA metabarcoding is a robust approach that could be useful to monitoring and assessment programs spanning the variety of conditions that exist throughout the conterminous United States.

Understanding the effects of phosphorus on diatom richness in rivers and streams using taxon-environment relationships.

Changes in phosphorus concentrations affect periphytic diatom composition in streams, yet we rarely observe strong relationships between diatom richness and phosphorus. In contrast, changes in conductivity are strongly associated with differences in both diatom composition and richness. We hypothesised that we could better understand the mechanisms that control the phosphorus-richness relationship by examining relationships between phosphorus and the occurrence of individual diatom taxa, comparing these with relationships between conductivity and taxon occurrence, and documenting how niche breadths of taxa affect richness patterns. We estimated relationships between phosphorus and taxon occurrence using DNA metabarcoding data of diatoms collected from 1,811 sites distributed across the conterminous U.S.A. and contrasted patterns in these relationships with those between conductivity and taxon occurrence. The distribution of taxon optima for phosphorus was bimodal, with most optima located at either the maximum or minimum observed phosphorus concentration. The distribution of taxon optima for conductivity was unimodal. Niche breadths of taxa for phosphorus and for conductivity both generally increased with optimum values. The distribution of conductivity optima gave rise to a prominent hump-shaped relationship between richness and conductivity. The relationship between richness and phosphorus was also slightly hump-shaped, but this relationship would not be expected from the bimodal distribution of optima. Instead, we determined that broad niche breadths caused the hump-shaped relationship between richness and phosphorus. Our results highlight the nuanced effects that increased P loadings exert on diatom assemblages in rivers and streams and identify reasons that weak relationships between taxon richness and increased phosphorus have been observed. These findings allow us to better describe how excess phosphorus and subsets of taxa and their niche breadths contribute to patterns of taxa richness in diatom assemblages, and to improve the tools used to manage phosphorus pollution.

Using Single-Species and Whole Community Stream Mesocosm Exposures for Identifying Major Ion Effects in Doses Mimicking Resource Extraction Wastewaters.

Wastewaters and leachates from various inland resource extraction activities contain high ionic concentrations and differ in ionic composition, which complicates the understanding and effective management of their relative risks to stream ecosystems. To this end, we conducted a stream mesocosm dose-response experiment using two dosing recipes prepared from industrial salts. One recipe was designed to generally reflect the major ion composition of deep well brines (DWB) produced from gas wells (primarily Na+, Ca2+, and Cl-) and the other, the major ion composition of mountaintop mining (MTM) leachates from coal extraction operations (using salts dissociating to Ca2+, Mg2+, Na+, SO42- and HCO3-)-both sources being extensive in the Central Appalachians of the USA. The recipes were dosed at environmentally relevant nominal concentrations of total dissolved solids (TDS) spanning 100 to 2000 mg/L for 43 d under continuous flow-through conditions. The colonizing native algal periphyton and benthic invertebrates comprising the mesocosm ecology were assessed with response sensitivity distributions (RSDs) and hazard concentrations (HCs) at the taxa, community (as assemblages), and system (as primary and secondary production) levels. Single-species toxicity tests were run with the same recipes. Dosing the MTM recipe resulted in a significant loss of secondary production and invertebrate taxa assemblages that diverged from the control at all concentrations tested. Comparatively, intermediate doses of the DWB recipe had little consequence or increased secondary production (for emergence only) and had assemblages less different from the control. Only the highest dose of the DWB recipe had a negative impact on certain ecologies. The MTM recipe appeared more toxic, but overall, for both types of resource extraction wastewaters, the mesocosm responses suggested significant changes in stream ecology would not be expected for specific conductivity below 300 µS/cm, a published aquatic life benchmark suggested for the region.

Evaluation of Nitrogen and Carbon Stable Isotopes in Filter Feeding Bivalves and Surficial Sediment for Assessing Aquatic Condition in Lakes and Estuaries.

Excessive inputs of nitrogen from anthropogenic activities in watersheds can cause detrimental effects to aquatic ecosystems, but these effects can be difficult to determine based solely on nitrogen concentrations because of their temporal variability and the need to link human activities to ecological responses. Here, we (1) tested the use of stable isotopes of nitrogen (δ15N) and carbon (δ13C) in benthic organic matter (BOM) as proxies for isotope ratios of filter feeding bivalves in lakes and estuaries, which can be used as indicators but are harder to sample and often spatially sparse, and (2) evaluated if stable isotope ratios in benthic organic matter could be used to assess impacts from anthropogenic land development of watersheds. The δ15N in BOM isolated from surficial sediment (δ15NBOM) was significantly correlated with δ15N in filter feeding unionid mussels (Elliptio complanata, δ15NUN) from lakes and with hard-shell clams (Mercenaria mercenaria, δ15NMM) from estuaries. In lakes, δ13CBOM was significantly correlated with δ13CUN, but δ13CBOM was not significantly correlated with δ13CMM in estuaries. Values of δ15NBOM and δ15NUN were significantly and positively correlated with increasing amounts of impervious surface, urban land cover, and human populations in watersheds surrounding lakes. In estuaries, δ15NBOM was only significantly and positively correlated with greater percent impervious surface in the watersheds. Correlations of δ13CBOM in lakes and estuaries, δ13CUN, and δ13CMM with land use and human population were mostly non-significant or weak. Overall, these results show that δ15NBOM can serve as a proxy for δ15N of filter feeding bivalves in lakes and estuaries and is useful for assessing anthropogenic impacts on aquatic systems and resources. Our study area was limited in size, but our results support further studies to test the application of this sediment stable isotope-based technique for assessing and ranking aquatic resources across broad geographical areas.

Quantifying spatial and temporal relationships between diatoms and nutrients in streams strengthens evidence of nutrient effects from monitoring data.

Observational data are frequently used to better understand the effects of changes in P and N on stream biota, but nutrient gradients in streams are usually associated with gradients in other environmental factors, a phenomenon that complicates efforts to accurately estimate the effects of nutrients. Here, we propose a new approach for analyzing observational data in which we compare the effects of changes in nutrient concentrations in time within individual sites and in space among many sites. Covarying relationships between other, potentially confounding environmental factors and nutrient concentrations are unlikely to be the same in both time and space, and, therefore, estimated effects of nutrients that are similar in time and space are more likely to be accurate. We applied this approach to diatom rbcL metabarcoding data collected from streams in the East Fork of the Little Miami River watershed, Ohio, USA. Changes in diatom assemblage composition were consistently associated with changes in the concentration of total reactive P in both time and space. In contrast, despite being associated with spatial differences in ammonia and urea concentrations, diatom assemblage composition was not associated with temporal changes in these nitrogen species. We suggest that the results of this analysis provide evidence of a causal effect of increased P on diatom assemblage composition. We further analyzed the effects of temporal variability in measurements of total reactive P and found that averaging periods greater than ~1 wk prior to sampling best represented the effects of P on the diatom assemblage. Comparisons of biological responses in space and time can sharpen insights beyond those that are based on analyses conducted on only 1 of the 2 dimensions.

Developing Indicators of Nutrient Pollution in Streams Using 16S rRNA Gene Metabarcoding of Periphyton-Associated Bacteria.

Indicators based on nutrient-biota relationships in streams can inform water quality restoration and protection programs. Bacterial assemblages could be particularly useful indicators of nutrient effects because they are species-rich, important contributors to ecosystem processes in streams, and responsive to rapidly changing conditions. Here, we sampled 25 streams weekly (12-14 times each) and used 16S rRNA gene metabarcoding of periphyton-associated bacteria to quantify the effects of total phosphorus (TP) and total nitrogen (TN). Threshold indicator taxa analysis identified assemblage-level changes and amplicon sequence variants (ASVs) that increased or decreased with increasing TP and TN concentrations (i.e., low P, high P, low N, and high N ASVs). Boosted regression trees confirmed that relative abundances of gene sequence reads for these four indicator groups were associated with nutrient concentrations. Gradient forest analysis complemented these results by using multiple predictors and random forest models for each ASV to identify portions of TP and TN gradients at which the greatest changes in assemblage structure occurred. Synthesized statistical results showed bacterial assemblage structure began changing at 24 µg TP/L with the greatest changes occurring from 110 to 195 µg/L. Changes in the bacterial assemblages associated with TN gradually occurred from 275 to 855 µg/L. Taxonomic and phylogenetic analyses showed that low nutrient ASVs were commonly Firmicutes, Verrucomicrobiota, Flavobacteriales, and Caulobacterales, Pseudomonadales, and Rhodobacterales of Proteobacteria, whereas other groups, such as Chitinophagales of Bacteroidota, and Burkholderiales, Rhizobiales, Sphingomonadales, and Steroidobacterales of Proteobacteria comprised the high nutrient ASVs. Overall, the responses of bacterial ASV indicators in this study highlight the utility of metabarcoding periphyton-associated bacteria for quantifying biotic responses to nutrient inputs in streams.

Characterizing temporal variability in streams supports nutrient indicator development using diatom and bacterial DNA metabarcoding.

Interest in developing periphytic diatom and bacterial indicators of nutrient effects continues to grow in support of the assessment and management of stream ecosystems and their watersheds. However, temporal variability could confound relationships between indicators and nutrients, subsequently affecting assessment outcomes. To document how temporal variability affects measures of diatom and bacterial assemblages obtained from DNA metabarcoding, we conducted weekly periphyton and nutrient sampling from July to October 2016 in 25 streams in a 1293 km2 mixed land use watershed. Measures of both diatom and bacterial assemblages were strongly associated with the percent agriculture in upstream watersheds and total phosphorus (TP) and total nitrogen (TN) concentrations. Temporal variability in TP and TN concentrations increased with greater amounts of agriculture in watersheds, but overall diatom and bacterial assemblage variability within sites-measured as mean distance among samples to corresponding site centroids in ordination space-remained consistent. This consistency was due in part to offsets between decreasing variability in relative abundances of taxa typical of low nutrient conditions and increasing variability in those typical of high nutrient conditions as mean concentrations of TP and TN increased within sites. Weekly low and high nutrient diatom and bacterial metrics were more strongly correlated with site mean nutrient concentrations over the sampling period than with same day measurements and more strongly correlated with TP than with TN. Correlations with TP concentrations were consistently strong throughout the study except briefly following two major precipitation events. Following these events, biotic relationships with TP reestablished within one to three weeks. Collectively, these results can strengthen interpretations of survey results and inform monitoring strategies and decision making. These findings have direct applications for improving the use of diatoms and bacteria, and the use of DNA metabarcoding, in monitoring programs and stream site assessments.

Urbanisation alters fatty acids in stream food webs.

Fatty acids are essential to macroinvertebrate growth and reproduction and can indicate food web structure and nutritional quality of basal resources. However, broad-scale examinations of how catchment land cover and associated stressors affect the proportions of fatty acids (FAs) in stream food webs are few.Here, we: (1) examine relationships among proportions of FAs among benthic periphyton and macroinvertebrate collector/gatherers, shredders, and predators; and (2) test if relationships between periphytic and macroinvertebrate FAs were altered due to the intensity of urban development in catchments.Proportions of the ≥20-C eicosapentaenoic acid (EPA 20:5ω3), arachidonic acid (ARA 20:4ω6), and docosahexaenoic acid (22:6ω3) indicated collector/gatherers had a diet richer in periphyton than in shredders, which had significantly lower proportions of these FAs. Collector/gatherers were in turn likely to be high-quality sources of ω3 and ≥ 20-C FAs for predators, which also had significantly greater EPA and ARA proportions than those in shredders. Linoleic (18:2ω6) and α-linolenic acid (18:3ω3) comprised the greatest proportions of FAs in shredders, which suggested a diet dominated by leaf litter and associated hyphomycetes.As catchment urbanisation increased, proportions of total ω3 FAs and EPA in periphyton were significantly greater. This pattern also was seen through macroinvertebrate consumers and predators, given that proportions of these FAs in macroinvertebrates also were significantly correlated with factors associated with catchment urbanisation. The significant increase in total ω3 FAs and EPA proportions within shredders indicated that periphyton growth, and their FAs, increased on leaf litter, probably due to greater nutrient concentrations associated with catchment urbanisation. Proportions of total ω6 FAs in biota were not significantly correlated with factors associated with urban development, which could indicate that they were of sufficient abundance for consumers regardless of urban intensity or possible changes in their sources.Our study provides an informative first step that identified notable differences in proportions of FAs among macroinvertebrates in urban streams and an increase in proportions of total ω3 FAs and EPA in periphyton, consumers, and predators as catchment urbanisation increases. Identifying how FA relationships within food webs change in response to catchment alterations and stressors could inform land use and management decisions by linking environmental changes to measures important to ecosystem outcomes.

Use of Stable Isotopes in Benthic Organic Material as a Baseline for Estimating Fish Trophic Positions in Lakes.

Estimating the trophic position of predators in food webs using stable isotopes requires establishing a 'baseline'; typically served by bivalves or snails. However, the frequent absence of such organisms in lakes leaves researchers in a difficult situation. Here we test the hypothesis that trophic position (TP) of largemouth bass (LB) in lakes can be estimated from δ15N of benthic organic material (BOM) in littoral surficial sediment. δ15NLB as a function of δ15NBOM was significant across 51 lakes with an R2 of 0.84, supporting our hypothesis. In a subset of six lakes, the mean TPLB did not differ significantly when calculated using baselines of δ15N in BOM, mussels (UN), gastropods (SN), or from a stable isotope mixing model. In a subset of 26 lakes, mean TPLB calculated from δ15NBOM and δ15NUN baselines were not significantly different, but TPLB calculated using δ15NBOM was not significantly correlated with TPLB calculated using δ15NUN. Although δ15NBOM should not be used interchangeably with δ15NUN as a baseline for estimating TPLB, δ15NBOM can be useful as a separate baseline where estimates of fish TP are needed in all study lakes given that mussels are frequently absent and BOM is always available for collection.

Performance of National Maps of Watershed Integrity at Watershed Scales.

Watershed integrity, the capacity of a watershed to support and maintain ecological processes essential to the sustainability of services provided to society, can be influenced by a range of landscape and in-stream factors. Ecological response data from four intensively monitored case study watersheds exhibiting a range of environmental conditions and landscape characteristics across the United States were used to evaluate the performance of a national level Index of Watershed Integrity (IWI) at regional and local watershed scales. Using Pearson's correlation coefficient (r), and Spearman's rank correlation coefficient (rs ), response variables displayed highly significant relationships and were significantly correlated with IWI and ICI (Index of Catchment Integrity) values at all watersheds. Nitrogen concentration and flux-related watershed response metrics exhibited significantly strong negative correlations across case study watersheds, with absolute correlations (|r|) ranging from 0.48 to 0.97 for IWI values, and 0.31 to 0.96 for ICI values. Nitrogen-stable isotope ratios measured in chironomids and periphyton from streams and benthic organic matter from lake sediments also demonstrated strong negative correlations with IWI values, with |r| ranging from 0.47 to 0.92, and 0.35 to 0.89 for correlations with ICI values. This evaluation of the performance of national watershed and catchment integrity metrics and their strong relationship with site level responses provides weight-of-evidence support for their use in state, local and regionally focused applications.