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.

National-scale remotely sensed lake trophic state from 1984 through 2020.

Lake trophic state is a key ecosystem property that integrates a lake's physical, chemical, and biological processes. Despite the importance of trophic state as a gauge of lake water quality, standardized and machine-readable observations are uncommon. Remote sensing presents an opportunity to detect and analyze lake trophic state with reproducible, robust methods across time and space. We used Landsat surface reflectance data to create the first compendium of annual lake trophic state for 55,662 lakes of at least 10 ha in area throughout the contiguous United States from 1984 through 2020. The dataset was constructed with FAIR data principles (Findable, Accessible, Interoperable, and Reproducible) in mind, where data are publicly available, relational keys from parent datasets are retained, and all data wrangling and modeling routines are scripted for future reuse. Together, this resource offers critical data to address basic and applied research questions about lake water quality at a suite of spatial and temporal scales.

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.

Natural and anthropogenic controls on lake water-level decline and evaporation-to-inflow ratio in the conterminous United States.

Lake water levels are integral to lake function, but hydrologic changes from land and water management may alter lake fluctuations beyond natural ranges. We constructed a conceptual model of multifaceted drivers of lake water-levels and evaporation-to-inflow ratio (Evap:Inflow). Using a structural equation modeling framework, we tested our model on 1) a national subset of lakes in the conterminous United States with minimal water management to describe natural drivers of lake hydrology and 2) five ecoregional subsets of lakes to explore regional variation in water management effects. Our model fit the national and ecoregional datasets and explained up to 47% of variation in Evap:Inflow, 38% of vertical water-level decline, and 79% of horizontal water-level decline (littoral exposure). For lakes with minimal water management, Evap:Inflow was related to lake depth (ß = -0.31) and surface inflow (ß = -0.44); vertical decline was related to annual climate (e.g., precipitation ß = -0.18) and water management (ß = -0.21); and horizontal decline was largely related to vertical decline (ß = 0.73) and lake morphometry (e.g., depth ß = -0.18). Anthropogenic effects varied by ecoregion and likely reflect differences in regional water management and climate. In the West, water management indicators were related to greater vertical decline (ß = 0.38), whereas in the Midwest, these indicators were related to more stable and full lake levels (ß = -0.22) even during drought conditions. National analyses show how human water use interacts with regional climate resulting in contrasting impacts to lake hydrologic variation in the US.

National framework for ranking lakes by potential for anthropogenic hydro-alteration.

Lakes face multiple anthropogenic pressures that can substantially alter their hydrology. Dams and land use in the watershed (e.g., irrigated agriculture) can modify lake water regimes beyond natural ranges, and changing climate may exacerbate anthropogenic stresses on lake hydrology. However, we lack cost-effective indicators to quantify anthropogenic hydrologic alteration potential in lakes at regional and national extents. We developed a framework to rank lakes by the potential for dams and land use to alter lake hydrology (HydrAP) that can be applied at a national scale. The HydrAP framework principles are that 1) dams are primary drivers of lake hydro-alteration, 2) land use activities are secondary drivers that alter watershed hydrology, and 3) topographic relief limits where land use and dams are located on the landscape. We ranked lakes in the United States Environmental Protection Agency National Lakes Assessment (NLA) on a HydrAP scale from zero to seven, where a zero indicates lakes with no potential for anthropogenic hydro-alteration, and a seven indicates large dams and/or intensive land use with high potential to alter lake hydrology. We inferred HydrAP population distributions in the conterminous US (CONUS) using the NLA probabilistic weights. Half of CONUS lakes had moderate to high hydro-alteration potential (HydrAP ranks 3-7), the other half had minimal to no hydro-alteration potential (HydrAP ranks 0-2). HydrAP ranks generally corresponded with natural and man-made lake classes, but >15% of natural lakes had moderate to high HydrAP ranks and ~10% of man-made lakes had low HydrAP ranks. The Great Plains, Appalachians, and Coastal Plains had the largest percentages (>50%) of high HydrAP lakes, and the West and Midwest had the lowest percentages (~30%). Water residence time (τ) and water-level change were associated with HydrAP ranks, demonstrating the framework's intended ability to differentiate anthropogenic stressors that can alter lake hydrology. Consistently across ecoregions high HydrAP lakes had shorter τ. But HydrAP relationships with water-level change varied by ecoregion. In the West and Appalachians, high HydrAP lakes experienced excessive water-level declines compared to low-ranked lakes. In contrast, high HydrAP lakes in the Great Plains and Midwest showed stable water levels compared to low-ranked lakes. These differences imply that water management in western and eastern mountainous regions may result in large water-level fluctuations, but water management in central CONUS may promote water-level stabilization. The HydrAP framework using accessible, national datasets can support large-scale lake assessments and be adapted to specific locations where data are available.

Exploring the potential value of satellite remote sensing to monitor chlorophyll-a for US lakes and reservoirs.

Assessment of chlorophyll-a, an algal pigment, typically measured by field and laboratory in situ analyses, is used to estimate algal abundance and trophic status in lakes and reservoirs. In situ-based monitoring programs can be expensive, may not be spatially, and temporally comprehensive and results may not be available in the timeframe needed to make some management decisions, but can be more accurate, precise, and specific than remotely sensed measures. Satellite remotely sensed chlorophyll-a offers the potential for more geographically and temporally dense data collection to support estimates when used to augment or substitute for in situ measures. In this study, we compare available chlorophyll-a data from in situ and satellite imagery measures at the national scale and perform a cost analysis of these different monitoring approaches. The annual potential avoided costs associated with increasing the availability of remotely sensed chlorophyll-a values were estimated to range between $5.7 and $316 million depending upon the satellite program used and the timeframe considered. We also compared sociodemographic characteristics of the regions (both public and private lands) covered by both remote sensing and in situ data to check for any systematic differences across areas that have monitoring data. This analysis underscores the importance of continued support for both field-based in situ monitoring and satellite sensor programs that provide complementary information to water quality managers, given increased challenges associated with eutrophication, nuisance, and harmful algal bloom events.

Lake Water Levels and Associated Hydrologic Characteristics in the Conterminous U.S.

Establishing baseline hydrologic characteristics for lakes in the U.S. is critical to evaluate changes to lake hydrology. We used the U.S. EPA National Lakes Assessment 2007 and 2012 surveys to assess hydrologic characteristics of a population of ~45,000 lakes in the conterminous U.S. based on probability samples of ~1,000 lakes/yr distributed across nine ecoregions. Lake hydrologic study variables include water-level drawdown (i.e., vertical decline and horizontal littoral exposure) and two water stable isotope-derived parameters: evaporation-to-inflow (E:I) and water residence time. We present 1) national and regional distributions of the study variables for both natural and man-made lakes and 2) differences in these characteristics between 2007 and 2012. In 2007, 59% of the population of U.S. lakes had Greater than normal or Excessive drawdown relative to water levels in ecoregional reference lakes with minimal human disturbances; while in 2012, only 20% of lakes were significantly drawn down beyond normal ranges. Water isotope-derived variables did not differ significantly between survey years in contrast to drawdown. Median E:I was 20% indicating that flow-through processes dominated lake water regimes. For 75% of U.S. lakes, water residence time was < 1 year and was longer in natural vs. man-made lakes. Our study provides baseline ranges to assess local and regional lake hydrologic status and inform management decisions in changing environmental conditions.

Functional and taxonomic biogeography of phytoplankton and zooplankton communities in relation to environmental variation across the contiguous USA.

For biomonitoring of aquatic ecosystems, the use of coarse group classifications, either taxonomic or functional, has been proposed as an alternative to more highly resolved taxonomic identification. We tested this proposition for phytoplankton and zooplankton using a pan-United States dataset, which also allows us to investigate biogeographic relationships between plankton groups and environmental variables. We used data from 1010 lakes composing the 2012 US National Lakes Assessment and compared relationships derived using genus-level, more aggregated taxonomic resolution and functional types. We examined responses nationally and by ecoregion. Differences in plankton assemblages among ecoregions were detected, especially at genus-level classification. Our analyses show a gradient of altitude and temperature influencing both phytoplankton and zooplankton, and another gradient of nutrients and anthropogenic activity influencing mostly phytoplankton. The overall variation in the planktonic communities explained by environmental variables ranged from 4 to 22%, but together indicated that aggregated taxonomic classification performed better for phytoplankton; for zooplankton, the performance of different classification types depended on the ecoregion. Our analyses also revealed linkages between particular phytoplankton and zooplankton groups, mainly attributable to similar environmental responses and trophic interactions. Overall, the results support the applicability of coarse classifications to infer general responses of plankton communities to environmental drivers.

Biogeography and co-occurrence of 16 planktonic species of Keratella Bory de St. Vincent, 1822 (Rotifera, Ploima, Brachionidae) in lakes and reservoirs of the United States.

We report current distributions of 16 species of Keratella Bory de St. Vincent, 1822 within lakes and reservoirs of the U.S. Specimens were identified from 988 lakes and reservoirs during spring and summer 2012 as part of the U.S. Environmental Protection Agency's National Lakes Assessment (NLA). We performed a co-occurrence analysis to determine correlations between species-pair occurrences and a niche-centroid analysis to predict optimal water quality conditions for each species. While a high degree of overlap in geographic range was observed among the species, distribution maps showed that K. quadrata was largely confined to more northern latitudes and K. americana was confined primarily to the eastern U.S. Other common species were either ubiquitously or sparsely distributed across ecoregions, suggesting that their distribution may be more related to inter-species competition or local water quality parameters. This study expands the limited pool of knowledge on rotifer biogeography within the U.S.

Assessing water-quality changes in US rivers at multiple geographic scales using results from probabilistic and targeted monitoring.

Two commonly used approaches for water quality monitoring are probabilistic and targeted. In a probabilistic approach like the US Environmental Protection Agency's National Rivers and Streams Assessment, monitoring sites are selected using a statistically representative approach. In a targeted approach like that used by many monitoring organizations, monitoring sites are chosen individually to answer specific questions. One important goal of both approaches is documenting long-term changes in water quality. Here, we compare chloride change results in US rivers and streams between the early 2000s and early 2010s from both approaches. The probabilistic approach provided an unbiased representation of change in all US rivers and streams, but was designed to measure low-streamflow conditions within a spring/summer index period during periodic survey years. The targeted approach was focused on larger, more developed watersheds but samples were collected frequently throughout the assessment period in different seasons and streamflows. The probabilistic results showed a small decrease in chloride concentrations in rivers and streams with the lowest concentrations, but no consistent increase or decrease in the remainder. The increased granularity of the targeted results showed that there was, in fact, a mix of changes occurring, with increases at 132 sites, decreases at 112 sites, and relatively stable conditions at 55 sites. The combined results suggest that chloride is not responding to a widespread, common driver across the USA and that management of chloride would be most effective when targeted regionally or locally.

Combining national and state data improves predictions of microcystin concentration.

Data collected from lakes at national (regional) scales and state (local) scales can provide different insights regarding relationships between environmental factors and biological responses, and combining these two types of data can potentially yield more precise and accurate understanding of ecological phenomena. National data can include many measures, cover large spatial areas, and span broad environmental gradients. Because of these characteristics, analyses of these data can yield accurate estimates of relationships among different lake characteristics. However, the number of samples in a national data set that is available for estimating a relationship specific to waterbodies within a smaller region, like a single state, is limited. Conversely, state monitoring data provide intensive sampling of lakes within a smaller area, but these data span a narrower range of conditions and may only include a subset of relevant measurements. Here, a Bayesian network model is described that represents the causal linkages between observations of chlorophyll a concentration, cyanobacterial biovolume, and microcystin concentration. This network model was fit to national data and provided a context for modeling observations of chlorophyll a and microcystin collected from lakes in Iowa. Using the knowledge inherent in the national network model improved the accuracy of predictions of microcystin concentrations in Iowa compared to a model based only on Iowa data.

Calanoid copepod zooplankton density is positively associated with water residence time across the continental United States.

Inherent differences between naturally-formed lakes and human-made reservoirs may play an important role in shaping zooplankton community structure. For example, because many reservoirs are created by impounding and managing lotic systems for specific human purposes, zooplankton communities may be affected by factors that are unique to reservoirs, such as shorter water residence times and a reservoir's management regime, compared to natural lakes. However, the environmental factors that structure zooplankton communities in natural lakes vs. reservoirs may vary at the continental scale and remain largely unknown. We analyzed data from the 2007 U.S. Environmental Protection Agency's National Lakes Assessment and the U.S. Army Corps of Engineers' National Inventory of Dams to compare large-bodied crustacean zooplankton communities (defined here as individuals retained by 0.243 mm mesh size) in natural lakes and reservoirs across the continental U.S. using multiple linear regressions and regression tree analyses. We found that large-bodied crustacean zooplankton density was overall higher in natural lakes compared to reservoirs when the effect of latitude was controlled. The difference between waterbody types was driven by calanoid copepods, which were also more likely to be dominant in the >0.243 mm zooplankton community in natural lakes than in reservoirs. Regression tree analyses revealed that water residence time was not a major driver of calanoid copepod density in natural lakes but was one of the most important drivers of calanoid copepod density in reservoirs, which had on average 0.5-year shorter water residence times than natural lakes. Reservoirs managed for purposes that resulted in shorter residence times (e.g., hydroelectric power) had lower zooplankton densities than reservoirs managed for purposes that resulted in longer residence times (e.g., irrigation). Consequently, our results indicate that water residence time may be an important characteristic driving differing large-bodied zooplankton dynamics between reservoirs and natural lakes.

PHYSICAL AND CHEMICAL CONNECTIVITY OF STREAMS AND RIPARIAN WETLANDS TO DOWNSTREAM WATERS: A SYNTHESIS.

Streams, riparian areas, floodplains, alluvial aquifers and downstream waters (e.g., large rivers, lakes, oceans) are interconnected by longitudinal, lateral, and vertical fluxes of water, other materials and energy. Collectively, these interconnected waters are called fluvial hydrosystems. Physical and chemical connectivity within fluvial hydrosystems is created by the transport of nonliving materials (e.g., water, sediment, nutrients, contaminants) which either do or do not chemically change (chemical and physical connections, respectively). A substantial body of evidence unequivocally demonstrates physical and chemical connectivity between streams and riparian wetlands and downstream waters. Streams and riparian wetlands are structurally connected to downstream waters through the network of continuous channels and floodplain form that make these systems physically contiguous, and the very existence of these structures provides strong geomorphologic evidence for connectivity. Functional connections between streams and riparian wetlands and their downstream waters vary geographically and over time, based on proximity, relative size, environmental setting, material disparity, and intervening units. Because of the complexity and dynamic nature of connections among fluvial hydrosystem units, a complete accounting of the physical and chemical connections and their consequences to downstream waters should aggregate over multiple years to decades.

Environmental factors influencing the quantitative distribution of microcystin and common potentially toxigenic cyanobacteria in U.S. lakes and reservoirs.

Many species of cyanobacteria are capable of producing toxins and causing nuisance blooms, however response to environmental conditions is likely taxon-specific. Environmental factors influencing cyanobacterial composition and toxin production in lakes have been examined in many studies; yet are often confined to individual water bodies, or to a small number of systems within the same region. Here, data from the 2012 USEPA National Lakes Assessment are used to examine relationships between biovolume of common potentially-toxigenic cyanobacteria (Aphanizomenon spp., Cylindrospermopsis spp., Dolichospermum spp., Microcystis spp. and Planktothrix spp.) and environmental variables across the entire conterminous United States, and results are compared across nine distinct ecoregions. Total phosphorus and water clarity were identified as the most influential environmental factors correlated with phytoplankton community composition. The Northern, Southern and Temperate Plains ecoregions displayed the highest biovolumes of potentially toxigenic taxa on average, as well as highest mean concentrations of microcystin. In those three ecoregions, samples with microcystin concentrations greater than 1 ppb were primarily dominated by Planktothrix spp. while in all other ecoregions Dolichospermum spp. was the dominant genus. Canonical Correlation Analysis revealed a strong association between high microcystin concentrations and high nutrient concentrations (total nitrogen and total phosphorus), and between high microcystin concentrations and low percentage of watershed forest cover. Results from this study indicate that the likely occurrence of potentially toxigenic taxa in lakes and reservoirs is predictable on a biogeographical basis, depending on morphological and water quality characteristics. Data from this study may be useful to regional managers attempting to prevent or mitigate nuisance cyanobacterial blooms.

BIOTA CONNECT AQUATIC HABITATS THROUGHOUT FRESHWATER ECOSYSTEM MOSAICS.

Freshwater ecosystems are linked at various spatial and temporal scales by movements of biota adapted to life in water. We review the literature on movements of aquatic organisms that connect different types of freshwater habitats, focusing on linkages from streams and wetlands to downstream waters. Here, streams, wetlands, rivers, lakes, ponds, and other freshwater habitats are viewed as dynamic freshwater ecosystem mosaics (FEMs) that collectively provide the resources needed to sustain aquatic life. Based on existing evidence, it is clear that biotic linkages throughout FEMs have important consequences for biological integrity and biodiversity. All aquatic organisms move within and among FEM components, but differ in the mode, frequency, distance, and timing of their movements. These movements allow biota to recolonize habitats, avoid inbreeding, escape stressors, locate mates, and acquire resources. Cumulatively, these individual movements connect populations within and among FEMs and contribute to local and regional diversity, resilience to disturbance, and persistence of aquatic species in the face of environmental change. Thus, the biological connections established by movement of biota among streams, wetlands, and downstream waters are critical to the ecological integrity of these systems. Future research will help advance our understanding of the movements that link FEMs and their cumulative effects on downstream waters.

Changes in the relationship between zooplankton and phytoplankton biomasses across a eutrophication gradient.

The relationship between zooplankton biomass and phytoplankton biomass can provide insight into the structure and function of lake biological communities. We used a Bayesian network model to analyze a continental-scale data dataset to estimate changes in the relationship between zooplankton (Z) and phytoplankton (P) biomasses along a eutrophication gradient. The Bayesian network model allowed us to combine two different measurements of phytoplankton biomass (chlorophyll a concentration and directly observed biovolume) to improve the precision of estimates of true biomass within each sample. The model also allowed us to estimate separate relationships between P and zooplankton abundance and between P and mean individual zooplankton biomass and then to combine these two relationships into an estimate of seasonal mean zooplankton biomass. The resulting analysis indicated that seasonal mean zooplankton biomass increased proportionally with phytoplankton biomass in oligotrophic lakes, yielding a constant ratio between Z and P and suggested that bottom-up forces determined zooplankton biomass in these systems. In eutrophic lakes, seasonal mean zooplankton biomass was nearly constant with increases in phytoplankton biomass, yielding a decrease in the ratio between Z and P with increasing eutrophication. Bottom-up forces, as quantified by an increase in the proportion of cyanobacteria, accounted for approximately one fifth of the residual variance in the model as the relationship between Z and P changed from direct proportionality in oligotrophic lakes to the nearly constant value of Z observed in eutrophic lakes, suggesting that a combination of both top-down and bottom-up forces likely determined zooplankton biomass in eutrophic lakes.

Fewer blue lakes and more murky lakes across the continental U.S.: Implications for planktonic food webs.

Elevated allochthonous inputs of organic matter are increasingly recognized as a driver of ecosystem change in lakes, particularly when concurrent with eutrophication. Evaluation of lakes in a nutrient-color paradigm (i.e., based on total phosphorus and true color) enables a more robust approach to research and management. To assess temporal and spatial patterns in nutrient-color status for U.S. lakes and associated food web attributes, we analyzed the U.S. Environmental Protection Agency's National Lakes Assessment (NLA) data. With 1000+ lakes sampled in 2007 and 2012 in a stratified random sampling design, the NLA enables rigorous assessment of lake condition across the continental U.S. We demonstrate that many U.S. lakes are simultaneously experiencing eutrophication and brownification to produce an abundance of "murky" lakes. Overall, "blue" lakes decreased by ~ 18% (46% of lakes in 2007 to 28% in 2012) while "murky" lakes increased by almost 12% (24% of lakes in 2007 to 35.4% in 2012). No statistical differences were observed in the proportions of "green" or "brown" lakes. Regionally, murky lakes significantly increased in the Northern Appalachian, Southern Plains, and Xeric ecoregions. Murky lakes exhibited the highest epilimnetic chlorophyll a concentrations, cyanobacterial densities, and microcystin concentrations. Total zooplankton biomass was also highest in murky lakes, primarily due to increased rotifer and copepod biomass. However, zooplankton : phytoplankton biomass ratios were low, suggesting reduced energy transfer to higher trophic levels. These results emphasize that many lakes in the U.S. are simultaneously "greening" and "browning", with potentially negative consequences for water quality and food web structure.

Using National-Scale Data To Develop Nutrient-Microcystin Relationships That Guide Management Decisions.

Quantitative models that predict cyanotoxin concentrations in lakes and reservoirs from nutrient concentrations would facilitate management of these resources for recreation and as sources of drinking water. Development of these models from field data has been hampered by the high proportion of samples in which cyanotoxin concentrations are below detection limits and by the high variability of cyanotoxin concentrations within individual lakes. Here, we describe a national-scale hierarchical Bayesian model that addresses these issues and that predicts microcystin concentrations from summer mean total nitrogen and total phosphorus concentrations. This model accounts for 69% of the variance in mean microcystin concentrations in lakes and reservoirs of the conterminous United States. Mean microcystin concentrations were more strongly associated with differences in total nitrogen than total phosphorus. A general approach for assessing this and similar types of models for their utility for guiding management decisions is also described.

Continental-Scale Increase in Lake and Stream Phosphorus: Are Oligotrophic Systems Disappearing in the United States?

We describe continental-scale increases in lake and stream total phosphorus (TP) concentrations, identified through periodic probability surveys of thousands of water bodies in the conterminous U.S. The increases, observed over the period 2000-2014 were most notable in sites in relatively undisturbed catchments and where TP was initially low (e.g., less than 10 µg L(-1)). Nationally, the percentage of stream length in the U.S. with TP ≤ 10 µg L(-1) decreased from 24.5 to 10.4 to 1.6% from 2004 to 2009 to 2014; the percentage of lakes with TP ≤ 10 µg L(-1) decreased from 24.9 to 6.7% between 2007 and 2012. Increasing TP concentrations appear to be ubiquitous, but their presence in undeveloped catchments suggests that they cannot be entirely attributed to either point or common non-point sources of TP.