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Persistence of pharmaceutical compounds and other organic wastewater contaminants in a conventional drinking-water-treatment plant

In a study conducted by the US Geological Survey and the Centers for Disease Control and Prevention, 24 water samples were collected at selected locations within a drinking-water-treatment (DWT) facility and from the two streams that serve the facility to evaluate the potential for wastewater-related organic contaminants to survive a conventional treatment process and persist in potable-water supplies. Stream-water samples as well as samples of raw, settled, filtered, and finished water were collected during low-flow conditions, when the discharge of effluent from upstream municipal sewage-treatment plants accounted for 37–67% of flow in stream 1 and 10–20% of flow in stream 2. Each sample was analyzed for 106 organic wastewater-related contaminants (OWCs) that represent a diverse group of extensively used chemicals. Forty OWCs were detected in one or more samples of stream water or raw-water supplies in the treatment plant; 34 were detected in more than 10% of these samples. Several of these compounds also were frequently detected in samples of finished water; these compounds include selected prescription and non-prescription drugs and their metabolites, fragrance compounds, flame retardants and plasticizers, cosmetic compounds, and a solvent. The detection of these compounds suggests that they resist removal through conventional water-treatment processes. Other compounds that also were frequently detected in samples of stream water and raw-water supplies were not detected in samples of finished water; these include selected prescription and non-prescription drugs and their metabolites, disinfectants, detergent metabolites, and plant and animal steroids. The non-detection of these compounds indicates that their concentrations are reduced to levels less than analytical detection limits or that they are transformed to degradates through conventional DWT processes. Concentrations of OWCs detected in finished water generally were low and did not exceed Federal drinking-water standards or lifetime health advisories, although such standards or advisories have not been established for most of these compounds. Also, at least 11 and as many as 17 OWCs were detected in samples of finished water. Drinking-water criteria currently are based on the toxicity of individual compounds and not combinations of compounds. Little is known about potential human-health effects associated with chronic exposure to trace levels of multiple OWCs through routes such as drinking water. The occurrence in drinking-water supplies of many of the OWCs analyzed for during this study is unregulated and most of these compounds have not been routinely monitored for in the Nation's source- or potable-water supplies. This study provides the first documentation that many of these compounds can survive conventional water-treatment processes and occur in potable-water supplies. It thereby provides information that can be used in setting research and regulatory priorities and in designing future monitoring programs. The results of this study also indicate that improvements in water-treatment processes may benefit from consideration of the response of OWCs and other trace organic contaminants to specific physical and chemical treatments.

Science of the Total Environment

Discharges of produced waters from oil and gas extraction via wastewater treatment plants are sources of disinfection by-products to receiving streams

Fluids co-produced with oil and gas production (produced waters) are often brines that contain elevated concentrations of bromide. Bromide is an important precursor of several toxic disinfection by-products (DBPs) and the treatment of produced water may lead to more brominated DBPs. To determine if wastewater treatment plants that accept produced waters discharge greater amounts of brominated DBPs, water samples were collected in Pennsylvania from four sites along a large river including an upstream site, a site below a publicly owned wastewater treatment plant (POTW) outfall (does not accept produced water), a site below an oil and gas commercial wastewater treatment plant (CWT) outfall, and downstream of the POTW and CWT. Of 29 DBPs analyzed, the site at the POTW outfall had the highest number detected (six) ranging in concentration from 0.01 to 0.09 μg L − 1 with a similar mixture of DBPs that have been detected at POTW outfalls elsewhere in the United States. The DBP profile at the CWT outfall was much different, although only two DBPs, dibromochloronitromethane (DBCNM) and chloroform, were detected, DBCNM was found at relatively high concentrations (up to 8.5 μg L − 1 ). The water at the CWT outfall also had a mixture of inorganic and organic precursors including elevated concentrations of bromide (75 mg L − 1 ) and other organic DBP precursors (phenol at 15 μg L − 1 ). To corroborate these DBP results, samples were collected in Pennsylvania from additional POTW and CWT outfalls that accept produced waters. The additional CWT also had high concentrations of DBCNM (3.1 μg L − 1 ) while the POTWs that accept produced waters had elevated numbers (up to 15) and concentrations of DBPs, especially brominated and iodinated THMs (up to 12 μg L − 1 total THM concentration). Therefore, produced water brines that have been disinfected are potential sources of DBPs along with DBP precursors to streams wherever these wastewaters are discharged.

Science of the Total Environment

Reconstructing the response of C3 and C4 plants to decadal-scale climate change during the late Pleistocene in southern Illinois using isotopic analyses of calcified rootlets

The ??13C and ??18O values of well-preserved carbonate rhizoliths (CRs) provide detailed insights into changes in the abundance of C3 and C4 plants in response to approximately decadal-scale changes in growing-season climate. We performed stable isotope analyses on 35-40 CRs sampled at 1-cm intervals from an 18-cm-thick paleosol formed in southern Illinois during Wisconsin interstadial 2. Minimum ??13C values show little variation with depth, whereas maximum values vary dramatically, and average values show noticeable variability; maximum ??18O values vary less than the minimum ??18O values. These findings indicate that a diverse and stable C3 flora with a limited number of C4 grass species prevailed during this interval, and suggest that the maximum growing-season temperatures were relatively stable, but minimum growing-season temperatures varied considerably. Two general patterns characterize the relationships between the ??13C and ??18O values obtained from the 1-cm samples. In some cases, low ??13C values correspond to low ??18O values and high ??13C values correspond to high ??18O values, suggesting that cooler growing-season temperatures favored C3 and warmer growing-season temperatures favored C4 plants. In other cases, low ??13C values correspond to high ??18O values, likely suggesting that wetter growing-season conditions were favorable to C3 plants. The high density of well-preserved CRs in this paleosol provides a unique opportunity to study detailed ecological responses to high-resolution variability in growing-season climate. ?? 2006 University of Washington.

Quaternary Research

A comparison of physiological indicators of sublethal cadmium stress in wetland plants

Physiological indices, including photosynthesis, chlorophyll fluorescence, adenylate energy charge (AEC) ratio, and leaf reflectance, were determined for Typha domingensis and Spartina alterniflora in response to increasing concentrations of Cd and compared with the growth responses of these species. Leaf expansion, the live/total ratio of plant aboveground biomass, and the aboveground regrowth rate after the initial harvests were significantly reduced with increasing Cd concentration in the growth medium. Of the four physiological responses measured, only photosynthesis and AEC responded to the Cd treatment before damage was visually apparent. Also, these indices were significantly correlated with leaf expansion rate and live/total ratio in most instances. Except at the end of the experiment, when the most stressed plants began to die, the Fv/Fm ratio was not significantly affected by the Cd treatment. The leaf spectral reflectance parameters showed no significant change during the entire treatment period. The significant correlation between the stress indicators and plant growth supported the findings that photosynthesis and AEC were the most responsive of the indicators tested, however, further research investigating other chlorophyll fluorescence and leaf reflectance parameters may demonstrate as well the value of these indicators in quantifying sublethal stress. ?? 2001 Elsevier Science B.V. All rights reserved.

Environmental and Experimental Botany

Carbon isotopic constraints on the contribution of plant material to the natural precursors of trihalomethanes

The δ 13 C values of individual trihalomethanes (THM) formed on reaction of chlorine with dissolved organic carbon (DOC) leached from maize (corn; Zea maize L. ) and Scirpus acutus (an aquatic bulrush), and with DOC extracted from agricultural drainage waters were determined using purge and trap introduction into a gas chromatograph-combustion-isotope ratio monitoring mass spectrometer. We observed a 16.8‰ difference between the δ 13 C values of THM produced from the maize and Scirpus leachates, similar to the isotopic difference between the whole plant materials. Both maize and Scirpus formed THM 12‰ lower in 13 C than whole plant material. We suggest that the low value of the THM relative to the whole plant material is evidence of distinct pools of THM-forming DOC, representing different biochemical types or chemical structures, and possessing different environmental reactivity.

Organic Geochemistry

Atmospheric radiocarbon for the period 1910 to 2021 recorded by annual plants

We present a timeseries of 14 CO 2 for the period 1910–2021 recorded by annual plants collected in the southwestern United States, centered near Flagstaff, Arizona. This timeseries is dominated by five commonly occurring annual plant species in the region, which is considered broadly representative of the southern Colorado Plateau. Most samples (1910–2015) were previously archived herbarium specimens, with additional samples harvested from field experiments in 2015–2021. We used this novel timeseries to develop a smoothed local record with uncertainties for “bomb spike” 14 C dating of recent terrestrial organic matter. Our results highlight the potential importance of local records, as we document a delayed arrival of the 1963–1964 bomb spike peak, lower values in the 1980s, and elevated values in the last decade in comparison to the most current Northern Hemisphere Zone 2 record. It is impossible to retroactively collect atmospheric samples, but archived annual plants serve as faithful scribes: samples from herbaria around the Earth may be an under-utilized resource to improve understanding of the modern carbon cycle.

Radiocarbon

Out of the frying pan and into the fire: Effects of volcanic heat and other stressors on the conservation of a critically endangered plant in Hawaiʻi

Loss of local biodiversity resulting from abrupt environmental change is a significant environmental problem throughout the world. Extinctions of plants are particularly important yet are often overlooked. Drawing from a case in Hawai‘i, a global hotspot for plant and other extinctions, we demonstrate an effort to better understand and determine priorities for the management of an endangered plant (‘Ihi makole or Portulaca sclerocarpa ) in the face of rapid and extreme environmental change. Volcanic heat emissions and biological invasions have anecdotally been suggested as possible threats to the species. We integrated P. sclerocarpa outplanting with efforts to collect geological and ecological data to gauge the role of elevated soil temperatures and invasive grasses in driving P. sclerocarpa mortality and population decline. We measured soil temperature, soil depth, surrounding cover and P. sclerocarpa survivorship over three decades. The abundance of wild P. sclerocarpa decreased by 99.7% from the 1990s to 2021. Only 51% of outplantings persisted through 3–4 years. Binomial regression and structural equation modelling revealed that, among the variables we analysed, high soil temperatures were most strongly associated with population decline. Finding the niche where soil temperatures are low enough to allow P. sclerocarpa survival but high enough to limit other agents of P. sclerocarpa mortality may be necessary to increase population growth of this species.

Hawaii

Herbivores and nutrients control grassland plant diversity via light limitation

Human alterations to nutrient cycles and herbivore communities are affecting global biodiversity dramatically. Ecological theory predicts these changes should be strongly counteractive: nutrient addition drives plant species loss through intensified competition for light, whereas herbivores prevent competitive exclusion by increasing ground-level light, particularly in productive systems. Here we use experimental data spanning a globally relevant range of conditions to test the hypothesis that herbaceous plant species losses caused by eutrophication may be offset by increased light availability due to herbivory. This experiment, replicated in 40 grasslands on 6 continents, demonstrates that nutrients and herbivores can serve as counteracting forces to control local plant diversity through light limitation, independent of site productivity, soil nitrogen, herbivore type and climate. Nutrient addition consistently reduced local diversity through light limitation, and herbivory rescued diversity at sites where it alleviated light limitation. Thus, species loss from anthropogenic eutrophication can be ameliorated in grasslands where herbivory increases ground-level light.

Nature

Differentiating moss from higher plants is critical in studying the carbon cycle of the boreal biome

The satellite-derived normalized difference vegetation index (NDVI), which is used for estimating gross primary production (GPP), often includes contributions from both mosses and vascular plants in boreal ecosystems. For the same NDVI, moss can generate only about one-third of the GPP that vascular plants can because of its much lower photosynthetic capacity. Here, based on eddy covariance measurements, we show that the difference in photosynthetic capacity between these two plant functional types has never been explicitly included when estimating regional GPP in the boreal region, resulting in a substantial overestimation. The magnitude of this overestimation could have important implications regarding a change from a current carbon sink to a carbon source in the boreal region. Moss abundance, associated with ecosystem disturbances, needs to be mapped and incorporated into GPP estimates in order to adequately assess the role of the boreal region in the global carbon cycle.

Nature Communications

Naturalized species drive functional trait shifts in plant communities

Despite decades of research documenting the consequences of naturalized and invasive plant species on ecosystem functions, our understanding of the functional underpinnings of these changes remains rudimentary. This is partially due to ineffective scaling of trait differences between native and naturalized species to whole plant communities. Working with data from over 75,000 plots and over 5,500 species from across the United States, we show that changes in the functional composition of communities associated with increasing abundance of naturalized species mirror the differences in traits between native and naturalized plants. We find that communities with greater abundance of naturalized species are more resource acquisitive aboveground and belowground, shorter, more shallowly rooted, and increasingly aligned with an independent strategy for belowground resource acquisition via thin fine roots with high specific root length. We observe shifts toward herbaceous-dominated communities but shifts within both woody and herbaceous functional groups follow community-level patterns for most traits. Patterns are remarkably similar across desert, grassland, and forest ecosystems. Our results demonstrate that the establishment and spread of naturalized species, likely in combination with underlying environmental shifts, leads to predictable and consistent changes in community-level traits that can alter ecosystem functions.

Proceedings of the National Academy of Sciences

Demographic stochasticity in small remnant populations of the declining distylous plant Primula veris

We studied ecological consequences of distyly for the declining perennial plant Primula veris in the Swiss Jura. Distyly favours cross-fertilization and avoids inbreeding, but may lead to pollen limitation and reduced reproduction if morph frequencies deviate from 50 %. Disassortative mating is promoted by the reciprocal position of stigmas and anthers in the two morphs (pin and thrum) and by intramorph incompatibility and should result in equal frequencies of morphs at equilibrium. However, deviations could arise because of demographic stochasticity, the lower intra-morph incompatibility of the pin morph, and niche differentiation between morphs. Demographic stochasticity should result in symmetric deviations from an even morph frequency among populations and in increased deviations with decreasing population size. If crosses between pins occurred, these would only generate pins, and this could result in a pin-bias of morph frequencies in general and in small populations in particular. If the morphs have different niches, morph frequencies should be related to environmental factors, morphs might be spatially segregated, and morphological differences between morphs would be expected. We tested these hypotheses in the declining distylous P. veris. We studied morph frequencies in relation to environmental conditions and population size, spatial segregation in field populations, morphological differences between morphs, and growth responses to nutrient addition. Morph frequencies in 76 populations with 1 - 80000 flowering plants fluctuated symmetrically about 50 %. Deviations from 50 % were much larger in small populations, and sixof the smallest populations had lost one morph altogether. In contrast, morph frequencies were neither related to population size nor to 17 measures of environmental conditions. We found no spatial segregation or morphological differences in the field or in the common garden. The results suggest that demographic stochasticity caused deviations of the morph ratiofrom unity in small populations. Demographic stochasticity was probably caused by the random elimination of plants during the fragmentation of formerly large continuous populations. Biased morph frequencies may be one of the reasons for the strongly reduced reproduction in small populations of P. veris.

Basic and Applied Ecology

Geochemical assessment of the suitability of converting a coal-fired power plant reservoir to a drinking-water reservoir

There is an increasing need for additional water storage in the United States, especially in arid regions. Alternatives like decommissioned power plant raw-water reservoirs would be cheaper to use than creating new reservoirs; however, the biogeochemical pollution risk of these reservoirs is not well understood. The San Juan Generating Station power plant and the associated reservoir will be used as a sediment-settling basin to store drinking water. To evaluate whether the reservoir is appropriate for this use, inorganic and organic constituents of concern were measured in reservoir sediment cores, pore water, and reservoir water in 2020. Forty-six percent of sediment arsenic concentrations measured in core subsamples (8 to 12 mg/kg) were slightly above the New Mexico residential cancer threshold for soil of 7.07 mg/kg. One sediment sample contained elevated total barium concentrations (6020 mg/kg). The organic compounds analyzed were either below detection limits or below regulatory thresholds. Reservoir water had one sample with arsenic greater than the drinking water standard (10 µg/L). Overall, the reservoir sediment inorganic and organic analyte concentrations are within acceptable ranges. The few samples that have elevated concentrations are not of a sufficient magnitude that dilution and/or treatment processes would preclude the use of the reservoir for water storage. Our findings show potential for use of former coal power station raw-water reservoirs as drinking-water reservoirs after plant closure.

New Mexico

Relations of alpine plant communities across environmental gradients: Multilevel versus multiscale analyses

Alpine plant communities vary, and their environmental covariates could influence their response to climate change. A single multilevel model of how alpine plant community composition is determined by hierarchical relations is compared to a separate examination of those relations at different scales. Nonmetric multidimensional scaling of species cover for plots in four regions across the Rocky Mountains created dependent variables. Climate variables are derived for the four regions from interpolated data. Plot environmental variables are measured directly and the presence of thirty-seven site characteristics is recorded and used to create additional independent variables. Multilevel and best subsets regressions are used to determine the strength of the hypothesized relations. The ordinations indicate structure in the assembly of plant communities. The multilevel analyses, although revealing significant relations, provide little explanation; of the site variables, those related to site microclimate are most important. In multiscale analyses (whole and separate regions), different variables are better explanations within the different regions. This result indicates weak environmental niche control of community composition. The weak relations of the structure in the patterns of species association to the environment indicates that either alpine vegetation represents a case of the neutral theory of biogeography being a valid explanation or that it represents disequilibrium conditions. The implications of neutral theory and disequilibrium explanations are similar: Response to climate change will be difficult to quantify above equilibrium background turnover.

Annals of the Association of American Geographers

A structural equation model of plant species richness and its application to a coastal wetland

Studies of plant species richness have often emphasized the role of either community biomass (as an indicator of density effects) or abiotic factors. In this article we present a general model that simultaneously examines the relative importance of abiotic and density effects. General and specific models were developed to examine the importance of abiotic conditions, disturbance, and community biomass on plant species richness. Models were evaluated using structural equation modeling based on data from 190 plots across a coastal marsh landscape. The accepted model was found to explain 45% of the observed variation in richness, 75% of biomass, and 65% of light penetration. Model results indicate that abiotic conditions have both direct effects on the species pool and indirect effects on richness mediated through effects on biomass and shading. Effects of disturbance were found to be indirect via biomass. Strong density effects on richness were indicated by the results, and canopy light penetration was found to be a better predictor of richness than was biomass. Overall, it appears that richness in this coastal landscape is controlled in roughly equal proportions by abiotic influences on the species pool and density effects, with disturbance playing a lesser role. The structure of the general model presented should be applicable to a wide variety of herbaceous plant communities.

Louisiana, Mississippi

Effects of prescribed fire timing on vigor of the invasive forb sericea lespedeza (Lespedeza cuneata), total forage biomass accumulation, plant-community composition, and native fauna on tallgrass prairie in the Kansas Flint Hills

The predominant grazing-management practice of the Kansas Flint Hills involves annual prescribed burning in March or April with postfire grazing by yearling beef cattle at a high stocking density from April to August. There has been a dramatic increase in sericea lespedeza ( Lespedeza cuneata [Dumont] G. Don) coincident with this temporally focused use of prescribed fire in the Flint Hills region. The species is an aggressive invader and a statewide noxious weed in Kansas. Control has generally been attempted using repeated herbicide applications. This approach has not limited proliferation of sericea lespedeza and resulted in collateral damage to nontarget flora and fauna. Alternative timing of prescribed fire has not been evaluated for its control. Our objectives for this 4-yr experiment were to (1) document the effects of prescribed burning during early April, early August, or early September on vigor of sericea lespedeza, standing forage biomass, and basal cover of native graminoids, forbs, and shrubs and (2) measure responses to fire regimes by grassland bird and butterfly communities. Whole-plant dry mass, basal cover, and seed production of sericea lespedeza were markedly less ( P < 0.01) in areas treated with prescribed fire in August or September compared with April. Forage biomass did not differ ( P ≥ 0.43) among treatments when measured during July; moreover, frequencies of bare soil, litter, and total basal plant cover were not different ( P ≥ 0.29) among treatments. Combined basal covers of C4 grasses, C3 grasses, annual grasses, forbs, and shrubs also did not differ ( P ≥ 0.11) between treatments. Densities of grasshopper sparrow ( Ammodramus savannarum ), dickcissel ( Spiza americana ), and eastern meadowlark ( Sturnella magna ) were not negatively affected ( P > 0.10) by midsummer or late-summer fires relative to early-spring fires. There were no differences ( P > 0.10) in densities of grassland-specialist butterfly species across fire regimes. Under the conditions of our experiment, prescribed burning during summer produced no detrimental effects on forage production, desirable nontarget plant species, grassland birds, or butterfly communities but had strong suppressive effects on sericea lespedeza. Additional research is warranted to investigate how to best incorporate late-summer prescribed fire into common grazing-management practices in the Kansas Flint Hills.

Kansas

Characterizing the marsh dieback spectral response at the plant and canopy level with hyperspectral and temporal remote sensing data

We describe newly developed remote sensing tools to map the localized occurrences and regional distribution of the marsh dieback in coastal Louisiana (Fig. 1). As a final goal of our research and development, we identified what spectral features accompanied the onset of dieback and could be directly linked to the optical signal measured at the satellite. In order to accomplish our research goal, we carried out two interlinked objectives. First, we determined the spectral features within the hyperspectral spectra of the impacted plant that could be linked to the spectral return. This was accomplished by measuring the differences in leaf optical properties of impacted and non impacted marsh plants in such a way that the measured differences could be linked to the dieback onset and progression. The spectral analyses were constrained to selected wavelengths (bands of reflectance data) historically associated with changes in leaf composition and structure caused by changes in the plant biophysical environment. Second, we determined what changes in the canopy reflectance (canopy signal sensed at the satellite) could be linked to dieback onset and progression. Third, we transformed a suite of six Landsat Thematic Mapper images collected before, during, and in the final stages of dieback to maps of dieback occurrences. ??2008 IEEE.

Conference Paper

Experimental warming in a dryland community reduced plant photosynthesis and soil CO 2 efflux although the relationship between the fluxes remained unchanged

1. Drylands represent our planet's largest terrestrial biome and, due to their extensive area, maintain large stocks of carbon (C). Accordingly, understanding how dryland C cycling will respond to climate change is imperative for accurately forecasting global C cycling and future climate. However, it remains difficult to predict how increased temperature will affect dryland C cycling, as substantial uncertainties surround the potential responses of the two main C fluxes: plant photosynthesis and soil CO 2 efflux. In addition to a need for an improved understanding of climate effects on individual dryland C fluxes, there is also notable uncertainty regarding how climate change may influence the relationship between these fluxes. 2. To address this important knowledge gap, we measured a growing season's in situ photosynthesis, plant biomass accumulation, and soil CO 2 efflux of mature Achnatherum hymenoides (a common and ecologically important C 3 bunchgrass growing throughout western North America) exposed to ambient or elevated temperature (+2°C above ambient, warmed via infrared lamps) for three years. 3. The 2°C increase in temperature caused a significant reduction in photosynthesis, plant growth, and soil CO 2 efflux. Of important note, photosynthesis and soil respiration appeared tightly coupled and the relationship between these fluxes was not altered by the elevated temperature treatment, suggesting C fixation's strong control of both above-ground and below-ground dryland C cycling. Leaf water use efficiency was substantially increased in the elevated temperature treatment compared to the control treatment. 4. Taken together, our results suggest notable declines in photosynthesis with relatively subtle warming, reveal strong coupling between above- and below-ground C fluxes in this dryland, and highlight temperature's strong effect on fundamental components of dryland C and water cycles.

Functional Ecology

Intraspecific variability and reaction norms of forest understory plant species traits

Trait-based models of ecological communities typically assume intraspecific variation in functional traits is not important, though such variation can change species trait rankings along gradients in resources and environmental conditions, and thus influence community structure and function. We examined the degree of intraspecific relative to interspecific variation, and reaction norms of 11 functional traits for 57 forest understory plant species, including: intrinsic water-use efficiency (iWUE), Δ 15 N, 5 leaf traits, 2 stem traits and 2 root traits along gradients in light, nitrogen, moisture and understory cover. Our results indicate that interspecific trait variation exceeded intraspecific variation by at least 50% for most, but not all traits. Intraspecific variation in Δ 15 N, iWUE, leaf nitrogen content and root traits was high (47-70%) compared with most leaf traits and stem traits (13-38%). Δ 15 N varied primarily along gradients in abiotic conditions, while light and understory cover were relatively less important. iWUE was related primarily to light transmission, reflecting increases in photosynthesis relative to stomatal conductance. Leaf traits varied mainly as a function of light availability, with some reaction norms depending on understory cover. Plant height increased with understory cover, while stem specific density was related primarily to light. Resources, environmental conditions and understory cover did not contribute strongly to the observed variation in root traits. Gradients in resources, environmental conditions and competition all appear to control intraspecific variability in most traits to some extent. However, our results suggest that species cross-over (i.e., trait rank reversals) along the gradients measured here are generally not a concern. Intraspecific variability in understory plant species traits can be considerable. However, trait data collected under a narrow range of environmental conditions appears sufficient to establish species rankings and scale between community and ecosystem levels using trait-based models. Investigators may therefore focus on obtaining a sufficient sample size within a single set of conditions rather than characterizing trait variation across entire gradients in order to optimize sampling efforts.

Oregon