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Patterns of diel variation in nitrate concentrations in the Potomac River

The Potomac River is a large source of N to Chesapeake Bay, where reducing nutrient loads is a focus of efforts to improve trophic status. Better understanding of NO 3 – loss, reflected in part by diel variation in NO 3 – concentrations, may refine model predictions of N loads to the Bay. We analyzed 2 y of high-frequency NO 3 – sensor data in the Potomac to quantify seasonal variation in the magnitude and timing of diel NO 3 – loss. Diel patterns were evident, especially during low flow, despite broad seasonal and flow-driven variation in NO 3 – concentrations. Diel variation was ~0.01 mg N/L in winter and 0.02 to 0.03 mg N/L in summer with intermediate values in spring and autumn, equivalent to <1% of the daily mean NO 3 – concentration in winter and ~2 to 4% in summer. Maximum diel NO 3 – values generally occurred in mid- to late morning, with more repeatable patterns in summer and wider variation in autumn and winter. Diel NO 3 – loss reduced loads by 0.7% in winter and 3% in summer. These losses were less than estimates of total in-stream NO 3 – load loss across the basin that averaged 33% of the annual groundwater contribution to the river. Water temperature and discharge had stronger relationships to the daily magnitude of diel NO 3 – variation than did photosynthetically active radiation. Estimated diel areal NO 3 – loss rates were generally >1000 mg N m –2 d –1 , greater than most published values because measurements in this large river integrate over a greater depth/unit stream bottom area than do those from smaller rivers. These diel NO 3 – patterns are consistent with the influence of photoautotrophic uptake and related denitrification, but we cannot attribute these patterns to assimilation alone because the magnitude and timing of diel dynamics were affected to an unknown extent by processes, such as evapotranspiration, transient storage, and hydrodynamic dispersion. Improvements to diel loss estimates will require additional high-frequency measures, such as dissolved O 2 , dissolved organic N, and NH 4 + , and deployment of 2 measurement stations.

Potomac River

A software tool to assess uncertainty in transient-storage model parameters using Monte Carlo simulations

Researchers and practitioners alike often need to understand and characterize how water and solutes move through a stream in terms of the relative importance of in-stream and near-stream storage and transport processes. In-channel and subsurface storage processes are highly variable in space and time and difficult to measure. Storage estimates are commonly obtained using transient-storage models (TSMs) of the experimentally obtained solute-tracer test data. The TSM equations represent key transport and storage processes with a suite of numerical parameters. Parameter values are estimated via inverse modeling, in which parameter values are iteratively changed until model simulations closely match observed solute-tracer data. Several investigators have shown that TSM parameter estimates can be highly uncertain. When this is the case, parameter values cannot be used reliably to interpret stream-reach functioning. However, authors of most TSM studies do not evaluate or report parameter certainty. Here, we present a software tool linked to the One-dimensional Transport with Inflow and Storage (OTIS) model that enables researchers to conduct uncertainty analyses via Monte-Carlo parameter sampling and to visualize uncertainty and sensitivity results. We demonstrate application of our tool to 2 case studies and compare our results to output obtained from more traditional implementation of the OTIS model. We conclude by suggesting best practices for transient-storage modeling and recommend that future applications of TSMs include assessments of parameter certainty to support comparisons and more reliable interpretations of transport processes.

Freshwater Science

Top-down control of invertebrates by Ninespine Stickleback in Arctic ponds

Despite their widespread presence in northern-latitude ecosystems, the ecological role of Ninespine Stickleback Pungitius pungitius is not well understood. Ninespine Stickleback can occupy both top and intermediate trophic levels in freshwater ecosystems, so their role in food webs as a predator on invertebrates and as a forage fish for upper level consumers probably is substantial. We introduced Ninespine Sticklebacks to fishless ponds to elucidate their potential effects as a predator on invertebrate communities in Arctic lentic freshwaters. We hypothesized that Ninespine Stickleback would affect freshwater invertebrate communities in a top-down manner. We predicted that the addition of Ninespine Sticklebacks to fishless ponds would: 1) reduce invertebrate taxonomic richness, 2) decrease overall invertebrate abundance, 3) reduce invertebrate biomass, and 4) decrease average invertebrate body size. We tested our hypothesis at 2 locations by adding Ninespine Stickleback to isolated ponds and compared invertebrate communities over time between fish-addition and fishless control ponds. Ninespine Sticklebacks exerted strong top-down pressure on invertebrate communities mainly by changing invertebrate taxonomic richness and biomass and, to a lesser extent, abundance and average invertebrate size. Our results supported the hypothesis that Ninespine Stickleback may help shape lentic food webs in the Arctic.

Alaska

Characterizing the early life history of an imperiled freshwater mussel ( Ptychobranchus jonesi ) with host-fish determination and fecundity estimation

Conservation of imperiled species is frequently challenged by insufficient knowledge of life history and environmental factors that affect various life stages. The larvae (glochidia) of most freshwater mussels in the family Unionidae are obligate ectoparasites of fishes. We described the early life history of the federally endangered Southern Kidneyshell Ptychobranchus jonesi and compared methods for estimating fecundity and conducting host trials on this conglutinate-producing mussel species. Glochidial inoculation baths and direct feeding of conglutinates to Percina nigrofasciata , Etheostoma edwini , and Etheostoma fusiforme resulted in successful metamorphosis to the juvenile life stage. Ptychobranchus jonesi glochidia did not metamorphose on 25 other species of fishes tested representing 11 families. Three juveniles were recovered from Gambusia holbrooki resulting in a metamorphosis rate <1%. We characterize P. jonesi as a host-fish specialist that fractionally releases conglutinates from late January to early June. Intact P. jonesi conglutinates resemble simuliid fly larvae attached to an egg-like structure, but most conglutinates were released as segments representing separate egg or larva mimics. Viability of glochidia encased within a conglutinate was >90% for ≥5 d. Feeding conglutinates directly to fishes allowed us to estimate seminatural infestation rates and calculate average numbers of juveniles produced per conglutinate, unlike the traditional approach of infesting fish hosts in an inoculation bath. Regressions based on the physical dimensions of each conglutinate or conglutinate segment were the most practical method used to estimate fecundity. Species distribution information, early life-history description, and methods developed for determining fecundity and conducting host trials may assist in the conservation of P. jonesi during recovery options that include captive propagation, augmentation, and reestablishment.

Alabama, Florida

Recruitment phenology and pelagic larval duration in Caribbean amphidromous fishes

Amphidromous fishes are major components of oceanic tropical island stream ecosystems, such as those of the Caribbean island, Puerto Rico. Fishes with this life history face threats related to the requirement for connectivity between freshwater and marine environments during early life stages. Pelagic larval duration and recruitment phenology are 2 early life-history processes that are crucial for the biology, ecology, conservation, and management of amphidromous fishes. However, these processes are understudied in the Caribbean in general and have never been quantified in Puerto Rico. We quantified recruit abundance, recruitment phenology, and pelagic larval duration of several Caribbean amphidromous fish species in multiple rivers in Puerto Rico and explored the effects of environmental variables on recruit abundances. Two fish taxa—sirajo goby ( Sicydium spp.) and River Goby ( Awaous banana )—were exceptionally abundant as postlarvae and recruited to Caribbean rivers in pulsed migration episodes that were periodic at annual and lunar scales. Sirajo goby and River Goby recruit abundances varied among rivers, were greater at sunrise than at sunset, and were positively related to river discharge. The pelagic larval duration of 4 fish taxa ranged from a minimum of 28 d to a maximum of 103 d with means between 43 ± 7 d (SD) and 65 ± 11 d. We identified the last-quarter moon phase during the months of June through January as periods of maximum amphidromous fish recruitment to freshwater streams. The results and conclusions of our study can be applied to identify critical times to maintain river–ocean connectivity and stream flow for the benefit of the amphidromous fish population dynamics, stream ecology, and natural resources of the Caribbean.

Puerto Rico

Thermal tolerances of fishes occupying groundwater and surface-water dominated streams

A thermal tolerance study mimicking different stream environments could improve our ecological understanding of how increasing water temperatures affect stream ectotherms and improve our ability to predict organism responses based on river classification schemes. Our objective was to compare the thermal tolerances of stream fishes of different habitat guilds among 3 exposure periods: critical thermal maximum (CTmax, increase of 2°C/h until loss of equilibrium [LOE] and death [D]), and 2 longer-term treatments (net daily increase of 1°C) that mimicked spring-fed (SF; 4°C daily increase) and non-spring-fed (NSF; 8°C daily increase) conditions. Fishes in the pelagic habitat guild had a 1°C higher average CTmax than benthic fishes. Thermal responses of species depended on exposure period with higher and increased variation in tolerances associated with the SF and NSF exposure periods. Logperch, Orangebelly Darter, Orangethroat Darter, and Southern Redbelly Dace were more sensitive to thermal increases regardless of SF or NSF treatment than were the 3 remaining species (Brook Silverside, Central Stoneroller, and Redspot Chub), which represented average thermal responses among the species tested. The 3 species that had a higher thermal response to CTmax-D (lethal endpoint of death) also were able to increase their tolerances more than other species in both SF and NSF treatments. Our data indicate finer guild designations may be useful for predicting thermal-response patterns. A diel thermal refuge increases the thermal responses of ectotherms to daily maxima, but the patterns across our SF and NSF treatments were similar suggesting minimum refuge temperatures may be more important than maximums. Nonetheless, stream temperature cooling over a 24-h period is important to ectotherm thermal tolerances, a result suggesting that sources of cooler water to streams might benefit from protection.

Freshwater Science

Predictability and selection of hydrologic metrics in riverine ecohydrology

The natural flow regime is critical to the health of riverine ecosystems. Many hydrologic metrics (HMs) have been developed to describe natural flow regimes, quantify flow alteration, and provide the hydrologic foundation for the development of environmental flow standards. Many applications require the use of models to predict expected natural values of HMs from basin characteristics at sites with no observed records of unimpaired flows. However, the error associated with HM estimation has not been evaluated. The primary goal of our study was to provide guidance for river scientists and managers in the selection, use, and interpretation of HMs for stream classification and hydroecological investigations of river ecosystems. We evaluated the predictability of a broad suite of HMs for the conterminous USA based on random forest statistical models. We also examined how the predictability of metrics varied among unique components of the flow regime. Roughly 40% of 612 HMs we examined could be predicted reliably from basin characteristics. The predictable metrics were disproportionately represented in 5 flow components: asymmetry, seasonality, magnitude, variability, and average monthly flows. Most metrics that represent extreme hydrological events (i.e., high and low flows) could not be reliably predicted. Roughly ⅔ of the evaluated HMs were incalculable or highly biased at intermittent/ephemeral streams because of the need for logarithmic transformations or scaling by other HMs, such as mean flows or percentile flow thresholds. Scaling metrics by drainage area tended to improve predictability. We recommend that the predictability of HMs be given greater consideration in studies and applications in which they are used to characterize and assess alteration of streamflow regimes.

Freshwater Science

Biological relevance of streamflow metrics: Regional and national perspectives

Protecting the health of streams and rivers requires identifying ecologically significant attributes of the natural flow regime. Streamflow regimes are routinely quantified using a plethora of hydrologic metrics (HMs), most of which have unknown relevance to biological communities. At regional and national scales, we evaluated which of 509 commonly used HMs were associated with biological indicators of fish and invertebrate community integrity. We quantified alteration of each HM by using statistical models to predict site-specific natural baseline values for each of 728 sites across the USA where streamflow monitoring data were available concurrent with assessments of invertebrate or fish community integrity. We then ranked HMs according to their individual association with biological integrity based on random forest models that included HMs and other relevant covariates, such as land cover and stream chemistry. HMs were generally the most important predictors of biological integrity relative to the covariates. At a national scale, the most influential HMs were measures of depleted high flows, homogenization of flows, and erratic flows. Unique combinations of biologically relevant HMs were apparent among regions. We discuss the implications of our findings to the challenge of selecting HMs for streamflow research and management.

Freshwater Science

Stream permanence is related to crayfish occupancy and abundance in the Ozark Highlands, USA

Crayfish use of intermittent streams is especially important to understand in the face of global climate change. We examined the influence of stream permanence and local habitat on crayfish occupancy and species densities in the Ozark Highlands, USA. We sampled in June and July 2014 and 2015. We used a quantitative kick–seine method to sample crayfish presence and abundance at 20 stream sites with 32 surveys/site in the Upper White River drainage, and we measured associated local environmental variables each year. We modeled site occupancy and detection probabilities with the software PRESENCE, and we used multiple linear regressions to identify relationships between crayfish species densities and environmental variables. Occupancy of all crayfish species was related to stream permanence. Faxonius meeki was found exclusively in intermittent streams, whereas Faxonius neglectus and Faxonius luteus had higher occupancy and detection probability in permanent than in intermittent streams, and Faxonius williamsi was associated with intermittent streams. Estimates of detection probability ranged from 0.56 to 1, which is high relative to values found by other investigators. With the exception of F. williamsi , species densities were largely related to stream permanence rather than local habitat. Species densities did not differ by year, but total crayfish densities were significantly lower in 2015 than 2014. Increased precipitation and discharge in 2015 probably led to the lower crayfish densities observed during this year. Our study demonstrates that crayfish distribution and abundance is strongly influenced by stream permanence. Some species, including those of conservation concern (i.e., F. williamsi , F. meeki ), appear dependent on intermittent streams, and conservation efforts should include consideration of intermittent streams as an important component of freshwater biodiversity.

Arkansas, Missouri

Hydrologic characteristics of freshwater mussel habitat: novel insights from modeled flows

The ability to model freshwater stream habitat and species distributions is limited by the spatially sparse flow data available from long-term gauging stations. Flow data beyond the immediate vicinity of gauging stations would enhance our ability to explore and characterize hydrologic habitat suitability. The southeastern USA supports high aquatic biodiversity, but threats, such as landuse alteration, climate change, conflicting water-resource demands, and pollution, have led to the imperilment and legal protection of many species. The ability to distinguish suitable from unsuitable habitat conditions, including hydrologic suitability, is a key criterion for successful conservation and restoration of aquatic species. We used the example of the critically endangered Tar River Spinymussel ( Parvaspina steinstansana ) and associated species to demonstrate the value of modeled flow data (WaterFALL™) to generate novel insights into population structure and testable hypotheses regarding hydrologic suitability. With ordination models, we: 1) identified all catchments with potentially suitable hydrology, 2) identified 2 distinct hydrologic environments occupied by the Tar River Spinymussel, and 3) estimated greater hydrological habitat niche breadth of assumed surrogate species associates at the catchment scale. Our findings provide the first demonstrated application of complete, continuous, regional modeled hydrologic data to freshwater mussel distribution and management. This research highlights the utility of modeling and data-mining methods to facilitate further exploration and application of such modeled environmental conditions to inform aquatic species management. We conclude that such an approach can support landscape-scale management decisions that require spatial information at fine resolution (e.g., enhanced National Hydrology Dataset catchments) and broad extent (e.g., multiple river basins).

North Carolina

Cyanobacteria reduce quagga mussel (Dreissena rostriformis bugensis) spawning and fertilization success

Quagga mussels ( Dreissena rostriformis bugensis ) are highly fecund broadcast spawners invasive to freshwaters of North America and western Europe. We hypothesized that environmental cues from phytoplankton can trigger gamete release in quagga mussels. Nutritious algae may stimulate dreissenid spawning, but less palatable food, such as bloom-forming cyanobacteria, could be a hindrance. The objective of our study was to test whether exposure to cyanobacteria can inhibit quagga mussel spawning and fertilization. We assessed spawning in the presence of serotonin, a known spawning inducer, where adult quagga mussels placed in individual vials were exposed to 13 cyanobacteria cultures and purified algal toxin (microcystin-LR) with artificial lake water as the control. Fertilization success was evaluated by combining eggs with sperm in conjunction with cyanobacteria, and enumerating zygote formation marked by cellular cleavage. Several cyanobacterial strains reduced spawning and fertilization success, but microcystin-LR had no effect. Fertilization was more sensitive to cyanobacteria than gamete release. Only 1 culture, Aphanizomenon flos-aquae , inhibited spawning, whereas 6 cultures consisting of Anabaena flos-aquae , Dolichospermum lemmermanii , Gloeotrichia echinulata , Lyngbya wollei , and 2 Microcystis aeruginosa isolates reduced fertilization rates by up to 44%. The effects of cyanobacteria on reproduction in invasive freshwater mussels in the wild have not yet been identified. However, our laboratory studies show that concentrations of cyanobacteria that are possible during bloom conditions probably limit reproduction. Reproductive consequences on wild populations may become more prevalent as cyanobacteria blooms occur earlier in the year, making overlap between blooms and mussel spawning more common. Describing the mechanism by which cyanobacteria inhibit spawning and fertilization could reveal novel control methods to limit reproduction of this invasive species.

Freshwater Science

Predicting biological conditions for small headwater streams in the Chesapeake Bay watershed

A primary goal for Chesapeake Bay watershed restoration is to improve stream health and function in 10% of stream miles by 2025. Predictive spatial modeling of stream conditions, when accurate, is one method to fill gaps in monitoring coverage and estimate baseline conditions for restoration goals. Predictive modeling can also monitor progress as additional data become available. We developed a random forests model to predict biological condition of small streams (<200 km 2 in drainage) in the Chesapeake Bay watershed. Biological condition was measured with the Chesapeake Bay Basin-wide Index of Biotic Integrity (Chessie BIBI), a stream macroinvertebrate index. Our goal was to predict biological condition in all unsurveyed small streams present in a 1:24,000 scale catchment layer as a 2004–2008 baseline. We reclassified the 5-category Chessie BIBI ratings into two categories, poor and fair/good, to align with management goals of the Chesapeake Bay Program. The model included 12 geospatial predictor variables including measures on spatial location, bioregion, land cover, soil, precipitation, and number of dams in local catchments. We trained the model with a random 75% subset of Chessie BIBI data ( n = 1449), and used the remaining 25% of Chessie BIBI data ( n = 484) as test data. The model performed well, correctly predicting 72% of samples in training data and 73% of samples in test data, but model accuracy varied among bioregions. We performed uncertainty analyses by adding bands of either ±0.05 or ±0.10 BIBI units to the cutoff between poor and fair/good. These uncertainty analyses resulted in 14.5% (±0.05 band) and 24.8% (±0.10 band) of samples in test data being classified as in uncertain condition. For 95,877 small stream reaches in the Chesapeake Bay watershed, the model predicted 64% in fair/good condition, the ±0.05 uncertainty analyses predicted 57% in fair/good condition, and the ±0.10 uncertainty analysis predicted 50% in fair/good condition. These reported values have different implications for the number of improved stream miles required to meet the goal of improving 10%. Incorporating uncertainty provides an assessment of model strength as well as confidence in predictions. We, therefore, suggest increased reporting of uncertainty in studies that spatially predict stream conditions.

Chesapeake Bay watershed

Phylogeny and foraging mode correspond with thiaminase activity in freshwater fishes: Potential links to environmental factors

Knowledge of the dietary components of fish species is important for understanding their growth, survival, and recruitment. Deficiency in thiamine (vitamin B1) leading to reproductive failure and physiological illness among freshwater fishes has been attributed to thiaminase activity in fish in the Great Lakes and the New York Finger Lakes, but the causes of variation in thiaminase activity among freshwater fishes is unclear. We characterized thiaminase activity in 29 species of freshwater fishes across 7 ray-finned and 1 jawless family. All fish were further categorized by phylogeny, trophic category (trophic level and feeding mode), and native or non-native status to evaluate how ecological processes correspond with thiaminase activity. Thiaminase activity varied significantly across species, families, trophic factors, phylogenetic groups, and sites. Teleosts that were more recently derived had higher thiaminase activity than more basal species. Thiaminase activity was also higher among herbivores than omnivores or carnivores. This trend was clearest in the Cyprinidae family, which had the greatest range in thiaminase activity and a wide range in trophic-level position and trophic categories (herbivores, omnivores, and carnivores). Variation in average thiaminase activity of Spotfin Shiners (Cyprinella spiloptera) among sites within a watershed was correlated with anthropogenic and natural components of land cover. Our study contributes much-needed quantitative ecological information related to thiaminase activity in a suite of fish species that vary in evolutionary history, trophic level, and foraging modes. However, more studies are needed to identify interacting causes of thiaminase variation and examine the implications of these findings on the overall health of aquatic populations and freshwater ecosystems.

Freshwater Science

Local scale spatial patterns of freshwater mussels in the Upper Mississippi River

Multiple physical and biological factors contribute to the structure of freshwater mussel communities in large rivers. Mussel distributions are frequently described as clumped or patchy. However, few surveys of mussel populations have been designed to quantify these spatial patterns. We used indicators of spatial autocorrelation to quantify spatial patterns of adult and juvenile (≤ 5 years of age) freshwater mussels at local scales (i.e., < 300 m), within 14 sites along ~700 km of the Upper Mississippi River, USA. Juveniles were patchily distributed in 43% of the sites, and adults were patchily distributed in 50%. Within sites, juveniles and adults displayed the same spatial pattern in 64% of the sites. At half of the sites, hotspots of adults and juveniles overlapped suggesting spatial and temporal persistence of habitat. Mussels have an important ecological role in rivers, and information on their spatial patterns aid our understanding of the spatial structure and function of riverine ecosystems, and the ecological consequences of population declines.

Illinois, Iowa, Minneapolis, Wisconsin

Applying the ecology of aquatic–terrestrial linkages to freshwater and riparian management

Global stressors such as climate change, invasive species, urbanization, agricultural practices, and pollution can alter aquatic resource subsidies to terrestrial consumers. The effects of these stressors on timing, quality, and quantity of aquatic subsidies, such as adult aquatic insects, to birds, herpetofauna, and mammals, have large implications for wildlife management (Baxter et al. 2004 , Saunders and Fausch 2007 , Walters et al. 2008 , Sullivan and Rodewald 2012 , Morrissey et al. 2015 , Kraus et al. 2016 , Larsen et al. 2016 , Sullivan et al. 2019 ). For example, insect-mediated contaminant transport from polluted rivers expose song bird nestlings and other protected birds to potentially toxic levels of persistent organic contaminants and pharmaceuticals (Walters et al. 2010 , Richmond et al. 2018 ). Recent declines in aquatic insect production caused by pollution and changes in land use have been tied to global declines in terrestrial insectivores such as birds and bats (Hallmann et al. 2014 , Morrissey et al. 2015 , Raby et al. 2018 ). These natural-resource impacts from aquatic–terrestrial exposure and loss of resource subsidies are leading to changes in monitoring protocols and how management agencies evaluate the effectiveness of corrective remedies (Muehlbauer et al. 2019 ). For researchers interested in the application of resource subsidy research, a logical next step is to help practitioners anticipate effects of global stressors on aquatic-terrestrial linkages and incorporate these principles into decision making.

Freshwater Science

Contaminants in linked aquatic–terrestrial ecosystems: Predicting effects of aquatic pollution on adult aquatic insects and terrestrial insectivores

Organisms that move across ecosystem boundaries connect food webs in apparently disparate locations. As part of their life cycle, aquatic insects transition from aquatic larvae to terrestrial adults, thereby linking freshwater ecosystem processes and terrestrial insectivore dynamics. These linkages are strongly affected by contamination of freshwater ecosystems, which can reduce production of adult aquatic insects (i.e., emergence), increase contaminant concentrations in adult insect tissues, and alter contaminant flux to terrestrial ecosystems. Despite the potential impact of contaminants on adult aquatic insects, little is known about predicting these effects. Here, I develop a heuristic model based on contaminant properties and ecotoxicological principles to predict the effects of various classes of aquatic contaminants on adult aquatic insects and discuss implications for terrestrial insectivores living near contaminated freshwaters. The main finding is that contaminant classes vary greatly in how their biologically-mediated effects on aquatic insects affect terrestrial insectivores. Highly bioaccumulative contaminants that are well retained during metamorphosis, like polychlorinated biphenyls (PCBs), are often non-toxic to aquatic insect larvae at concentrations commonly found in the environment. Such contaminants flux from aquatic ecosystems in large quantities in the bodies of emerging adult aquatic insects and expose terrestrial insectivores to toxic levels of pollution. On the other hand, contaminants that are less bioaccumulative, excreted during metamorphosis, and more toxic to insects, like trace metals, tend to affect terrestrial insectivores by reducing production of adult aquatic insects on which they prey. Management applications of this model illustrate type and severity of risk of aquatic contaminants to consumers of adult aquatic insects.

Freshwater Science

Diatom enumeration method influences biological assessments of southeastern USA streams

Current fixed-count enumeration methods for benthic diatoms are likely inadequate for most research and monitoring objectives. These methods underestimate taxa richness and may fail to detect losses of species caused by human impacts. Consequently, the full potential of diatoms is not realized in current assessments of biological integrity or species diversity. In this study, we hypothesize that alternative enumeration methods differ in their ability to quantify species composition. Furthermore, we hypothesize that an alternative to the traditional fixed-count method will improve both performance of observed/expected (O/E) indices derived from River Inver- tebrate Prediction and Classification System models and the discrimination of reference-quality and human-modified sites by other standard metrics used in biological assessments. To test these hypotheses, we assessed 1) how well 3 counting methods characterized species richness in a subset of 15 samples of stream benthic diatoms and 2) how counting method affected the performance of O/E indices and metrics by comparing the traditional fixed- count method against the best-performing alternative method. These latter comparisons were based on samples collected from 68 reference-quality streams and 20 streams located along an urban disturbance gradient. We dem- onstrate that traditional fixed counts failed to detect >1⁄2 of species present in most of the 68 reference-quality sites. Instead, timed-presence data produced the O/E index with the best performance and a level of precision similar to published invertebrate O/E indices. Furthermore, the O/E index based on the timed-presence data allowed us to determine which species are most often lost with urbanization. We found that traditional fixed-count and alter- native timed-presence data produce metrics that are nearly equally able to discriminate between reference and dis- turbed sites. This study demonstrates that alternative counting methods improve species detection and require up to ∼30% less effort.

Arkansas, Alabama, Georgia, Louisiana, Maryland, M

Spatial population structure of a widespread aquatic insect in the Colorado River Basin: Evidence for a Hydropsyche oslari species complex

Structural connectivity and dispersal ability are important constraints on functional connectivity among populations. For aquatic organisms that disperse among stream corridors, the regional structure of a river network can, thus, define the boundaries of gene flow. In this study, we used mitochondrial DNA (mtCO1 barcoding gene) to examine the genetic diversity and population structure of a caddisfly with strong dispersal capabilities, Hydropsyche oslari (Trichoptera:Hydropsychidae), in the topologically-diverse Colorado River Basin. We expected to find less genetic differentiation among populations of H . oslari within the Upper Basin, which has a dense dendritic network of perennial tributaries that allow for greater potential dispersal and gene flow, than among populations within the arid and sparse river network of the Lower Basin. We also expected to find genetic differentiation among H. oslari in the Upper and Lower Basins because contemporary populations are geographically distant from each other and have been separated by a >300-km-long reservoir (Lake Powell) for ½ a century. Consistent with these predictions, we found that populations of H . oslari within the Upper Basin had more shared haplotypes and less nucleotide diversity (π = 0.001–0.008) than H . oslari within the Lower Basin ( F ST = 0.01, π = 0.014–0.028). However, populations were genetically more structured in the Upper Basin ( F ST = 0.47) than in the Lower Basin ( F ST = 0.01). We also found that populations in the Upper and Lower Basin are entirely genetically differentiated ( S nn = 1), suggesting that these 2 populations were isolated thousands of years before the 1963 closure of Glen Canyon Dam and subsequent filling of Lake Powell. The most similar haplotypes among the 2 basins represent a 5.4% difference, which indicates the presence of a species complex within H. oslari .

Colorado River Basin