Karen A. Blocksom

METHODS DEVELOPMENT AND USE OF MACROINVERTEBRATES AS INDICATORS OF ECOLOGICAL CONDITIONS FOR STREAMS IN THE MID-ATLANTIC HIGHLANDS REGION

The Mid-Atlantic Highlands Assessment (MAHA) included the sampling of macroinvertebrates from 424 wadeable stream sites to determine status and trends, biological conditions, and water quality in first through third order streams in the Mid-Atlantic Highlands Region (MAHR) of the United States in 1993–1995. We identified reference and impaired sites using water chemistry and habitat criteria and evaluated a set of candidate macroinvertebrate metrics using a stepwise process. This process examined several metric characteristics, including ability of metrics to discriminate reference and impaired sites, relative scope of impairment, correlations with chemical and habitat indicators of stream disturbance, redundancy with other metrics, and within-year variability. Metrics that performed well were compared with metrics currently being used by three states in the region: Pennsylvania, Virginia, and West Virginia. Some of the metrics used by these states did not perform well when evaluated using regional data, while other metrics used by all three states in some form, specifically number of taxa, number of EPT taxa, and Hilsenhoff Biotic Index, performed well overall. Reasons for discrepancies between state and regional evaluations of metrics are explored. We also provide a set of metrics that, when used in combination, may provide a useful assessment of stream conditions in the MAHR.

Development and evaluation of the Lake Macroinvertebrate Integrity Index (LMII) for New Jersey lakes and reservoirs.

In response to the recent focus by the U.S. EnvironmentalProtection Agency on bioassessment of lakes, a multimetric index was developed for New Jersey lakes and reservoirs using benthicmacroinvertebrates. Benthic samples were collected fromreference and impaired lakes with muck and intermediate sedimentsin central and northern New Jersey during summer 1997. We used astepwise process to evaluate properties of candidate metrics andselected five for the Lake Macroinvertebrate Integrity Index(LMII): Hilsenhoff Biotic Index (HBI), percent chironomidindividuals, percent collector-gatherer taxa, percentoligochaetes/leeches, and number of Diptera taxa. We scoredmetrics as the fraction of the best expected value (based on allsites) achieved at a site and summed them into the LMII. Evaluation of the LMII showed that it discriminated well betweenreference and impaired lakes and was strongly related to severalpotential stressors. Chemical and physical gradients distinguished between reference and impaired lakes, and the LMIIsummarized these gradients well. The LMII corresponded stronglywith land use, but some lakes with more urban land use stillachieved high scores. Based on a power analysis, the ability ofthe LMII to detect differences in condition was sensitive to thenumber of samples from each lake.

Persistent organic pollutants in fish tissue in the mid-continental great rivers of the United States

Great rivers of the central United States (Upper Mississippi, Missouri, and Ohio rivers) are valuable economic and cultural resources, yet until recently their ecological condition has not been well quantified. In 2004-2005, as part of the Environmental Monitoring and Assessment Program for Great River Ecosystems (EMAP-GRE), we measured legacy organochlorines (OCs) (pesticides and polychlorinated biphenyls, PCBs) and emerging compounds (polybrominated diphenyl ethers, PBDEs) in whole fish to estimate human and wildlife exposure risks from fish consumption. PCBs, PBDEs, chlordane, dieldrin and dichlorodiphenyltrichloroethane (DDT) were detected in most samples across all rivers, and hexachlorobenzene was detected in most Ohio River samples. Concentrations were highest in the Ohio River, followed by the Mississippi and Missouri Rivers, respectively. Dieldrin and PCBs posed the greatest risk to humans. Their concentrations exceeded human screening values for cancer risk in 27-54% and 16-98% of river km, respectively. Chlordane exceeded wildlife risk values for kingfisher in 11-96% of river km. PBDE concentrations were highest in large fish in the Missouri and Ohio Rivers (mean>1000 ng g(-1) lipid), with congener 47 most prevalent. OC and PBDE concentrations were positively related to fish size, lipid content, trophic guild, and proximity to urban areas. Contamination of fishes by OCs is widespread among great rivers, although exposure risks appear to be more localized and limited in scope. As an indicator of ecological condition, fish tissue contamination contributes to the overall assessment of great river ecosystems in the U.S.

Temporal Dynamics of Periphyton Exposed to Tetracycline in Stream Mesocosms

Significant amounts of antibiotics enter the environment via point and nonpoint sources. We examined the temporal dynamics of tetracycline exposure to stream periphyton and associated organisms across a logarithmically dosed-series of experimental mesocosms, designed to mimic natural conditions. Target in-stream tetracycline exposures were based on environmentally relevant concentrations in aquatic ecosystems throughout the United States (<1-100 μg L(-1)). Significant changes in the stream biotic community were observed within 7 days with in-stream tetracycline concentrations as low as 0.5 μg L(-1), including significant changes in antibiotic resistance, bacteria abundance and productivity, algae biomass, cyanobacteria, organic biomass, and nematodes. These effects were magnified with increased exposure time and dosing concentration. Recovery of the periphyton community after 28 days of exposure was dependent upon the tetracycline dose. At the highest doses, 10 and 100 μg L(-1), bacteria productivity recovered; however, bacteria, algae, and nematode abundance did not recover at the same rate and remained low even after a 28-day recovery period (of nondosing). This study demonstrates that tetracycline exposure under near-natural conditions and at concentrations currently observed in aquatic environments may have important consequences for the structure and function of stream periphyton and, potentially, public health via increasing resistance of naturally occurring bacteria.

Proteomic analysis of a model fish species exposed to individual pesticides and a binary mixture

Pesticides are nearly ubiquitous in surface waters of the United States, where they often are found as mixtures. The molecular mechanisms underlying the toxic effects of sub-lethal exposure to pesticides as both individual and mixtures are unclear. The current work aims to identify and compare differentially expressed proteins in brains of male fathead minnows (Pimephales promelas) exposed for 72 h to permethrin (7.5 μg/L), terbufos (57.5 μg/L) and a binary mixture of both. Twenty-four proteins were found to be differentially expressed among all three treatments relative to the control using an ANOVA followed by a Dunnetts post hoc test (p ≤0.05). One protein was found to be differentially expressed among all treatment groups and one protein was in common between the terbufos and the mixture group. Fifteen spots were successfully sequenced using LC-MS/MS sequencing. Proteins associated with the ubiquitin-proteasome system, glycolysis, the cytoskeleton and hypoxia were enriched. As a second objective, we attempted to establish protein expression signatures (PES) for individual permethrin and terbufos exposures. We were unable to generate a useable PES for terbufos; however, the permethrin PES was able to distinguish between control and permethrin-exposed individuals in an independent experiment with an accuracy of 87.5%. This PES also accurately classified permethrin exposed individuals when the exposure occurred as part of a mixture. The identification of proteins differentially expressed as a result of pesticide exposure represent a step forward in the understanding of mechanisms of toxicity of permethrin and terbufos. They also allow a comparison of molecular responses of the binary mixture to single exposures. The permethrin PES is the first step in establishing a method to determine exposures in real-world scenarios.

Mercury Contamination in Fish in Midcontinent Great Rivers of the United States: Importance of Species Traits and Environmental Factors

We measured mercury (Hg) concentrations in whole fish from the Upper Mississippi, Missouri, and Ohio Rivers to characterize the extent and magnitude of Hg contamination and to identify environmental factors influencing Hg accumulation. Concentrations were generally lower (80% of values between 20-200 ng g(-1) wet weight) than those reported for other regions (e.g., upper Midwest and Northeast U.S.). Mercury exceeded the risk threshold for belted kingfisher (Ceryle alcyon, the most sensitive species considered) in 33-75% of river length and 1-7% of river length for humans. Concentrations were lower in the Missouri than in the Mississippi and Ohio Rivers, consistent with continental-scale patterns in atmospheric Hg deposition. Body size and trophic guild were the best predictors of Hg concentrations, which were highest in large-bodied top predators. Site geochemical and landscape properties were weakly related with fish Hg. Moreover, relationships often ran contrary to conventional wisdom, and the slopes of the relationships (positive or negative) were inconsistent among fish guilds and rivers. For example, sulfate is positively associated with fish Hg concentrations but was negatively correlated with Hg in five of six regression models of tissue concentrations. Variables such as pH, acid neutralizing capacity, and total phosphorus did not occur at levels associated with high fish Hg concentrations, partially explaining the relatively low Hg values we observed.

Sampling effort needed to estimate condition and species richness in the Ohio river, USA

The level of sampling effort required to characterize fish assemblage condition in a river for the purposes of bioassessment may be estimated via different approaches. However, the goal with any approach is to determine the minimum level of effort necessary to reach some specific level of confidence in the assessment. In the Ohio River, condition is estimated and reported primarily at the level of pools defined by lock and dam structures. The goal of this study was to determine the minimum level of sampling effort required to adequately characterize pools in the Ohio River for the purpose of bioassessment. We followed two approaches to estimating required sampling effort using fish assemblage data from a long-term intensive survey across a number of Ohio River pools. First, we estimated the number of samples beyond which variation in the multimetric Ohio River Fish Index (ORFIn) leveled off. Then, we determined the number of samples necessary to collect approximately 90% of the fish species observed across all samples collected within the pool. For both approaches, approximately 15 samples were adequate to reduce variation in IBI scores to acceptable levels and to capture 90% of observed species in a pool. The results of this evaluation provide a basis not only for the Ohio River Valley Water Sanitation Commission (ORSANCO) but also states and other basin commissions to develop sampling designs for bioassessment that ensure adequate sampling of all assessment units.

An assessment of stressor extent and biological condition in the North American mid-continent great rivers (USA)

We assessed the North American mid-continent great rivers (Upper Mississippi, Missouri, and Ohio). We estimated the extent of each river in most- (MDC) or least-disturbed condition (LDC) based on multiple biological response indicators: fish and macroinvertebrate, trophic state based on chlorophyll a, macrophyte cover, and exposure of fish-eating wildlife to toxic contaminants in fish tissue (Hg, total chlordane, total DDT, PCBs). We estimated the extent of stressors on each river including nutrients, suspended solids, sediment toxicity, invasive species, and land use (agriculture and impervious surface). All three rivers had a greater percent of their river length in MDC than in LDC based on fish assemblages. The Upper Mississippi River had the greatest percent of river length with eutrophic status. The Ohio River had the greatest percent of river length with fish with tissue contaminant levels toxic to wildlife. Overall, condition indices based on fish assemblages were more sensitive to stress than macroinvertebrate indices. Compared to the streams in its basin, more of the Upper Mississippi and Missouri Rivers were in MDC for nutrients than the Ohio River. Invasive species (Asian carp and Dreissenid mussels) were less widespread and less abundant on the Missouri River than on the other great rivers. The Ohio River had the most urbanized floodplains (greatest percent impervious surface). The Missouri River had the most floodplain agriculture. The effect of large urban areas on river condition was apparent for several indicators. Ecosystem condition based in fish assemblages, trophic state, and fish tissue contamination was related to land use on the floodplain and at the subcatchment scale. This is the first unbiased bioassessment of the mid-continent great rivers in the United States. The indicators, condition thresholds, results, and recommendations from this program are a starting point for improved future great river assessments.

Evaluation of an alternate method for sampling benthic macroinvertebrates in low-gradient streams sampled as part of the National Rivers and Streams Assessment

Benthic macroinvertebrates are sampled in streams and rivers as one of the assessment elements of the US Environmental Protection Agency’s National Rivers and Streams Assessment. In a 2006 report, the recommendation was made that different yet comparable methods be evaluated for different types of streams (e.g., low gradient vs. high gradient). Consequently, a research element was added to the 2008–2009 National Rivers and Streams Assessment to conduct a side-by-side comparison of the standard macroinvertebrate sampling method with an alternate method specifically designed for low-gradient wadeable streams and rivers that focused more on stream edge habitat. Samples were collected using each method at 525 sites in five of nine aggregate ecoregions located in the conterminous USA. Methods were compared using the benthic macroinvertebrate multimetric index developed for the 2006 Wadeable Streams Assessment. Statistical analysis did not reveal any trends that would suggest the overall assessment of low-gradient streams on a regional or national scale would change if the alternate method was used rather than the standard sampling method, regardless of the gradient cutoff used to define low-gradient streams. Based on these results, the National Rivers and Streams Survey should continue to use the standard field method for sampling all streams.

An interlaboratory comparison of sediment elutriate preparation and toxicity test methods

Elutriate bioassays are among numerous methods that exist for assessing the potential toxicity of sediments in aquatic systems. In this study, interlaboratory results were compared from 96-h Ceriodaphnia dubia and Pimephales promelas static-renewal acute toxicity tests conducted independently by two laboratories using elutriate samples prepared from the same sediment. The goal of the study was to determine if the results from the elutriate tests were comparable between two U.S. Environmental Protection Agency (USEPA) laboratories when different elutriate preparation procedures were employed by each lab. Complete agreement in site characterization was attained in 22 of the 25 samples for both bioassays amongst each lab. Of the 25 samples analyzed, 10 were found to be toxic to at least one of the species tested by either laboratory. The C. dubia elutriate tests conducted by the National Exposure Research Laboratory (NERL) indicated that 7 of the 25 sediment samples were toxic, while 8 sediment samples were characterized as such in testing conducted by USEPA Region 6 (Region 6). The P. promelas elutriate tests conducted by NERL determined 8 samples as toxic, while Region 6 tests displayed toxicity in 5 of the samples. McNemars test of symmetry for C. dubia (S = 0.33, p = 0.5637) and P. promelas (S = 3.0, p = 0.0833) tests indicated no significant differences in designating a site toxic between NERL and Region 6 laboratories. Likewise, Cohens kappa test revealed significant agreement between NERL and Region 6 C. dubia (K = 0.7148, p < 0.01) and P. promelas (K = 0.6939, p < 0.01) elutriate tests. The authors conclude that differences in interlaboratory elutriate preparation procedures have no bearing on the ability of either the C. dubia or P. promelas bioassay testing methods to detect toxicity while yielding similar results.