Ryan R. Otter

THE DARK SIDE OF SUBSIDIES: ADULT STREAM INSECTS EXPORT ORGANIC CONTAMINANTS TO RIPARIAN PREDATORS

Aquatic insects provide a critical energy subsidy to riparian food webs, yet their role as vectors of contaminants to terrestrial ecosystems is poorly understood. We investigated relationships between aquatic resource utilization and contaminant exposure for a riparian invertivore assemblage (spiders and herptiles) along a stream contaminated with polychlorinated biphenyls (PCBs). Stable carbon (delta13C) and nitrogen (delta15N) isotopes indicated that aquatic insect utilization varied among predators, with progressive enrichment of delta13C and depletion of delta15N as predators shifted from aquatic to terrestrial prey. PCB concentrations significantly increased along these isotopic gradients; delta13C and delta15N explained 65% and 15% of the variance in predator sigmaPCBs, respectively. PCBs in predators were high, exceeding 2000 ng/g wet mass (the human-health advisory prohibiting any consumption of fish tissue) in three species. Greater consideration should be given to streams as lateral exporters rather than simply as longitudinal conduits for contaminants. Persistent contaminants are underutilized for addressing landscape-level questions in subsidy research, but our results demonstrate they are an ideal in situ tracer of stream-derived energy because they label stream organic matter and invertebrates over large distances. Likewise, riparian predators such as tetragnathid spiders have great potential as biological monitors of stream condition and as an assessment tool for risk management of contaminated aquatic sediments.

Mercury toxicity in livers of northern pike (Esox lucius) from Isle Royale, USA

Many laboratory studies have documented that mercury can be toxic to fish, but it is largely unknown if mercury is toxic to fish in their natural environments. The objective of our study was to investigate the toxic effects of mercury on northern pike (Esox lucius) at Isle Royale, Michigan. In 124 northern pike from eight inland lakes, concentrations of total mercury in skin-on fillets ranged from 0.069 to 0.622 microg/g wet mass (wet wt). Concentrations of total mercury in livers increased exponentially compared with concentrations in fillets, to a maximum of 3.1 microg/g wet wt. Methylmercury constituted a majority of the mercury in livers with total mercury concentrations <0.5 microg/g wet wt, but declined to 28-51% of the mercury in livers with total mercury concentrations >0.5 microg/g wet wt. Liver color (absorbance at 400 nm) varied among northern pike and was positively related to liver total mercury concentration. The pigment causing variation in liver color was identified as lipofuscin, which results from lipid peroxidation of membranous organelles. An analysis of covariance revealed lipofuscin accumulation was primarily associated with mercury exposure, and this association obscured any normal accumulation from aging. We also documented decreased lipid reserves in livers and poor condition factors of northern pike with high liver total mercury concentrations. Our results suggest (i) northern pike at Isle Royale are experiencing toxicity at concentrations of total mercury common for northern pike and other piscivorous fish elsewhere in North America and (ii) liver color may be useful for indicating mercury exposure and effects in northern pike at Isle Royale and possibly other aquatic ecosystems and other fish species.

Effects of silver nanoparticles on zebrafish (Danio rerio) and Escherichia coli (ATCC 25922): a comparison of toxicity based on total surface area versus mass concentration of particles in a model eukaryotic and prokaryotic system.

Silver nanoparticles (Ag NPs) have been classified as the most abundant NP found in commercial products. In the present study, zebrafish (Danio rerio) and bacteria (Escherichia coli; ATCC 25922) were used to test the size-dependent toxicological effects of Ag NPs, the effects of ionic silver versus Ag NPs, and Ag NP effects on mortality using mass concentration (mg/L) compared with total surface area (nm(2) /L). Several diameters of Ag NPs (20, 50, 110 nm) as well as AgNO(3) were chosen as experimental treatments. Treated zebrafish embryos exhibited anomalies of the heart, namely, slower heart rates and pericardial edema. A size-dependent response was not observed in zebrafish when viewing mortality across all Ag NP treatments, although 20 nm elicited the highest incidence of abnormal motility and induced slower development. An Ag NP dose- and size-dependent response was observed in treated bacteria using mass concentration, with 20-nm Ag NP producing the highest mortality rate. In both zebrafish and bacteria, AgNO(3) was shown to be more toxic than Ag NPs at equivalent concentrations. When total surface area of Ag NPs was used to gauge bacterial mortality, a total surface area-dependent, but not size-dependent, response was observed for all three Ag NPs used in the present study, with nearly 100% mortality observed once a total surface area of approximately 1E + 18 nm(2) /L was reached. This trend was not apparent, however, when measuring total surface area for zebrafish mortality.

A BIOMARKER APPROACH TO MEASURE BIOLOGICAL EFFECTS OF CONTAMINANT EXPOSURE IN LARGEMOUTH BASS FROM LAKE CONESTEE, SOUTH CAROLINA, USA

Sediments from Lake Conestee, a former reservoir now filled with pollutant-enriched sediments, located south of Greenville, South Carolina, USA, and other nearby reservoirs were collected and analyzed for lead and polycyclic aromatic hydrocarbons (PAHs). Hepatic ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), UDP-glucuronosyltransferase (UGT), sulfotransferase (SULT), and erythrocyte delta-aminolevulinic acid dehydratase (ALAD) were measured in largemouth bass (Micropterus salmoides) to characterize biological effects of these contaminants over three seasons. Results showed that total PAH concentrations in Lake Conestee sediments were significantly greater than the control during each season. An average 10-fold induction in EROD activity was observed at Lake Conestee compared with the control over all three seasons, indicating that PAHs present in sediment were bioavailable to fish. Significant gender effects were observed in EROD activity during the spring, in which activity in reproductively active female fish was significantly suppressed compared with the male fish. Sediments from Lake Conestee had elevated lead concentrations, but the lack of ALAD inhibition in bass indicated that lead was not biologically available. Total GST activity, UGT activity, and SULT activity were not significantly induced in fish from any of the affected sites compared with the reference site. Both EROD and UGT activities were highest during the winter, as were sediment PAH concentrations in Lake Conestee, possibly linked to seasonal resuspension events. The biomarkers measured in this study were useful as a first investigation into the biological effects of contaminant exposure, as well as in determining the bioavailability of contaminants in Lake Conestee.

The use of tetragnathid spiders as bioindicators of metal exposure at a coal ASH spill site

On 22 December 2008, a dike containing coal fly ash from the Tennessee Valley Authority Kingston Fossil Fuel Plant (TN, USA) failed, resulting in the largest coal ash spill in US history. The present study was designed to determine sediment metal concentrations at multiple site locations and to determine whether site-specific bioaccumulation of metals existed in tetragnathid spiders. Selenium and nickel were the only 2 metals to exceed the US Environmental Protection Agency sediment screening levels. Selenium concentrations in spiders were significantly higher at ash-affected sites than in those from reference sites. The ratio of methylmercury to total mercury in spiders was found to be similar to that in other organisms (65-75%), which highlights the potential use of tetragnathid spiders as an indicator species for tracing contaminant transfer between the aquatic and terrestrial ecosystems.

Trophic status and metal bioaccumulation differences in multiple fish species exposed to coal ash-associated metals.

On December 22, 2008 a dike containing coal fly ash from the Tennessee Valley Authority Kingston Fossil Plant near Kingston Tennessee USA failed and resulted in the largest coal ash spill in U.S. history. Coal ash, a by-product of coal combustion, is known to contain multiple contaminants of concern, including arsenic and selenium. The purpose of this study was to investigate species differences in the bioaccumulation of arsenic and selenium and potential factors contributing to these differences (i.e., trophic dynamics and gut pH) in the vicinity of the Kingston coal ash spill. Elevated levels of arsenic and selenium were observed in various tissues of largemouth bass, white crappie, bluegill and redear sunfish from sites associated with the Kingston coal ash spill. Highest concentrations of selenium were found in redear sunfish with liver concentrations as high as 24.83mg/kg dry weight and ovary concentrations up to 10.40mg/kg dry weight at coal ash-associated sites. Investigations into the gut pH and trophic dynamics of redear sunfish and bluegill demonstrated a large difference in the gut physiology between these two species. Redear sunfish stomach and intestinal pH was found to be 1.1 and 0.16 pH units higher than in bluegill, respectively. In addition, fish from coal ash-associated sites showed enrichment differences ((15)N and (13)C) compared to no ash sites, indicating differences in food web dynamics between sites. These results imply the incorporation of coal ash-associated compounds into local food webs and/or a shift in diet at ash sites compared to the no ash reference sites. Based on these results, further investigation into a broader food web at ash-associated sites is warranted.

Consistency of morphological endpoints used to assess developmental timing in zebrafish (Danio rerio) across a temperature gradient

Zebrafish (Danio rerio) are model organisms for testing developmental toxicity at the morphological level. In this study, influence of temperature (24.5-28.5°C) and silver nanoparticles on developmental staging, ear-eye distance, and ratio of ear-eye distance to inner ear diameter was investigated. As temperature decreased, all endpoints showed developmental delay, with differences between endpoints in amount and type of delay measured. Differences in developmental delay patterns were observed, with rate delays increasing over time when staging endpoints were utilized and rates remaining constant when using ear-eye measurements. Integrated predictive equations were created to normalize each endpoint for temperature. Influence of image rotation on ear-eye distance accuracy showed that more than 75% eye overlap during analysis is necessary to minimize error. Exposure to silver nanoparticles demonstrated a lack of consistency between developmental endpoints and highlighted the usefulness of a multi-endpoint approach when measuring changes to developmental timing.

The effects of urbanization on Lepomis macrochirus using the comet assay.

Urbanization has been linked to increased concentrations of polycyclic aromatic hydrocarbons in natural waterways. This study was designed to examine the impact of urbanization and a wastewater treatment plant by investigating the impact on field-collected bluegill (Lepomis macrochirus). Results show a significant increase in DNA strand breaks in blood cells (comet assay) linked to urbanization and a reduction in DNA strand breaks downstream of the WWTP, likely the result of dilution. A laboratory study exposing L. macrochirus to the known mutagen, methyl methanesulfonate, was performed to validate the comet assay endpoints in this species. Results of the laboratory study showed that the comet assay endpoints of tail length and tail extent moment responded in a dose- and time-dependent manner. Habitat quality assessments, along with chemical concentrations of polycyclic hydrocarbons in sediments showed that habitat quality between all sites were similar and that hydrocarbons likely contributed to the DNA strand breaks observed.

Stepwise Information-Filtering Tool (SIFT): A method for using risk assessment metadata in a nontraditional way

Tools exist to evaluate large ecotoxicity databases for risk assessment purposes, but these tools are less useful for alternative analytical purposes. In the present study, the authors developed the Stepwise Information-Filtering Tool (SIFT), a strategic method to select relevant, reliable data from a large ecotoxicity database; demonstrated utility in a case study of chronic toxicity data for statistical endpoint comparison purposes; and evaluated SIFT by comparison with 2 existing data evaluation methods.

A molecular method for assessing the effects of potential contaminants on the rate of zebrafish (Danio rerio) development

Monitoring expression of the developmentally regulated genes shh, sox2, and tnnt1 by reverse transcriptase polymerase chain reaction (RT-PCR) allows determination of the rate of embryogenesis in zebrafish (Danio rerio) embryos without direct visual observation. The utility of combining this approach and morphological methods during toxicity studies was demonstrated with embryos developing at either 28.5 °C or 24.5 °C and with embryos exposed to sublethal doses of silver nanoparticles.