Effects of Military activity and habitat quality on DNA damage and oxidative stress in the largest population of the Federally threatened gopher tortoise.

Department of Defense lands are essential for providing important habitat for threatened, endangered, and at-risk species (TER-S). However, there is little information on the effects of military-related contaminants on TER-S on these lands in field situations. Thus, this study examined genotoxicity and oxidative stress in gopher tortoises (Gopherus polyphemus) on Camp Shelby, MS-the largest known population of this species, which is listed as an "endangered species" in Mississippi and a "threatened species" by the U.S. government. Blood was collected from tortoises at 19 different sites on the base with different levels of habitat quality (high-quality and low-quality habitat) and military activity (high, low, and no military activity). Oxidative stress was quantified as lipid peroxidation and GSSG/GSH ratios, while DNA damage was determined using flow cytometry. Our results suggest that: (1) for tortoises residing in low-quality habitats, oxidative stress and DNA damage increased with increasing military activity, while in high-quality habitats, oxidative stress and DNA damage decreased with increasing military activity; (2) in the absence of military activity, tortoises in high-quality habitat had higher levels of oxidative stress and DNA damage than those in low-quality habitat, and (3) there were interactions between military activity, habitat quality, and landuse in terms of the amount of observable DNA damage and oxidative stress. In particular, on high-quality habitat, tortoises from areas with high levels of military activity had lower levels of oxidative stress and DNA damage biomarkers than on reference sites. This may represent a compensatory or hormetic response. Conversely, on low-quality habitats, the level of oxidative stress and DNA damage was lower on the reference sites. Thus, tortoises on higher-quality habitats may have a greater capacity for compensatory responses. In terms of management implications, it is suggested that low quality habitats should be a higher priority for remediation, and lower priority for conducting military activities.

Influence of metal(loid) bioaccumulation and maternal transfer on embryo-larval development in fish exposed to a major coal ash spill.

In December 2008, an earthen retaining wall at the Tennessee Valley Authority (TVA) Kingston Fossil Fuel Plant failed and released 4.1 millionm(3) of coal ash to rivers flowing into Watts Bar Reservoir in east Tennessee, United States (U.S.). As part of a comprehensive effort to evaluate the risks to aquatic resources from this spill - the largest in U.S. history - we compared bioaccumulation and maternal transfer of selenium (Se), arsenic (As), and mercury (Hg) in adult redear sunfish (Lepomis macrolophus), collected two years after the spill from both coal-ash exposed and non-exposed areas of the Emory and Clinch Rivers, with the success of embryo-larval development in their offspring. Whole body and ovary concentrations of Se in female sunfish at three study sites downstream of the spill were significantly elevated (site means=4.9-5.3 and 6.7-9.0mg/kg d.w. whole body and ovary concentrations, respectively) compared with concentrations in fish from reference sites upstream of the spill site (2.2-3.2mg/kg d.w. for whole bodies and 3.6-4.8mg/kg d.w. for ovaries). However, Se concentrations in coal ash-exposed areas remain below proposed U.S. Environmental Protection Agency (USEPA) criteria for the protection of aquatic life. Site-to-site variation in fish concentrations of As and Hg were not well-correlated with ash-exposure, reflecting the multiple sources of these metal(loid)s in the affected watersheds. In 7-day laboratory tests of embryos and larvae derived from in vitro crosses of eggs and sperm from these field-collected sunfish, fertilization success, hatching success, embryo-larval survival, and incidences of developmental abnormalities did not differ significantly between ash-exposed and non-exposed fish. Furthermore, these developmental endpoints were not correlated with whole body or ovary concentrations of Se, As, or Hg in the maternal fish, or with fish size, ovary weight, or gonadal-somatic indices. Results from this and related studies associated with the Kingston coal ash spill are consistent with proposed USEPA fish-based water quality criteria for Se, and to date continue to suggest that long-term exposures to sediment containing residual ash may not present a significant chronic risk to fish populations exposed to this major coal ash release.

Using ordination and clustering techniques to assess multimetric fish health response following a coal ash spill.

The effect of coal ash exposure on fish health in freshwater communities is largely unknown. Given the large number of possible pathways of effects (e.g., toxicological effect of exposure to multiple metals, physical effects from ash exposure, and food web effects), measurement of only a few health metrics is not likely to give a complete picture. The authors measured a suite of 20 health metrics from 1100+ fish collected from 5 sites (3 affected and 2 reference) near a coal ash spill in east Tennessee over a 4.5-yr period. The metrics represented a wide range of physiological and energetic responses and were evaluated simultaneously using 2 multivariate techniques. Results from both hierarchical clustering and canonical discriminant analyses suggested that for most species × season combinations, the suite of fish health indicators varied more among years than between spill and reference sites within a year. In a few cases, spill sites from early years in the investigation stood alone or clustered together separate from reference sites and later year spill sites. Outlier groups of fish with relatively unique health profiles were most often from spill sites, suggesting that some response to the ash exposure may have occurred. Results from the 2 multivariate methods suggest that any change in the health status of fish at the spill sites was small and appears to have diminished since the first 2 to 3 yr after the spill.

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.

Long-term biological monitoring of an impaired stream: synthesis and environmental management implications.

The long-term ecological recovery of an impaired stream in response to an industrial facility's pollution abatement actions and the implications of the biological monitoring effort to environmental management is the subject of this special issue of Environmental Management. This final article focuses on the synthesis of the biological monitoring program's components and methods, the efficacy of various biological monitoring techniques to environmental management, and the lessons learned from the program that might be applicable to the design and application of other programs. The focus of the 25-year program has been on East Fork Poplar Creek, an ecologically impaired stream in Oak Ridge, Tennessee with varied and complex stressors from a Department of Energy facility in its headwaters. Major components of the long-term program included testing and monitoring of invertebrate and fish toxicity, bioindicators of fish health, fish contaminant accumulation, and instream communities (including periphyton, benthic macroinvertebrate, and fish). Key parallel components of the program include water chemistry sampling and data management. Multiple lines of evidence suggested positive ecological responses during three major pollution abatement periods. Based on this case study and the related literature, effective environmental management of impaired streams starts with program design that is consistent across space and time, but also adaptable to changing conditions. The biological monitoring approaches used for the program provided a strong basis for assessments of recovery from remedial actions, and the likely causes of impairment. This case study provides a unique application of multidisciplinary and quantitative techniques to address multiple and complex regulatory and programmatic goals, environmental stressors, and remedial actions.

Application of biochemical and physiological indicators for assessing recovery of fish populations in a disturbed stream.

Recovery dynamics in a previously disturbed stream were investigated to determine the influence of a series of remedial actions on stream recovery and to evaluate the potential application of bioindicators as an environmental management tool. A suite of bioindicators, representing five different functional response groups, were measured annually for a sentinel fish species over a 15 year period during which a variety of remedial and pollution abatement actions were implemented. Trends in biochemical, physiological, condition, growth, bioenergetic, and nutritional responses demonstrated that the health status of a sentinel fish species in the disturbed stream approached that of fish in the reference stream by the end of the study. Two major remedial actions, dechlorination and water flow management, had large effects on stream recovery resulting in an improvement in the bioenergetic, disease, nutritional, and organ condition status of the sentinel fish species. A subset of bioindicators responded rather dramatically to temporal trends affecting all sites, but some indicators showed little response to disturbance or to restoration activities. In assessing recovery of aquatic systems, application of appropriate integrative structural indices along with a variety of sensitive functional bioindicators should be used to understand the mechanistic basis of stress and recovery and to reduce the risk of false positives. Understanding the mechanistic processes involved between stressors, stress responses of biota, and the recovery dynamics of aquatic systems reduces the uncertainty involved in environmental management and regulatory decisions resulting in an increased ability to predict the consequences of restoration and remedial actions for aquatic systems.

The role of periphyton in mediating the effects of pollution in a stream ecosystem.

The effects of pollutants on primary producers ramify through ecosystems because primary producers provide food and structure for higher trophic levels and they mediate the biogeochemical cycling of nutrients and contaminants. Periphyton (attached algae) were studied as part of a long-term biological monitoring program designed to guide remediation efforts by the Department of Energy's Y-12 National Security Complex on East Fork Poplar Creek (EFPC) in Oak Ridge, Tennessee. High concentrations of nutrients entering EFPC were responsible for elevated periphyton production and placed the stream in a state of eutrophy. High rates of primary production at upstream locations in EFPC were associated with alterations in both invertebrate and fish communities. Grazers represented >50% of the biomass of invertebrates and fish near the Y-12 Complex but <10% at downstream and reference sites. An index of epilithic periphyton production accounted for 95% of the site-to-site variation in biomass of grazing fish. Analyses of heavy metals in EFPC periphyton showed that concentrations of zinc, cadmium, copper and nickel in periphyton decreased exponentially with distance downstream from Y-12. Zinc uptake by periphyton was estimated to reduce the concentration of this metal in stream water approximately 60% over a 5-km reach of EFPC. Management options for mitigating eutrophy in EFPC include additional reductions in nutrient inputs and/or allowing streamside trees to grow and shade the stream. However, reducing periphyton growth may lead to greater downstream transport of contaminants while simultaneously causing higher concentrations of mercury and PCBs in fish at upstream sites.

Assessing effects of stress across levels of biological organization using an aquatic ecosystem health index.

Multimetric approaches typically consider only one level of biological organization to assess the effects of environmental stressors on the health of aquatic ecosystems. The present study applied integrative star-plot analysis to evaluate effects of stressors over several levels of biological organization ranging from the sub-organism to the community level at study sites subjected to different levels of contaminant stress. An aquatic ecosystem health index (AEHI), based on the sum of all the star-plot areas over these levels of biological organization, was developed to reflect an integrative and holistic assessment of stressors on ecosystem health. Star-plot areas ranged from 1.96 at a reference site to 0.79-1.08 at sites located at increasing distances downstream from a pulp mill discharge. The values of the AEHI were positively correlated with the index of biotic integrity (IBI) scores (Pearson's r=0.824). The AEHI can be used to evaluate the overall health status of aquatic systems, to identify those levels of biological organization that have the greatest response to environmental stressors, and to help identify possible causes of observed effects. Results from this study suggest that the AEHI approach can avoid false-negative responses that can occur from assessing effects at only one level of biological organization.

Evidence of altered gene flow, mutation rate, and genetic diversity in redbreast sunfish from a pulp-mill-contaminated river.

To determine effects of pulp mill effluent on population genetic structure, redbreast sunfish (Lepomis auritus) were collected from several sites along the Pigeon River, NC, as well as from reference sites. Previous studies found effects on molecular, biochemical, physiological, population, and community level endpoints in these populations. The population genetic structure of these fish was determined using the randomly amplified polymorphic DNA (RAPD) technique. The level of genetic diversity was higher in the Pigeon River populations than in the reference populations. Genetic distances among populations could not be explained by drainage patterns and may have been altered by contaminant exposure. Phylogeographic analysis, maximum likelihood analysis, and assignment tests suggested that there were fewer emigrants and more immigrants in the contaminated sites than in the reference sites, suggesting that the contaminated sites may harbor "sinklike" populations. Finally, a "terminal branch amplitype" analysis (neighbor-joining and minimum-spanning trees) and maximum likelihood analysis indicated that there may be an elevated mutation rate in the polluted sites. Thus, the genetic diversity (within and among populations) in the Pigeon River populations may have been affected by altered gene flow and mutational processes as a result of pulp mill effluent discharge.

Assessing cause and effect of multiple stressors on marine systems.

An operational framework is developed to serve as a guideline for investigating causal relationships between environmental stressors and effects on marine biota. Because of the complexity and variability of many marine systems, multiple lines of evidence are needed to understand relationships between stressors and effects on marine resources. Within this framework, a weight of evidence approach based on multiple lines of evidence are developed and applied in a sequential manner by (1) characterizing the study system which involves determining if target biota are impaired, assessment of food and habitat availability, and measuring contaminant levels in the environment, (2) assessing direct effects of contaminant exposure on target biota using biomarkers and assessing indirect effects of exposure using suites of bioindicators, and (3) applying standard causal criteria based on epidemiological principles and diagnostic health profiling techniques to assess potential causes. Use of multiple lines of evidence should also reduce the risk of false positives (Type I error or falsely concluding that there is a causal relationship when there is none) and false negatives (Type II error or falsely concluding there is not a causal relationship when there actually is). Understanding causal relationships and the mechanistic processes between environmental stressors and effects on biota is important in the effective management and restoration of impaired marine ecosystems.