Legacy Contaminants in Aquatic Biota in a Stream Associated with Nuclear Weapons Material Production on the Savannah River Site.

Former nuclear weapons material production at the U.S. Department of Energy's Savannah River Site (SRS) has resulted in contamination of certain terrestrial and aquatic ecosystems on site with legacy wastes such as radiocesium (137Cs), tritium (3H), and metals. We collected fish and invertebrates from five beaver ponds (sites) above, adjacent, and downgradient of three SRS facilities (H-, F-, and C-Areas) to evaluate whether the accumulation of metals and radionuclides in biota were associated with specific facility operations and if the measured levels could pose risks to aquatic organisms. We compared concentrations of various metals, 137Cs, and 3H in fish, as well as in water (3H only), among sites along the stream gradient. Fish collected from sites adjacent to H-Area had significantly higher 137Cs concentrations compared to fish from other sites. Both biota and water samples indicated significantly greater levels of 3H in sites adjacent to and downstream of C-Area. Concentrations of zinc (Zn), copper (Cu), and mercury (Hg) in some samples exceeded effects levels reported for fish and may pose a risk to fish populations. This study reported fish tissue concentrations of 137Cs and 3H, which have not been documented extensively in ecotoxicological studies. Our results suggested that industrial operations such as nuclear material production at SRS could have long-lasting impact on the aquatic ecosystem via the release of radionuclides and metals, and long-term monitoring of physiological effects and population level impact in biota exposed to these contaminants are recommended.

Evidence of niche differentiation for two sympatric vulture species in the Southeastern United States.

BACKGROUND: As obligate scavengers utilizing similar habitats, interspecific competition undoubtedly occurs between resident black (Coragyps atratus) and turkey (Cathartes aura) vultures. In the interest of exploring how sympatric species coexist through habitat segregation, we examined resource selection of resident black and turkey vultures in the southeastern United States (US) for evidence of niche differentiation. METHODS: Using fine-scale movement data, we assessed interspecific seasonal differences in monthly roost reuse frequency and roost site fidelity, as well as monthly flight, roost, and diurnal rest site resource selection based on > 2.8 million locations of 9 black vultures and 9 turkey vultures tracked from September 2013 to August 2015 using Groupe Spécial Mobile/Global Positioning System (GSM/GPS) transmitters. RESULTS: Black vultures generally exhibited greater roost fidelity as well as a greater maximum number of nights spent at a single roost than turkey vultures. Patterns of flight, roost, and resting habitat selection within the home range varied monthly as well as between species, providing evidence for habitat segregation and niche differentiation by sympatric vultures. In particular, our results indicate the importance of wooded wetlands for resting and roosting locations for both species, and revealed clear differences in the use of forested habitats between species during flight, resting, and roosting behavioral states. CONCLUSIONS: By examining differences in resource selection and spatial ecology of black and turkey vultures across a range of behaviors, this study demonstrates mechanisms of niche differentiation in these ecologically similar species, and enhances potential for conservation and informed management of this important group of birds.

Bioaccumulation of 137Cs in anuran larvae utilizing a contaminated effluent canal on the U.S. Department of Energy's Savannah River Site.

As a result of activities such as nuclear weapons testing, nuclear power generation and waste disposal, and nuclear accidents, radiocesium (137Cs) is a widely distributed radio-contaminant of concern that readily accumulates in exposed wildlife. Although bioaccumulation of 137Cs is an important factor for understanding its fate within the environment, there are currently limited data available on bioaccumulation patterns of 137Cs in amphibians, despite their widespread distribution and potential to transport contaminants between aquatic and terrestrial ecosystems. Therefore, the objective of this study was to determine the amount of time necessary for anuran larvae experimentally placed in a contaminated system to reach a steady-state whole-body 137Cs concentration, and to determine the threshold at which that steady-state 137Cs concentration occurred for tadpoles within our study system. By restricting uncontaminated bullfrog (Lithobates catesbeianus) larvae to three experimental enclosures located along a137Cs contaminated effluent canal on the U.S. Department of Energy's Savannah River Site, we modeled 137Cs uptake through time using the von Bertalanffy modification of the Richards Model. The results of our modified Richards Model indicate that bullfrog tadpoles achieved steady-state 137Cs concentrations of 3.68-4.34 Bq/g137Cs dry whole-body weight after 11.63-15.50 days of exposure among sampling sites, with an average of 3.94 Bq/g after 14.07 days exposure. Radiocesium accumulation in bullfrog tadpoles was more rapid than that reported for other biota studied from other contaminated systems, likely due to incidental ingestion of sediments and a diet consisting of periphyton and other items that accumulate high levels of 137Cs. Given their rapid accumulation of 137Cs and inability to leave aquatic environments prior to metamorphosis, our data suggest amphibian larvae may be useful indicators for monitoring 137Cs distributions and bioavailability within aquatic systems.

Radiocesium (137Cs) accumulation by fish within a legacy reactor cooling canal system on the Savannah River Site.

The aquatic cooling canal system associated with a nuclear reactor built in the early 1950s received accidental releases of radiocesium (137Cs) from the reactor between 1954 and 1964, resulting in the dispersion of ~8.2 × 1012 Bq of 137Cs into the associated canals and ponds. The primary purpose of this study was to document concentrations of 137Cs in littoral zone fish currently occupying components of the cooling canal system, 3 canals and 2 impoundments, to determine how concentrations varied among these various components. Secondarily, we examined for potential influence of weirs within the canal system on concentrations in fish as well as the potential relationship between fish species and body size and on 137Cs concentrations in fish. We collected samples of sediment, biofilm, and fish from each component of the R-Reactor cooling system and compared 137Cs among sites and species in individual sites. Concentrations of 137Cs in sediments, biofilms and mosquitofish varied significantly among sampling areas with higher concentrations in RCAN1, a canal segment that was the closest to the reactor and received reactor effluent for a longer period than other components. Comparisons among other components of the cooling system, and species comparisons relative to presumed trophic positions and fish length were not consistent. However, littoral zone fish in the cooling canal system continue to bioaccumulate 137Cs >50 years after the original releases of contamination.

Parasitaemia data and molecular characterization of Haemoproteus catharti from New World vultures (Cathartidae) reveals a novel clade of Haemosporida.

BACKGROUND: New World vultures (Cathartiformes: Cathartidae) are obligate scavengers comprised of seven species in five genera throughout the Americas. Of these, turkey vultures (Cathartes aura) and black vultures (Coragyps atratus) are the most widespread and, although ecologically similar, have evolved differences in morphology, physiology, and behaviour. Three species of haemosporidians have been reported in New World vultures to date: Haemoproteus catharti, Leucocytozoon toddi and Plasmodium elongatum, although few studies have investigated haemosporidian parasites in this important group of species. In this study, morphological and molecular methods were used to investigate the epidemiology and molecular biology of haemosporidian parasites of New World vultures in North America. METHODS: Blood and/or tissue samples were obtained from 162 turkey vultures and 95 black vultures in six states of the USA. Parasites were identified based on their morphology in blood smears, and sequences of the mitochondrial cytochrome b and nuclear adenylosuccinate lyase genes were obtained for molecular characterization. RESULTS: No parasites were detected in black vultures, whereas 24% of turkey vultures across all sampling locations were positive for H. catharti by blood smear analysis and/or PCR testing. The phylogenetic analysis of cytochrome b gene sequences revealed that H. catharti is closely related to MYCAMH1, a yet unidentified haemosporidian from wood storks (Mycteria americana) in southeastern USA and northern Brazil. Haemoproteus catharti and MYCAMH1 represent a clade that is unmistakably separate from all other Haemoproteus spp., being most closely related to Haemocystidium spp. from reptiles and to Plasmodium spp. from birds and reptiles. CONCLUSIONS: Haemoproteus catharti is a widely-distributed parasite of turkey vultures in North America that is evolutionarily distinct from other haemosporidian parasites. These results reveal that the genetic diversity and evolutionary relationships of avian haemosporidians are still being uncovered, and future studies combining a comprehensive evaluation of morphological and life cycle characteristics with the analysis of multiple nuclear and mitochondrial genes will be useful to redefine the genus boundaries of these parasites and to re-evaluate the relationships amongst haemosporidians of birds, reptiles and mammals.

Fine-scale assessment of home ranges and activity patterns for resident black vultures (Coragyps atratus) and turkey vultures (Cathartes aura).

Knowledge of black vulture (Coragyps atratus) and turkey vulture (Cathartes aura) spatial ecology is surprisingly limited despite their vital ecological roles. Fine-scale assessments of space use patterns and resource selection are particularly lacking, although development of tracking technologies has allowed data collection at finer temporal and spatial resolution. Objectives of this study were to conduct the first assessment of monthly home range and core area sizes of resident black and turkey vultures with consideration to sex, as well as elucidate differences in monthly, seasonal, and annual activity patterns based on fine-scale movement data analyses. We collected 2.8-million locations for 9 black and 9 turkey vultures from June 2013 -August 2015 using solar-powered GSM/GPS transmitters. We quantified home ranges and core areas using the dynamic Brownian bridge movement model and evaluated differences as a function of species, sex, and month. Mean monthly home ranges for turkey vultures were ~50% larger than those of black vultures, although mean core area sizes did not differ between species. Turkey vulture home ranges varied little across months, with exception to a notable reduction in space-use in May, which corresponds with timing of chick-rearing activities. Black vulture home ranges and core areas as well as turkey vulture core areas were larger in breeding season months (January-April). Comparison of space use between male and female vultures was only possible for black vultures, and space use was only slightly larger for females during breeding months (February-May). Analysis of activity patterns revealed turkey vultures spend more time in flight and switch motion states (between flight and stationary) more frequently than black vultures across temporal scales. This study reveals substantive variability in space use and activity rates between sympatric black and turkey vultures, providing insights into potential behavioral mechanisms contributing to niche differentiation between these species.

The effects of low-level ionizing radiation and copper exposure on the incidence of antibiotic resistance in lentic biofilm bacteria.

Environmental reservoirs of antibiotic resistant bacteria are poorly understood. Understanding how the environment selects for resistance traits in the absence of antibiotics is critical in developing strategies to mitigate this growing menace. Indirect or co-selection of resistance by environmental pollution has been shown to increase antibiotic resistance. However no attention has been given to the effects of low-level ionizing radiation or the interactions between radiation and heavy metals on the maintenance or selection for antibiotic resistance (AR) traits. Here we explore the effect of radiation and copper on antibiotic resistance. Bacteria were collected from biofilms in two ponds - one impacted by low-level radiocesium and the other an abandoned farm pond. Through laboratory controlled experiments we examined the effects of increasing concentrations of copper on the incidence of antibiotic resistance. Differences were detected in the resistance profiles of the controls from each pond. Low levels (0.01 mM) of copper sulfate increased resistance but 0.5 mM concentrations of copper sulfate depressed the AR response in both ponds. A similar pattern was observed for levels of multiple antibiotic resistance per isolate. The first principal component response of isolate exposure to multiple antibiotics showed significant differences among the six isolate treatment combinations. These differences were clearly visualized through a discriminant function analysis, which showed distinct antibiotic resistance response patterns based on the six treatment groups.

Radiocesium in migratory aquatic game birds using contaminated U.S. Department of Energy reactor-cooling reservoirs: A long-term perspective.

Low-level releases of radiocesium into former nuclear reactor cooling-reservoirs on the U.S. Department of Energy's Savannah River Site (SRS) in South Carolina, USA, dating primarily to the late 1950s and early 1960s, have allowed examination of long-term contaminant attenuation in biota occupying these habitats. Periodic collections of migratory game birds since the 1970s have documented 137Cs (radiocesium) activity concentrations in birds of SRS reservoirs, including mainly Par Pond and Pond B. In this study, during 2014 and 2015 we released wild-caught American coots (Fulica americana) and ring-necked ducks (Aythya collaris) onto Pond B. We made lethal collections of these same birds with residence times ranging from 32 to 173 days to examine radiocesium uptake and estimate the rate of natural attenuation. The two species achieved asymptotic whole-body activity concentrations of radiocesium at different times, with ring-necked ducks requiring almost three times longer than the 30-35 days needed by coots. We estimated ecological half-life (Te) for Pond B coots over a 28-yr period as 16.8 yr (95% CI = 12.9-24.2 yr). Pond B coot Te was nearly four times longer than Te for coots at nearby Par Pond where radiocesium bioavailability had been constrained for decades by pumping of potassium-enriched river water into that reservoir. Te could not be estimated from long-term data for radiocesium in Pond B diving ducks, including ring-necked ducks, likely because of high variability in residence times of ducks on Pond B. Our results highlight the importance: (1) for risk managers to understand site-specific bio-geochemistry of radiocesium for successful implementation of countermeasures at contaminated sites and (2) of residence time as a critical determinant of observed radiocesium activity concentrations in highly mobile wildlife inhabiting contaminated habitats.

The effects of a stannous chloride-based water treatment system in a mercury contaminated stream.

We assessed the impacts of an innovative Hg water treatment system on a small, industrially-contaminated stream in the southeastern United States. The treatment system, installed in 2007, removes Hg from wastewater using tin (Sn) (II) chloride followed by air stripping. Mercury concentrations in the receiving stream, Tims Branch, decreased from >100 to ∼10 ng/L in the four years following treatment, and Hg body burdens in redfin pickerel (Esox americanus) decreased by 70% at the most contaminated site. Tin concentrations in water and fish increased significantly in the tributary leading to Tims Branch, but concentrations remain below levels of concern for human health or ecological risks. While other studies have shown that Sn may be environmentally methylated and methyltin can transfer its methyl group to Hg, results from our field studies and sediment incubation experiments suggest that the added Sn to the Tims Branch watershed is not contributing to methylmercury (MeHg) production or bioaccumulation in this system. The stannous chloride treatment system installed at Tims Branch was effective at removing Hg inputs and reducing Hg bioaccumulation in the stream, but future studies are needed to assess longer term impacts of Sn on the environment.

Testing for associations between hematozoa infection and mercury in wading bird nestlings.

Several wading bird species in the southeastern US have a history of infection by hematozoa/avian malaria as well as mercury accumulation through their diet, and thus may be exposed to two, generally sublethal, yet chronic, stressors. We analyzed nestling wading birds (n = 171) of varying size and trophic position from the southeastern US, and a smaller sample (n = 23) of older, free-ranging birds, to look for potential interrelationships between infection by hematozoa and mercury (Hg) uptake. Only one nestling was PCR positive for hematozoa (Plasmodium/Haemoproteus) whereas nine (39%) of the older wading birds were positive. Sequencing indicated that both nestling and adult positives were infected with Plasmodium. Given the low infection rate of the nestlings, there was no association between Hg and malaria. The older birds exhibited a possible malaria/Hg association, but it may be confounded by their greater potential exposure period and large-scale movements.

Trophic dynamics of U, Ni, Hg and other contaminants of potential concern on the Department of Energy's Savannah River Site.

The Department of Energy's Savannah River Site is a former nuclear weapon material production and current research facility located in South Carolina, USA. Wastewater discharges from a fuel and nuclear reactor target manufacturing facility released depleted and natural U, as well as other metals into the Tims Branch-Steed Pond water system. We investigated the current dynamics of this system for the purposes of environmental monitoring and assessment by examining metal concentrations, bioavailability, and trophic transfer of contaminants in seven ponds. Biofilm, detritus, and Anuran and Anisopteran larvae were collected and analyzed for stable isotopes (δ (15)N, δ (13)C) and contaminants of potential concern (COPC) with a focus on Ni, U, and Hg, to examine metal mobility. Highest levels of Ni and U were found in biofilms U (147 and 332 mg kg(-1) DW, respectively), while highest Hg levels were found in tadpoles (1.1 mg kg(-1) DW). We found intraspecific biomagnification of COPCs as expressed through stable isotope analysis. Biofilms were the best indicators for contamination and Anuran larvae with the digestive tract removed were the best indicators of the specific bioavailability of the focal metals. Monitoring data showed that baseline δ (15)N values differed between ponds, but within a pond, values were stable throughout tadpole Gosner stage, strengthening the case to use this species for monitoring purposes. It is likely that there still is risk to ecosystem integrity as COPC metals are being assimilated into lower trophic organisms and even low levels of this mixture has shown to produce deleterious effects to some wildlife species.

Mercury concentrations in nestling wading birds relative to diet in the southeastern United States: a stable isotope analysis.

Mercury (Hg) is a ubiquitous environmental contaminant that is transferred trophically through aquatic and terrestrial food webs. To better understand the routes of Hg uptake in organisms that rely on both aquatic and terrestrial food resources, we analyzed feather and down samples from nestling wading birds of varying trophic positions in both inland and coastal colonies. We used stable nitrogen and carbon isotope analyses to evaluate trophic positions of individual species (δ(15)N) and differences in foraging habitat use (δ(13)C). Inland, aquatic species had higher trophic status than the single terrestrial species examined, and the expected positive relationship between δ(15)N and Hg content of feathers was observed. However, the same was not true for all species from coastal colonies. Feathers from species that primarily consumed saltwater prey were relatively high in δ(15)N value and low in Hg content, which is opposite of the trend expected due to Hg biomagnification in food chains. In contrast, coastal species foraging in freshwater or a combination of freshwater and saltwater habitats displayed greater Hg contents in feathers. The apparent differential use of the two aquatic systems (freshwater and saltwater) in coastal environments by wading bird species results in variations in δ(15)N values and Hg contents in nestling feathers not found in species associated with only freshwater systems.

Blood parasites in wood storks (Mycteria americana) from the southeastern United States.

Wood storks (Mycteria americana) are endangered throughout their breeding range in the United States. Because of this, researchers have had little opportunity to thoroughly examine the continental wood stork population for endoparasites. The blood protozoan Haemoproteus crumenium has been identified from several populations of wood storks in North America. However, there have been no reports of wood storks being infected with species of Plasmodium or Leucocytozoon. During 2003, 42 nestling wood storks were captured at sites in Georgia and 27 free-ranging wood storks were captured at postbreeding dispersal sites in Mississippi and Louisiana. Two thin blood smears were made from each bird, and they were examined for parasites. Haemoproteus crumenium was found in 5 wood storks (7.2%), whereas Leucocytozoon sp., Plasmodium sp., and microfilariae were not observed. Mean intensity of H. crumenium was 4.4 +/- 0.7 (SE) per 2,000 erythrocytes counted. All infected wood storks were subadult or adult and were from postbreeding dispersal sites in Mississippi.

Concentrations of p,p'-DDE in plasma of nestling Wood Storks from Georgia.

We determined the concentrations of organochlorine pesticides and total PCBs in plasma of nestling Wood Storks (Mycteria americana) from two colonies in Georgia in 2000. Of 20 compounds analyzed for, only p,p'-DDE was quantified in reportable concentrations, ranging from <2.0-174.0 ng/g wet weight (ww). Concentrations of p,p'-DDE were significantly greater (P<0.0001) in plasma of nestlings from an inland colony, Chew Mill (x 16.9 ng/g,ww) than a coastal colony, Harris Neck (x 1.2 ng/g,ww). Concentrations from nestlings from Chew Mill were significantly different among sampling weeks (P=0.01), with week 8 being significantly greater than weeks 1, 2, and 5. While the Chew Mill colony had much greater concentrations of p,p'-DDE in plasma of nestlings than the Harris Neck colony, reproduction was greater there (1.8+/-1.0 SD fledged young per nesting attempt) than Harris Neck (1.4+/-1.0 SD fledged young per nesting attempt). While concentrations of p,p'-DDE determined in plasma of nestling Wood Storks are an indirect measure of adult exposure to environmental toxicants, concentrations reported would not be considered detrimental to reproduction in these colonies.

Potential risk to wood storks (Mycteria americana) from mercury in Carolina Bay fish.

Carolina bays are freshwater wetlands that serve as important feeding habitats for the endangered wood stork (Mycteria americana). Water levels in these bays fluctuate greatly and tend to be acidic and rich in dissolved organic carbon (DOC), factors that favor mercury (Hg) methylation and bioaccumulation in fish. To assess potential risks to wood storks consuming mercury contaminated fish in bays, we sampled fish from 10 bays on the Savannah River Site (SRS), South Carolina, an area with documented use by wood storks. Whole body mercury concentrations in 258 fishes of three species (Erimyzon sucetta, Acantharchuspomotis and Esox americanus) commonly consumed by wood storks were determined. Risk factors for nestlings and free-ranging adults were calculated using published no and lowest observable adverse effect concentration (NOAEC and LOAEC) values for birds. Fish from higher trophic levels and those from wetlands with relatively shallow maximum depths and fluctuating water levels were more likely to exceed NOAEC and LOAEC values. Calculation of exposure rates of nestling wood storks indicated they are at highest risk during the first 10 days of the nestling period. These calculations suggest that there is potential concern for wood storks foraging in relatively shallow bays with fluctuating water levels, even though there is no obvious local source of mercury to these wetlands.

Mercury in bald eagle nestlings from South Carolina, USA.

Bald eagles (Haliaeetus leucocephalus) may be at risk from contaminants in their diet and young birds may be particularly sensitive to contaminant exposure. To evaluate potential risks from dietary mercury exposure to eagle nestlings in South Carolina (USA), we surveyed mercury concentrations in 34 nestlings over two breeding seasons (1998 and 1999). Samples were also obtained from several post-fledging eagles in the region. Nestling feather mercury ranged from 0.61-6.67 micrograms Hg/g dry weight, nestling down mercury from 0.50-5.05 micrograms Hg/g dry weight, and nestling blood mercury from 0.02-0.25 microgram Hg/g wet weight. We did not detect significant differences in tissue mercury between nestlings from coastal and inland regions in contrast to some other studies of piscivorous birds. Mercury concentrations were much higher in the post fledging birds we sampled. Our data show that nestling eagles in South Carolina are accumulating mercury, and that concentrations in older birds may exceed regulatory guidelines.