Progress 07/01/06 to 06/30/12
OUTPUTS: 1. Characterize the effects of antidepressants on fish behaviors that affect ecological fitness. We developed a predator-prey bioassay using hybrid striped bass and fathead minnows. This assay was calibrated using diazinon (acetylcholinesterase inhibitor) that was expected to interrupt neuromuscular function and reduce the ability of a predator to capture its prey. Results of the calibration effort demonstrated a dose-dependent decrease in acetylcholinesterase activity with increasing exposure to diazinon. There was a dose-dependent increase in the time required for prey-capture by hybrid striped bass. This assay was used to investigate the change in prey capture behavior and brain chemistry as a function of exposure to antidepressant pharmaceuticals. Fluoxetine exposure in hybrid striped bass caused a decrease in brain serotonin concentration and an increase in the time to capture prey. Similar results were seen with another antidepressant, venlafaxin. Exposure to both of these antidepressants in combination resulted in an additive response for both decreases in brain serotonin and increases in the time to capture prey. We demonstrated that another antidepressant, buproprion, did not alter brain serotonin levels, acting instead on dopamine and norepinephrine. It also did not alter the ability of hybrid striped bass to capture prey. This suggests that brain serotonin levels play a major role in prey-capture behavior in fish. 2. Characterize effects of episodic metal exposure on aquatic organisms. Our work quantifying copper toxicity to Daphnia magna and Pimephales promelas was used to develop the Biotic Ligand Model that is now used by USEPA and state agencies to set site-specific water quality criteria for copper. These data have facilitated the development of a predictive model to integrate the influence of metal concentration, duration of exposure, and frequency of exposure on toxicity. Results of this research have laid the scientific foundation for the development of National Pollutant Discharge Elimination System (NPDES) permits by federal and state agencies. 3. Characterize the bioavailability and food chain transfer of PCBs from contaminated sediments in Lake Hartwell, SC. Our initial efforts focused on bioavailability of sediment-bound PCBs. We have documented uptake into benthic invertebrates including Chironomus sp. A significant amount of PCBs move from contaminated sediments in first order streams through emergent insects into the riparian zone through spiders. 4. Characterize the environmental fate and effect of engineering nanomaterials. We began working with carbon, titanium, and gold nanoparticles (NP) and were the first to demonstrate that organisms can modify NP and alter their physical/chemical characteristics. We quantified the physical effects of carbon nanotubes in clogging the gut tracts of Daphnia magna and interrupting food processing. Uptake, accumulation, and elimination of Au NP by D. magna was size dependent. We also discovered that carbon dots migrated out of the gut tract and into the body of D.magna. We also reported the potential for carbon nanotubes to change the bioavailabiity and toxicity of copper. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Regulatory agencies, other scientists PROJECT MODIFICATIONS: Not relevant to this project.
1. Characterize the effects of antidepressants on fish behaviors that affect ecological fitness. Specifically, we will quantify the effects of these contaminants on predator-prey behavior, swimming performance, and reproduction. In addition, we will attempt to correlate changes in behavior with biochemical changes in the fish caused by the antidepressant (e.g. brain monoamine levels). This research has furthered our knowledge of the effects of antidepressants on fish behavior and brain chemistry. This knowledge will help us begin to develop chemical management plans to reduce the risk of antidepressants. In addition, results of this research have increased our knowledge about the connections between fish brain chemistry and behavior. The connection between fish brain serotonin levels and predatory behavior is novel. 2. Characterize effects of episodic metal exposure on aquatic organisms. Specifically, we will characterize the response of aquatic organisms to metal exposures as a function of metal concentration, duration of exposure, and frequency of exposure. Results of this research are forming the foundation for the development of a predictive model that will be used to develop effluent guidelines for metals. 3. Characterize the bioavailability and food chain transfer of PCBs from contaminated sediments in Lake Hartwell, SC. Specifically, we will characterize the influence of sediment organic carbon quality on uptake of PCBs by benthic invertebrates; quantify the transfer of PCBs from the aquatic food chain to the terrestrial food chain; and estimate the risk to human health from these processes. Results of this work have been incorporated into PCB ecological risk assessments and are currently being used in follow-up assessments of the Lake Hartwell PCB superfund site. 4. Characterize the environmental fate and effect of engineering nanomaterials. Our research has demonstrated the need to quantitatively describe nanoparticle exposures when assessing toxicity in order to better explain effects. Further, our results have demonstrated the need for future research to quantify chronic effects, nanoparticle uptake, translocation, and food chain transfer.
- Klaine SJ, Horne N, Diamond S, Cornelius G, Rogers K, Fernandes T, Koelmans AA. 2012. Paradigms for the Environment Assessment of Manufactured Nanomaterials. Environmental Toxicology and Chemistry 31(1): 3-14.
- Seda BC, Ke PC, Mount AS, Klaine SJ. 2012. Toxicity of aqueous C70-gallic acid suspension in Daphnia magna. Environmental Toxicology and Chemistry 31(1): 215-220.
- Glenn JB, White SA, Klaine SJ. 2012. Uptake of Gold Nanoparticles in Aquatic Plants is Size and Species Dependent. Environmental Toxicology and Chemistry 31(1):194-201
- Bone AJ, Colman BP, Gondikas AP, Newton KM, Harrold KH, Cory RM, Unrine JM, Klaine SJ, Matson CW, Di Giulio RT. 2012. Biotic and abiotic interactions in aquatic microcosms determine fate and toxicity of Ag nanoparticles: Part 2:Toxicity and Ag Speciation. Environ. Sci. Technol. 46 (13):6925-6933.
- Nowack B, Ranville JF, Diamond S, Gallego-Urrea J, Metcalfe C; Rose J, Horne N, Koelmans AA, Klaine SJ. 2012. Nanoparticle Release, Aging and Transformation in the Environment. Environmental Toxicology and Chemistry 31(1): 50-59.
- Handy RD, Cornelis G, Fernandes T, Tsyusko O, Decho A, Sabo-Attwood T, Metcalfe C, Steevens JA, Klaine SJ, Koelmans AA, Horne N. 2012. Ecotoxicity Test Methods for Engineered Nanomaterials: Practical Experiences and Recommendations from the Bench. Environmental Toxicology and Chemistry 31(1):15-31.
- von der Kammer F, Ferguson PL, Holden PA, Masion A, Rogers K, Klaine SJ, Koelmans AA , Horne N , Unrine JM. 2012. Analysis of Nanomaterials in Complex Matrices (Environment and Biota): General Considerations and Conceptual Case Studies. Environmental Toxicology and Chemistry 31(1): 32-49.
Progress 01/01/11 to 12/31/11
OUTPUTS: Research during the past year has focused on the toxicity of arsenic and selenium in coal fired power plant effluents (there are several such facilities in South Carolina and many in the Southeast and the rest of the US), the potential additive effects of two pharmaceuticals commonly found in municipal effluents on the ability of freshwater fish to capture their prey, and the effects of nano materials on the aquatic filter-feeding invertebrate, Daphnia magna. The toxicity of coal-fired power plant effluents was accomplished through bioassays with fish and invertebrates, including freshwater mussels. This work was performed in conjunction with an industry consortium. Two pharmaceuticals, Prozac and Effexor were tested using our predator-prey bioassay that we developed previously. Carbon nanotubes, carbon dots, quantum dots, gold and silver nano particles were tested for their effects and potential uptake in food chains. Three doctoral students conducting research in this area successfully defended their Ph.D. Analysis and compilation of the work from last year's workshop on nano materials in the environment was also performed. PARTICIPANTS: Stephen J. Klaine, PI Research partner organizations include the US EPA and the Electric Power Research Institute. Research collaborators from Clemson University include Thompson Mefford, Chris Kitchens, Christina Wells, Nishaanth Tharayil, Ya-Ping Sun, and Pu-Chun Ke. Research collaborators outside of Clemson University include Aaron Robetts, University of North Texas, G. Allen Burton, University of Michigan, Pedro Alvarez, Rice University, Sam Luoma, University of California - Davis, Jamie Leeds, University of Birmingham Training opportunities for this program include several doctoral and master's students as well as several undergraduates. TARGET AUDIENCES: Industries, municipalities, farmers, and regulatory agencies. Efforts include over 20 research presentations at scientific meetings, service on the Environmental Protection Agencies, Federal Insecticide, Fungicide, and Rodenticide Act Science Advisory Panel, and presentations at farm days. PROJECT MODIFICATIONS: The only modification to this research program is the addition of nano materials to our efforts.
Several outcomes/impacts have resulted from our efforts this year: 1. We learned that results of Daphnia magna bioassays could be used to predict the results from juvenile mussel bioassays. This is important since many mussels are threatened or endangered and regulatory agencies were considering requiring all industries to perform these bioassays. Our work indicates that results of the standard daphnia bioassay can be used to predict the sensitivity of freshwater mussels thus saving significant resources. 2. We learned that the two pharmaceuticals are additive not only on the ability of a fish to capture prey but also on changes in the brain chemistry of that fish. This is important since it suggests that we need to better understand the effects of multiple pharmaceuticals in wastewater effluents as well as runoff from livestock areas. This is a very understudied area. 3. We learned that carbon dots are much less toxic than quantum dots. Quantum dots have been the preference for medical imaging research but this research has been held back due to the inherent toxicity of these particles. 4. We learned that the toxicity of carbon nanotubes was due to clogging of the gut tract and the resulting inability of an organism to process and assimilate food. This non-biochemical mechanism of toxicity is understudied but may be the primary mechanism of toxic effects. 5. We have suggestive evidence that the toxic effects mentioned in (4) are related to gut tract clearance time and that this time might be related to certain particle characteristics. If this continues we will be able to develop a predictive model for nano particle toxicity. 6. The analyses from the nanomaterials workshop held in August, 2010 was completed and manuscripts submitted for publication (to be published in January, 2012).
- Stojak AR, Raftery T, Klaine SJ, McNealy TL. 2011. Morphological responses of Legionella pneumophila biofilm to nanoparticle exposure. Nanotoxicology 5(4):730-742.
- Bielmyer GK, Ray, AW, Tomasso JR, Isely JJ, Klaine, SJ. 2011. Effects of roof and rainwater characteristics on copper concentrations in runoff. Environmental Monitoring and Assessment. DOI 10.1007/s10661-011-2152-1.
Progress 01/01/10 to 12/31/10
OUTPUTS: 2010 was a very productive year in terms of outputs. We presented over 20 research presentations at regional, national, and international scientific meetings. We presented three talks to lay audiences on nanotechnology. We hosted the 5th International Conference on the Environmental Effects of Nanoparticles and Nanomaterials bringing together 200 scientists from five continents to discuss the environmental, health, and safety (EHS) of nanomaterials. We hosted an intensive workshop with 27 invited scientists to discuss needed research to make the next significant advances in nanomaterials EHS. We have five manuscripts submitted from these efforts. PARTICIPANTS: Graduate students have been trained and are continuing to be trained on this project. TARGET AUDIENCES: This research is of importance to scientific and regulatory audiences. PROJECT MODIFICATIONS: Not relevant to this project.
Change in Knowledge 1. We discovered that the antidepressant, Venlafaxine, caused fish to reduce food consumption. This drug is a common contaminant in surface waters impacted by domestic sewage effluents. 2. We discovered that nanomaterial uptake from the gut tract of a daphnid was size dependent. 3. We discovered that citrate-coated gold nanomaterials had an impact on biofilms. 4. We discovered that carbon dots migrated out of the gut tract and into the body a daphnid. Change in Action 1. We developed a consistent method for suspending carbon nanomaterials in solutions of natural organic matter. Changes in Condition 1. Our metals toxicity research was used to develop the Biotic Ligand Model (BLM) that accurately predicts the toxicity of copper in various water quality scenarios. This tool is now being used by US EPA and states to determine site-specific water quality criteria.
- Robinson SE, Capper NA, Klaine SJ. 2010. The Effects of continuous and pulsed exposures of suspended clay on the survival, growth, and reproduction of Daphnia magna. Environ. Toxicol. Chem. 29(1): 168-175.
- Kim KT, Klaine SJ, Cho J, Kim SH, Kim SD. 2010. Oxidative responses of Daphnia magna exposed to titanium dioxide nanoparticles. Science of the Total Environment 408 (10): 2268-2272.
- Edgington AJ, Roberts AP, Taylor LM, Alloy MM, Reppert J, Rao, AM, Mao J, Klaine SJ. 2010. The influence of natural organic matter on the toxicity of multiwalled carbon nanotubes. Environ. Toxicol. Chem. 29(11): 2511-2518.
- Kim KT, Klaine SJ, Lin S, Ke PC, Kim SD. 2010. Acute toxicity of a mixture of copper and single-walled carbon nanotubes to Daphnia magna. Environ. Toxicol. Chem. 29(1): 122-126.
Progress 01/01/09 to 12/31/09
OUTPUTS: We have made significant progress quantifying the impacts of anthropogenic contaminants on the ecosystem, This work has included studying the movement of PCBs from the aquatic to the terrestrial environment by way of the aquatic/riparian food chain; quantifying the effects of the pharmaceutical, Prozac, on fish behavior and development; characterizing the influence of water quality on metal bioavailability; and quantifying the interactions of emerging nanomaterials with aquatic organisms. Results have been communicated to scientists through presentations at national and international meetings, peer-reviewed publications, presentations to local and national regulatory agencies, participation on National Research Council review panels, and interviews in scientific magazines. PARTICIPANTS: Graduate students were trained this past year and continue to be trained on this project. During 2009 the following students graduated: Holly Zahner, Ph.D. Response of Whole Body Sodium in larval Pimephales promelas to Episodic Copper Exposure; 5/09 Anthony Sowers, Ph.D., Effects of Wastewater Treatment Effluents on fish and amphibian development 5/09 Current Employer: US Environmental Protection Agency Hung Vu, MS Testing the Individual Effective Dose (IED) hypothesis; 12/09 Amber Stojak, MS Effects of gold nanoparticles on microbial biofilms; 12/09 Matt Osborne-Koch MS Bioaccumulation of gold nanoparticles in an algal-daphnid Foodchain 12/09 Sarah Robinson, MS. Toxicity of suspended clay to Daphnia magna; 12/09 TARGET AUDIENCES: This research effort is important to both scientific and regulatory personnel. PROJECT MODIFICATIONS: Not relevant to this project.
1. A significant amount of PCBs move from contaminated sediments in first order streams through emergent insects into the riparian zone through spiders. 2. Pharmaceutics designed to alter brain monoamines in humans alter them in fish and this correlates with a loss in ability to capture prey. 3. Pharmaceuticals designed to serve as synthetic hormones in humans alter development in fish. 4. The bioavailability of metals in surface waters is controlled by several water quality parameters including dissolved organic carbon, pH, water hardness, and alkalinity. 5. Toxicity of nanoparticles in aquatic systems is influenced by particle characteristics. While a diverse set of projects, the commonality is the focus on insuring good water quality for aquatic organisms as well as humans. Results of this past year and those of future years will serve as the foundation to develop better chemical management strategies to minimize pollution of critical water resources.
- Gaworecki KM, Roberts AP, Ellis N, Sowers AD, Klaine SJ. 2009. Correlating Biochemical and Behavioral Effects of Diazinon Exposure in Hybrid Striped Bass. Environmental Toxicology and Chemistry 28(1): 105-112
- Parmenter KJ, Bisesi JH, Young SP, Klaine SJ, Atwod HL, Browdy CL, Tomasso JR. 2009. The culture of pacific white shrimp Litopenaeus vannamei in a mixed ion solution. North American Journal of Aquaculture 71(2):134-137.
- Kim, K., Edgington, A., Klaine, S., Cho, J., Kim, S. 2009. Influence of Multiwalled Carbon Nanotubes Dispersed in Natural Organic Matter on Speciation and Bioavailability of Copper. Environmental Science and Technology. 43:8979-8984.
- Ryan AC, Tomasso JR, Klaine S. 2009. Influence of pH, hardness, dissolved organic carbon concentration, and dissolved organic matter source on the acute toxicity of copper to Daphnia magna in soft waters: implications for the biotic ligand model. Environ. Toxicol. Chem. 28(8):1663-1670.
- Sowers AD, Mills MA, Klaine SJ. 2009. The developmental effects of a municipal wastewater effluent on the northern leopard frog, Rana pipens. Aquatic Toxicology 94:145-152.
- Klaine SJ. 2009. Considerations for Research on the Environmental Fate and Effects of Nanoparticles. Environ. Toxcol. Chem. 29(9): 1787-1788.
- Sowers AD, Gaworecki KM, Mills MA, Roberts AP, Klaine SJ. 2009. Developmental Effects of a Municipal Wastewater Effluent on two generations of the Fathead Minnow, Pimephales promelas. Aquatic Toxicol. 95(3): 173-181.
Progress 01/01/08 to 12/31/08
OUTPUTS: This research characterizes the environmental impacts of emerging and legacy contaminants. In the past year we have made progress on several fronts. First, we have completed our study on the food chain transport of Polychlorinated Biphenyls (PCBs). We have characterized movement from contaminated sediment through the aquatic ecosystem and into the riparian zone. Manuscripts are in preparation. Second we have continued characterizing the impacts of episodic metal exposure on aquatic organisms. This work is facilitating the development of a predictive model for developing National Pollutant Discharge Elimination System (NPDES) permits. Third, we have characterized the effects of the drug Prozac on the ability of hybrid striped bass to capture live prey. This work is some of the first to demonstrate a connection between brain chemistry and behavior in fish. Fourth, we have initiated research to better understand the behavior and effects of nanomaterials in aquatic ecosystems. In particular we are working with carbon nanoparticles and have begun to work with titanium and gold nanoparticles. PARTICIPANTS: We have collaborated with USEPA and SC DHEC on much of this research. This research provides education and training opportunities for 5 graduate students and 1 undergraduate student. TARGET AUDIENCES: This research impacts regulatory agencies (EPA and DHEC) by providing the scientific foundation for establishing contaminant limits in water and soil. Results can and will be used to better educate the public in order to create more knowledgeable voters. PROJECT MODIFICATIONS: We have added nanoparticles to list of emerging contaminants that we are studying.
The impacts of our work are several-fold. (1) Our metals research is being used to develop regulatory tools to protect aquatic ecosystems from metal pollution. (2) The results of the PCB research is demonstrating the need to examine PCB movement out of contaminated systems when considering remediation of a site. (3) Our work with Prozac and fish behavior has garnered the attention of EPA as they struggle with how to regulate these wastes in effluents. (4) Our nanomaterial research \has begun to lay the groundwork for assessing the environmental risk of these materials.
- Sciera, K.L., J. Smink, J. Morse, C. Post, J. Pike, W. English, TZ. Karanfil, M. Schlautman, S.J. Klaine. 2008. Impacts of Land Disturbance on Aquatic Ecosystem Health: Quantifying the Cascade of Events. Integrated Environmental Assessment and Management 4(4):431-442.
- Gaworecki, K.M., A.P. Roberts, N. Ellis, A.D. Sowers and S.J. Klaine. 2009. Correlating Biochemical and Behavioral Effects of Diazinon Exposure in Hybrid Striped Bass. Environmental Toxicology and Chemistry 28(1): 105-112
- Van Genderen, E.J., J.R. Tomasso, and S.J. Klaine. 2008. Influence of Cu exposure on whole-body sodium levels in larval fathead minnows (Pimephales promelas). Environmental Toxicology and Chemistry 27(6):1442-1449.
- Van Genderen, E.J. and S.J. Klaine. 2008. Demonstration of a spatial approach for predicting acute copper toxicity to larval fathead minnows (Pimephales promelas) in surface waters. Integrated Environmental Assessment and Management 4(2):237-245.
- Gaworecki, K.M. and S.J. Klaine. 2008. Behavioral and Biochemical Responses of Hybrid Striped Bass During and After Fluoxetine Exposure. Aquatic Toxicology 88:207-213.
- Klaine, S. J., P. J.J. Alvarez, G. E. Batley, T. F. Fernandes, R. D. Handy, D. Lyon, S. Mahendra, M. J. McLaughlin, and J. R. Lead. 2008. Nanomaterials in the environment: Behavior, fate, bioavailability, and effects. Environmental Toxicology and Chemistry 27(9):1825-1851.
Progress 01/01/07 to 12/31/07
OUTPUTS: This research characterizes the environmental impacts of emerging and legacy contaminants. In the past year we have characterized the response of fish to pharmaceuticals, the response of fish to episodic metal exposures, and food chain transport of PCBs in aquatic and terrestrial animals. Our research with pharmaceuticals demonstrates the impact of these contaminants on brain chemistry and the consequential impact on the ability of fish to capture prey. This research has captured the attention of the public given the recent report of pharmaceuticals in drinking water. Our research on episodic metal exposure has laid the scientific foundation for the development of National Pollutant Discharge Elimination System (NPDES) permits by federal and state agencies. Our PCB research has documented movement of PCBs from the aquatic food chain to the terrestrial food chain through emerging insects and spiders. To this set of research we have added an initiative to characterize the
environmental fate and effect of nanoparticles since these emerging contaminants are used extensively yet we have little environmental information on them.
PARTICIPANTS: We have collaborated with USEPA and SC DHEC on much of this research. This research procvides education and training opportunities for 5 graduate students and 2 undergraduate students.
TARGET AUDIENCES: This research impacts regulatory agencies (EPA and DHEC) by providing the scientific foundation for establishing contaminant limits in water and soil. Results can and will be used to better educate the public in order to create more knowledgeable voters.
PROJECT MODIFICATIONS: We have added nanoparticles to list of emerging contaminants that we are studying.
Results of our research during the 2007 have facilitated the establishment of site specific water quality criteria for metals, drawn attention to sublethal behavioral changes in fish exposed to pharmaceuticals, and has begun to explain how PCBs are migrating from contaminated aquatic ecosystems.
- Lin, S., D. Keskar, Y. Wu, X. Wang, A.S. Mount, S.J. Klaine, J.M. More, A.M. Rao, and P.C. Ke. 2007. Detection of Phospholipid-Carbon Nanotube Translocation Using Fluorescence Energy Transfer. Applied Physics Letters 89:143118.
- Qiao, R., A.P. Roberts, A.S. Mount, S.J. Klaine, P.C. Ke. 2007. Translocation of C60 and Its Derivatives across a Lipid Bilayer. Nano Letters 7(3): 614-619.
- Roberts, A.P., A.S. Mount, B. Seda, J. Souther, R. Qiao, S. Lin, P.C. Ke, and S.J. Klaine. 2007. In vivo Biomodification of Lipid-Coated Carbon Nanotubes by Daphnia magna. Environmental Science & Technology. 41:3025-3029.
- Hoang, T.C., J.R. Tomasso, S.J. Klaine. 2007. An integrated model describing the toxic responsesof Daphnia magna to pulsed exposures of three metals. Environmental Toxicology and Chemistry 26(1): 132-138.
- Hoang, T.C., J.R. Tomasso, S.J. Klaine. 2007. Influence of organism age on metal toxicity to Daphnia magna. Environmental Toxicology and Chemistry 26(6):1198-1204.
- Hoang, T.C., J.S. Gallagher, S.J. Klaine. 2007. Responses of Daphnia magna to pulsed exposures of arsenic. Environmental Toxicology 22(3):308-317.
- Hoang, T.C., G.S. Jallagher, T.R. Tomasso, S.J. Klaine. 2007. Relationship between the toxicity of two pulsed metal exposures in Daphnia magna: Role of pulsed duration and concentration and interval between pulsed. Archives of Environmental Contamination and Toxicology 53:579-589.
Progress 01/01/06 to 12/31/06
This research focuses on the assessment of the behavior and effects of pharmaceuticals, metals and PCBs in aquatic ecosystems. In preparation for the pharmaceutical research we have developed a predator-prey bioassay using hybrid striped bass and fathead minnows. We have made significant progress characterizing the response of aquatic organisms to pulsed metal exposure. These data have facilitated the development of a predictive model to integrate the influence of metal concentration, duration of exposure, and frequency of exposure on toxicity. Our current PCB research is quanitfying the role of sediment organic matter on the accumulation of PCBs in aquatic food chains. We are also quantifying the movementl of PCBs from the aquatic food chain to the terrestrial food chain.
Pharmaceutical research will ultimately be used in ecological risk assessment. Metal research will be used to develop more scientifically defensible effluent permits. PCB research will facilitate better management of superfund sites.
- Hoang, T.C., J.R. Tomasso, S.J. Klaine. 2007. An Integrated Model Describing the Toxic Responses of Daphnia magna to Pulsed Exposures of Three Metals. Environmental Toxicology and Chemistry 26(1):132-138.