Final Report: Influence of Water Quality on the Bioavailability and Food Chain Transport of Carbon Nanoparticles

EPA Grant Number: R834092
Title: Influence of Water Quality on the Bioavailability and Food Chain Transport of Carbon Nanoparticles
Investigators: Klaine, Stephen J. , Burton, Jr., G. Allen , Ke, Pu-Chun , Mukhopadhyay, Sharmila , Roberts, Aaron
Institution: Clemson University
EPA Project Officer: Shapiro, Paul
Project Period: October 1, 2008 through September 30, 2011
Project Amount: $400,000
RFA: Exploratory Research: Nanotechnology Research Grants Investigating Fate, Transport, Transformation, and Exposure of Engineered Nanomaterials: A Joint Research Solicitation - EPA, NSF, & DOE (2007) RFA Text |  Recipients Lists
Research Category: Nanotechnology , Safer Chemicals

Objective:

The overall goal of this research was to characterize the influence of natural organic matter (NOM) on the bioavailability and food chain transfer of carbon nanoparticles. During the course of this research, it became apparent that carbon nanoparticles do not absorb from the gut tract into the body cavity. These results are discussed in detail below. However, due to this, the focus of this research switched from carbon to gold nanoparticles. The specific objectives of this work include the following:
  1. Characterize the stability of carbon nanoparticles in NOM.
  2. Characterize the bioavailability of carbon nanoparticles to aquatic organisms.
  3. Characterize the bioavailability of gold nanoparticles to Daphnia magna.
  4. Characterize the uptake and food chain transport of gold nanoparticles.

Summary/Accomplishments (Outputs/Outcomes):

In order to test the influence of NOM on nanoparticle stability single-walled carbon nanotubes (SWNTs), multi-walled carbon nanotubes (MWNTs), C70 , and C60 were each suspended in 5 mg/L of each Suwannee River NOM (International Humic Substances Society) dissolved in EPA moderately hard water (MHW: pH = 8.0; Water Hardness = 8 mg/L as CaC03)(Lewis 1994). NOM was dissolved in EPA synthetic moderately hard water and filtered using 0.22 µm cellulose membrane filter (Millipore, Bellerica, MA). The total organic carbon (TOC) composition of the SO mg/L NOM in MHW was measured to be 42% (i.e., 21 (1 mg C/L) using TOC analyzer (Sievers, Boulder, CO).
 
 
 
Further research was conducted to determine if any nanotubes moved out of the gut tract and into the organism and, if so, under what conditions this happened. Four different functionalized SWCNTs were used in this study. Hydroxylated SWCNTs (OH-SWCNTs) were purchased from cheaptubes.com (Brattleboro, Vermont, USA). A sample of this material was then further functionalized by Dr. Mukhopadhyay’'s laboratory at Wright State University (Dayton, Ohio, USA) with silicon dioxide (SiO2-SWCNTs). Poly aminobenzenesulfonic acid (PABS-SWCNTs) and polyethylene glycol (PEG-SWCNTs) functionalized SWCNTs were purchased from Carbon Solutions, Inc. (Riverside, California, USA). These functional groups were chosen because of their differences in polarity and size. Transmission electron micrographs of the SWCNTs were taken after sonication to determine material length and diameter. These particles were suspended in SR-NOM as described previously and D. magna was exposed to these suspensions as described previously. The conclusion of all of these studies was that neither SWNTs nor MWNTs migrated outside the gut tract and that toxicity was due to gut tract clogging and interference with food assimilation.
 
The behavior of suspended MWNTs, C70 and C60 in the three different sources of NOM was characterized using several different analyses. There was no difference in particle sizes (hydrodynamic diameter) as measured by dynamic light scattering for either fullerene or MWNTs among the three NOM sources. Zeta potential did not show a significant difference for MWNTs or either fullerene among the three NOM sources. Other attempts to discern differences among particles suspended in the three sources were also not successful. Further research focused on the bioavailability of MWNTs to pelagic invertebrates, D. magna and Ceriodaphnia dubia.
 
Dose-dependent mortality was found in D. magna exposed to MWNT suspended in NOM (5 mg/L DOC) (96hLC50 = 2.06 ± 0.41; 95% CI). However, 96-h LCSO values did not vary as a function of SR-NOM concentration as measured by DOC. D. magna growth over 96 h was inversely related to MWNT concentrations (Figure 2; r2 = 57%; p < 0.01). Results of the elimination bioassays suggested that gut tract elimination of MWNTs increased in the presence of food, but that the presence of N OM had no effect on time-to-elimination. MWNTs were not lethal to C. dubia at the tested concentrations (0-1mg MWNT/L) during the 7-d exposure. Mean survival was greater than 85% in all treatments. Reproduction was significantly reduced at all MWNT concentrations by at least 20% (Fig. l; p < 0.05). Furthermore, a decrease in growth occurred after the 7-d exposure (Fig. 2; p = 0.045).
 
 
 
 
This study focused on the movement of gold nanomaterials (AuNM) through an aquatic food chain in a recirculating flume system. Periphyton (both multi- and single-species), Lymnaea stagnalis, and Hyalella azteca were chosen to simulate a simple lotic food chain. It was hypothesized that periphyton would accumulate AuNM rapidly and little AuNM would be detected in the water column after just a few hours. In addition, we hypothesized that, after feeding on the periphyton, we would detect Au in the tissues of macroinvertebrates. However, we expected tissue concentrations to be lower than in periphyton suggesting no bioaccumulation of AuNM.
 
Experiments were conducted in which AuNM were added to recirculating flumes containing tiles seeded with periphyton and the tiles were then moved to a beaker to investigate trophic transfer to macroinvertebrates. Two separate experiments were performed using different periphyton communities: a single species monoculture and a field-collected mixed community. Once exposure to AuNM was complete, two aquatic macroinvertebrate species, Lymnaea stagnalis and Hyalella Azteca, were allowed to feed on the tiles and then depurated before processing for total gold analysis.
 
 
Flumes were constructed using PVC downspout extensions (24 cm long), plastic sheets, flexible PVC tubing (3/16 inch inner diameter), 2000 mL plastic bottles, and submersible pumps. Plastic bottles serve as a “reservoir” at the end of each flume and a submersible pump returned water to the head of the flume (flow rate = 200 L/h). A small piece of notched plastic was glued to the end of each channel to retain about 1500 mL of water within each flume. Flumes were randomly arranged on tables (Figure 3).
 
Total Au concentrations in water were similar to nominal concentrations at time zero and then declined exponentially. There was no significant difference between the mixed periphyton (Huron River) community compared to single strain (Oscillatoria) species treatments (p = 0.14). However, Au concentrations declined significantly through time (p < 0.001) and the rate of decline differed between high and low Au treatments (p < 0.05). Around 48 h into the experiment in both high and low treatments almost no Au was detectible in the water column.
 
The Huron River periphyton community make-up was dominated by the cyanobacteria Limnothrix and diatoms. No significant differences were seen between periphyton biomass (as chlorophyll a) for different periphyton communities (p = 0.28), nominal Au treatment type (p = 0.89), or the interaction between periphyton community and treatment type (p = 0.84). Gold was found in periphyton in both algae types and in both high and low treatments. Results of the 2-way ANOVA indicated a significant difference between treatment type (p = 0.02). Nominal Au treatment had strong significant effect on periphyton Au content (p < 0.001) with low and high treatments differing from one another for both Huron and Oscillatoria treatments. In addition, no significant differences (p = 0.29) were reported in AuNM concentration between mixed-culture and single species periphyton.
 
We measured differences in Au concentrations between our two grazers as we measured Au in L. stagnalis but no Au was detected H. azteca. No significant mortality was reported in either species during 24h exposure period or 24h depuration period. Au was found in the tissues of L. stagnalis in both the Huron high and low Au treatments (2.4 and 2.1µg/g dry weight, respectively) as well as in the Oscillatoria high and low treatments (2.2 and 1.45 µg/g, respectively). Between different nominal Au treatments, L. stagnalis tissues differed in Au concentration (p < 0.001) with control treatments having significantly lower Au body burden than low and high treatments. However, we found no significant difference between low and high treatments, no interaction between nominal Au concentration and periphyton community (p = 0.8551), and no significant difference was seen between periphyton type (p = 0.25). No statistical analysis was done for H. azteca as all results were returned as non-detectable.
 
A mass balance was used to estimate the fate of AuNM for three compartments: water, periphyton, and attached to the flume. We estimated that the majority of AuNM spiked during experiments was not measured in the water and periphyton, and likely ended up in flume tubing and reservoir. For the low and high treatments, on average 79.7% and 80.3%, respectively, of the Au was unaccounted for and likely aggregated to physical structures in the mesocosm. Assuming even settling of AuNM, every square centimeter of the flume surfaces should have 0.66 µg Au/cm2 for high treatments and 0.13µg Au/cm2 for low treatments. Results from Oscillatoria and Huron River periphyton high treatments were 0.17 µg Au/cm2 and 0.38 µg Au/cm2, respectively, while low treatment results were 0.04 µg Au/cm2 and 0.07 µg Au/cm2, respectively.
 
Nanomaterials are known to have unique properties that affect their behavior, partitioning, and exposure in aquatic systems (Handy et. al. 2012; Holbrook et. al. 2010; Keller et. al. 2010; Kennedy et. al. 2008; Klaine et. al. 2012). Based on results of the current study, we suggest that suspended AuNM quickly aggregated and precipitated out of the water column. The highest Au concentrations occurred at 1 hour after spiking and then declined exponentially until no Au was measured in any treatment after 48 hours. This may be attributed to nanomaterial agglomeration, aggregation, or precipitation (Tourinho et. al. 2012). Rapid aggregation and precipitation of Au NM was noted in this study, similar to other NM studies (Glenn et. al. 2012; Manusadzian et. al. 2012; Nason et. al. 2012). Upon flume examination during the experiments, it was evident AuNM had attached to the inside of plastic tubing, reservoirs, and possibly flume pumps. However, AuNM aggregated throughout each mesocosm including on periphyton tiles, which allowed for examination of dietary exposure. Surface area calculations indicate uneven settling of AuNM within each system, specifically proportionally less settling on periphyton tiles. This may be explained by tile location in fast moving water of flumes, in addition to AuNM affinity for plastic surfaces. Due to rapid AuNM aggregation, organism exposure to dissolved Au is unlikely. In the environment, even without thick biofilm communities, AuNM may settle out just as quickly as in the current study as it is dominated by physical aggregation.

Conclusions:

Regardless of community composition, periphyton communities exposed to AuNM in closed flume systems displayed elevated Au concentrations, most likely due to physical aggregation and settling of AuNM. Dietary exposure to contaminated periphyton led to elevated Au tissue concentrations in L. stagnalis while Au was not detected in H. Azteca tissues. Differences between L. stagnalis and H. Azteca body burden may be attributable to feeding mechanisms. These results suggest selective feeding by macroinvertebrates, NM settling, and NM aggregation are important when considering NM fate in the environment and their movement throughout the food chain.

References:

Glenn, J. B., White, S. A. & Klaine, S. J. Interactions of gold nanoparticles with freshwater aquatic macrophytes are size and species dependent. Environ. Toxicol. Chem. 2012.;31:194-201.
 
Handy, R.D., Cornelis, G., Fernandes, T., Tsyusko, O., Decho, A., Sabo-Attwood, T., Metcalfe, C., Steevens, J.A., Klaine, S.J., Koelmans, A.A, and Horne, N. Ecotoxicity test methods for engineered nanomaterials: Practical experiences and recommendations from the bench. Environmental Toxicology and Chemistry. 2012;31:15-31.
 
Holbrook, R.D., Kline, C.N., and Filliben, J. J. Impact of source water quality on multiwall carbon nanotube coagulation. Environmental Science and Technology. 2010;44:1386-91.
 
Keller, A.A., Wang, H., Zhou, D., Lenihan, S., Cherr, G., Cardinale, B.J., Miller, R., and Ji, Z. Stability and aggregation of metal oxide nanoparticles in natural aqueous matrices. Environmental Science and Technology. 2010;44:1962-7.
 
Kennedy, A.J., Hull, M.S., Steevens, J.A., Dontsova, K.M., Chappell, M.A., Gunter, J.C., and Weiss, C.A. Factors influencing the partitioning and toxicity of nanotubes in the aquatic environment. Environmental Toxicology and Chemistry. 2008;27:1932-41.
 
Klaine, S.J., Koelmans, AA, Horne, N., Carley, S., Handy, R.D., Kapustka, L., Nowack, B., and von der kammer, F. Paradigms to assess the environmental impact of manufactured nanomaterials. Environmental Toxicology and Chemistry. 2012;31:1823-30.
 
Manusadzianas, L., Caillet, C., Fachetti, L., Gylyte, B., Grigutyte, R., Jurkoniene, S., Karitonas, R., Sadauskas, K., Thomas, F., Vitkus, R., Ferard, J.F. Toxicity of copper oxide nanoparticle suspensions to aquatic biota. Environmental Toxicology and Chemistry. 2012;31:108-114.
 
Nason, J.A, McDowell, S.A., and Callahan, T.W. Effects of natural organic matter type and concentration on the aggregation of citrate-stabilized gold nanoparticles. Journal of Environmental Monitoring. 2012;14:1885.
 
Tourinho, P.S., van Gestel, C.A.M., Lofts, S., Svendsen, C., Soares, A.M.V.M., and Loureiro, S. Metal-based nanoparticles in soil: fate, behavior, and effects on soil invertebrates. Environmental Toxicology and Chemistry. 2012;31:1679-92.


Journal Articles on this Report : 11 Displayed | Download in RIS Format

Other project views: All 11 publications 11 publications in selected types All 11 journal articles
Type Citation Project Document Sources
Journal Article Bhattacharya P, Lin S, Turner JP, Ke PC. Physical adsorption of charged plastic nanoparticles affects algal photosynthesis. Journal of Physical Chemistry C 2010;114(39):16556-16561. R834092 (Final)
  • Abstract: ACS Publications-Abstract
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  • Journal Article Bhattacharya P, Chen P, Spano MN, Zhu L, Ke PC. Copper detection utilizing dendrimer and gold nanowire-induced surface plasmon resonance. Journal of Applied Physics 2011;109:014911. R834092 (Final)
  • Abstract: AIP-Abstract
    Exit
  • Journal Article Chen P, Yang Y, Bhattacharya P, Wang P, Ke PC. A tris-dendrimer for hosting diverse chemical species. Journal of Physical Chemistry C 2011;115(26):12789-12796. R834092 (Final)
  • Abstract: ACS Publications-Abstract
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  • Journal Article Chen R, Huang G, Ke PC. Calcium-enhanced exocytosis of Au nanoparticles. Applied Physics Letters 2010;97:093706. R834092 (Final)
  • Abstract: AIP-Abstract
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  • Journal Article Edgington AJ, Roberts AP, Taylor LM, Alloy MM, Reppert J, Rao AM, Mao J, Klaine SJ. The influence of natural organic matter on the toxicity of multiwalled carbon nanotubes. Environmental Toxicology and Chemistry 2010;29(11):2511-2518. R834092 (Final)
  • Abstract from PubMed
  • Abstract: Wiley Online-Abstract
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  • Journal Article Edgington AJ, Petersen EJ, Herzing AA, Podila R, Rao A, Klaine SJ. Microscopic investigation of single-wall carbon nanotube uptake by Daphnia magna. NanoToxicology 2014;8(Suppl 1):2-10. R834092 (Final)
    R834575 (Final)
  • Abstract from PubMed
  • Full-text: SemanticScholar-Full Text PDF
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  • Abstract: Taylor&Francis-Abstract
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  • Journal Article Glenn JB, White SA, Klaine SJ. Interactions of gold nanoparticles with freshwater aquatic macrophytes are size and species dependent. Environmental Toxicology and Chemistry 2012;31(1):194-201. R834092 (Final)
    R834575 (2013)
    R834575 (Final)
  • Abstract from PubMed
  • Full-text: ResearchGate-Full Text PDF
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  • Abstract: Wiley-Abstract
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  • Journal Article Glenn JB, Klaine SJ. Abiotic and biotic factors that influence the bioavailability of gold nanoparticles to aquatic macrophytes. Environmental Science & Technology 2013;47(18):10223-10230. R834092 (Final)
    R834575 (2013)
    R834575 (Final)
  • Abstract from PubMed
  • Full-text: ACS-Full Text HTML
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  • Abstract: ACS-Abstract
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  • Other: ACS-Full Text PDF
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  • Journal Article Lard M, Kim SH, Lin S, Bhattacharya P, Ke PC, Lamm MH. Fluorescence resonance energy transfer between phenanthrene and PAMAM dendrimers. Physical Chemistry Chemical Physics 2010;12(32):9285-9291. R834092 (Final)
  • Abstract from PubMed
  • Abstract: RSC Publishing-Abstract
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  • Journal Article Seda BC, Ke P-C, Mount AS, Klaine SJ. Toxicity of aqueous C70-gallic acid suspension in Daphnia magna. Environmental Toxicology and Chemistry 2012;31(1):215-220. R834092 (Final)
    R834575 (2013)
    R834575 (Final)
  • Abstract from PubMed
  • Full-text: ResearchGate-Full Text PDF
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  • Abstract: Wiley Online-Abstract
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  • Journal Article Wray AT, Klaine SJ. Modeling the influence of physicochemical properties on gold nanoparticle uptake and elimination by Daphnia magna. Environmental Toxicology and Chemistry 2015;34(4):860-872. R834092 (Final)
    R834575 (Final)
  • Abstract from PubMed
  • Abstract: Wiley-Abstract
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  • Supplemental Keywords:

    Bioconcentration, bioaccumulation, nanoparticle suspension characterization

    Progress and Final Reports:

    Original Abstract
  • 2009
  • 2010