, Volume 19, Issue 1, pp 185–195 | Cite as

The effects of silver nanoparticles on fathead minnow (Pimephales promelas) embryos

  • Geoff Laban
  • Loring F. Nies
  • Ronald F. Turco
  • John W. Bickham
  • Maria S. Sepúlveda


Nanoparticles are being used in many commercial applications. We describe the toxicity of two commercial silver (Ag) nanoparticle (NP) products, NanoAmor and Sigma on Pimephales promelas embryos. Embryos were exposed to varying concentrations of either sonicated or stirred NP solutions for 96 h. LC50 values for NanoAmor and Sigma Ag NPs were 9.4 and 10.6 mg/L for stirred and 1.25 and 1.36 mg/L for sonicated NPs, respectively. Uptake of Ag NPs into the embryos was observed after 24 h using Transmission Electron Microscopy and Ag NPs induced a concentration-dependent increase in larval abnormalities, mostly edema. Dissolved Ag released from Ag NPs was measured using Inductively Coupled-Mass Spectrometry and the effects tested were found to be three times less toxic when compared to Ag nitrate (AgNO3). The percentage of dissolved Ag released was inversely proportional to the concentration of Ag NPs with the lowest (0.625 mg/L) and highest (20 mg/L) concentrations tested releasing 3.7 and 0.45% dissolved Ag, respectively and percent release was similar regardless if concentrations were stirred or sonicated. Thus increased toxicity after sonication cannot be solely explained by dissolved Ag. We conclude that both dissolved and particulate forms of Ag elicited toxicity to fish embryos.


Ecotoxicology Fish early life stages Nanoparticles Silver 



We would like to thank Bob Rode, laboratory manager for the aquatic research facility and many students (Nathan Barton, Sonia Johns, Lynn Henneberger, Brett Lowry, Aaron McAlexander, Reid Morehouse, and Brian Sanchez) for their help with the maintenance of the fathead minnow colony. The authors would also like to thank Gary Ankley and Dan Villeneuve (USEPA, Mid-Continent Ecology Division, Duluth, MN) and James Lazorchack and Marke Smith (USEPA, Molecular Indicator Research Branch, Cincinnati, OH) for donating fathead minnow adults that were used to establish our breeding colony. We also thank Steve Sassman and Dr. Nicole Ramlachan (Purdue University) for assistance with LC-MS and data analysis. This work was supported by a grant from the Lilly Endowment, Inc. awarded through Purdue University Center for the Environment at Discovery Park.


  1. Ankley GT, Jensen KM, Kahl MD et al (2001) Description and evaluation of a short-term reproduction test with the fathead minnow (Pimephales promelas). Environ Toxicol Chem 20:1276–1290CrossRefGoogle Scholar
  2. Asharani PV, Wu YL, Gong Z, Valliyaveettil S et al (2008) Toxicity of silver nanoparticles in zebrafish models. Nanotechnol 19:255102CrossRefGoogle Scholar
  3. Baker C, Pradhan A, Pakstis L et al (2005) Synthesis and antibacterial properties of silver nanoparticles. J Nanosci Nanotechnol 5:244–249CrossRefGoogle Scholar
  4. Benn TM, Westerhoff P (2008) Nanoparticle silver released into water from commercially available sock fabrics. Environ Sci Technol 42:4133–4139CrossRefGoogle Scholar
  5. Blaser SA, Scheringer M, MacLeod M, Hungerbuhler K et al (2008) Estimation of cumulative aquatic exposure and risk due to silver: contribution of nano-functionalized plastics and textiles. Sci Total Environ 360:396–409Google Scholar
  6. Braydich-Stolle L, Hussain S, Schlager JJ et al (2005) In vitro cytotoxicity of nanoparticles in mammalian germline stem cells. Toxicol Sci 88:412–419CrossRefGoogle Scholar
  7. Brayner R, Ferrari-Iliou R, Brivois N et al (2006) Toxicological impact studies based on Escherichia coli bacteria in ultrafine ZnO nanoparticles colloidal medium. Nano Lett 6:866–870CrossRefGoogle Scholar
  8. Brooking J, Davis SS, Illum L (2001) Transport of nanoparticles across the rat nasal mucosa. J Drug Target 9:267–279CrossRefGoogle Scholar
  9. Bury NR, Galvez F, Wood CM (1999) Effects of chloride, calcium and dissolved organic carbon on silver toxicity: comparison between rainbow trout and fathead minnows. Environ Toxicol Chem 18:56–62CrossRefGoogle Scholar
  10. Chang JS, Chang KL, Hwang DF et al (2007) In vitro cytotoxicitiy of silica nanoparticles at high concentrations strongly depends on the metabolic activity type of the cell line. Environ Sci Technol 41:2064–2068CrossRefGoogle Scholar
  11. Chen X, Schluesener HJ (2008) Nanosilver: a nanoproduct in medical application. Toxicol Lett 176:1–12CrossRefGoogle Scholar
  12. Cheng D, Yang J, Zhao Y (2004) Antibacterial materials of silver nanoparticles applications in medical appliances and appliances for daily use. Chin Med Equip J 4:26–32Google Scholar
  13. Chithrani BD, Chan WC (2007) Elucidating the mechanism of cellular uptake and removal of protein-coated gold nanoparticles of different sizes and shapes. Nano Lett 7:1542–1550CrossRefGoogle Scholar
  14. Chithrani BD, Ghazani AA, Chan WC (2006) Determining the size and shape dependence of gold nanoparticle uptake into mammalian cells. Nano Lett 6:662–668CrossRefGoogle Scholar
  15. Cohen MS, Stern JN, Vanni AJ et al (2007) In vitro analysis of nanocrystalline silver-coated surgical mesh. Surg Infect 8:397–403CrossRefGoogle Scholar
  16. Dethloff GM, Naddy RB, Gorsuch JW (2007) Effects of sodium chloride on chronic silver toxicity to early life stages of rainbow trout (Oncorhynchus mykiss). Environ Toxicol Chem 26:1717–1725CrossRefGoogle Scholar
  17. Dreher KL (2004) Health and environmental impact of nanotechnology: toxicological assessment of manufactured nanoparticles. Toxicol Sci 77:3–5CrossRefGoogle Scholar
  18. Fabrega J, Fawcett SR, Renshaw JC, Lead JR (2009) Silver nanoparticle impact on bacterial growth: effect of pH, concentration, and organic matter. Environ Sci Technol (in press)Google Scholar
  19. Ferin J, Oberdorster G, Penney DP (1992) Pulmonary retention of ultrafine and fine particles in rats. Am J Respir Cell Mol Biol 6:535–542Google Scholar
  20. Glover CN, Wood CM (2005) Accumulation and elimination of silver in Daphnia magna and the effect of natural organic matter. Aquat Toxicol 73:406–417CrossRefGoogle Scholar
  21. Greulich C, Kittler S, Epple M, Muhr G, Koller M et al (2009) Studies on the biocompatibilty and interaction of silver nanoparticles with human mesnchymal stem cells (hMSCs). Langenbecks Arch Surg 394:495–502CrossRefGoogle Scholar
  22. Griffitt RJ, Lu J, Gao J, Bonzongo JC, Barber DS et al (2008) Effects of particle composition and species on toxicity of metallic nanomaterials in aquatic organisms. Environ Toxicol Chem 27:1972–1978CrossRefGoogle Scholar
  23. Gustafsson O, Long CM, Macfarlane J et al (2001) Fate of linear alkylbenzenes released to the coastal environment near Boston harbor. Environ Sci Technol 35:2040–2048CrossRefGoogle Scholar
  24. Guzman KA, Taylor MR, Banfield JF (2006) Environmental risks of nanotechnology: national nanotechnology initiative funding, 2000–2004. Environ Sci Technol 40:1401–1407CrossRefGoogle Scholar
  25. Gwinn MR, Vallyathan V (2006) Nanoparticles: health effects—pros and cons. Environ Health Perspect 114:1818–1825Google Scholar
  26. Handy RD, Shaw BJ (2007) Ecotoxicity of nanomaterials to fish: challenges for ecotoxicity testing. Integr Environ Assess Manag 3:458–460CrossRefGoogle Scholar
  27. Hillyer JF, Albrecht RM (2001) Gastrointestinal persorption and tissue distribution of differently sized colloidal gold nanoparticles. J Pharm Sci 90:1927–1936CrossRefGoogle Scholar
  28. Hoet PH, Bruske-Hohlfeld I, Salata OV (2004) Nanoparticles—known and unknown health risks. J Nanobiotechnol 2:12CrossRefGoogle Scholar
  29. Hogstrand C, Wood CM (1998) Toward a better understanding of the bioavailability, physiology and toxicity of silver in fish: indications for water quality criteria. Environ Toxicol Chem 17:547–561CrossRefGoogle Scholar
  30. Hostynek JJ (2003) Factors determining percutaneous metal absorption. Food Chem Toxicol 41:327–345CrossRefGoogle Scholar
  31. Huang XL, Zhang B, Ren L et al (2008) In vivo toxic studies and biodistribution of near infrared sensitive Au–Au(2)S nanoparticles as potential drug delivery carriers. J Mater Sci Mater Med 19:2581–2588CrossRefGoogle Scholar
  32. Hussain SM, Hess KL, Gearhart JM et al (2005) In vitro toxicity of nanoparticles in BRL 3A rat liver cells. Toxicol In Vitro 19:975–983CrossRefGoogle Scholar
  33. Ji JH, Jung JH, Kim SS et al (2007) Twenty-eight-day inhalation toxicity study of silver nanoparticles in Sprague–Dawley rats. Inhal Toxicol 19:857–871CrossRefGoogle Scholar
  34. Kane MD, Sringer JA, Iannotti NV et al (2008) Identification of development and tissue-specific gene expression in the fathead minnow Pimephales promelas, rafinesque using computational and DNA microarray methods. J Fish Biol 72:2341–2353CrossRefGoogle Scholar
  35. Karn B, Roco MC, Masciangioli T et al (2003) Nanotechnology and the environment. American Chemical Society, ArlingtonGoogle Scholar
  36. Kashiwada S (2006) Distribution of nanoparticles in the see-through medaka (Oryzias latipes). Environ Health Perspect 114:1697–1702Google Scholar
  37. Kim JS, Kuk E, Yu KN et al (2007) Antimicrobial effects of silver nanoparticles. Nanomedicine 3:95–101Google Scholar
  38. Kim YS, Kim JS, Cho HS et al (2008) Twenty-eight-day oral toxicity, genotoxicity, and gender-related tissue distribution of silver nanoparticles in Sprague–Dawley rats. Inhal Toxicol 20:575–583CrossRefGoogle Scholar
  39. Lansdown AB (2006) Silver in healthcare: antimicrobial effects and safety in use. Curr Probl Dermatol 33:17–34CrossRefGoogle Scholar
  40. Larkin P, Villeneuve DL, Knoebl I et al (2007) Development and validation of a 2,000-gene microarray for the fathead minnow (Pimephales promelas). Environ Toxicol Chem 26:1497–1506CrossRefGoogle Scholar
  41. LeBlanc GA, Mastone JD, Paradice AP et al (1984) The influence of speciation on the toxicity of silver to fathead minnow (Pimephalies promelas). Environ Toxicol Chem 3:37–47CrossRefGoogle Scholar
  42. Lee HY, Park HK, Lee WM et al (2007a) A practical procedure for producing silver nanocoated fabric and its antibacterial evaluation for biomedical applications. Chem Commun 28:2959–2961CrossRefGoogle Scholar
  43. Lee KJ, Nallathamby PD, Browning LM, Osgood CJ, Xu XH et al (2007b) In vivo imaging of transport and biocompatibility of single silver nanoparticles in early development of zebrafish embryos. ACS Nano 1:133–143CrossRefGoogle Scholar
  44. Lemke AE (1981) Interlaboratory comparison acute testing set. United States Environmental Protection Agency, Washington, DCGoogle Scholar
  45. Limbach LK, Li Y, Grass RN et al (2005) Oxide nanoparticle uptake in human lung fibroblasts: effects of particle size, agglomeration, and diffusion at low concentrations. Environ Sci Technol 39:9370–9376CrossRefGoogle Scholar
  46. Lok CN, Ho CM, Chen R et al (2007) Silver nanoparticles: partial oxidation and antibacterial activities. J Biol Inorg Chem 12:527–534CrossRefGoogle Scholar
  47. Luoma SN (2008) Silver nanotechnologies and the environment: old problemds or new challenges? edn. Woodrow Wilson International Center for Scholars, Project on Emerging Nanotechnologies, The PEW Charitable Trusts, Washington, DCGoogle Scholar
  48. Luoma SN, Ho YB, Bryan GW et al (1995) Fate, bioavailability and toxicity of silver in estuarine environments. Mar Pollut Bull 31:44–54CrossRefGoogle Scholar
  49. Masciangioli T, Zhang WX (2003) Environmental technologies at the nanoscale. Environ Sci Technol 37:102A–108ACrossRefGoogle Scholar
  50. Maynard AD, Baron PA, Foley M et al (2004) Exposure to carbon nanotube material: aerosol release during the handling of unrefined single walled carbon nanotube material. J Toxicol Environ Health 67:87–107CrossRefGoogle Scholar
  51. Morel F, Hering J (1993) Principles and applications of aquatic chemistry. Wiley, New YorkGoogle Scholar
  52. Morgan IJ, Henry RP, Wood CM (1997) The mechanism of acute silver nitrate toxicity in freshwater rainbow trout Oncorhynchus mykiss is inhibition of gill Na+ and Cl transport. Aquat Toxicol 38:145–163CrossRefGoogle Scholar
  53. Morones JR, Elechiguerra JL, Camacho A, Holt K, Kouri JB, Ramirez JT, Yacaman MJ et al (2005) The bactericidal effects of silver nanoparticles. Nanotechnology 16:2346–2353CrossRefGoogle Scholar
  54. Murdock RC, Braydich-Stolle L, Schrand AM et al (2008) Characterization of nanomaterial dispersion in solution prior to in vitro exposure using dynamic light scattering technique. Toxicol Sci 101:239–253CrossRefGoogle Scholar
  55. Naddy RB, Gorsuch JW, Rehner AB et al (2007a) Chronic toxicity of silver nitrate to Ceriodaphnia dubia and Daphnia magna, and potential mitigating factors. Aquat Toxicol 84:1–10CrossRefGoogle Scholar
  56. Naddy RB, Rehner AB, McNerney GR et al (2007b) Comparison of short-term chronic and chronic silver toxicity to fathead minnows in unamended and sodium chloride-amended waters. Environ Toxicol Chem 26:1922–1930CrossRefGoogle Scholar
  57. Navarro E, Piccapietra F, Wagner F et al (2008) Toxicity of silver nanoparticles to Chlamydomonas reinhardtii. Environ Sci Technol 42:8959–8964CrossRefGoogle Scholar
  58. Nebeker AV, McAuliffe CK, Mshar R et al (1983) Toxicity of silver to steelhead and rainbow trout, fathead minnows and Daphnia magna. Environ Toxicol Chem 2:95–104CrossRefGoogle Scholar
  59. Nowack B, Bucheli TD (2007) Occurrence, behavior and effects of nanoparticles in the environment. Environ Pollut 150:5–22CrossRefGoogle Scholar
  60. Owen R, Handy RD (2007) Viewpoint: formulating the problems for environmental risk assessment of nanomaterials. Environ Sci Technol 40:5582–5588Google Scholar
  61. Pan Y, Neuss S, Leifert A et al (2007) Size-dependent cytotoxicity of gold nanoparticles. Small 11:1941–1949CrossRefGoogle Scholar
  62. Rejman J, Oberle V, Zuhorn IS et al (2004) Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis. Biochem J 377:159–169CrossRefGoogle Scholar
  63. Roberts ES, Malstrom SE, Dreher KL (2007) In situ pulmonary localization of air pollution particle-induced oxidative stress. J Toxicol Environ Health A 70:1929–1935CrossRefGoogle Scholar
  64. Roco MC (2003) Nanotechnology: convergence with modern biology and medicine. Curr Opin Biotechnol 14:337–346CrossRefGoogle Scholar
  65. Roco MC (2005) Environmentally responsible development of nanotechnology. Environ Sci Technol 39:106A–112ACrossRefGoogle Scholar
  66. Salata O (2004) Applications of nanoparticles in biology and medicine. J Nanobiotechnol 2:3CrossRefGoogle Scholar
  67. Schins RP (2002) Mechanisms of genotoxicity of particles and fibers. Inhal Toxicol 14:57–78CrossRefGoogle Scholar
  68. Schnizler MK, Bogdan R, Bennert A et al (2007) Short-term exposure to waterborne free silver has acute effects on membrane current of Xenopus oocytes. Biochim Biophys Acta 1768:317–323CrossRefGoogle Scholar
  69. Service RF (2003) Nanomaterials show signs of toxicity. Science 300:243CrossRefGoogle Scholar
  70. Sondi I, Salopek-Sondi B (2004) Silver nanoparticles as antimicrobial agent: a case study on E. coli as a model for gram-negative bacteria. J Colloid Interface Sci 275:177–182CrossRefGoogle Scholar
  71. Suzuki H, Toyooka T, Ibuki Y (2007) Simple and easy method to evaluate uptake potential of nanoparticles in mammalian cells using a flow cytometric light scatter analysis. Environ Sci Technol 41:3018–3024CrossRefGoogle Scholar
  72. Takenaka S, Karg E, Roth C et al (2001) Pulmonary and systemic distribution of inhaled ultrafine silver particles in rats. Environ Health Perspect 109:547–551CrossRefGoogle Scholar
  73. Terharr C, Ewell W, Dziuba S et al (1972) Toxicity of photographic processing chemicals to fish. Photogr Sci Eng 16:370–377Google Scholar
  74. USEPA (1994) Users guide for probit analysis of data from acute and short-term chronic toxicity tests with aquatic organisms. Biological Methods Branch, USEPA, Cincinnati, OHGoogle Scholar
  75. Wood C, Hogstrand C, Galvez F et al (1996) The physiology of waterborne silver toxicity in freshwater rainbow trout (Oncorhynchus mykiss): the effect of ionic Ag+. Aquat Toxicol 35:93–109CrossRefGoogle Scholar
  76. Yu C, Zhao J, Guo Y et al (2008) A novel method to prepare water-dispersible magnetic nanoparticles and their biomedical applications: magnetic capture probe and specific cellular uptake. J Biomed Mater Res 2:364–372CrossRefGoogle Scholar
  77. Zhu S, Oberdorster E, Haasch ML (2006) Toxicity of an engineered nanoparticle (fullerene, C60) in two aquatic species, Daphnia and fathead minnow. Mar Environ Res 62(Suppl):S5–S9CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Geoff Laban
    • 1
    • 4
  • Loring F. Nies
    • 2
  • Ronald F. Turco
    • 3
  • John W. Bickham
    • 1
    • 4
  • Maria S. Sepúlveda
    • 1
    • 2
  1. 1.Department of Forestry & Natural ResourcesPurdue UniversityWest LafayetteUSA
  2. 2.School of Civil EngineeringPurdue UniversityWest LafayetteUSA
  3. 3.Department of AgronomyPurdue UniversityWest LafayetteUSA
  4. 4.Center for the EnvironmentPurdue UniversityWest LafayetteUSA

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