Effects of nano-silver particles on some vital tissues of Zebra fish (Danio rerio) fed via oral administration


1 Faculty of Fishery, Islamic Azad University of Science and Research Branch, Tehran, Iran

2 Department of Aquatic Animal Health and Diseases, Iranian Fisheries Science Research Institute, Agricultural Research, Education and Extension Organization, Tehran, Iran

3 Department of Aquatic Animal Health, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran


BACKGROUND: This study was initiated to improve our understanding  of the health and environmental impact of silver nanoparticles (Ag-np). OBJECTIVES: The purpose of the study is application and direct effects of silver nanoparticles on Zebra fish (Danio rerio). METHODS: After characterizing the AgNPs using TEM, EDX, UV-Vis Spectroscopy, XRF and SEM methods, their effects on some vital tissues have been tested successfully in vitro. In this study, 540 fish (2±0.05 g) were randomly divided into 9 groups in triplicate for acute tests (0,10, 50, 100, 200, 400, 600, 800 and 1000 mg/kgfood). After short term (96h), chronic toxicity tests were evaluated using under lethal concentration (100, 400, 600 and 800 mg/kgfood) fed with experimental diet for 8 weeks. Fish in groups 1 to 4 were fed by food supplemented with 100, 400, 600 and 800 mg/kg food, respectively. Group 5 was fed with basal food without supplementation. After histopathology, heavy metals were measured by spectrum photometry reveal. RESULTS: According to the results of acute tests, the 96h LC50 values in 24, 48, 72 and 96h were 804.601, 486.637, 323.696 and 195.208 mg/kgfood AgNPs for the Zebra fishes respectively. According to the results of chronic toxicity tests, fed via oral administration of AgNPs produced significant histopathological effects. Also, the most important histopathological effects of AgNPs were observed in the liver (vasculature and exposure, degeneration of some hepatocytes), intestine (increase in the submucosa layer, narrowing of the intestinal lumen  and reduced intestinal absorption), gills (clubbing of gill secondary lamaleas, hyperplasia, hyperemia and shortening of the primary lamaleas gills) and kidney (degeneration, high increase in interstitial cells and dilatation of Bowman’s space of glomeruli), respectively. The greatest bioaccumulation of silver occurred in the liver, gills and muscle of fish respectively (p<0.05). CONCLUSIONS: The release of untreated nanoparticle waste to the environment should be restricted for the well-being of human and aquatic species.


Alishahi, A., Mirvaghefi, A., Tehrani, M.R., Farahmand, H., Shojaosadati, S.A., Dorkoosh, F.A (2011) Shelf  life and delivery enhancement of vitamin C using chitosan nanoparticles. Food Chem. 126: 935-940.
Asharani, Y.L., Wu, Z.Y., Gong and Valiyaveettil, S. (2008) Toxicity of silver nanoparticles in zebrafish models. Nanotechnology. Volume 24, (9)-255102.Doi:10.1088/0957-4484/19/25/255102.
Bhui, D.K., Bar, H., Sarkar, P., Sahoo, G.P., De, S.P., Misra, A. (2009) Synthesis and UV-vis spectroscopic study of silver nanoparticles in aqueous SDS solution. J Mol Liq. 145: 33-37.
Braydich-Stolle, L., Saber, H., Schlager, J.J., Hofmann, M-C. (2005) In vitro cytotoxicity of nanoparticles in mammalian germline stem cells Toxicol Sci. 88: 412-9.
Chen, R., Lok, C.N., Ho, C.M., , He, Q.Y., Yu, W.Y., Sun, H. (2007) Silver nanoparticles: partial oxidation and antibacterial activities. J Biol Inorg Chem. 12: 527-34.
Federici, G., Shaw, B.J., Handy, R.D. (2007) Toxicity of titanium dioxide nanoparticles to rainbow trout (Oncorhynchus mykiss): Gill injury, oxidative stress, and other physiological effects. Aquat Toxicol. 84: 415-430.
Fu-ping, T., Zhi-guo, Z., Dun-fu, W., Dun-hu, W. (2009) Study on the effect of bionano-selenium feed additive on the breed of freshwater fish. Studies of Trace Elements and Health. 5: 31-34.
Gajjar, P., Pettee, B., Britt, D.W., Huang, W. (2009) Antimicrobial activities of commercial nanoparticles against an environmental soil microbe, Pseudomonas putida KT2440. J Biol Eng. 3: 26-9.
Griffitt, R.J., Griffitt, R., Weil, K.A., Hyndman. N.D., Denslow, K., Taylor, D. (2007) Exposure to copper nanoparticles causes gill injury and acute lethality in zebrafish (Danio rerio). Environ Sci Technol.  41: 8178-8186.
Griffitt, R.J., Griffitt, J., Luo, J., Gao, J.C., Bonzongo, D., Barber, S. (2008)  Effects of particle composition and species on toxicity of metallic nanomaterials in aquatic organisms. Environ Toxicol Chem,  27: 1972-1978.
Griffitt, R.J., Griffitt, K., Hyndman, N.D., Denslow, D., Barber, S. (2009) Comparison of molecular and histological changes in zebrafish gills exposed to metallic nanoparticles. Toxicol Sci. 107: 404-415.
Grosell, M., Nielsen, C., Bianchini, A. (2002) Sodium turnover rate determines sensitivity to acute copper and silver exposure in freshwater animals. Comparative Biochemistry and physiology, Part C: Toxicology & Pharmacology. 133: 287-303.
Handy, R.D., Shaw, B.J. (2011)  Ecotoxicity of nanomaterials to fish: challenges for ecotoxicity testing. Integr Environ Assess Manag. 3: 458-460.
Handy, R.D., Henry, T.B., Scown, T.M., Johnston, B.D., Tyler, C.R. (2008) Manufactured nanoparticles: their uptake and effects on fish: a mechanistic analysis. Ecotoxicology. 17: 396-409.
Kim, J.S., Kuk, E., Yu, K.N., Kim, J.H., Park, S.J., Lee, H.J. (2007) Antimicrobial effect of silver nanoparticles. Nanomed: Nanotechnology, Biology and Medicine. 3: 95-101.
Kim, S.H., Woo, K.S., Liu, B.Y.H., Zachariah, M.R. (2005) Method of measuring charge distribution of nanosized aerosols. J Colloid Interface Sci. 282: 46-57, 1-23.
Kittler, S., Greulich, C., Diendorf, J., Keoller, M., Epple, M. (2010) Toxicity of silver nanoparticles increases during storage because of slow dissolution under release of silver ions. Chemistry of Materials. 22: 4548-4554.
Kong, H., Jang, J. (2006) One-step fabrication of silver nanoparticle embedded polymer nanofibers by radical-mediated dispersion polymerization. Chem Commun. 28: 3010-3012.
Kumar, V., Cotran, R., Robbins, S.L. (1992) Basic Pathology. (5th ed.) W.B. The University of Chicago, Chicago, Illinois.
Lanno, R.P., Hicks, B., Hilton, J.W. (1987) Histological observations on intrahepatocytic coppercontaining granules in rainbow trout reared on diets containing elevated levels of copper. Aquat Toxicol. 12: 291-376.
Liu, S., Huang, W., Chen, S., Avivi, S., Gedanken, A. (2001) Synthesis of X-ray amorphous silver nanoparticles by the pulse sonoelectrochemical method. J Non Cryst Solids. 283: 231-236.
MOOPAM. (1993) Manual of  oceanographic and pollutant Analysis Methods. (3rd ed.) Kuwait,
Regional Organization for the Protection of the Marine Environment, (OCoLC) 646961227.
Obserdorster, E. (2004) Manufactured nanomaterials (fullerenes, C60) induce oxidative stress in the brain of juvenile largemouth bass Environ. Health Perspect. 112: 1058-62.
Petit, C., Lixon, P., Pileni, M.P. (1993) In-situ synthesis of silver nanocluster in AOT reverse micelles. J Phys Chem. 97: 12974-12983.
Reynolds, G.H. (2001) Environmental regulation of nanotechnology: some preliminary observations. Nano Archive. 31: 10681-10688.
Richard, R.O., Handy, R., Eugenia, V.J. (2008) The ecotoxicology of nanoparticles and nanomaterials: current status, knowledge gaps, challenges, and future needs. Ecotoxicology. 17: 315-325.
Rushton, E.K., Jiang, J., Leonard, S.S., Eberly, S., Castranova, V., Biswas, P. (2010) Concept of assessing nanoparticle hazards considering nanoparticle dosemetric and chemical/biological response metrics. J Toxicol Environ Health Part A. 73: 445-461.
Scown, T.M., Santos, E.M., Johnston, B.D., Gaiser, B., Baalousha, M., Mitov, S. (2010) Effects of aqueous exposure to silver nanoparticles of different sizes in rainbow trout. Toxicol Sci. 115: 521-534.
Shahverdi, A.R., Fakhimi, A., Shahverdi, H.R., Minaian, S. (2007) Synthesis and effect of silver Nanoparticles on the antibacterial activity of different antibiotics against Staphylococcus aureus and Escherichia coli. Nanomedicine: Nanotechnology, Biology and Medicine. 3: 168-171.
Sharma, V.K., Yngard, R.A., Lin, Y. (2009) Silver nanoparticles: Green synthesis and their antimicrobial activities. Adv Colloid Interface Sci. 145: 83-96.
Skebo, J.E., Grabinski, C.M., Schrand, A.M., Schlager, J.J., Hussain, S.M. (2007) Assessment of metal nanoparticle agglomeration, uptake, and interaction using high-illuminating system Int. J Toxicol. 26: 135-41.
Smith, C.J., Shaw, B.J., Handy, R.D. (2007) Toxicity of single walled carbon nanotubes on rainbow trout (Oncorhynchus mykiss): respiratory toxicity, organ pathologies, and other physiological effects. Aquat Toxicol. 82: 94-109.
Snell, T.W., Hicks, D.G. (2009) Assessing Toxicity of Nanoparticles Using Brachionus manjavacas (Rotifera). Environ Toxicol. 26:146-152.
Soltani, M., Esfandiary, M., Sajadi, M.M., Khazraeenia, S., Bahonar, A.R., Ahari. H. (2011) Effect of nanosilver particles on hatchability of rainbow trout (Oncorhynchus mykiss) egg and survival of the produced larvae. Iran J Fish Sci. 10: 167-176.
Tian, J., Wong, K.K., Ho, C.M., Lok, C.N., Yu, W.Y., Che, C.M. (2007) Topical delivery of silver nanoparticles promotes wound healing. J  Med Chem. 2: 129-36.
Wang, Y., Li, J. (2011) Effects of chitosan nanoparticles on survival, growth and meat quality of tilapia, Oreochromis nilotica. Nanotoxicology. 5: 425-431.
Webb, N.A., Wood, C.M. (1998) Physiological analysis of the stress response associated with acute silver nitrate exposure in freshwater rainbow trout (Oncorhynchus mykiss). Environ Toxicol Chem. 17: 579-88.
Yoon, K.Y., Hoon-Byeon, J., Park, J.H., Hwang, J. (2007) Susceptibility constants of Escherichia coli and Bacillus subtilis to silver and copper nanoparticles. J Sci Total Environ. 373: 572-5.
Zhuo, P.F., Yu, X.C., Du, Q., Song, K.S., He, Z.H. (2011) Photocatalytic degradation of ammonia nitrogen in aquaculture wastewater by using nano-TiO2. Adv Mat Res. 197-198: 774-779.