The Protective effect of vitamin C on DNA breakage of gills and liver cells in Rainbow trout (Oncorhynchus mykiss) exposed to different concentrations of zinc oxide nanoparticles (ZnO)

Document Type : Research Paper



This study was to evaluate the effect of emerging pollutant nanoparticles of zinc oxide (ZnO) and safety effects of vitamin C on the DNA breakage in the liver and gill cells of rainbow trout (Oncorhynchus mykiss). Fish were exposed to nanoparticles on three levels (0, 40 and 80 mg/L) and three levels of vitamin C (0, 400 and 800 mg/Kg) in 9 treatments with 3 replications. Fish with average weight 170±10g were treated in 10 days. Liver and gill tissues were sampled in the fifth and tenth days. After the isolation, target samples were stored in microtubes contain alcohol 96%. DNA extraction was run with gel electrophoresis. At the end of the test, it was photographed with Gel-doc. Breakage of DNA was determined using the weighted average. According to the results, in liver and gill, effect of zinc oxide nanoparticle, vitamin C and interaction with time-concentration were quite significant on the level of DNA damage. Most damage on liver and gill tissues in 80 mg/L of zinc oxide nanoparticles were on the 5th day. Also in both tissues, DNA breakage rate was on the 10th day.  However, vitamin C was reduced DNA breakage.


خدابخشی ل. و پورباقر ه. 1392. اثر نانوذره اکسید روی (ZnO) بر بافت  کبد و آبشش ماهی قزل­آلای رنگین­کمان (Oncorhynchus mykiss). پروژه عملی، دانشگاه تهران. 54ص.
رسولی س. و عبدی ک. 1384. مبانی ایمنی‌شناسی ماهیان. انتشارات سیمرغ. 251ص.
غفاری ح.، پورباقر ه. و فرحمند ح. 1395. اثر سم مالاتیون بر شکستگی DNA در بافت کبد و آبشش ماهی قزل‌آلای رنگین‌کمان (Oncorhynchus mykiss) با استفاده از میانگین وزنی. نشریه شیلات (مجله منابع طبیعی ایران)، 69(1): 99-89.
یگانی م. 1377. نگاهی تازه به ویتامین ث. انتشارات سپهر. 180ص.
 Ahmed  M.K., Kundu  G.K., Al-Mamun  M.H., Sarkar  S.K., Akter  M.S. and Khan M.S. 2013. Chromium (VI) induced acute toxicity and genotoxicity in freshwater stinging catfish, Heteropneustes fossilis. Journal of Ecotoxicology and environmental Safety, 92: 64–70.
Ali D., Nagpure N.S., Kumar S., Kumar R. and Kushwaha B. 2008. Genotoxicity assessment of acute exposure of chlorpyrifos to freshwater fish Channa punctatus (Bloch) using micronucleus assay and alkaline single-cell gel electrophoresis. Journal of Chemosphere, 71(10): 1823–1831.
Azewedo P.A., Leeson S., Cho C.Y. and Bureau D.P. 2004. Growth and feed utilization of large size rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) reared in freshwater: Diet and species effects, and responses over time. Aquaculture Nutrition, 10(6): 401–411.
Banaee M., Mirvaghefi A.R., Rafei  G.R. and Majazi Amiri B. 2008. Effect of sub-lethal diazinon concentration on blood plasma biochemistry. International Journal of Environmental Research, 2(2): 189–198.
Banu B.S., Danadevi K., Rahman M.F., Ahuja Y.R. and Kaiser J. 2001. Genotoxic effect of mono-crotophos to sentinel species using comet assay. Food and Chemical Toxicology, 39(4): 361–366.
Borm P.J., Robbins D., Haubold S., Kuhlbusch T., Fissan H. Donaldson K., Schins R., Stone V., Kreyling W., Lademann J., Krutmann J., Warheit D. and Oberdorster E. 2006. The potential risks of nanomaterials: A review carried out for ECETOC. Particle and Fiber Toxicology, 3(11): 48–57.
Chae Y.J., Pham C.H., Lee J., Bae E., Yi J. and Gu M.B. 2009. Evaluation of the toxic impac of silver nanoparticles on Japanese medaka (Oryzias latipes). Aquatic Toxicology, 94(4): 320–327.
Chapman P.M., Dexter R.N. and Long E.R. 1998. Synoptic measures of sediments contamination, toxicity and infaunal-community composition (the Sediment Quality Triad) in San Francisco Bay. Marine Ecological Progress Series, 37: 75–93.
Dabrowski K. 2001. Ascorbic Acid in Aquatic Organisms Status and Perspectives. CRC Press, USA. 280P.
De Menezes G.C., Tavares-Dias M., Ono E.A., De Andrade J.I.A., Brasil E.M., Roubach R., Urbinati E.C., Marcon J.L. and Affonso E.G. 2006. The influence of dietary vitamin C and E supplementation on the physio-logical response of pirarucu, Arapaima gigas, in net culture. Comparative Biochemistry and Physiology A, 145(2): 9–24.
Devaux A., Pesonen M. and Monod G. 1997. Alkaline comet assay in rainbow trout hepatocytes. Toxicology in Vitro, 11(1-2): 71–79.
Farkas J., Christian P., Urrea J.A.G. and Roos N. 2010. Effect of silver and gold nanoparticles on rainbow trout (Oncorhynchus mykiss) hepatocyteses. Aquatic Toxicology, 96: 44–52.
Fassbender C. and Braunbeck T. 2013. Assessment of genotoxicity in gonads, liver and gills of zebrafish (Danio rerio) by use of the comet assay and micronucleus test after in vivo exposure to methyl methanesulfonate. Bulletin of Environmental Contamination and Toxicology, 91(1): 89–95.
Ferrari M. 2005. Cancer nanotechnology: Opportunities and challenge. Nature Reviews, Cancer, 5: 161–171.
Frenzilli G., Nigro M. and Lyons B.P. 2009. The comet assay for the evaluation of genotoxic impact in aquatic environment. Mutation Research, Reviews in Mutation Research, 681: 80–92.
Guilherme S., Gaivao I., Santos M.A. and Pacheco M. 2010. European eel (Anguilla anguilla) genotoxic and pro-oxidant responses following short-term exposure to Roundup®- a glyphosate-based herbicide. Mutagenesis, 25(5): 523–530.
Halver J.E. 1980. Vitamin require-ments of finfish. P: 8–191. In: Santos W., Lopes N., Barbosa J.J., Chaves D. and Valente J.V. (Eds.). Nutrition and Food Science, Present Knowledge and Utilization. Plenum Press, New York.
Handy R.D., Al-Bairuty G., Al-Jubory A., Ramsden C.S., Boyle D. and Shaw B.J. 2006. Effects of manufactured nanomaterials on fishes: a target organ and body systems physiology approach. J Fish Biol. 79(4): 821-853.
Henao B., Palacio J.A. and Camargo M. 2005. Evaluacion genotoxica de los plaguicidas cipermetrina y diazinon en tilapia roja (Oreochromis sp.). Journal of  Actualidades Biologicas, 27(82): 43–55.
Lee R.F. and Steinert S. 2003. Use of single cell gel electro-phoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. Journal of Mutation Research, 544: 43–64.
Li M.H. and Robinson E.H. 1999. Dietary ascorbic acid requirement for growth and health in fish. Journal of Applied Aquaculture, 9(2): 53–79.
Mitchelmore C.L. and Chipman J.K. 1997. DNA strand breakage in aquatic organisms and the potential value of the Comet assay in environmental monitoring. Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis, 399(2): 135–147.
Muntro D., Marrero M., Izquierdo M.S., Robaina L., Vergara J.M. and Tort L. 1999. Effect of vitamin E and C dietary supplementation on some immune parameters of gilthead seabream (Sparus aurata) juveniles subjected to crowding stress. Aquaculture, 171(3-4): 269–278.
N.R.C. 2011. Nutrient Requirements of Fish and Shrimp. National Academies Press, Washington DC, USA. 392P.
Safahieh A., Hedayati A., Savari A. and Movahedinia A. 2012. Effect of sublethal dose of mercury toxicity on liver cells and tissue of yellowfin seabream. Journal of Toxicology and Industrial Health, 28(7): 583–592.
Sharma S., Nagpure N.S., Kumar R., Pandey S., Srivastava S.K., Singh P.J. and Mathur P.K. 2007. Studies on the genotoxicity of endosulfan in different tissues of fresh water fish Mystus vittatus using the comet assay. Journal of Archives of Environmental Contamination and Toxicology, 53(4): 617–623.
Sies H. and Stahl W. 1995. Vitamins E and C, betacarotene, and other carotenoids as antioxidants. The American Journal of Clinical Nutrition, 62(6): 1315–1321.
Singh N., Monshion B., Jenkis G.I.S., Griffiths S.M., Williams P.M. and Moffeis T.O. 2009. Nanogenotoxicology: the DNA damaging potential engineered nanomaterials. Biomaterials, 30: 3897–3974.
Treves-Brown K. 2000. Applied Fish Pharmacology. Master of Arts, Cambridge, UK. 302P.
Verlhac V., Obach A., Gabaudan J., Schuep W. and Hole R. 1998. Immunomodulation by dietaryvitaminC and glucan in rainbow trout (Oncorhynchus mykiss). Fish and Shellfish Immunology, 8: 409–424.
Winston G.W., Regoli  F., Dugas A.J., Fong J.H. and Blanchard K.A. 1998. A rapid G.C. assay for determining oxyradical scavenging capacity of antioxidants and biological fluids, free radical. Free Radical Biology and Medicine, 24(3): 461–480.
Wise J.P., Goodale B.C., Wise S.S., Craig G.A., Pongan A.F., Walter R.B., Thompson W.D., Ng A.K., Aboueissa A.M., Mitani H., Spalding M.J. and Mason M.D. 2010. Silver nanospheres are cytotoxic and genotoxic to fish cells. Aquatic Toxicology, 97(1): 34–41.