Korean Journal of Ecology and Environment (생태와환경)

The Korean Society of Limnology (한국수생태학회)
권호 표지

Comet Assay as a New DNA-Level Approach for Aquatic Ecosystem Health Assessments

  • Sung, Min-Sun (School of Bioscience and Biotechnology Chungnam National University) ;
  • Lee, Sang-Jae (School of Bioscience and Biotechnology Chungnam National University) ;
  • Lee, Jae-Hoon (School of Bioscience and Biotechnology Chungnam National University) ;
  • Park, Sun-Young (Department of Food and Nutrition, Chungnam National University) ;
  • Ly, Sun-Yung (Department of Food and Nutrition, Chungnam National University) ;
  • An, Kwang-Guk (School of Bioscience and Biotechnology Chungnam National University)
  • Published : 2008.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Little is known about DNA-level and physiological levels for health assessments of stream or river environments. Recently, comet assay, so called Single Cell Gel Electrophoresis (SCGE) is introduced for assessments of DNA damage in the medical science, food science and mammal toxicology. The comet assay is known as a biomarker which is one of the best barometers in assessing the DNA damage by oxidative stress. In this study, we conducted the comet assay using sentinel species, Zacco platypus, as one of the pre-warning alarm systems for the aquatic ecosystem health assessments and also applied it to Gap Stream as a model system. Tail extent moments in the S1 and S2 were 5.20 and 9.90 respectively and the moment was 19.89 in the S3. Statistical ANOVA in the tail moments showed a significant difference (n=75, p<0.05) between S1 and S3. Also, the proportions of DNA in the tail were 14.47, 23.64, and $30.04{\mu}m$ in the upstream (control site), midstream, downstream sites, respectively. Our results in the downstream were accord with previous studies of individual-level, population-level, and community-level in Gap Stream. Our results suggest that the comet assay may be used as an important tool for diagnosing ecological health of aquatic ecosystems in the level of DNA.

Keywords

References

  1. Adams, S.M., A.M. Brown and R.W. Goede. 1993. A quantitative health assessment index for rapid evaluation of fish condition in the field. Transactions of the American Fisheries Society 122: 63-73 https://doi.org/10.1577/1548-8659(1993)122<0063:AQHAIF>2.3.CO;2
  2. Ali, D. and S. Kumar. 2008. Long-term genotoxic effect of monocrotophos in different tissues of freshwater fish Channa punctatus (Bloch) using alkaline single cell gel electrophoresis. Science of The Total Environment 405: 345-350 https://doi.org/10.1016/j.scitotenv.2008.05.037
  3. An, J.H., M.H. Back, J.S. Kim, J.H. Jeong and S.T. Kwon. 2004. Secondary metabolite/biochemistry/ metabolic engineering: Assessment of nucleus- DNA damage in red pepper cells treated with $\gamma$-Radiation through comet Assay. Korean Journal of Plant Biotechnology 31: 225-230 https://doi.org/10.5010/JPB.2004.31.3.225
  4. An, K.G., D.S. Kim, D.S. Kong and S.D. Kim. 2004. Integrative assessments of a temperate stream based on a multimetric determination of biological integrity, physical habitat evaluations, and toxicity tests. Bulletin of Environmental Contamination and Toxicology 73: 471-478
  5. An, K.G., J.H. Kim, D.Y. Bae and J.Y. Lee. 2006. Preliminary studies on the necropsy-based health assessment Index (HAI) using a ecological indicator species. Korean Jounal of Limnology 39: 62-72
  6. Anderson, R.O. and S.J. Gutreuter. 1983. Length, weight, and associated structural indices, p. 283- 300. In: Fisheries techniques (L.A. Nielsen and D.L. John, eds.). American Fisheries Society, Bethesda, Maryland
  7. Barton, B.A. 1994. Monitoring the health and condition of mountain whitefish populations using the autopsy-based assessment method: a case study. p. 459-463. In: High performance fish (D.D. Mackinlay ed.). High performance: Proceedings of the International Fish Physilolgy Symposium, Vancouver, British Columbia
  8. Bony, S., C. Gillet, A. Bouchez, C. Margoum and A. Devaux. 2008. Genotoxic pressure of vineyard pesticides in fish: Field and mesocosm surveys. Journal of Aquatic Toxicology 89: 197-203 https://doi.org/10.1016/j.aquatox.2008.06.017
  9. Cotelle, S. and J.F. Ferard. 1999. Comet assay in genetic ecotoxicology: A Review. Environmental and Molecular Mutagenesis 34: 246-255 https://doi.org/10.1002/(SICI)1098-2280(1999)34:4<246::AID-EM4>3.0.CO;2-V
  10. Frenzilli, G., M. Nigro and B.P. Lyons. 2008. The Comet assay for the evaluation of genotoxic impact in aquatic environments Journal of Mutation Research/ Reviews in Mutation Research, In Press, Corrected Proof, Available online 13 March 2008
  11. Goede, R.W. and B.A. Barton. 1990. Organismic indices and autopsy-based assessment as indicators of health and condition in fish, p. 93-108. In: Biological indicator of stress in fish (S.M. Adams, ed.). American Fisheries Society, Symposium 8, Bethesda, Maryland
  12. Harris, J.H. 1995. The use of fish in ecological assessments. Australian Journal of Ecology 20: 65-80 https://doi.org/10.1111/j.1442-9993.1995.tb00523.x
  13. Huguency, B., S. Camara, B. Samoura and M. Magassouba. 1996. Applying and index of biotic integrity based on communities in a west african. Hydrobiologia 331: 71-78 https://doi.org/10.1007/BF00025409
  14. Isani, G., G. Andreani, F. Cocchioni, D. Fedeli, E. Carpene and G. Falcioni. 2008. Cadmium accumulation and biochemical responses in Sparus aurata following sub-lethal Cd exposure. Ecotoxicology and Environmental Safety 72: 224-230 https://doi.org/10.1016/j.ecoenv.2008.04.015
  15. Jeon, E.J., Y.K. Park., T.S. Kim, J.S. Kim and M.H. Kang. 2003. Protective effect of yellow-green vegetable juices on DNA damage in Chinese hamster lung cell using comet assay. Journal of Korean Nutrient Society 36: 24-31
  16. Karr, JR. 1981. Assessment of biotic integrity using fish communities. Fisheries 6: 21-27 https://doi.org/10.1577/1548-8446(1981)006<0021:AOBIUF>2.0.CO;2
  17. Kim, D.S., K.G. An and K.H. Kim. 2003a. Heavy metal pollution in the of various land use types based on physicochemical characteristics. Journal of Environmental Science and Health 33: 839-853
  18. Kim, G.B., R.F. Lee and K.A. Maruya. 2003b. Application of single cell gel electrophoresis for detection of DNA single strand breaks in DNA of fish blood cell. Journal of the Korea Fisheries Society 36: 346-351 https://doi.org/10.5657/kfas.2003.36.4.346
  19. Koizumi, N. and Y. Matsumiya. 1997. Assessment of stream fish habitat based on index of biotic Integrity. Bulletin of Japanese Society Oceanography 61: 144-156
  20. Li, Y., Y. Wu, Y. Chen and Z. Kong. 2006. Genotoxicity evaluation and a primary risk assessment of organic pollutants in the drinking water sources of Nanjing, China. Journal of Environmental Sciences 18: 983-988 https://doi.org/10.1016/S1001-0742(06)60026-0
  21. Lyon, J., S. Navarro-Perez, P.A. Cochran, E. Snatana and C.M. Guzman-Arroyo. 1995. Index of biotic integrity based on fish assemblages for the conservation of stream and rivers in West-central Mexico. Conservation Biology 9: 569-584 https://doi.org/10.1046/j.1523-1739.1995.09030569.x
  22. Munkittrick, K.R., M.E. McMaster, G. Van Der Kraak, C. Portt, W.N. Gibbons, A. Farwell and M. Gray. 2000. Development of methods for effects-driven cumulative effects assessment using fish populations: moose river project. A Technical Publication of SETAC, Florida. USA
  23. Nehls, S. and H. Segner. 2005. Comet assay with the fish cell line rainbow trout gonad-2 for invitro genotoxicity testing of xenobiotics and surface waters. Journal of Environmental Toxicology and Chemistry 24: 2078-2087 https://doi.org/10.1897/04-301R.1
  24. Oberdorff, T. and R.M. Hughes. 1992. Modification of an index of biotic integrity based on fish assemblages to characterize rivers of the Seine Basin, France. Hydrobiologia 228: 117-130 https://doi.org/10.1007/BF00006200
  25. Ohio EPA. 1989. Biological criteria for the protection of aquatic life. Vol. III, Standardized biological field sampling and laboratory method for assessing fish and macroinvertebrate communities. USA
  26. Park, E.J. and M.H. Kang. 2002. Application of the alkaline comet assay for detecting oxidative DNA damage in human biomonitoring. Journal of Korean Nutrient Society 35: 213-222
  27. Sanders, R.E., R.J. Miltner, C.O. Yoder and E.T. Rankin. 1999. The use of external deformities, erosion, lesions, and tumors (DELT Anomalies) in fish assemblages for characterizing aquatic resouurces: A case study of seven Ohio streams. In: Assessing the sustainability and biological intergrity of water resources using fish communities. (T.P. Simon ed.). CRC press LLC, Florida
  28. Seo, J.W. and S.A. Lee. 2006. Determination of DNA damage in crucian carp (Carassius auratus) influenced by discharge of municipal sewage treatment plant in the Gap Stream, Korea. Journal of Korean Society of Environmental Engineers, Fall Symposium. p. 648-650
  29. Singh, N.P., M.T. McCoy, R.R. Tice and E.L. Schneider. 1998. A simple technic for quantitation of low levels of DNA damage in individual cells. Journal of Experimental Cell Research 175: 184-191
  30. US EPA. 2002. Summary of biological assessment programs and biocriteria development for states, tribes, territories, and interstate commissions: streams and wadable river. EPA-822-R-02-048. US EPA, USA