Effects on Biomarkers and Endocrine in Muddy Loach (Misgurnus anguillicaudatus) under 21 day Exposure to Methomyl

21일간 methomyl에 노출한 미꾸리의 생물지표 및 내분비계 영향

  • Han, Sun-Young (Environmental Toxicology Research Center, Korea Institute of Toxicology) ;
  • Kim, Ja-Hyun (Environmental Toxicology Research Center, Korea Institute of Toxicology) ;
  • Gwon, Ga-Young (Environmental Toxicology Research Center, Korea Institute of Toxicology) ;
  • Yeom, Dong-Hyuk (Environmental Toxicology Research Center, Korea Institute of Toxicology)
  • 한선영 (안전성평가연구소 환경독성연구센터) ;
  • 김자현 (안전성평가연구소 환경독성연구센터) ;
  • 권가영 (안전성평가연구소 환경독성연구센터) ;
  • 염동혁 (안전성평가연구소 환경독성연구센터)
  • Received : 2011.12.28
  • Accepted : 2012.02.20
  • Published : 2012.03.31


To evaluate the effect of endocrine disruption chemicals (EDCs) to aquatic organisms, muddy loach (Misgurnus anguillicaudatus) was exposed to low concentration methomyl for 21 days in order to identify the effect of biomarkers and endocrine. Vitellogenin (VTG) in blood plasma, which used widely as validated biomarker for endocrine disruption, was significantly greater in male fish exposed to 0.4 mg/L and 2 mg/L methomyl, and in female fish exposed to 0.08 mg/L, 0.4 mg/L, and 2 mg/L methomyl for 21 days (p<0.05). This results suggest that methomyl have probability of endocrine disruption to organism on aquatic system. While inhibition of Acetylcholinesterase (AChE) activity and increase of DNA damage in comet assay were verified by fish exposed to methomyl, change of ethoxyresorufin-O-deethylase (EROD) activity was not occurred, comparing the control group (p<0.05). Indicators at the level of organism such as condition factor (CF), hepato-somatic index (HSI), and gonado-somatic index (GSI) were not influenced by exposure of methomyl. In conclusion, these results showed the possibility of methomyl in regard to not only endocrine disruption but also impacts on biochemical biomarkers to aquatic organisms.


Supported by : 농촌진흥청


  1. Adrien Albert (1981) Selective Toxicity: The physico- chemical basis of therapy. p. 455, Chapman & Hall, the UK
  2. Anderson, M. J., H. Olsen, F. Matsumura and D. E. Hinton (1996) In vivo modulation of 17$\beta$-estradiol-induced vitellogenin synthesis and estrogen receptor in rainbow trout(Oncorhynchus mykiss) liver cells by $\beta$-naphthoflavone. Toxicol. Appl. Pharmacol. 137:210-218.
  3. Boenke, A. C., C. Searle and T. Karjalainen (2002) Contribution of European research to endocrine disruptors. Analytica. Chimica. Acta. 473:161-165.
  4. Bolognesi, C., M. Peluso, P. Degan, R. Rabboni, A. Munnia and A. Abbondandolo (1994) Genotoxic effects of the carbamate insecticide, methomyl. II. In vivo studies with pure compound and the technical formulation, "Lannate 25". Environ. Mol. Mutagen. 24(3):235-242.
  5. Bonatti, S., C. Bolognesi, P. Degan and A. Abbondandolo (1994) Genotoxic effect of the carbamate insecticide methomyl: I. In vitro studies with pure compound and the technical formulation "Lannate 25". Environ. Mol. Mutagen. 23(4): 306-311.
  6. Bucheli, T. D. and K. Fent (1995) Induction of cytochrome P450 as a biomarker for environmental contamination in aquatic ecosystem. Crit. Rev. Environ. Sci. Technol. 25: 201-268.
  7. David Pimentel (1992) Ecological Effects of Pesticides on Nontarget Species in Terrestrial Ecosystems. In Methods to Assess Adverse Effects of Pesticides on Non-target Organisms Vol. SCOPE 49 (ed. Robert G. Tardiff), Scientific Committee on Problems of the Environment (SCOPE), France.
  8. Ellman G. L., K. D. Courtney, V. Andres, Jr. and R. M. Feather- Stone (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol 7:88 -95.
  9. Fujimoto Y., Y. Ouchi, T. Hakuba, H. Chiba and M. Iwata (2008) Influence of modern irrigation, drainage system and water management on spawning migration of mud loach, Misgurnus anguillicaudatus C. Environ. Biol. Fish 81:185 -194.
  10. Giesy, J. P., K. Hilscherova, P. D. Jones, K. Kannan, and M. Machala (2002) Cell bioassays for detection of aryl hydrocarbon (AhR) and estrogen receptor (ER) mediated activity in environmental samples. Mar. Pollut. Bull. 45:3-16.
  11. Ham, K. D., S. M. Adams and M. J. Peterson (1997) Application of multiple bioindicators to differentiate spatial and temporal variability from the effects of contaminant exposure on fish. Ecotoxicol. Environ. Saf. 37:53-61.
  12. Hashimoto, Y. and Y. Nishiuchi (1981) Establishment of Bioassay Methods for the Evaluation of Acute Toxicity of Pesticides to Aquatic Organisms. Journal of Pesticide Science 6:257- 264 (Japanese, with English abstract).
  13. Hayes, T., K. Haston, M. Tsui, A. Hoang, C. Haeffele and A. Vonk (2002) Feminization of male frogs in the wild. Nature 419, 895-896.
  14. Hayes, T. B., P. Case, S. Chui, D. Chung, C. Haeffele, K. Haston, M. Lee, V. P. Mai, Y. Marjuoa, J. Parker and M. Tsui (2006) Pesticide Mixtures, Endocrine Disruption, and Amphibian Declines: Are We Underestimating the Impact? Environ. Health Perspect. 114:40-50.
  15. Horng, S. B., H. H. Kuo, M. Y. Lin, W. W. Lin and T. C. Wang (2007) Human gastric cells resistant to (-)-epigallocatechin gallate show cross-resistance to several environmental pollutants. Food Chem. Toxicol. 45:2171-2178.
  16. Hutchinson, T. H., G. T. Ankley, H. Segner and C. R. Tyler (2006) Screening and testing for endocrine disruption in fish-biomarker as "signposts," not "traffic light," in risk assessment. Environ. Health Perspect. 114(Suppl 1):106-114.
  17. Jung J. H., S. J. Kim, T. K. Lee, W. J. Shim, S. Woo, D. J. Kim and C. H. Han (2008) Biomarker responses in caged rockfish (Sebastes schlegeli) from Masan Bay and Haegeumgang, South Korea. Mar. Pollut. Bull. 57:599-606.
  18. Kennedy S. W. and S. P. Jones (1994) Simultaneous measurement of cytochrome P4501A catalytic activity and total concentration with a fluorescence plate reader. Anal. Biochem. 222:217- 223.
  19. Kim W. K., S. K. Lee and J. Jung (2010) Integrated assessment of biomarker responses in common carp (Cyprinus carpio) exposed to perfluorinated organic compounds. J. Hazard. Mater. 180:395-400.
  20. Koltz D. M., B. S. Beckman, S. M. Hill, J. A. Mclachlan, M. R. Walters and S. F. Arnold (1996) Identification of environmental chemicals with estrogenic activity using a combination of in vitro assays. Environ. Health Perspect. 104(10):1084-1089.
  21. Li, H., H. Jiang, X. Gao, X. Wang, W. Qu, R. Lin and J. Chen (2008) Acute toxicity of the pesticide methomyl on the topmouth gudgeon (Pseudorasbora parva): mortality and effects on four biomarkers. Fish Physiol. Biochem. 34:209-216.
  22. OECD (2008) Draft report of the validation of the 21-day fish endocrine screening assay for the detection of endocrine active substances (Phase 2- Testing Negative Substance). April 2007.
  23. OECD (2009) OECD guideline for the testing of chemical No. 230. 21-day Fish Assay: A Short-Term Screening for Oestrogenic and Androgenic Activity, and Aromatase Inhibition. Adopted: 7 September 2009.
  24. Ozdemir, A., M. Duran and A. Sen (2010) Potential use of the oligochaete Limnodrilus profundicola V., as a bioindicator of contaminant exposure. Environ. Toxicol. Published online 22 July 2010 in Wiley Online Library: DOI 10.1002/tox.20527.
  25. Stegeman, J. J. and M. E. Hahn (1994) Biochemistry and molecular biology of monooxygenases: current perspectives on forms, functions, and regulation of cytochrome P450 in aquatic species. pp. 87-206, In Aquatic Toxicology: Molecular, Biochemical, and Cellular Perspectives (ed. DC Malins and GK Ostrander), Lewis Publishers, U. S. A.
  26. Sun, X. Y., Y. T. Jin, B. Wu, W. Q. Wang, X. L. Pang and J. Wang (2010) Study on genotoxicity of aldicarb and methomyl. Huan Jing Ke Xue 31(12):2973-2980 (Chinese, with English abstract).
  27. Tiemann, U. (2008) In vivo and in vitro effects of the organochlorine pesticides DDT, TCPM, methoxychlor, and lindane on the female reproductive tract of mammals: A review. Reprod. Toxicol. 25:316-326.
  28. Tomlin, C. D. S. (2003) The pesticide manual(13th Ed.), British Crop Protection Council, th UK.
  29. US EPA (1997) EPA special report on endocrine disruption.
  30. US EPA (1998) Final report of endocrine disruptor screening and testing advisory committee.
  31. US Geological Survey (USGS) (1999) Biomonitoring of Environmental Status and Trends (BEST) program: Field Procedures for Assessing the Exposure of Fish to Environmental Contaminants, USGS/BRD/ITR-1999-0007.
  32. US Geological Survey (USGS) (2000) Biomonitoring of Environmental Status and Trends (BEST) Program: selected methods for monitoring chemical contaminants and their effects in aquatic ecosystems. USGS/BRD/ITR-2000-0005.
  33. Wei, L. Y., J. S. Chao and C. C. Hong (1997) Assessment of the ability of propoxur, methomyl and aldicarb, three carbamate insecticides, to induce micronuclei in vitro in cultured Chinese hamster ovary cells and in vivo in BALB/c mice. Environ. Mol. Mutagen. 29:386-393.<386::AID-EM7>3.0.CO;2-8
  34. Whyte J. J., R. E. Jung, C. J. Schmitt and D. E. Tillitt (2000) Ethoxyresorufin-O-deethylase (EROD) activity in fish as a biomarker of chemical exposure. Crit. Rev. Toxicol. 30(4): 347-570.
  35. Woods, M. and A. Kumar (2011) Vitellogenin induction by 17 $\beta$-estradiol and 17$\alpha$-ethynylestradiol in male Murray rainbowfish (Melanotaenia fluviatilis). Environ. Toxicol. Chem. 30(11): 2620-2627.
  36. Xuereb, B., E. Lefévre, J. Garric and O. Geffard (2009) Acetylcholinesterase activity in Gammarus fossarum (Crustacea Amphipoda): Linking AChE inhibition and behavioural alteration. Aqua. Toxicol. 94:114-122.
  37. Yi, M. Q., H. X. Liu, X. Y. Shi, P. Liang and X. W. Gao (2006) Inhibitory effects of four carbamate insecticides on acetylcholinesterase of male and female Carassius auratus in vitro. Comp. Biochem. Physiol. C 143:113-116.
  38. Doopedia 두산백과, "미꾸리"
  39. 국립환경연구원 (1999) 내분비계 장애물질 이해와 대응.
  40. 국립환경연구원 (2007) 내분비계장애물질에 의한 생태영향 정밀조사.
  41. 김정곤, 박예나, 김우근, 김지원, 이성규, 최경호 (2010) 유해물질 노출로 인한 분자 . 생화학적 바이오마커와 담수 어류에 대한 현장 적용성, 한국환경보건학회지 36(5):418-434.
  42. 농촌진흥청 (2010) 농촌진흥청 고시 제2010-29호 농약의 등록기준 (2010. 10. 13.)
  43. 유홍일, 이해근, 전성환 (1991) 농약잔류분석법. pp. 119-127, 동화기술, 대한민국.
  44. 이병천, 김수진, 윤준헌, 김은주, Duong, N. C., 엄익춘, Shiraishi, F., 최경희 (2010a) 에스트로겐과 다이옥신 수용체 효모를 이용한 내분비계장애 영향 평가, 한국환경분석학회지 13(2):109-115.
  45. 이제봉, 신진섭, 이희동, 정미혜, 유아선, 강규영 (2004) 내분비계 장애추정농약에 대한 에스트로겐성 영향검색 및 위해성 평가, 농약과학회지 8(2):95-102.
  46. 이주영, 이희정, 김우성, 최동미, 채갑용, 강찬순, 신용운, 도정아, 최원조 (2010b) 2009년 유통 농산물 중 잔류농약 실태조사, 한국식품위생안전성학회지 25(2);192-202.
  47. 장정희, 봉영훈, 김동규, 김미경, 정갑수, 손성완 (2010a) 2008-2009 년 국내 폐사 야생조류 및 동물 체내의 잔류농약 분석, 대한수의학회지 50(3):197-203.
  48. 장제헌, 김정희, 최재원, 오은전, 노혜란 (2010b) 한강수계 하천수의 카바메이트계 농약 모니터링(2007-2009), 한국환경분석학회지 13(1):33-39.
  49. 정영호외 (2000) (최신) 농약학. p. 100, 시그마프레스, 대한민국
  50. 한국브리태니커, 전상인, "미꾸리"
  51. 한국작물보호협회, 농림수산식품부 친환경농업팀 (2009)