Toxicity and Uptake of 2,4,6-Trinitrotoluene (TNT) in Contaminated Soils to Eisenia fetida

토양내 오염된 2,4,6-trinitrotoluene (TNT)의 Eisenia fetida에 대한 독성 및 생물흡수

  • Received : 2015.10.06
  • Accepted : 2015.11.19
  • Published : 2015.11.30


Toxicity and uptake of 2,4,6-trinitrotoluene (TNT) in three different soils (OECD soil, natural soil and loess) to earthworm (Eisenia fetida) were investigated at several different spiked concentrations of TNT (0 to 200 mg/kg for OECD and natural soils, and 0 to 35 mg/kg for loess) and for different exposure periods (7, 14, 21, and 28 d). The LC50 values for 7 d exposure were 160.1, 159.4, and 28.81 mg/kg for OECD soil, natural soil, and loess, respectively. The LC50 values for 14, 21, and 28 d exposure were almost the same as those for 7 d exposure, showing that 7 d exposure time was enough to decide the toxicity (LC50) of TNT to Eisenia fetida, because the highest concentration of TNT in earthworm body was observed within around 5 d. The LC50 and uptake of TNT in loess were higher than those in OECD and natural soil. The uptake of TNT to the earthworm were correlated well with the initial concentration of TNT in the soil and TNT porewater concentration (R2> 0.9 in OECD, natural, and loess). The concentration of TNT in earthworm body decreased after 5 d, possibly caused by natural degradation of TNT by soil bacteria as well as earthworm.


2,4,6-trinitrotoluene;Eisenia fetida;Soil;Toxicity;Uptake


  1. Bae, B., Kim, S.-Y., Lee, I.-S., and Chang, Y.-Y., 2001, A study on the screening of 2,4,6-trinitrotoluene tolerant indigenous herbaceous plants, Journal of KoSSGE, 6, 3-11.
  2. Belden, J.B., Lotufo, G.R., Chambliss, C.K., Fisher, J.C., Johnson, D.R., and Boyd, R.E., Sims, J.G, 2011, Accumulation of 14C-trinitrotoluene and related nonextracteble (bound) residues in Eisenia fetida, Environ. Pollut., 159, 1363-1368.
  3. Best, E.P.H., Tatem, H.E., Geter, K.N., Wells, M.L., and Lane, B.K., 2008, Effects, uptake, and fate of 2,4,6-trinitrotoluene aged in soil in plants and worms, Environ. Toxicol. Chem., 27, 2539-2547.
  4. Brinch, U.C., Ekelund, F., and Jacobsen, C.S., 2002, Method for spiking soil samples with organic compounds, Appl. Environ. Microb., 68, 1808-1816.
  5. Conder, J.M., La Point, T.W., Steevens, J.A., and Lotufo, G.R., 2004, Recommendations for the assessment of TNT toxicity in sediment, Environ. Toxicol. Chem. 23, 141-149.
  6. Dalby, P.R., Baker, G.H., and Smith, S.E., 1996, Filter paper method to remove soil from earthworm intestines and to standardize the water content of earthworm tissue, Soil Biol. Biochem., 28, 685-687.
  7. Dodard, S.G., Powlowski, J., and Sunahara, G.I., 2004, Biotransformation of 2,4,6-trinitrotoluene (TNT) by enchytraeids (Enchytraeus albidus) in vivo and in vitro, Environ. Pollut., 131, 263-273.
  8. Gong, P., Siciliano, S.D., Greer, C.W., Paquet, L., Hawari, J., and Sunahara, G.I., 1999, Effects and bioavailability of 2,4,6-trinit rotoluene in spiked and field-contaminated soils to indigenous microorganisms, Environ. Toxicol. Chem., 18, 2681-2688.
  9. IBM, 2010, SPSS Statistics for Windows, Version 19.0, Armonk, NY: IBM Corp., USA.
  10. ISO (International Standard Organization), 2012, Soil quality - Effects of pollutants on earthworms - Part 2: Determination of effects on reproduction of Eisenia fetida/Eisenia andrei, ISO 11268-2:2012, Geneve.
  11. KMOE 2013, ES.07302.1a, pH-glass electrode method, Korean Ministry of Environment standard, Sejong, Korea.
  12. Kuperman, R.G., Simini, M., Siciliano, S., and Gong, P., Effects of energetic materials on soil organisms, In:Sunahara, G.I., Hawari, J., Lotufo, G., Kuperman, R.G., editors, Ecotoxicology of Explosives, Boca Raton: FL, CRC Press; 2009. p.35-76.
  13. Lachance, B., Renoux, A.Y., Sarrazin, M., Hawari, J., and Sunahara, G.I., 2004, Toxicity and bioaccumulation of reduced TNT metabolites in the earthworm Eisenia andrei exposed to amended forest soil. Chemosphere, 55, 1339-1348.
  14. Liu, K., Pan, X., Han, Y., Tang, F., and Yu, Y., 2012, Estimating the toxicity of the weak base carbendazim to the earthworm (Eisenia fetida) using in situ pore water concentrations in different soils, Sci. Total Environ., 438, 26-32.
  15. OECD, 1984, OECD Guideline for testing of chemicals: Earthworm, acute toxicity test, OECD Publishing: Paris, p. 1-9.
  16. OECD, 2008, OECD Guidlines for the testing of chemicals, section 2: Effects on biotic systems, test no. 226: Predatory mite (Hypoaspis (Geolaelaps) aculeifer) reproduction test in soil, OECD Publishing: Paris, p. 4-5.
  17. OECD, 2010, OECD Guidlines for the testing of chemicals, section 3: Degradation and accumulation, test no. 317: Bioaccumulation in terrestrial oligochaetes, OECD Publishing: Paris, p. 21.
  18. Nelson, D.W. and Sommers, L.E., Total carbon, organic carbon,and organic matter. In: Black, C.A., editors, Methods of soil analysis; Part 3. Chemical methods, Madison:WI, Soil Science of America and American Society of Agronomy; 1996, p. 961-1010.
  19. Renoux, A.Y., Sarrazin, M., Hawari, J., and Sunahara, G.I., 2000, Transformation of 2,4,6-trinitrotoluene in soil in the earthworm Eisenia andrei, Environ. Toxicol. Chem., 19, 1473-1480.
  20. Robidoux, P.Y., Hawari, J., Bardai, G., Paquet, L., Ampleman, G., Thiboutot, S., and Sunahara, G.I., 2002, TNT, RDX, and HMX decrease earthworm (Eisenia andrei) life-cycle responses in a spiked natural forest soil, Environ. Contam. Toxicol., 43, 379-388.
  21. Sarrazin, M., Dodard, S.G., Savard, K., and Lachance, B., 2009, Accumulation of hexahydro-1,3,5-trinitro-1,3,5-triazine by the earthworm Eisenia Andrei in a sandy loam soil, Environ. Toxicol. Chem., 28, 2125-2133.
  22. USEPA, 2007, Method 8330A: Nitroaromatics and nitroamines by high performance liquid chromatography (HPLC), Test Methods for the Evaluation of Solid Waste: Laboratory Manual Physical Chemical Methods. SW 846, Washington, DC, USA, Office of Solid Waste.
  23. Sims, J.G. and Steevens, J.A., 2008, The role of metabolism in the toxicity of 2,4,6-trinitrotoluene and its degradation products to the aquatic amphipod Hyalella azteca, Ecotoxicol. Environ. Safety 70, 38-46.
  24. Ter laak, T.L, Agbo, S.O., Barendregt, A., and Hermens, J.L.M., 2006, Freely dissolved concentration of PAHs in soil pore water: measurements via solid-phase extraction and consequences for soil tests, Environ. Sci. Technol., 40, 1307-1313.
  25. USEPA, 1986, Method 9081: Cation-Exchange Capacity of Soils (Sodium Acetate), Test Methods for the Evaluation of Solid Waste: Laboratory Manual Physical Chemical Methods. SW 846, Washington, DC, USA, Office of Solid Waste.
  26. Widianarko, B. and Straalen, N.V., 1996, Toxicokinetics-based survival analysis in bioassays using nonpersistent chemicals, Environ. Toxicol. Chem., 15, 402-406.