Bioremediation of Tetrachloroethylene-Contaminated Groundwater in a Model Aquifer: Effects on Green Frogs (Rana clamitans) and Xenopus laevis as Potential Wetland Receptors

  • Tana V. McDaniel
  • Nathalie Ross
  • Pamela A. Martin
  • Helena Steer
  • Ann-Marie Irwin Abbey
  • Suzanne Lesage


Recent regulations require that the ecological effects of microorganisms introduced into the environment, such as for groundwater bioremediation, be assessed prior to their utilization. A native anuran (Rana clamitans) and a model anuran (Xenopus laevis) were used as potential wetland receptors of tetrachloroethylene (PCE)-contaminated groundwater, undergoing three bioremediation treatments: natural attenuation (NA), biostimulation (ST), and bioaugmentation (AU). Eggs of both species were exposed acutely (96 h) to remediated effluents. Xenopus tadpoles were chronically exposed to the effluents for 100 days and were screened for the presence of bacterial pathogens. There was no impact on the survivorship of the frogs exposed either acutely or chronically to the NA, ST, or AU effluents; nor was there any evidence of bacterial infection found, with the exception of control individuals. The results of these exposures suggest that bioremediation with KB-1™ culture poses a minimal threat to anuran development and survivorship.


Chloroethylenes Natural Attenuation Xenopus Embryo Green Frog Model Aquifer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This project was funded by the Environmental Management of Biotechnology Regulation and Research (EMBRR) program of Environment Canada. The authors thank Sandra Toquica Diaz, Gregg Ferris, Liam Kelly, and Sue Brown for their advice and technical support.


  1. Alford R (1999) Ecology: resource use, competition and predation. In: McDiarmid RW, Altig RA (eds) Tadpole: Biology of anuran larvae. University of Chicago Press, Chicago, IL, p 240–278Google Scholar
  2. Anver MR, Pond CL (1984) Biology and diseases of amphibians. In: Fox J G, Cohen BJ, Loew FM (eds) Laboratory animal medicine. Academic Press, New York, NY, p 434–445Google Scholar
  3. ASTM (1998) Standard Guide for Conducting the Frog Embryo Teratogenesis Assay-Xenopus (FETAX) ASTM E1439-98. ASTM International, West Conshohocken, PAGoogle Scholar
  4. Baker P, Futamata H, Harayama S, Watanabe K (2001) Bacterial populations occurring in a trichloroethylene-contaminated aquifer during methane injection. Environ Microbiol 3:187–193CrossRefGoogle Scholar
  5. Baldassi L, Hipolito M, DeSouza FL, DeSouza CWO (1995) Clostridia presence in the bullfrog (Rana catesbeiana Shaw, 1802) mouth myiasis lesion. Bol Inst Pesca Sau Paulo 22:95–102Google Scholar
  6. Bantle JA, Dumont JN, Finch RA, Linder G, Fort DJ (1998) Atlas of abnormalities. Oklahoma State University Press, OklahomaGoogle Scholar
  7. Bergey DH, Holt JG, Kriet NR, Sneath P (1984) Bergey’s manual of systematic bacteriology. Williams and Wilkins, Baltimore, MDGoogle Scholar
  8. Call DJ, Brooke LT, Ahmad N, Richter JE (1983) Toxicity and metabolism studies with EPA priority pollutants and related compounds in freshwater organisms. U.S. Environmental Protection Agency, Duluth, MNGoogle Scholar
  9. Crawshaw GJ (2000) Diseases and pathology of amphibians and reptiles. In: Sparling DW, Linder G, Bishop CA (eds) Ecotoxicology of amphibians and reptiles. SETAC Press, Pensacola, FL, pp 199–231Google Scholar
  10. Devinny JS, Chang S H (2000) Bioaugmentation for soil bioremediation. In: Wise D L, (ed) Bioremediation of contaminated soils. Marcel Dekker, New York, NY, pp 465–488Google Scholar
  11. Duhamel M, Wehr SD, Yu L, Rizvi H, Seepersad D, Dworatzek S, Cox EE, Edwards EA (2002) Comparison of anaerobic dechlorinating enrichment cultures maintained on tetrachloroethene, trichloroethene, cis-dichloroethene and vinyl chloride. Water Res 36:4193–4202CrossRefGoogle Scholar
  12. Duhamel M, Mo K, Edwards EA (2004) Characterization of a highly enriched dehalococcoides-containing culture that grows on vinyl chloride and trichloroethene. Appl Environ Microbiol 70:5538–5545CrossRefGoogle Scholar
  13. Environment Canada (1994) Tetrachloroethylene. Canadian Environmental Protection Act. Priority substances list supporting document. Environment Canada, Ottawa, ONGoogle Scholar
  14. Environment Canada (1997) Canadian Environmental Protection Act - New Substances Notification Regulation, Environment Canada, Ottawa, ONGoogle Scholar
  15. Fort DJ, Stover EL, Rayburn JR, Hull M, Bantle JA (1993) Evaluation of the developmental toxicity of trichloroethylene and detoxification metabolites using Xenopus. Teratogen Carcinogen Mutagen 13:35–45CrossRefGoogle Scholar
  16. Fuller ME, Scow KM, Lau S, Ferris H (1997) Trichloroethylene (TCE) and toluene effects on the structure and function of the soil community. Soil Biol Biochem 29:75–89CrossRefGoogle Scholar
  17. Hird DW, Diesch S L, McKinnell RG, Gorham E, Martin FB, Meadows CA, Gasiorowski M (1983) Enterobacteriaceae and Aeromonas hydrophila in Minnesota frogs and tadpoles (Rana pipiens). Appl Env Microbiol 46:1423–1425Google Scholar
  18. Iwamoto T, Tani K, Nakamura K, Suzuki Y, Kitagawa M, Eguchi M, Nasu M (2000) Monitoring impact of in situ biostimulation treatment on groundwater bacterial community by DGGE. FEMS. Microbiol Ecol 32:129–141CrossRefGoogle Scholar
  19. Kaplan HM (1953) The care and diseases of the frog. Proc Anim Care Panel 4:74–92Google Scholar
  20. Lendvay JM, Loeffler FE, Dollhopf M, Aiello MR, Daniels G, Fathepure BZ, Gebhard M, Heine R, Helton R, Shi J, Krajmalnik-Brown R, Major CL Jr, Barcelona MJ, Adriaens P (2003) Bioreactive barriers: a comparison of bioaugmentation and biostimulation for chlorinated solvent remediation. Environ Sci Technol 37:1422–1431CrossRefGoogle Scholar
  21. Lorah MM, Olsen LD (1999) Natural attenuation of chlorinated volatile organic compounds in a freshwater tidal wetland: Field evidence of anaerobic biodegradation. Water Resources Res 35:3811–3827CrossRefGoogle Scholar
  22. Major DW, McMaster ML, Cox EE, Edwards EA, Dworatzek SM, Hendrickson ER, Starr MG, Payne JA, Buonamici LW (2002) Field demonstration of succesful bioaugmentation to acheive dechlorination of tetrachloroethene to ethene. Environ Sci Technol 36:5106–5116CrossRefGoogle Scholar
  23. McDaniel T, Martin P, Ross N, Brown S, Lesage S, Pauli B (2004) Effects of chlorinated solvents on four species of North American amphibians. Arch Environ Contam Toxicol 47:101–107CrossRefGoogle Scholar
  24. Meteyer CU, Loeffler IK, Fallon JF, Converse KA, Green E, Helgen JC, Kersten S, Levey R, Eaton-Poole L, Burkhart JG (2000) Hind limb malformations in free-living northern leopard frogs (Rana pipiens) from Maine, Minnesota and Vermont suggest multiple etiologies. Teratology 62:151–171CrossRefGoogle Scholar
  25. Négrel P, Petelet- Giraud E, Barbier J, Gautier E (2003) Surface water-groundwater interactions in an alluvial plain: Chemical and isotopic systematics. J Hydrology 277:248–267CrossRefGoogle Scholar
  26. Nieuwkoop, Faber (1994) Normal table of Xenopus laevis (Daudin). Garland Publishing Inc, New YorkGoogle Scholar
  27. Ouellet M (2000) Amphibian deformities: Current state of knowledge. In: Sparling DW, Linder G, Bishop CA (eds) Ecotoxicology of amphibians and reptiles. SETAC Press, Pensacola FL pp 617–661Google Scholar
  28. Parichy DM, Kaplan RH (1992) Maternal effects on offspring growth and development depend on environmental quality in the frog Bombina orientalis. Oecologia 91:579–586CrossRefGoogle Scholar
  29. Pierce BA, Sikand N (1985) Variation in acid tolerance in wood frogs: Genetic and maternal effects. Can J Zool 63:1647–1651CrossRefGoogle Scholar
  30. Richards CM (1958) The inhibition of growth in crowded Rana pipiens tadpoles. Physiol Zool 31:138–151Google Scholar
  31. Ross N, Deschenes L, Bureau J, Clément B, Comeau Y, Samson R (1998) Ecotoxicological assessment and effects of physicochemical factors on biofilm development in groundwater conditions. Environ Sci Technol 32:1105–1111CrossRefGoogle Scholar
  32. Ross N, Millar K, Brown S, Steer H, Robinson C, Grande P, McDaniel T, Martin P, Pauli B, Lesage S (2002) Assessing the biosafety of three bioremediation approaches in a tetrachloroethylene-contaminated environment, Paper 1C-04. In: Proceedings of the Third International Conference on Remediation of Chlorinated and Recalcitrant Compounds, Monterey, CA, May 20–23, 2002Google Scholar
  33. Sokal RR, Rohlf FL (1999) Biometry, 2nd ed. WH Freeman and Co. New York, NYGoogle Scholar
  34. StatSoft Inc. (2003) STATISTICA (data analysis software system). Tulsa, OKGoogle Scholar
  35. Sun Y, Bartlett C, Johnson CD, Klecka GM, Clement TP (2000) Natural attenuation of chlorinated ethene compounds: Model development and field-scale application at the Dover site. J Contam Hydrol 42:113–140CrossRefGoogle Scholar
  36. Tobias CR, Harvey JW, Anderson IC (2001) Quantifying groundwater discharge through fringing wetland to estuaries. Seasonal variability, methods comparison, and implications for wetland-estuary exchange. Limnol Oceanog 46:604–615CrossRefGoogle Scholar
  37. Todd D K (1980) Groundwater hydrology. John Wiley & Sons, New York, NYGoogle Scholar
  38. US EPA (1996) U.S. EPA Test Method 82 60B: Volatile organic compounds (VOCs)-GC/MS. EPA report # SW-846 ch 4.3.2. United States Environmental Protection Agency, Washington, DCGoogle Scholar
  39. US EPA (1997) Point to consider in the preparation of TSCA biotechnology submissions for microorganisms. Office of Pollution Prevention and Toxics, Washington, DCGoogle Scholar
  40. Westrick JJ, Mello JW, Thomas RF (1984) The groundwater supply survey. J Am Water Works Assoc 76:52–59Google Scholar

Copyright information

© Springer Science+Business Media, Inc. 2007

Authors and Affiliations

  • Tana V. McDaniel
    • 1
  • Nathalie Ross
    • 2
  • Pamela A. Martin
    • 1
  • Helena Steer
    • 2
  • Ann-Marie Irwin Abbey
    • 2
  • Suzanne Lesage
    • 2
  1. 1. Canadian Wildlife Service, Environment CanadaBurlingtonCanada
  2. 2.National Water Research Institute, Environment CanadaBurlingtonCanada

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