Skip to main content
Log in

Determination of Urease Activity in Soils by Carbon Dioxide Release for Ecotoxicological Evaluation of Contaminated Soils

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

A method for the quantification of urease enzyme activity has been set up, which is based on the quantification of carbon dioxide set free into the head space of gastight vessels. The method can be applied for ecotoxicological characterisation of contaminated soil samples besides other methods like soil respiration measurements or nitrification inhibition tests. The sieved soil sample can be incubated under nearly natural conditions with an adjusted water content of about 50% of the water holding capacity. Ammonia or urea do not need to be extracted, since carbon dioxide release is correlated to urease activity. Thus carbon dioxide release is a direct result of urease activity which can be measured in the head space using gastight syringes and gaschromatographic equipment. The urease activity is determined by comparing the carbon dioxide release of incubation vessels with and without urea supply. The applicability of this method has been demonstrated by experiments with N -(n -butyl)phosphoric triamide (NBPT), copper ions and zinc ions as known inhibitors of urease activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alef, K. (1991). Ureaseaktivität. In Methodenhandbuch Bodenmikrobiologie; Aktivitäten—Biomasse—Differenzierung . pp. 216–26. Landsberg: Ecomed Verlagsgesellschaft mbH.

    Google Scholar 

  • Antil, R.S., Gupta, A.P. and Narwal, R.P. (1993). Effect of substrate concentration, soil moisture and organic amendments on urease activity of soil containing variable nickel amounts. Arid. Soil. Res. Rehab. 7 (4), 381–7.

    Google Scholar 

  • Brack, W., Paschke, A., Segner, H., Wennrich, R. and Schüürmann, G. (2000). Urease inhibition: A tool for toxicity identification in sediment elutriates. Chemosphere 40 , 829–34.

    Google Scholar 

  • Bremner, J.M. and Mulvaney, R.L. (1978). Urease activity in soils. In R.G. Burns (ed.) Soil Enzymes . pp. 149–96. New York: Academic Press.

    Google Scholar 

  • Bremner, J.M. (1982). Nitrogen-Urea. In A.L. Page, R.H. Miller and D.R. Keeney (eds). Methods of Soil Analysis, Part 2. Chemical and Microbial Properties, Agronomy Monograph no. 9 , (2nd edn), pp. 699–709. ISBN 0–89118–072–9 (Teil 2). American Society of Agronomy und Soil Science Society of America.

  • Byrnes, B.H. and Freney, J.R. (1995). Recent developments on the use of urease inhibitors in the tropics. Fert. Res. 42 , 251–9.

    Google Scholar 

  • Bogomolov, D.M., Chen, S.-K., Parmelee, R.W., Subler, S. and Edwards, C.A. (1996). An ecosystem approach to soil toxicity testing: A study of copper contamination In laboratory soil microcosms. Appl. Soil Ecol. 4 , 95–105.

    Google Scholar 

  • Cervelli, S., Nannipieri, B. and Sequi, P. (1978). Interactions between agrotechnicals and soil enzymes. In R.G. Burns (ed.). pp. 251–93. London: Academic Press.

    Google Scholar 

  • Douglas, L.A. and Bremner, J.M. (1970). Extraction and colorimetric determination of urea in soils. Soil Sci. Soci. Am. Proc. 34 , 859–62.

    Google Scholar 

  • DIN 38 405 Teil 19 (1988). Deutsche Einheitsverfahren zur Wasser-, Abwasser-und Schlammuntersuchung: Bestimmung der Anionen Fluorid, Chlorid, Nitrit, Phosphat (ortho-), Bromid, Nitrat und Sulfat in wenig belasteten Wässern mit der Ionenchromatographie. Weinheim: VCH Verlagsgesellschaft mbH.

  • DIN 38 406 Teil 6, 7, 8, 10, 19 (1981–1993). Deutsche Einheitsverfahren zur Wasser-Abwasser-und Schlammuntersuchung. Bestimmung von Schwermetallen. Weinheim: VCH Verlagsgesellschaft mbh.

  • DIN 38 414 S4 (1984). Deutsche Einheitsverfahren zur Wasser-, Abwasser-und Schlammuntersuchung. Bestimmung der Eluierbarkeit von Wasser (S4).

  • Freney, J.R., Keerthisinghe, D.G., Chaiwanakupt, P. and Phongpan, S. (1993). Use of urease inhibitors to reduce ammonia loss following application of urea to flooded rice fields. Plant and Soil 155/156 , 371–3.

    Google Scholar 

  • Grant, C.A., Jia, S., Brown, K.R. and Bailey, L.D. (1996). Volatile losses of NH3 from surface-applied urea and urea ammonium nitrate with and without the urease inhibitors NBPT or ammonium thiosulphate. Canadian J. Soil. Sci. 76 , 417–19.

    Google Scholar 

  • Hamida, S.K., Omar, S.A. and Abdel-Mallek, A.Y. (1997). Microbial populations and enzyme activity in soil treated with heavy metals. Water, Air and Soil Pollution 95 , 13–22.

    Google Scholar 

  • Jung, K., Bitton, G. and Koopman, B. (1995). Assessment of urease assays for measuring toxicity of environmental samples. Water Res. 29 (8), 1929–33.

    Google Scholar 

  • Kandeler, E. and Gerber, H. (1988). Short-term assay of soil urease activity using colorimetric determination of ammonium. Biol. Fertility Soils 6 , 68–72.

    Google Scholar 

  • Kolb, B., Auer, M. and Pospisil, P. (1981). Multiple headspace extraction. A procedure for quantitative analysis of volatile compounds in soil samples. Appl. Chromat. 35E , 2–32.

    Google Scholar 

  • Kolb, B., Pospisil, P. and Auer, M. (1984). Quantitative headspace analysis of solid samples; a classification for various sample types. Chromatographia 19 , 113–22.

    Google Scholar 

  • Prell-Swaid, A. and Schwedt, G. (1994). A screening procedure for heavy metals in soil extracts by alcohol dehydrogenase and urease. Acta Hydrochem. Hydrobiol. 22 (2), 70–75.

    Google Scholar 

  • Öhlinger, R. (1993). Bestimmung der Korngrößenverteilung. In F. Schinner, R. Dhlinger, E. Kandeler and R. Margesin (eds). Bodenbiologische Arbeitsmethoden , ISBN 3–540–56206–0. Berlin: Springer-Verlag.

    Google Scholar 

  • Qui-Xiang, L., Freney, J.R., Keerthisinghe, D.G. and Peoples, M.B. (1994). Inhibition of urease activity in flooded soils by phenylphosphordimidate and N(-n-butyl) thiophosphoric triamide. Soil Biol. Biochem. 26 (8), 1059–65.

    Google Scholar 

  • Rheinheimer, G., Hegemann, W., Raff, J. and Sekoulov, I. (1988). Stickstoffkreislauf im Wasser, Stickstoffumsetzung in natürlichen Gewässern, in der Abwasserreinigung und Wasserversorgung R. Oldenburger Verlag München Wien. ISBN 3–486–26296–3.

  • Rotini, O.T. (1935). La transformazione enzymatica dell'-urea nell terreno. Ann. Labor. Ric. Ferm. Spallanzani. 3 , 143–54.

    Google Scholar 

  • Skujin, J.J. and McLaren, A.D. (1968). Persistance of encymatic activity in stored and geologically preserved soils. Enzymologia 34 , 213–25.

    Google Scholar 

  • Skujin, J.J. and McLaren, A.D. (1969). Assay of urease activity using 14C-urea in stored, geological preserved and irradiated soils. Soil Biol. Biochem. 1 , 89–99.

    Google Scholar 

  • Tabatai, M.A. and Bremner, J.M. (1972). Assay of urease activity in soils. Soil Biol. Biochem. 4 , 479–87.

    Google Scholar 

  • US-Environmental Protection, Agency (1984). Part VIII, 49, 209, method 610.

    Google Scholar 

  • van, Beelen P. and Doelman, P. (1997). Significance and application of microbial toxicity tests in assessing ecotoxicological risks of contaminants in soil and sediment. Chemosphere 34 (3), 455–99.

    Google Scholar 

  • Vittori Antisari, L., Marzadori, C., Gioacchini, P., Ricci, S. and Gessa, C. (1995). Effects of the urease inhibitor N-(n-butyl) phosphorothioic triamide in low concentrations an ammonia volatilization and evaluation of mineral nitrogen. Biol. Fertility Soils 22 , 196–201.

    Google Scholar 

  • Waid, J.S. and Pugh, K.B. (1967). Acetohydroxamate inhibition of the urease activity of an acid soil. Chemy. Ind. 71 (3), 71–3.

    Google Scholar 

  • Zeien, H. and Brümmer, G.W. (1991). Chemische Extraktion zur Bestimmung der Bindungsformen von Schwermetallen in Böden. In Berichte aus der ökologischen Forschung. Bd 6 Auswirkungen von Siedlungsabfällen auf Böden, Bodenorganismen und Pflanzen, Jülich KFA, 62–91.

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Adolf Eisentraeger.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guettes, R., Dott, W. & Eisentraeger, A. Determination of Urease Activity in Soils by Carbon Dioxide Release for Ecotoxicological Evaluation of Contaminated Soils. Ecotoxicology 11, 357–364 (2002). https://doi.org/10.1023/A:1020509422554

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1020509422554

Navigation