Influence of the soil storage method on soil enzymatic activities

M. Andrés Abellan, C. Wic Baena, F.A. García Morote, M.I. Picazo Cordoba, D. Candel Pérez, M.E. Lucas-Borja


Soil storage method may alter enzymatic activity being storage conditions of the soil samples prior to analysis decisive for the results. Studies made on freshly collected soils are generally preferred. However it is always not possible due to practical reasons since for example sampling is often restricted to short period of the year or because a great quantity of microbiological analyses must be made on time and by few people. On this context, soil storage methods are needed, being cold at 4°C the most widely used although sometimes alternative storage methods are also utilized. The aim of this study is to evaluate the effect of two alternative storage methods of soil samples (freezing at –20°C and air drying conservation methods) in comparison to cold at 4°C on the enzymatic activities (dehydrogenase, phosphatase, β-glucosidase and urease soil enzymes). Samples of two forest ecosystems (pine and holm oak forest stand) were taken in two different season of the year (winter and spring 2009). Results showed that enzymatic activities differed when freezing or air drying conservation methods were used in comparison with cold soil samples. Generally, alternative soil storage methods presented lower enzymatic activity than cold at 4°C. However, these changes depend on season and sampling location.


soil enzyme; soil storage technique; soil conservation; enzymatic analysis.

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Ahmed A., Daniels L.B., Nelson T.S., Piper E.L., Beasley J.N. Bolt D., 1982. Fescue toxicity as influenced by nitrogen fertilization. J Dairy Sc 65(Suppl 1), 167.

Allué J.L., 1990. Atlas fitoclimático de España. Taxonomías. Mº de Agricultura, IIIA, Madrid. 221 pp. PMid:2121804

Arias A., Leirós C., Gil-Sotres F., Trasarcepeda C., 1997. Comparison of methods for conserving soil samples pending biological and biochemical analysis. Abstracts of 9th International Symposium on Environmental Pollution and its impact on Life in the Mediterranean Region, S Agnello di Sorrento, Italy. pp.145.

Bastida F., Moreno J.L., Hernández T., García C., 2007. Microbial activity in nonagricultural degraded soils exposed to semiarid climate. Sci Total Environ 378, 183-186. PMid:17306863

Bastida F., Kandeler E., Moreno J.L., Ros M., García C., Hernández T., 2008. Application of fresh and composted organic wastes modifies structure, size and activity of soil microbial community under semiarid climate. Appl Soil Ecol 40, 318-329.

Breitenbeck G.A., Bremner J.M., 1987. Effects of storing soils at various temperatures on their capacity for denitrification. Soil Biology & Biochemistry 4, 377-380.

Brohon B., Delolme C., Gourdon R., 1999. Qualification of soils though microbial activities measurements: influence of the storage period on INT-reductase, phosphatase and respiration. Chemosphere 38, 1973-1984.

Caldwell B., 2005. Enzyme activities as a component of soil biodiversity: a review. Pedobiologia 49, 637- 644.

Ceccanti B., García C., 1994. Coupled chemical and biochemical methodologies to characterize a composting process and the humic substances. In: Humic substances in the global environment and implications on human health (Senesi N., Miano, T.M., eds). Elsevier, Amsterdam, Holanda. pp. 1279-1284.

Coxson D., Parkinson D., 1987. Winter respiratory activity in aspen woodland forest floor litter and soils. Soil Biology & Biochemistry 19, 49-59.

De Nobili M., Contin M., Brookes P.C., 2006. Microbial biomass dynamics in recently air dried and rewetted soils compared to others stored air-dry for up to 103 years. Soil Biology & Biochemistry 38, 2871-2881.

Dick W., Tabatabai M., 1993. Significance and potential uses of soil enzymes. In: Soil microbial ecology: appli cations in agricultural and environmental management (Blaine, Meeting F., eds). Ed Marcel Dekker, New York. pp. 95-127.

Faccendini N., Benintende M., Benintende S., 2003. Biomasa microbiana del suelo: almacenaje de muestras en freezer. Actas de la IV Reunión Nacional CientíficoTécnica de Biología de Suelos IV Encuentro de Fijación Biológica de Nitrógeno, Santiago del Estero, Argentina (CD-ROM).

García C., Hernández T., Costa F., Ceccanti B., Masciandaro G., 1993. The dehydrogenase activity of soil as an ecological marker in processes of perturbed system regeneration. In: Proceedings of the XI International Symposium of Environmental Biochemistry (Gallardo-Lancho J., ed). CSIC, Salamanca, España. pp. 89-100. PMCid:45605

Horwath W., Paul E.,1994. Microbial Biomass. In: Methods of soil analysis. Part 2: Microbiological & biochemical properties (Weaver W. et al., eds) . Ed SSSA Book Series No 5, USA., pp. 735-772.

ISO 10381-6. 1993. Soil quality-sampling. Guidance on the collection, handling and storage of soil for the assessment of aerobic microbial processes in the laboratory.

Jenkinson D., 1992. La materia orgánica del suelo y desarrollo de las plantas. Ed Mundi-Prensa, Madrid.

Kandeler E., Gerber H., 1988. Short-term assay of soil urease activity using colorimetric determination or ammonium. Biology & Fertility of Soils 6, 68-72.

Kandeler E., Stemmer M., Klimanek E.M., 1999. Response of soil microbial biomass, urease and xylanase within particle size fraction to long-term soil management. Soil Biology & Biochemistry 31, 205-211.

Lucas-Borja M.E., Bastida F., Moreno J.L., Nicolás C., Andrés M., López F.R., Del Cerro A., 2010 a. The effects of human trampling on the microbiological properties of soil and vegetation in Mediterranean Mountain areas. Land Degradation & Development. doi: 10.1002/ldr.1014.

Lucas-Borja M.E., Bastida F., Moreno J.L., Nicolás C., Andres M., López F.R., Del Cerro A. 2010 b. Influence of forest cover and herbaceous vegetation on the microbiological and biochemical properties of soil under Mediterranean humid climate. European Journal of Soil Biology 46, 273-279

Macleod R., Calcott P. 1976. Cold shock and freezing damage to microbes. In: The survival of vegetative microbes (Gray T., Postgate J., eds). Ed Cambridge University Press, Cambridge. pp. 81-109.

Mondini C., Contin M., Leita L., De Nobili M., 2002. Reponse of microbial to air-drying and rewetting in soils and compost. Geoderma 105, 111-124.

Nannipieri P., Greco S., Ceccanti B., 1990. Ecological significance of the biological activity in soil. In: Soil biochemestry, Vol 6 (Bollag J.M., Stozk G., eds). Ed Marcel Dekker, New York.

OCDE, 1995. Final Report of the OCDE Workshop on selection of Soils/Sediments, Belgirate, Italy, 18-20 January 1995.

Palma R.M., Conti M.E., 1990. Urease activity in Argentine soils. In: Field studies and influence of simple treatment (Speir T.W., Ross D.J., eds). pp. 105-108.

Pancholy S.K., Rice E.L., 1972. Effect of storage conditions on activities of urease, invertase, amylase and dehydrogenase in soil. Soil Science Society of America Proceedings 36, 536-537.

Ros M., Hernández M.T., García C., 2003. Bioremediation of soil degraded by sewage sludge: effects on soil properties and erosion losses. Environmental Management 31, 741-747. PMid:14565694

Ros M., García C., Hernández T., Andrés M., Barja A., 2004. Short-term effects of human trampling on vegetation and soil microbial activity. Communications in Soil Science and Plant Analysis 35, 1591-1603.

Ross D., 1970. Effects of storage on dehydrogenase activities of soils. Soil Biology & Biochemistry 2, 759-765.

Ross D., Tate K.R., Cairns A., Meyrick K., 1980. Influence of storage on soil microbial biomass estimated by three biochemical procedures. Soil Biology & Biochemistry 4, 369-374.

Ross D., 1991. Microbial biomass in a stored soil: a comparison of different estimation procedures. Soil Biology & Biochemistry 23, 1005-1007.

Rugber D., Helveg A., 1989. 14C-labelled TCA and Atrazine degradation in soil: Influence of freezing and air-drying. Presented at the 6th Danish Plant Protection Conference, Lyngby, Denmark, February 28.

Shen S.M., Brookes P. C., Jenkison D. S., 1987. Soil respiration and the measurement of microbial biomass C by fumigation technique in fresh and in air-dried soil. Soil Biology & Biochemistry 19, 153-158.

Speir T.W., Ross D.J., 1975. Effects of storage on the activities of protease, urease, phosphatase and sulphatase in three soils under pasture. New Zealand Journal of Soil Science 18, 231-237.

Speir T.W., Ross D.J., 1981. A comparison of the effects of air drying and acetone dehydration on soil enzyme activities. Soil Biology & Biochemistry 13, 225-230.

Stenberg B., Johansson M., Pell M., Sjodahl S., Stenstrom J., Torstensson L., 1998. Microbial biomass and activities in soil as affected by frozen and cold storage. Soil Biology & Biochemistry 3, 393-402.

Stephen C.L., Whittaker D.G., Gillis D., Cox L., Rhodes O., 2005. Genetic consequences of reintroductions: an example from Oregon prong horn antelope (Antilocapra americana). Journal of Wildlife Management 69(4), 1463-1474.[1463:GCORAE]2.0.CO;2

Tabatabai M.A., Bremner J.M., 1969. Use of p-nitrophenyl phosphate for assay of soil phosphatase activity. Biology & Biochemistry 1, 301-307.

Tabatabai M.A., 1982. Soil Enzymes. In: Methods of soil análysis. Part 2. Chemical and microbiologycal properties (Page A.L., Miller E.M., Keeney D.R. eds). Ed Soil Science Society of America, INC, Madison. pp. 903-947.

Trasar-Cepeda C., Leirós M.C., Gil-Sotres F., Seoane S., 1998. Towards a biochemical quality index for soils: an expression relating several biological and biochemical properties. Biology & Fertility of Soils 26, 100-106.

Trabue S.L., Palmquist D.E., Lydick T.M., Koch S., 2006. Effects of soil storage no the microbial community and degradation of mersulfuron-methyl. Journal of Agriculture & Food Chemistry 54, 142-151. PMid:16390191

Vance N.C., Entry J.A., 2000. Soil properties important to the restoration of Shasta red fir barrens in the Siskiyou Mountains. Forest Ecology & Management 138, 427-434.

Zantua M.I., Bremner J.M., 1975a. Comparison of methods of assaying urease activity in soils. Soil Biology & Biochemistry 7, 291-295.

Zantua M.I., Bremner J.M., 1975b. Preservation of soil samples for assay of urease activity in soils. Soil Biology & Biochemistry 7, 297-299.

Zelles L., Adrian P., Bai Q. Y.,Stepper K., Adria M.V., Fischer K., Maier A., Ziegler A., 1991. Microbial activity measured in soils stored under different temperature and humidity conditions. Soil Biology & Biochemistry 23, 955-962.

Zornoza R., Guerrero C., Mataix S.J., Arcenegui V., García F., Mataix B.J., 2006. Assesing air-drying and rewetting pre-treatment effect on some enzyme activities under Mediterranean conditions. Soil Biology & Biochemistry 38, 2125-2134.

DOI: 10.5424/fs/20112003-11081