Biology and Fertility of Soils

, Volume 19, Issue 2–3, pp 87–99

Current methods for measuring microbial biomass C in soil: Potentials and limitations

  • R. Martens
Review Article

Abstract

Methods for measuring soil microbial biomass C were reviewed. The basic ideas behind the fumigation-incubation method, the fumigation-extraction method, the substrate-induced respiration method, and the ATP method were examined together with the advantages, disadvantages, and limitations as reported in the literature and those found by our own recent investigations.

The fumigation-incubation method is the basic technique which is also used for calibration of the three other methods. It is characterized by simple performance without the need of expensive equipment. Its application is limited to soils with a pH above 5 and to soils that do not contain easily degradable C sources. If these limitations are not considered, too low or even negative biomass values will be obtained. These restrictions are largely overcome by the fumigation-extraction method. However, the kEC factor applied to calculate microbial biomass C from the C additionally made extractable by the fumigation is still controversial. The substrate-induced respiration requires expensive equipment for the hourly measurement of soil respiration. This method is also susceptible to amendment of soils with C sources, leading to an overestimate of biomass C. Although a few authors disagree with some basic assumptions behind the methods described, they are widely used and accepted. The use of ATP to measure biomass C in soil is far more uncertain. A high diversity of applied techniques for the extraction and measurement of ATP has led to biomass C : ATP ratios which vary between about 150 and 1 000. Our own current investigations are expected to shed more light on the problems of ATP extraction. Preliminary results indicate that a constant biomass C : ATP ratio of about 200 may be more realistic.

Key words

Mirobial biomass C Estimation methods Fumigation-incubation method Fumigation extraction method Substrate induced respiration method 

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References

  1. Alef K (1993) Bestimmung mikrobieller Biomasse im Boden: Eine kritische Betrachtung. Z Pflanzenernähr Bodenkd 156:109–114Google Scholar
  2. Anderson JPE, Domsch KH (1973) Quantification of bacterial and fungal contributions to soil respiration. Arch Mikrobiol 93:113–127Google Scholar
  3. Anderson JPE, Domsch KH (1975) Measurement of bacterial and fungal contributions to respiration of selected agricultural and forest soils. Can J Microbiol 21:314–322Google Scholar
  4. Anderson JPE, Domsch KH (1978a) Mineralisation of bacteria and fungi in chloroform-fumigated soils. Soil Biol Biochem 10:207–213Google Scholar
  5. Anderson JPE, Domsch KH (1978b) A physiological method for the quantitative measurement of microbial biomass in soils. Soil Biol Biochem 10:215–221Google Scholar
  6. Bai QY, Zelles L, Scheunert I, Korte F (1988) A simple effective procedure for the determination of adenosine triphosphate in soils. Chemosphere 17:2461–2470Google Scholar
  7. Bremer E, van Kessel C (1990) Extractability of microbial 14C and 15N following addition of variable rates of labelled glucose and (NH4)2SO4 to soil. Soil Biol Biochem 22:707–713Google Scholar
  8. Brookes PC, Newcombe AD, Jenkinson DS (1987) Adenylate energy charge measurements in soil. Soil Biol Biochem 19: 211–217Google Scholar
  9. Chaussod R, Nicolardot B (1982) Measure de la biomass microbienne dans les sols cultivés. I. Approche cinétique et estimation simplifiée du carbone facilement minéralisable. Rev Ecol Biol Sol 19:501–512Google Scholar
  10. Ciardi C, Nannipieri P (1990) A comparison of methods ofr measuring ATP in soil. Soil Biol Biochem 22:725–727Google Scholar
  11. Coûteaux MM, Henkinet R, Pitta P, Bottner P, Billès G, Palka L, Vannier G (1989) Native carbon mineralisation of an acid organic soil after use of the chloroform-fumigation method to estimate microbial biomass. Biol Fertil Soils 8:172–177Google Scholar
  12. Coûteaux MM, Henkinet R, Botter P (1990) Anomalies in microbial biomass measurements in acid organic soils using extractable carbon following chloroform fumigation. Soil Biol Biochem 22:955–957Google Scholar
  13. Dumontet S, Mathur SP (1989) Evaluation of respiration-based methods for measuring microbial biomass in metal-contaminated acidic mineral and organic soils. Soil Biol Biochem 21:431–436Google Scholar
  14. Eiland F (1983) A simple method for quantitative determination of ATP in soil. Soil Biol Biochem 15:665–670Google Scholar
  15. Heinemeyer O, Insam H, Kaiser EA, Walenzik G (1989) Soil microbial biomass and respiration measurements: An automated technique based on infra-red gas analysis. Plant and Soil 116:191–195Google Scholar
  16. Ingham ER, Horten KA (1987) Bacterial, fungal and protozoan responses to chloroform fumigation in stored soil. Soil Biol Biochem 19:545–550Google Scholar
  17. Ingham ER, Griffiths RP, Cromack K, Entry JA (1991) Comparison of direct vs fumigation incubation microbial biomass estimates from ectomycorrhizal mat and non-mat soils. Soil Biol Biochem 23:465–471Google Scholar
  18. Inubushi K, Brookes PC, Jenkinson DS (1991) Soil microbial biomass C, N and nonhydrin-N in aerobic and anaerobic soils measured by the fumigation-extraction method. Soil Biol Biochem 27:737–741Google Scholar
  19. Jenkinson DS (1966) Studies on the decomposition of plant material in soil. II. J Soil Sci 17:280–302Google Scholar
  20. Jenkinson DS (1976) The effects of biocidal treatment on metabolism in soil. IV. The decomposition of fumigated organisms in soil. Soil Biol Biochem 8:203–208Google Scholar
  21. Jenkinson DS (1977) The soil biomass. NZ Soil News 25:213–218Google Scholar
  22. Jenkinson DS (1987) This week's citation classic. Curr Contents 23:18Google Scholar
  23. Jenkinson DS (1988) Determination of microbial biomass carbon and nitrogen in soil. In: Wilson JR (ed) Advances in nitrogen cycling in agricultural ecosystems. CAB, Wallingford, pp 368–386Google Scholar
  24. Jenkinson DS, Ladd JN (1981) Microbial biomass in soil: Measurement and turnover. In: Paul EA, Ladd JN (eds) Soil Biochemistry, vol 5. Marcel Dekker, New York, Basel, pp 415–471Google Scholar
  25. Jenkinson DS, Oades JM (1979) A method for measuring adenosine triphosphate in soil. Soil Biol Biochem 11:193–199Google Scholar
  26. Jenkinson DS, Powlson DS (1976a) The effects of biocidal treatment on metabolism in soil. I. Fumigation with chloroform. Soil Biol Biochem 8:167–177Google Scholar
  27. Jenkinson DS, Powlson DS (1976b) The effects of biocidal treatment of metabolism in soil. V. A. method for measuring soil biomass. Soil Biol Biochem 8:209–213Google Scholar
  28. Jenkinson DS, Powlson DS, Wedderburn RWM (1976) The effects of biocidal treatment on metabolism in soil. III. The relationship between soil biovolume, measured by optical microscopy, and the flush of decomposition caused by fumigation. Soil Biol Biochem 8:189–202Google Scholar
  29. Kaiser EA, Mueller T, Joergensen RG, Insam H, Heinemeyer O (1992) Evaluation of methods to estimate the soil microbial biomass and the relationship with soil texture and organic matter. Soil Biol Biochem 24:675–683Google Scholar
  30. Karl DM (1980) Cellular nucleotide measurements and application in microbial ecology. Microb Rev 44:739–796Google Scholar
  31. Knowles CJ (1977) Microbial metabolic regulation by adenine nucleotide pools. Symp Soc Gen Microbiol 27:241–283Google Scholar
  32. Lynch JM, Panting LM (1980) Cultivation and the soil biomass. Soil Biol Biochem 12:29–33Google Scholar
  33. McGill WB, Cannon KR, Robertson JA, Cook FD (1986) Dynamics of soil microbial biomass and water-soluble organic C in Breton L after 50 years of cropping to two rotations. Can J Soil Sci 66:1–19Google Scholar
  34. Martikainen PJ, Palojärvi A (1990) Evaluation of the fumigation-extraction method for the determination of microbial C and N in a range of forest soils. Soil Biol Biochem 22:792–802Google Scholar
  35. Martens R (1985) Limitations in the application of the fumigation technique for biomass estimations in amended soils. Soil Biol Biochem 17:57–63Google Scholar
  36. Martens R (1987) Estimation of microbial biomass in soil by the respiration method: Importance of soil pH and flushing methods for the measurement of respired CO2. Soil Biol Biochem 19:77–81Google Scholar
  37. Mueller T, Joergensen RG, Meyer B (1992) Estimation of soil microbial biomass C in the presence of living roots by fumigation-extraction. Soil Biol Biochem 24:179–181Google Scholar
  38. Oades JM, Jenkinson DS (1979) Adenosine triphosphate content of the soil microbial biomass. Soil Biol Biochem 11:201–204Google Scholar
  39. Ocio JA, Brookes PC (1990) An evaluation of methods for measuring the microbial biomass in soils following recent additions of wheat straw and the characterization of the biomass that develops. Soil Biol Biochem 22:685–694Google Scholar
  40. Powlson DS, Jenkinson DS (1976) The effects of biocidal treatment on metabolism in soil. II. Gamma irradiation, autoclaving, airdrying and fumigation. Soil Biol Biochem 8:179–188Google Scholar
  41. Ross DJ (1990) Estimation of soil microbial C by a fumigation-extraction method: Influence of seasons, soils and calibration with the fumigation-incubation procedure. Soil Biol Biochem 22:295–300Google Scholar
  42. Ross DJ (1991) Microbial biomass in a stored soil: A comparison of different estimation procedures. Soil Biol Biochem 23:1005–1007Google Scholar
  43. Ross DJ, Tate KR, Cairns A, Pansier EA (1980) Microbial biomass estimations in soils from tussock grasslands by three biochemical procedures. Soil Biol Biochem 12:375–383Google Scholar
  44. Schnürer J, Clarholm M, Rosswall T (1985) Microbial biomass and activity in an agricultural soil with different organic matter contents. Soil Biol Biochem 17:611–618Google Scholar
  45. Shen SM, Brookes PC, Jenkinson DS (1987) Soil respiration and the measurement of microbial biomass C by the fumigation technique in fresh and air-dried soil. Soil Biol Biochem 19:153–158Google Scholar
  46. Shields JA, Paul EA, Lowe WE (1974) Factors influencing the stability of labelled microbial materials in soils. Soil Biol Biochem 6:31–37Google Scholar
  47. Sparling GP (1981) Microcalorimetry and other methods to assess biomass and activity in soil. Soil Biol Biochem 13:93–98Google Scholar
  48. Sparling GP, West AW (1988) A direct extraction method to estimate soil microbial C: Calibration in situ using microbial respiration and 14C labelled cells. Soil Biol Biochem 20:337–343Google Scholar
  49. Sparling GP, Ord BG, Vaughan D (1981) Microbial biomass and activity in soils amended with glucose. Soil Biol Biochem 13:99–104Google Scholar
  50. Sparling GP, Feltham CW, Reynolds J, West AW, Singleton P (1990) Estimation of soil microbial C by a fumigation-extraction method: Use on soils of organic matter content, and a reassessment of the k EC-factor. Soil Biol Biochem 22:301–307Google Scholar
  51. Störmer K (1908) Über die Wirkungen des Schwefelkohlenstoffs und ähnlicher Stoffe auf den Boden. Zentralbl Bakteriol Parasitenkd Infektionskr Hyg Abt 2 20:282–288Google Scholar
  52. Tate KR, Jenkinson DS (1982) Adenosine triphosphate measurements in soil: An improved method. Soil Biol Biochem 14:331–335Google Scholar
  53. Tate KR, Ross DJ, Feltham CW (1988) A direct extraction method to estimate soil microbial C: Effects of experimental vaiables and some different calibration procedures. Soil Biol Biochem 20:329–335Google Scholar
  54. Vance ED, Brookes PC, Jenkinson DS (1987a) Microbial biomass measurements in forest soils: The use of the chloroform fumigation-incubation method in strongly acid soils. Soil Biol Biochem 19:697–702Google Scholar
  55. Vance ED, Brookes PC, Jenkinson DS (1987b) An extraction method for measuring soil microbial biomass. Soil Biol Biochem 19:703–707Google Scholar
  56. Voroney RP (1983) Decomposition of crop residues. PhD thesis, University of SaskatchewanGoogle Scholar
  57. Voroney RP, Paul EA (1984) Determination of k C and k N in situ for calibration of the chloroform fumigation-incubation method. Soil Biol Biochem 16:9–14Google Scholar
  58. Wardle DA, Parkinson D (1990) Comparison of physiological techniques for estimating the response of the soil microbial biomass to soil moisture. Soil Biol Biochem 22:817–823Google Scholar
  59. Wardle DA, Parkinson D (1991) A statistical evaluation of equations for predicting total microbial biomass carbon using physiological and biochemical methods. Agric Ecosyst Environ 34:75–86Google Scholar
  60. Webster JJ, Hampton GJ, Leach FR (1984) ATP in soil: A new extractant and extraction procedure. Soil Biol Biochem 16: 335–342Google Scholar
  61. West AW, Sparling GP, Grant WD (1986) Correlation between four methods to estimate total microbial biomass in stored, air-dried and glucose-amended soils. Soil Biochem 18:569–576Google Scholar
  62. Wu J, Joergensen RG, Pommerening B, Chaussod R, Brookes PC (1990) Measurement of soil microbial biomass C by fumigation-extraction — an automated procedure. Soil Biol Biochem 22:1167–1169Google Scholar
  63. Zagal E (1993) Measurement of microbial biomass in rewetted airdried soil by fumigation-incubation and fumigation-extraction techniques. Soil Biol Biochem 25:553–559Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • R. Martens
    • 1
  1. 1.Institut für BodenbiologieBundesforschungsanstalt für LandwirtschaftBraunschweigGermany

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