Comparison and improvement of methods for determining soil dehydrogenase activity by using triphenyltetrazolium chloride and iodonitrotetrazolium chloride

Abstract

The triphenyltetrazolium chloride (TTC) method described by Thalmann (1968) and the iodonitrotetrazolium chloride (INT) method described by Spothelfer-Magaña and Thalmann (1992), used for measuring soil dehydrogenase activity, have been modified to overcome some methodical short-comings. Absorption maxima of 485 nm for triphenylformazan dissolved in acetone, 491 nm for iodonitrotetrazolium formazan (INTF) dissolved in tetrahydrofuran and 455 nm for INTF dissolved in N,N-dimethylformamide are recommended for measuring wavelengths. Extracting triphenylformazan twice with acetone is less toxic and proved to be at least as efficient as extraction with a mixture of 90% acetone and 10% carbon tetrachloride (Thalmann 1968 method). Tetrahydrofuran and dimethylformamide were equally good in extracting INTF from soils, but the former was less toxic. Anaerobic incubation resulted in the formation of higher amounts of triphenylformazan and INTF as well as reduced standard error. Both TTC and INT reduction showed high reproducibility and good differentiation of the microbial activity of six soils. For several reasons (more easily determined substrate dose depending on different soil types, better reduction, shorter incubation time), INT reduction seems to be a more suitable method of measuring soil microbial activity than TTC reduction.