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Appl. Environ. Microbiol., Jun 1995, 2186-2192, Vol 61, No. 6
Copyright © 1995, American Society for Microbiology

An Inhibitor-Based Method To Measure Initial Decomposition of Naturally Occurring Polysaccharides in Sediments

HTS Boschker, SA Bertilsson, EMJ Dekkers and TE Cappenberg
Center for Limnology, Netherlands Institute of Ecology, 3631 AC Nieuwersluis, The Netherlands

A method that can be used to measure the initial decomposition rates of polysaccharides in sediment samples was developed. It uses toluene to specifically inhibit microbial uptake of carbohydrates without affecting extracellular hydrolysis of polysaccharides. Accumulating carbohydrates were determined by high-performance liquid chromatography. Field-sampled litter from the common reed (Phragmites australis), which contains cellulose and arabinoxylan as its main polysaccharides, was used as a model system. Toluene concentrations of between 1 and 10% resulted in the accumulation of similar amounts of monomeric carbohydrates, which was linear over time for most neutral sugars. Toluene (3%) did not have an effect on extracellular enzyme activities, and microbial sugar uptake was completely inhibited, as demonstrated with (sup14)C-labelled xylose and glucose. Experiments with enhancement cultures and fixed reed litter suggested that enzymatic hydrolysis of polysaccharides in reed litter was the main source of glucose, xylose, arabinose, and galactose accumulation. In contrast, the accumulation of high amounts of the alditols mannitol and glucitol was probably caused by lysis of the microbial population in toluene-treated reed litter. Glucose accumulated at rates of 1.3 and 0.10 (mu)mol (middot) g of dry matter content(sup-1) (middot) h(sup-1) under aerobic and anaerobic conditions, respectively, whereas xylose accumulation rates were only 10% of the glucose accumulation rates.