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Dehydrogenases are exclusively intracellular enzymes, which play an important role in the initial stages of oxidation of soil organic matter. One of the most frequently used methods to estimate dehydrogenase activity in soil is based on the use of triphenyltetrazolium chloride as an artificial electron acceptor. The purpose of this study was to compare the activity of dehydrogenases of forest soils with varied physicochemical properties using different triphenyltetrazolium chloride assays. The determination was carried out using the original procedure by Casida et al., a modification of the procedure which involves the use of Ca(OH)2 instead of CaCO3, the Thalmann method, and the assay by Casida et al. without addition of buffer or any salt. Soil dehydrogenase activity depended on the assay used. Dehydrogenase determined by the Casida et al. method without addition of buffer or any salt correlated with the pH values of soils. The autoclaved strongly acidic samples of control soils showed high concentrations of triphenylformazan, probably due to chemical reduction of triphenyltetrazolium chloride. There is, therefore, a need for a sterilization method other than autoclaving, ie a process that results in significant changes in soil properties, thus helping to increase the chemical reduction of triphenyltetrazolium chloride.
The aim of this research was to develop a suitable method of succinate dehydrogenase activity assay in situ for different industrial yeast strains. For this purpose different compounds: EDTA, Triton X-100, sodium deoxycholate, digitonin, nystatin and β-mercaptoethanol were used. The permeabilization process was controlled microscopically by primuline staining. Enzyme assay was conducted in whole yeast cells with Na-succinate as substrate, phenazine methosulfate (PMS) as electron carrier and in the presence one of two different tetrazolium salts: tetrazolium blue chloride (BT) or cyanoditolyl tetrazolium chloride (CTC) reduced during the assay. In comparabile studies of yeast vitality the amount of intracellular ATP was determined according to luciferin/luciferase method. During the succinate dehydrogenase assay in intact yeast cells without permeabilization, BT formazans were partially visualized in the cells, but CTC formazans appeared to be totally extracellular or associated with the plasma membrane. Under these conditions there was no linear relationship between formazan color intensity signal and yeast cell density. From all chemical compounds tested, only digitonin was effective in membrane permeabilization without negative influence on cell morphology. Furthermore, with digitonin-treated cells a linear relationship between formazan color intensity signal and yeast cell number was noticed. Significant decreasing of succinate dehydrogenase activity and ATP content were observed during aging of the tested yeast strains.
Microbiological characteristics of sewage sludge from a mechanical-and-biological sewage treatment plant composted in controlled conditions with straw and sawdust are presented. Prepared composts were placed in four bioreactors with airflow of 4 l air·min⁻¹. In bioreactor K1, K2 and K3 the composted mass consisted of 65% sewage sludge (K1–sewage sludge 1, K2 – sludge 2, K3 – sludge 3) + 30% sawdust + 5% straw; while in bioreactor K4 the proportion was: 45% sludge 2 + 50% sawdust + 5% straw. Compost samples were taken from all chambers at the same time, depending on actual temperature. Microbiological analyses consisted in the determination (by plate method) on selective media of the numbers of mesophilic and thermophilic bacteria, fungi and pathogenic bacteria Salmonella sp. Clostridium perfringens and Enterobacteriaceae. Furthermore, in the experiment, the activity levels of dehydrogenase were determined using 1% triphenyltetrazole chloride as substratum. Studies have indicated that the composting process caused a decrease of the number of fungi and pathogenic bacteria from Enterobacteriaceae family and Clostridium perfringens in all composted matters, as well as an increase in the number of thermophilic bacteria. Changes in the number of mesophilic bacteria depended on the compost type. In composts K1 and K2, the composting process caused an increased proliferation of cells, while in the composts K3 and K4 the number of mesophilic bacteria decreased. On the basis of the obtained results, it was also found that in the majority of analysis terms, the lowest activity of dehydrogenases occurred in compost K3, while their level of its activity, in the majority of the studied composts, correlated most intensively with the number of thermophilic bacteria.
Torlińska T., Ożegowski S., Paluszak J., and Hryniewiecki T.: In vivo effect of 2-deoxy-D-glucose on glucose-6-phosphate dehydrogenase activity in the cytosol of liver, heart and skeletal muscle of rats. Acta Physiol. Pol. 1990, 41 (6): 137-143. 2-deoxy-D-glucose (2-DG), the unmetabolizable analogue of glucose induces a series of metabolic, hormonal and behavioral responses, causing cellular glucoprivation. According to in vitro studies, 2-DG inhibits phosphofructokinase in cultured human cells. The present investigations deal with changes in the cytosolic glucocse-6-phosphate dehydrogenase activity following in vivo 2-DG administration. A single dose of 2-DG (600 mg/kg) has no influence on the activity of glucose-6-phosphate dehydrogenase in the cytosol of liver, heart and skeletal muscle of the rat. The concommitant increase in serum glucose, lactate and FFA concentrations observed in the study indicates indirectly a stimulation of adrenergic system. After three days of successive administration of 2-DG to rats, dehydrogenase activity decreased in the liver by approx 57% and in the skeletal muscle by approx 82% in comparison with control animals. Moreover the in vivo effect of 2-DG was found to be fully reversible, probably when the total amount of the inhibitor was excreted.
Most of the processes occurring in soil are catalysed by enzymes. As a result of their sensitivity towards heavy metals, enzymes in contaminated soils are usually less active. The purpose of this paper was to assess the influence of bioavailable forms of Cd, Cu, Pb and Zn on the activity of dehydrogenases, urease, acid and alkaline phosphatase, and to compare the results obtained from naturally and artificially contaminated soils. A pot experiment was carried out on two loamy sand soils, naturally and artificially contaminated with Cd, Cu, Pb and Zn. The total content of heavy metals classified these soils as very heavily contaminated with Cu, heavily contaminated with Pb and contaminated with Cd and Zn, all according to the IUNG system (1995). One of the following organic materials: swine manure or triticale straw, was added to the soil batches. The experiment was carried out in three replications, in two pH ranges: slightly acid and acid. Soil samples for analyses were taken after 14, 28, 165 and 450 days of incubation. The results of the experiment showed that the activity of soil enzymes depended on the content of bioavailable heavy metals; the total concentration of trace elements and H+ were less important. However, considerable differences were found in enzyme activity between naturally and artificially contaminated soils. This indicates that results obtained from other research conducted on freshly contaminated soils cannot be easily transferred to field conditions. The analysed enzymes responded differently to the concentration of bioavailable forms of heavy metals. Alkaline phosphatase was the least tolerant to bioavailable forms of heavy metals, unlike urease, which was the most tolerant soil enzyme. A similar pattern of sensitivity toward trace elements, which could be ordered as Zn > Cd > Cu > Pb, was noticed for dehydrogenases, acid and alkaline phosphatases. Urease was found to be more tolerant to Zn.
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