R. Cord-Ruwisch

A quick method for the determination of dissolved and precipitated sulfides in cultures of sulfate-reducing bacteria

Dissolved sulfide was determined spectrophotometrically as a colloidal solution of copper sulfide. Calibration curves were linear. Maximal deviation error was below 5%. Sulfide precipitated as FeS was determined after acidification of the medium.

The capacity of hydrogenotrophic anaerobic bacteria to compete for traces of hydrogen depends on the redox potential of the terminal electron acceptor

The effect of different electron acceptors on substrate degradation was studied in pure and mixed cultures of various hydrogenotrophic homoacetogenic, methanogenic, sulfate-reducing, fumarate-reducing and nitrate-ammonifying bacteria. Two different species of these bacteria which during organic substrate degradation produce and consume hydrogen, were cocultured on a substrate which was utilized only by one of them. Hydrogen, which was excreted as intermediate by the first strain (and reoxidized in pure culture), could, depending on the hydrogen acceptor present, also be used by the second organism, resulting in interspecies hydrogen transfer. The efficiency of H2 transfer was similar when methanol, lactate or fructose were used as organic substrate, although the free energy changes of fermentative H2 formation of these substrates are considerably different. In coculture experiments nitrate or fumarate>sulfate> CO2/CH4>sulfur or CO2/acetate were the preferred electron acceptors, and an increasing percentage of H2 was transferred to that bacterium which was able to utilize the preferred electron acceptor. In pure culture the threshold values for hydrogen oxidation decreased in the same order from ≤1,100 ppm for homoacetogenic bacteria to about 0.03 ppm for nitrate or fumarate reducing bacteria. The determined H2-threshold values as well as the percentage of H2 transfer in cocultures were related to the Gibbs free energy change of the respective hydrogen oxidizing reaction.

Sulfate-reducing bacteria and their activities in oil production

This paper presents an overview of the microbiology of sulfate-reducing bacteria (SRB) and their detrimental effects in oil technology and summarizes a study on SRB in an oil field. SRB are a group of specialized microorganisms that occur in aqueous environments in the absence of oxygen. The main nutrients for SRB are simple organic acids and molecular hydrogen (H//2) from decomposing natural organic matter. The nutrients are oxidized, with sulfate being reduced to sulfide (hydrogen sulfide, H//2S). The formed H//2S is the principal agent in the disastrous effects caused by SRB. It contaminates gas and stored oil, precipitates ferrous sulfide that plugs injection wells, and promotes corrosion of iron and steel in the absence of oxygen (anaerobic corrosion). Another principal mechanism by which SRB are involved in corrosion is their ability to depolarize iron surfaces by consumption of cathodically formed hydrogen.

Effect of pre-aeration and inoculum on the start-up of batch thermophilic anaerobic digestion of municipal solid waste.

In this study, a short pre-aeration step was investigated as pre-treatment for thermophilic anaerobic digestion of the organic fraction of municipal solid waste (OFMSW). It was found that pre-aeration of 48 h generated enough biological heat to increase the temperature of bulk OFMSW to 60 degrees C. This was sufficient self-heating of the bulk OFMSW for the start-up of thermophilic anaerobic digestion without the need for an external heat source. Pre-aeration also reduced excess easily degradable organic compounds in OFMSW, which were the common cause of acidification during the start-up of the batch system. Careful consideration however must be taken to avoid over aeration as this consumes substrate, which would otherwise be available to methanogens to produce biogas. To accelerate methane production and volatile solids destruction, the anaerobic digestion in this study was operated as a wet process with the anaerobic liquid recycled through the OFMSW. Appropriate anaerobic liquid inoculum was found to be particularly beneficial. It provided high buffer capacity as well as suitable microbial inoculum. As a result, acidification during start-up was kept to a minimum. With volatile fatty acids (VFAs-acetate in particular) and H2 accumulation typical of hydrolysis and fermentation of the easily degradable substrates during start-up, inoculum with high numbers of hydrogenotrophic methanogens was critical to not only maximise CH4 production but also reduce H2 partial pressure in the system to allow VFAs degradation. In a lab-scale bioreactor, the combined pre-aeration and wet thermophilic anaerobic digestion was able to stabilise the OFMSW within a period of only 12 days. The stabilised inert residual material can be used as a soil amendment product.

Desulfotomaculum geothermicum sp. nov., a thermophilic, fatty acid-degrading, sulfate-reducing bacterium isolated with H2 from geothermal ground water

A strictly anaerobic, thermophilic, fatty acids-degrading, sporulating sulfate-reducing bacterium was isolated from geothermal ground water. The organism stained Gram-negative and formed gas vacuoles during sporulation. Lactate, ethanol, fructose and saturated fatty acids up to C18 served as electron donors and carbon sources with sulfate as external electron acceptor. Benzoate was not used. Stoichiometric measurements revealed a complete oxidation of part of butyrate although growth with acetate as only electron donor was not observed. The rest of butyrate was oxidized to acetate. The strain grew chemolithoautotrophically with hydrogen plus sulfate as energy source and carbon dioxide as carbon source without requirement of additional organic carbon like acetate. The strain contained a c-type cytochrome and presumably a sulfite reductase P582. Optimum temperature, pH and NaCl concentration for growth were 54°C, pH 7.3–7.5 and 25 to 35 g NaCl/l. The G+C content of DNA was 50.4 mol %. Strain BSD is proposed as a new species of the spore-forming sulfate-reducing genus Desulfotomaculum, D. geothermicum.

Characterization of Desulfovibrio fructosovorans sp. nov.

Desulfovibrio strain JJ isolated from estuarine sediment differed from all other described Desulfovibrio species by the ability to degrade fructose. The oxidation was incomplete, leading to acetate production. Fructose, malate and fumarate were fermented mainly to succinate and acetate in the absence of an external electron acceptor. The pH and temperature optima for growth were 7.0 and 35° C respectively. Strain JJ was motile by means of a single polar flagellum. The DNA base composition was 64.13% G+C. Cytochrome c3 and desulfoviridin were present. These characteristics established the isolate as a new species of the genus Desulfovibrio, and the name Desulfovibrio fructosovorans is proposed.

The role of iron-oxidizing bacteria in stimulation or inhibition of chalcopyrite bioleaching

A series of bacterial and chemical leaching experiments were conducted to clarify contradictory reports in the literature regarding the role of bacteria in the bioleaching of chalcopyrite. Tests containing a high bacterial concentration showed inhibited leaching, even lower than non-inoculated controls. However, when bacterial cells were washed before inoculation, it was apparent that it was not the bacterial cells but rather the chemical species introduced with them that influenced the leaching rate. In addition, the results of comparative tests with 0.1 M ferrous sulphate or ferric sulphate showed that copper was leached from the ore 2.7 times faster in leach solutions containing ferrous ion, suggesting that ferric ions inhibit chalcopyrite dissolution. The results indicated that the chalcopyrite dissolution rate is strongly dependent on the reduction potential (Eh) in solution, and that this parameter is far more influential than the number or activity of bacterial cells. These results imply that the role of bacteria may only be stimulatory when the prevailing electrochemical conditions are also favourable.

Corroding iron as a hydrogen source for sulphate reduction in growing cultures of sulphate-reducing bacteria

SummaryIn anaerobic corrosion experiments, hydrogenase-positiveDesulfovibrio strains, grown with limiting lactate concentrations in the presence of steel wool, formed more sulphide than expected or observed with lactate alone. The additional sulphide obviously originated from sulphate reduction with cathodically formed hydrogen from the steel surface. The hydrogenasenegativeD. sapovorans did not produce additional sulphide. The observations agree with the theory of von Wolzogen Kühr and van der Vlugt (1934) that explains anaerobic corrosion as a cathodic depolarization of iron surfaces by hydrogen-consuming sulphate-reducing bacteria. The influence of the iron surface area, the salt concentration and the pH-value on the utilization of cathodically formed hydrogen was investigated. The significance of an additional organic electron donor for the corrosion of iron in aqueous environments is discussed.

Ammonium as a sustainable proton shuttle in bioelectrochemical systems.

This work examines a pH control method using ammonium (NH(4)(+)) as a sustainable proton shuttle in a CEM-equipped BES. Current generation was sustained by adding NH(3) or ammonium hydroxide (NH(4)OH) to the anolyte, controlling its pH at 7. Ammonium ion migration maintained the catholyte pH at approximately 9.25. Such NH(4)(+)/NH(3) migration accounted for 90±10% of the ionic flux in the BES. Reintroducing the volatilized NH(3) from the cathode into the anolyte maintained a suitable anolyte pH for sustained microbial-driven current generation. Hence, NH(4)(+)/NH(3) acted as a proton shuttle that is not consumed in the process.

A practical kinetic model that considers endproduct inhibition in anaerobic digestion processes by including the equilibrium constant

The classical Michaelis‐Menten model is widely used as the basis for modeling of a number of biological systems. As the model does not consider the inhibitory effect of endproducts that accumulate in virtually all bioprocesses, it is often modified to prevent the overestimation of reaction rates when products have accumulated. Traditional approaches of model modification use the inclusion of irreversible, competitive, and noncompetitive inhibition factors. This article demonstrates that these inhibition factors are insufficient to predict product inhibition of reactions that are close the dynamic equilibrium. All models investigated were found to violate thermodynamic laws as they predicted positive reaction rates for reactions that were endergonic due to high endproduct concentrations. For modeling of biological processes that operate close to the dynamic equilibrium (e.g., anaerobic processes), it is critical to prevent the prediction of positive reaction rates when the reaction has already reached the dynamic equilibrium. This can be achieved by using a reversible kinetic model. However, the major drawback of the reversible kinetic model is the large number of empirical parameters it requires. These parameters are difficult to determine and prone to experimental error. For this reason, the reversible model is not practical in the modeling of biological processes.