Sulfidogenic Bacteria Research Articles

Competition of methanogenic and sulfidogenic bacteria

The competition of acetotrophic methanogenic bacteria (AMB) and acetotrophic sulfidogenic bacteria (ASRB) was studied by assessing growth rates, activities and acetate and sulfate affinities at different pH levels and sulfide concentrations in batch reactors. Both anaerobic granular sludge and suspended anaerobic sludge were tested. The results indicate that at pH-levels below 6.9 AMB will outcompete ASRB, whereas above a pH of 7.7, ASRB will win the competition. If ASRB and AMB are present in granular sludge growth will be found in a wider pH-range than if they are present as suspended sludge. The affinities for acetate and the sulfide toxicity are dependent on the sludge form as well. In granular sludge the acetate affinities of ASRB and AMB are comparable, whereas in suspended sludge ASRB show a lower affinity than AMB. With respect to sulfide toxicity, the results indicate that above pH 7 sulfide inhibition in granular sludge is caused by the total sulfide concentration, while in suspended sludge the free H2S-concentration determines the toxicity. At high pH-levels growth is stronger inhibited than the activity.

Effects of sulfur-containing compounds on the growth and methane production of acclimated-mixed methanogens

A variety of sulfur-containing compounds were investigated for use as medium reductant and sulfur sources for growth of the acclimated-mixed methanogens. In Hungate tube cultures, the optimum concentrations of sulfur compounds for growth and CH 4 formation were sulfide, 0.9 mmol; cysteine, 3.6 mmol; thiosulfate, 0.675 mmol; elemental sulfur (S 0), 0.45 gl −1; glutathione, 2.5 mmol; SO 4 2−, 3.6 mmol; SO 3 2−, 0.9 mmol; and S 2O 5 2−, 0.09 g l −1. In comparison with sulfide as sole sulfur source, CH 4 formation was about the same while higher increments of cell density were obtained by using cysteine, thiosulfate, S 0 and glutathione. Other sulfur-containing compounds tested resulted in decreased CH 4 formation but with a higher increment of cell density. When contaminating sulfidogenic bacteria were inhibited by addition of antibiotics to the medium which are known to affect eubacteria but not methanogens, only a slight amount of hydrogen sulfide was produced by assimilatory reduction of methanogens, and the growth of the methanogens was poor, especially in the case of growing on SO 4 2−, SO 3 2−, thiosulfate or glutathione, indicating that evolved hydrogen sulfide was mainly due to a sulfide-producer that existed symbiotically with predominant methanogens in the cultures. However, in the case of cysteine or S 0 as sulfur sources, in spite of inhibition of contaminating bacteria, sufficient sulfide was reduced by methanogens to support growth and methane formation effectively. These sulfur compounds were also tested for their ability to enhance and restore growth and CH 4 production with sulfur-depleted cells in culture tubes. The results of the present study show that, for the growth of the acclimated-mixed methanogens, volatile and malodorous sulfide can be replaced with the sulfur-containing compounds described above.

Isolation and characterization of a novel bacterium growing via reductive dehalogenation of 2-chlorophenol.

A bacterium capable of anaerobic growth via reductive dehalogenation of 2-chlorophenol was isolated from a culture enriched from sediment taken from a small stream near Lansing, Mich. The organism, designated strain 2CP-1, is a gram-negative rod ca. 3 by 0.5 micron in size and is a catalase-negative, oxidase-negative, facultative anaerobe that forms small red colonies in anaerobic media. The organism grew in reduced anaerobic mineral medium supplemented with 2-chlorophenol, acetate, and vitamins, producing phenol as a product. It did not grow when either 2-chlorophenol or acetate was omitted. The growth yield was about 3 g of protein per mol of 2-chlorophenol dechlorinated, and the doubling time was 3.7 days. Only the ortho position was dehalogenated, and additional chlorines at other positions decreased or blocked ortho dechlorination. The organism also grew with fumarate as its electron acceptor. Dechlorination activity is inducible, since cultures grown in fumarate containing medium with 2-chlorophenol rapidly dechlorinated additional 2-chlorophenol, while cultures grown in the same medium without 2-chlorophenol did not. Analysis of the organism's 16S rRNA sequence revealed that it is a member of the delta proteobacteria, more closely related to the myxobacteria than to the sulfidogenic bacteria.

Physiological characterization of strain DCB-1, a unique dehalogenating sulfidogenic bacterium.

Strain DCB-1 is an obligately anaerobic bacterium which carries out the reductive dehalogenation of halobenzoates and was previously known to grow only on pyruvate plus 20% ruminal fluid. When various electron acceptors were supplied, thiosulfate and sulfite were found to stimulate growth. Sulfide was produced from thiosulfate. Cytochrome c and desulfoviridin were detected. The mol% G+C was 49 (at the thermal denaturation temperature). Of 55 carbon sources tested, only pyruvate supported growth as the sole carbon source in mineral medium. Lactate, acetate, L- and D-malate, glycerol, and L- and D-arabinose stimulated growth when supplemented with 10% ruminal fluid and 20 mM thiosulfate. In mineral medium, pyruvate was converted to acetate and lactate, with small amounts of succinate and fumarate accumulating transiently. During growth with thiosulfate, all of these products accumulated transiently. Addition of excess hydrogen to pyruvate-grown cultures resulted in diversion of carbon to formate, lactate, and butyrate, which caused a decrease in cell yield. We conclude that strain DCB-1 is a new type of sulfidogenic bacterium.

Bioenergetic Conditions of Butyrate Metabolism by a Syntrophic, Anaerobic Bacterium in Coculture with Hydrogen-Oxidizing Methanogenic and Sulfidogenic Bacteria

The butyrate-oxidizing, proton-reducing, obligately anaerobic bacterium NSF-2 was grown in batch cocultures with either the hydrogen-oxidizing bacterium Methanospirillum hungatei PM-1 or Desulfovibrio sp. strain PS-1. Metabolism of butyrate occurred in two phases. The first phase exhibited exponential growth kinetics (phase a) and had a doubling time of 10 h. This value was independent of whether NSF-2 was cultured with a methanogen or a sulfate reducer and likely represents the maximum specific growth rate of NSF-2. This exponential growth phase was followed by a second phase with a nearly constant rate of degradation (phase b) which dominated the time course of butyrate degradation. The specific activity of H(2) uptake by the hydrogen-oxidizing bacterium controlled the bioenergetic conditions of metabolism in phase b. During this phase both the Gibbs free energy (DeltaG') and the butyrate degradation rate (v) were greater for NSF-2-Desulfovibrio sp. strain PS-1 (DeltaG' = -17.0 kJ/mol; v = 0.20 mM/h) than for NSF-2-M. hungatei PM-1 (DeltaG' = -3.8 kJ/mol, v = 0.12 mM/h). The DeltaG' value remained stable and characteristic of the two hydrogen oxidizers during phase b. The stable DeltaG' resulted from the close coupling of the rates of butyrate and H(2) oxidation. The addition of 2-bromoethanesulfonate to a NSF-2-methanogen coculture resulted in the total inhibition of butyrate degradation; the inhibition was relieved when Desulfovibrio sp. strain PS-1 was added as a new H(2) sink. When the specific activity of H(2) consumption was increased by adding higher densities of the Desulfovibrio sp. to 2-bromoethanesulfonate-inhibited NSF-2-methanogen cocultures, lower H(2) pool sizes and higher rates of butyrate degradation resulted. Thus, it is the kinetic parameters of H(2) consumption, not the type of H(2) consumer per se, that establishes the thermodynamic conditions which in turn control the rate of fatty acid degradation. The bioenergetic homeostasis we observed in phase b was a result of the kinetics of the coculture members and the feedback inhibition by hydrogen which prevents butyrate degradation rates from reaching their theoretical V(max).

Aerobic and anaerobic microbial consumption of hydrogen in geothermal spring water

Hydrogen consumption was measured in hot geothermal water from two ponds of the San Federigo solfatara, Tuscany, Italy, where emanation gases contained approx. 4% H2. H2 consumption was completely inhibited by NaOH and partially by HgCl2 indicating microbial utilization. Aerobic and anaerobic H2 consumption activities coexisted in the same water with aerobic activity being higher in one pond and anaerobic activity in the other. The kinetics of H2 consumption were consistent with those of ‘Knallgas’, methanogenic or sulfidogenic bacteria.

Physiological function of hydrogen metabolism during growth of sulfidogenic bacteria on organic substrates.

Desulfovibrio vulgaris Madison and Thermodesulfobacterium commune contained functionally distinct hydrogenase activities, one which exchanged 3H2 into 3H2O and was inhibited by carbon monoxide and a second activity which produced H2 in the presence of CO. Cell suspensions of D. vulgaris used either lactate, pyruvate, or CO as the electron donor for H2 production in the absence of sulfate. Both sulfidogenic species produced and consumed hydrogen as a trace gas during growth on lactate or pyruvate as electron donors and on thiosulfate or sulfate as electron acceptors. Higher initial levels of hydrogen were detected during growth on lactate-sulfate than on pyruvate-sulfate. D. vulgaris but not T. commune also produced and then consumed CO during growth on organic electron donors and sulfate or thiosulfate. High partial pressures of exogenous H2 inhibited growth and substrate consumption when D. vulgaris was cultured on pyruvate alone but not when it was metabolizing pyruvate plus sulfate or lactate plus sulfate. The data are discussed in relation to supporting two different models for the physiological function of H2 metabolism during growth of sulfidogenic bacteria on organic electron donors plus sulfate. A trace H2 transformation model is proposed for control of redox processes during growth on either pyruvate or lactate plus sulfate, and an obligate H2 cycling model is proposed for chemiosmotic energy coupling during growth on CO plus sulfate.