Sulfate reduction from phosphogypsum using a mixed culture of sulfate-reducing bacteria

Phosphogypsum (CaSO₄), a primary by-product of phosphoric acid production, is accumulated in large stockpiles and occupies vast areas of land. It poses a severe threat to the quality of water and land in countries producing phosphoric acid. In this study, the potential of sulfate-reducing bacteria for biodegradation of this sulfur-rich industrial solid waste was assessed. The effect of phosphogypsum concentration, carbon and nitrogen sources, temperature, pH and stirring on the growth of sulfate-reducing bacteria was investigated. Growth of sulfate-reducing bacteria was monitored by measuring sulfide production. Phosphogypsum was shown to be a good source of sulfate, albeit that the addition of organic carbon was necessary for bacterial growth. Biogenic sulfide production occurred with phosphogypsum up to a concentration of 40 g L⁻¹, above which no growth of sulfate-reducing bacteria was observed. Optimal growth was obtained at 10 g L⁻¹ phosphogypsum. Both the gas mixture H₂/CO₂ and lactate supported high amounts of H₂S formation (19 and 11 mM, respectively). The best source of nitrogen for sulfate-reducing bacteria was yeast extract, followed by ammonium chloride. The presence of nitrate had an inhibitory effect on the process of sulfate reduction. Stirring the culture at 150 rpm slightly stimulated H₂S formation, probably by improving sulfate solubility.     

Sulfate Reduction Relative to Methane Production in High-Rate Anaerobic Digestion: Technical Aspects

The effect of different substrates and different levels of sulfate and sulfide on methane production relative to sulfate reduction in high-rate anaerobic digestion was evaluated. Reactors could be acclimated so that sulfate up to a concentration of 5 g of sulfate S per liter did not significantly affect methanogenesis. Higher levels gave inhibition because of salt toxicity. Sulfate reduction was optimal at a relatively low level of sulfate, i.e., 0.5 g of sulfate S per liter, but was also not significantly affected by higher levels. Both acetoclastic and hydrogenotrophic methane-producing bacteria adapted to much higher levels of free H₂S than the values reported in the literature (50% inhibition occurred only at free H₂S levels of more than 1,000 mg/liter). High levels of free H₂S affected the sulfate-reducing bacteria only slightly. Formate and acetate supported the sulfate-reducing bacteria very poorly. In the high-rate reactors studied, intensive H₂S formation occurred only when H₂ gas or an H₂ precursor such as ethanol was supplied.     

Pathways and Microbiology of Thiosulfate Transformations and Sulfate Reduction in a Marine Sediment (Kattegat, Denmark)

Reductive and oxidative pathways of the sulfur cycle were studied in a marine sediment by parallel radiotracer experiments with ³⁵SO₄^2-, H₂³⁵S, and ³⁵S₂O₃^2- injected into undisturbed sediment cores. The distributions of viable populations of sulfate- and thiosulfate-reducing bacteria and of thiosulfate-disproportionating bacteria were concurrently determined. Sulfate reduction occurred both in the reducing sediment layers and in oxidized and even oxic surface layers. The population density of sulfate-reducing bacteria was >10⁶ cm^-3 in the oxic layer, high enough that it could possibly account for the measured rates of sulfate reduction. The bacterial numbers counted in the reducing sediment layers were 100-fold lower. The dominant sulfate reducers growing on acetate or H₂ were gas-vacuolated motile rods which were previously undescribed. The products of sulfide oxidation, which took place in both oxidized and reduced sediment layers, were 65 to 85% S₂O₃^2- and 35 to 15% SO₄^2-. Thiosulfate was concurrently oxidized to sulfate, reduced to sulfide, and disproportionated to sulfate and sulfide. There was a gradual shift from predominance of oxidation toward predominance of reduction with depth in the sediment. Disproportionation was the most important pathway overall. Thiosulfate disproportionation occurred only as cometabolism in the marine acetate-utilizing sulfate-reducing bacteria, which could not conserve energy for growth from this process alone. Oxidative and reductive cycling of sulfur thus occurred in all sediment layers with an intermediate “thiosulfate shunt” as an important mechanism regulating the electron flow.     

End Products of Anaerobic Chitin Degradation by Salt Marsh Bacteria as Substrates for Dissimilatory Sulfate Reduction and Methanogenesis

The anaerobic pathway of chitin decomposition by chitinoclastic bacteria was examined with an emphasis on end product coupling to other salt marsh bacteria. Actively growing chitinoclastic bacterial isolates produced primarily acetate, H₂, and CO₂ in broth culture. No sulfate-reducing or methanogenic isolates grew on chitin as sole carbon source or produced any measurable degradation products. Mixed cultures of chitin degraders with sulfate reducers resulted in positive sulfide production. Mixed cultures of chitin-degrading isolates with methanogens resulted in the production of CH₄ with reductions in headspace CO₂ and H₂. The combination of all three metabolic types resulted in the simultaneous production of methane and sulfide, with more methane being produced in mixed cultures containing CO₂-reducing methanogens and acetoclastic sulfate reducers because of less interspecific H₂ competition.     

Gas phase H<Subscript>2</Subscript>S product recovery in a packed bed bioreactor with immobilized sulfate-reducing bacteria

An N₂ strip gas was used in a packed bed sulfate-reducing bioreactor to recover the dissolved sulfide product and improve sulfate conversion. The highest volumetric productivity obtained was 261 mol H₂S m⁻³ d⁻¹. Lowering the initial pH of the medium from 7 to 6 increased the H₂S content of the strip gas from 3.6 to 5.8 mol%. The ratio of strip gas to liquid flow rates (G/L) was found be to a suitable basis for scaling the process. Calculations indicated that modest G/L values (<10²) were required to recover the residual dissolved sulfide in a downstream stripping column.     

Methanol utilizing <Emphasis Type="Italic">Desulfotomaculum</Emphasis> species utilizes hydrogen in a methanol-fed sulfate-reducing bioreactor

A sulfate-reducing bacterium, strain WW1, was isolated from a thermophilic bioreactor operated at 65°C with methanol as sole energy source in the presence of sulfate. Growth of strain WW1 on methanol or acetate was inhibited at a sulfide concentration of 200 mg l⁻¹, while on H₂/CO₂, no apparent inhibition occurred up to a concentration of 500 mg l⁻¹. When strain WW1 was co-cultured under the same conditions with the methanol-utilizing, non-sulfate-reducing bacteria, Thermotoga lettingae and Moorella mulderi, both originating from the same bioreactor, growth and sulfide formation were observed up to 430 mg l⁻¹. These results indicated that in the co-cultures, a major part of the electron flow was directed from methanol via H₂/CO₂ to the reduction of sulfate to sulfide. Besides methanol, acetate, and hydrogen, strain WW1 was also able to use formate, malate, fumarate, propionate, succinate, butyrate, ethanol, propanol, butanol, isobutanol, with concomitant reduction of sulfate to sulfide. In the absence of sulfate, strain WW1 grew only on pyruvate and lactate. On the basis of 16S rRNA analysis, strain WW1 was most closely related to Desulfotomaculum thermocisternum and Desulfotomaculum australicum. However, physiological properties of strain WW1 differed in some aspects from those of the two related bacteria.     

Biofilm Dynamics and Kinetics during High-Rate Sulfate Reduction under Anaerobic Conditions

The sulfate kinetics in an anaerobic, sulfate-reducing biofilm were investigated with an annular biofilm reactor. Biofilm growth, sulfide production, and kinetic constants (K_m and V_max) for the bacterial sulfate uptake within the biofilm were determined. These parameters were used to model the biofilm kinetics, and the experimental results were in good agreement with the model predictions. Typical zero-order volume rate constants for sulfate reduction in a biofilm without substrate limitation ranged from 56 to 93 μmol of SO²₄-cm⁻³ h⁻¹ at 20°C. The temperature dependence (Q₁₀) of sulfate reduction was equivalent to 3.4 at between 9 and 20°C. The measured rates of sulfate reduction could explain the relatively high sulfide levels found in sewers and wastewater treatment systems. Furthermore, it has been shown that sulfate reduction in biofilms just a few hundred micrometers thick is limited by sulfate diffusion into biofilm at concentrations below 0.5 mM. This observation might, in some cases, be an explanation for the relatively poor capacity of the sulfate-reducing bacteria to compete with the methanogenic bacteria in anaerobic wastewater treatment in submerged filters.     

A sulfate-reducing bacterium from the oxic layer of a microbial mat from Solar Lake (Sinai), Desulfovibrio oxyclinae sp. nov.

In an investigation on the oxygen tolerance of sulfate-reducing bacteria, a strain was isolated from a 10⁷-fold dilution of the upper 3-mm layer of a hypersaline cyanobacterial mat (transferred from Solar Lake, Sinai). The isolate, designated P1B, appeared to be well-adapted to the varying concentrations of oxygen and sulfide that occur in this environment. In the presence of oxygen strain P1B respired aerobically with the highest rates [260 nmol O₂ min^–1 (mg protein)^–1] found so far among marine sulfate-reducing bacteria. Besides H₂ and lactate, even sulfide or sulfite could be oxidized with oxygen. The sulfur compounds were completely oxidized to sulfate. Under anoxic conditions, it grew with sulfate, sulfite, or thiosulfate as the electron acceptor using H₂, lactate, pyruvate, ethanol, propanol, or butanol as the electron donor. Furthermore, in the absence of electron donors the isolate grew by disproportionation of sulfite or thiosulfate to sulfate and sulfide. The highest respiration rates with oxygen were obtained with H₂ at low oxygen concentrations. Aerobic growth of homogeneous suspensions was not obtained. Additions of 1% oxygen to the gas phase of a continuous culture resulted in the formation of cell clumps wherein the cells remained viable for at least 200 h. It is concluded that strain P1B is oxygen-tolerant but does not carry out sulfate reduction in the presence of oxygen under the conditions tested. Analysis of the 16S rDNA sequence indicated that strain P1B belongs to the genus Desulfovibrio, with Desulfovibrio halophilus as its closest relative. Based on physiological properties strain P1B could not be assigned to this species. Therefore, a new species, Desulfovibrio oxyclinae, is proposed. Received: 7 August 1996 / Accepted: 29 January 1997     

Performance of a thermophilic sulfate and sulfite reducing high rate anaerobic reactor fed with methanol

Thermophilic sulfate and sulfite reduction was studied in lab-scale Expanded Granular Sludge Bed (EGSB) reactors operated at 65°C and pH 7.5 with methanol as the sole carbon and energy source for the sulfate- and sulfite-reducing bacteria. At a hydraulic retention time (HRT) of 10 h, maximum sulfite and sulfate elimination rates of 5.5 gSO₃ ^2- L^-1 day^-1 (100 % elimination) and 5.7 gSO₄ ^{2- -1} day^-1 (55% elimination) were achieved, resulting in an effluent sulfide concentration of approximately 1800 mgS L^-1. Sulfate elimination was limited by the sulfide concentration, as stripping of H₂S from the reactor with nitrogen gas was found to increase the sulfate elimination rate to 9.9 gSO₄ ^2- L^-1 day^-1 (100 % elimination). At a HRT of 3 h, maximum achievable sulfite and sulfate elimination rates were even 18 gSO₃ ^2- L^-1 day^-1 (100% elimination) and 11 gSO₄ ^2- L^-1 day^-1(50% elimination). At a HRT of 3 h, the elimination rate was limited by the biomass retention of the system. 5.5 ± 1.8% of the consumed methanol was converted to acetate, which was not further degraded by sulfate reducing bacteria present in the sludge. The acetotrophic activity of the sludge could not be stimulated by cultivating the sludge for 30 days under methanol-limiting conditions. Omitting cobalt as trace element from the influent resulted in a lower acetate production rate, but it also led to a lower sulfate reduction rate. Sulfate degradation in the reactor could be described by zeroth order kinetics down to a threshold concentration of 0.05 g L^-1, while methanol degradation followed Michaelis-Menten kinetics with a K_m of 0.037 gCOD L^-1.     

Terminal Processes in the Anaerobic Degradation of an Algal-Bacterial Mat in a High-Sulfate Hot Spring

The algal-bacterial mat of a high-sulfate hot spring (Bath Lake) provided an environment in which to compare terminal processes involved in anaerobic decomposition. Sulfate reduction was found to dominate methane production, as indicated by comparison of initial electron flow through the two processes, rapid conversion of [2-¹⁴C]acetate to ¹⁴CO₂ and not to ¹⁴CH₄, and the lack of rapid reduction of NaH¹⁴CO₃ to ¹⁴CH₄. Sulfate reduction was the dominant process at all depth intervals, but a marked decrease of sulfate reduction and sulfate-reducing bacteria was observed with depth. Concurrent methanogenesis was indicated by the presence of viable methanogenic bacteria and very low but detectable rates of methane production. A marked increased in methane production was observed after sulfate depletion despite high concentrations of sulfide (>1.25 mM), indicating that methanogenesis was not inhibited by sulfide in the natural environment. Although a sulfate minimum and sulfide maximum occurred in the region of maximal sulfate reduction, the absence of sulfate depletion in interstitial water suggests that methanogenesis is always severely limited in Bath Lake sediments. Low initial methanogenesis was not due to anaerobic methane oxidation.     

The relationship between hydrogen metabolism,sulfate reduction and nitrogen fixation in sulfate reducers

Summary Hydrogenase and nitrogenase activities of sulfate-reducing bacteria allow their adaptation to different nutritional habits even under adverse conditions. These exceptional capabilities of adaptation are important factors in the understanding of their predominant role in problems related to anaerobic metal corrosion. Although the D₂–H⁺ exchange reaction indicated thatDesulfovibrio desulfuricans strain Berre-Sol andDesulfovibrio gigas hydrogenases were reversible, the predominant activity in vivo was hydrogen uptake. Hydrogen production was restricted to some particular conditions such as sulfate or nitrogen starvation. Under diazotrophic conditions, a transient hydrogen evolution was followed by uptake when dinitrogen was effectively fixed. In contrast, hydrogen evolution proceeded when acetylene was substituted as the nitrogenase substrate. Hydrogen can thus serve as an electron donor in sulfate reduction and nitrogen metabolism.     

Biotransformation of phosphogypsum in media containing different forms of nitrogen

Studies on the biotransformation of phosphogypsum (a waste product formed in the course of the production of phosphorous fertilizers) with the use of sulfate reducing bacteria (SRB) demonstrated that it is a good source of sulfates and biogenic elements for these bacteria, though the addition of organic carbon and nitrogen is necessary. The aim of this study was to investigate the form of nitrogen and C:N ratio in the medium on the growth of SRB community in cultures containing phosphogypsum. Batch community cultures of sulfate reducing bacteria were maintained in medium with phosphogypsum (5.0 g/l), different concentrations of sodium lactate (1.6 - 9.4 g/l) and different forms (NH4CI, CO(NH2)2, KNO3) and concentrations (0 - 250 mg/l) of nitrogen. The growth of SRB was studied in the C:N ratio of from 2:1 to 300:1. It was found that: 1 - the best source of nitrogen for SRB is urea, followed by ammonium, the worst were nitrates; 2 - the bacteria were also able to grow in medium without nitrogen but their activity was then by approximately 15% lower than in optimal growth conditions; 3 - in medium with KNO3 inhibition of sulfate reduction by approx. 50% was observed; 4 - the highest reduction of nitrates (removal of nitrate) in media with phosphogypsum and nitrates was at limiting concentrations of sodium lactate. This is probably caused by the selection under these conditions (low concentration of hydrogen sulfide) of denitrifying bacteria or sulfate reducing bacteria capable of using nitrates as an electron acceptor.     

Planktonic nitrate-reducing bacteria and sulfate-reducing bacteria in some western Canadian oil field waters

Oil fields that use water flooding to enhance oil recovery may become sour because of the production of H₂S from the reduction of sulfate by sulfate-reducing bacteria (SRB). The addition of nitrate to produced waters can stimulate the activities of nitrate-reducing bacteria (NRB) and control sulfide production. Many previous studies have focused on chemolithotrophic bacteria that can use thiosulfate or sulfide as energy sources while reducing nitrate. Little attention has been given to heterotrophic NRB in oil field waters. Three different media were used in this study to enumerate various types of planktonic NRB present in waters from five oil fields in western Canada. The numbers of planktonic SRB and bacteria capable of growth under aerobic conditions were also determined. In general, microbial numbers in the produced waters were very low (<10 ml⁻¹) in samples taken near or at wellheads. However, the numbers increased in the aboveground facilities. No thiosulfate-oxidizing NRB were detected in the oil field waters, but other types of NRB were detected in 16 of 18 produced water samples. The numbers of heterotrophic NRB were equal to or greater than the number of sulfide-oxidizing, chemolithotrophic NRB in 12 of 15 samples. These results showed that each of the oil fields contained NRB, which might be stimulated by nitrate amendment to control H₂S production by SRB. Journal of Industrial Microbiology & Biotechnology (2002) 29, 83–92 doi:10.1038/sj.jim.7000274 Received 20 February 2002/ Accepted in revised form 14 May 2002     

Effect of nitrate and nitrite on sulfide production by two thermophilic,sulfate-reducing enrichments from an oil field in the North Sea

Thermophilic sulfate-reducing bacteria (tSRB) can be major contributors to the production of H₂S (souring) in oil reservoirs. Two tSRB enrichments from a North Sea oil field, NS-tSRB1 and NS-tSRB2, were obtained at 58°C with acetate-propionate-butyrate and with lactate as the electron donor, respectively. Analysis by rDNA sequencing indicated the presence of Thermodesulforhabdus norvegicus in NS-tSRB1 and of Archaeoglobus fulgidus in NS-tSRB2. Nitrate (10 mM) had no effect on H₂S production by mid-log phase cultures of NS-tSRB1 and NS-tSRB2, whereas nitrite (0.25 mM or higher) inhibited sulfate reduction. NS-tSRB1 did not recover from inhibition, whereas sulfate reduction activity of NS-tSRB2 recovered after 500 h. Nitrite was also effective in souring inhibition and H₂S removal in upflow bioreactors, whereas nitrate was similarly ineffective. Hence, nitrite may be preferable for souring prevention in some high-temperature oil fields because it reacts directly with sulfide and provides long-lasting inhibition of sulfate reduction.     

Bacterial Disproportionation of Elemental Sulfur Coupled to Chemical Reduction of Iron or Manganese

A new chemolithotrophic bacterial metabolism was discovered in anaerobic marine enrichment cultures. Cultures in defined medium with elemental sulfur (S⁰) and amorphous ferric hydroxide (FeOOH) as sole substrates showed intense formation of sulfate. Furthermore, precipitation of ferrous sulfide and pyrite was observed. The transformations were accompanied by growth of slightly curved, rod-shaped bacteria. The quantification of the products revealed that S⁰ was microbially disproportionated to sulfate and sulfide, as follows: 4S⁰ + 4H₂O → SO₄^2- + 3H₂S + 2H⁺. Subsequent chemical reactions between the formed sulfide and the added FeOOH led to the observed precipitation of iron sulfides. Sulfate and iron sulfides were also produced when FeOOH was replaced by FeCO₃. Further enrichment with manganese oxide, MnO₂, instead of FeOOH yielded stable cultures which formed sulfate during concomitant reduction of MnO₂ to Mn²⁺. Growth of small rod-shaped bacteria was observed. When incubated without MnO₂, the culture did not grow but produced small amounts of SO₄^2- and H₂S at a ratio of 1:3, indicating again a disproportionation of S⁰. The observed microbial disproportionation of S⁰ only proceeds significantly in the presence of sulfide-scavenging agents such as iron and manganese compounds. The population density of bacteria capable of S⁰ disproportionation in the presence of FeOOH or MnO₂ was high, > 10⁴ cm^-3 in coastal sediments. The metabolism offers an explanation for recent observations of anaerobic sulfide oxidation to sulfate in anoxic sediments.     

Reduction of chromate by fixed films of sulfate-reducing bacteria using hydrogen as an electron source

The ability of sulfate-reducing bacteria (SRB) to reduce chromate, Cr(VI), was evaluated using fixed-film growth systems and H₂ as the electron source. A main objective of the experiment was to distinguish between direct enzymatic reduction and indirect reduction by hydrogen sulfide, in order to subsequently verify and control the synergy of these two mechanisms. In batch experiments with the sulfate-reducing consortium CH10 selected from a mining site, 50 mg l⁻¹ Cr(VI) was reduced in 15 min in the presence of 500 mg l⁻¹ hydrogen sulfide compared to 16 mg l⁻¹ reduced in 1 h without hydrogen sulfide. Fixed films of a CH10 population and Desulfomicrobium norvegicum were fed-batch grown in a column bioreactor. After development of the biofilm, hydrogen sulfide was removed and the column was fed continuously with a 13-mg l⁻¹ Cr(VI) solution. Specific Cr(VI) reduction rates on pozzolana were close to 90 mg Cr(VI) h⁻¹ per gram of protein. Exposure to Cr(VI) had a negative effect on the subsequent ability of CH10 to reduce sulfate, but the inhibited bacteria remained viable. Journal of Industrial Microbiology & Biotechnology (2002) 28, 154–159 DOI: 10.1038/sj/jim/7000226 Received 20 September 2000/ Accepted in revised form 13 November 2001     

Sulfate reduction with methanol by a thermophilic consortium obtained from a methanogenic reactor

 An enrichment culture obtained from anaerobic granular sludge of a bench-scale anaerobic reactor degraded methanol at 65°C via sulfate reduction and acetogenesis. Sulfate reduction was the dominant process (S^2-/acetate=2.5). No methane formation was observed. Approximately 30% of the methanol was converted by acetogenic bacteria to acetate, while the remainder was degraded by these bacteria to H₂ and CO₂ in syntrophy with hydrogen-consuming sulfate-reducing bacteria. Pure cultures of sulfate-reducing and acetogenic bacteria were isolated and characterized. Received: 4 December 1995 / Received revision: 15 April 1996 / Accepted: 22 April 1996     

Sulfate-Reducing Bacteria Release Barium and Radium from Naturally Occurring Radioactive Material in Oil-Field Barite

Scale and sludge deposits formed during oil production can contain elevated levels of Ra, often coprecipitated with barium sulfate (barite). The potential for sulfate-reducing bacteria to release ²²⁶Ra and Ba (a Ra analog) from oil-field barite was evaluated. The concentration of dissolved Ba increased when samples containing pipe scale, tank sludge, or oil-field brine pond sediment were incubated with sulfate-reducing bacteria Desulfovibrio sp., Str LZK1, isolated from an oil-field brine pond. However, Ba release was not stoichiometric with sulfide production in oil-field samples, and &lt; 0.1% of the Ba was released. Potential for the release of ²²⁶Ra was demonstrated, and the ²²⁶Ra release associated with sulfate-reducing activity was predictable from the amount of Ba released. As with Ba, only a fraction of the ²²⁶Ra expected from the amount of sulfide produced was released, and most of the Ra remained associated with the solid material.     

Oxidation of H2, organic compounds and inorganic sulfur compounds coupled to reduction of O2 or nitrate by sulfate-reducing bacteria

All of fourteen sulfate-reducing bacteria tested were able to carry out aerobic respiration with at least one of the following electron donors: H₂, lactate, pyruvate, formate, acetate, butyrate, ethanol, sulfide, thiosulfate, sulfite. Generally, we did not obtain growth with O₂ as electron acceptor. The bacteria were microaerophilic, since the respiration rates increased with decreasing O₂ concentrations or ceased after repeated O₂ additions. The amounts of O₂ consumed indicated that the organic substrates were oxidized incompletely to acetate; only Desulfobacter postgatei oxidized acetate with O₂ completely to CO₂. Many of the strains oxidized sulfite (completely to sulfate) or sulfide (incompletely, except Desulfobulbus propionicus); thiosulfate was oxidized only by strains of Desulfovibrio desulfuricans; trithionate and tetrathionate were not oxidized by any of the strains. With Desulfovibrio desulfuricans CSN and Desulfobulbus propionicus the oxidation of inorganic sulfur compounds was characterized in detail. D. desulfuricans formed sulfate during oxidation of sulfite, thiosulfate or elemental sulfur prepared from polysulfide. D. propionicus oxidized sulfite and sulfide to sulfate, and elemental sulfur mainly to thiosulfate. A novel pathway that couples the sulfur and nitrogen cycles was detected: D. desulfuricans and (only with nitrite) D. propionicus were able to completely oxidize sulfide coupled to the reduction of nitrate or nitrite to ammonia. Cell-free extracts of both strains did not oxidize sulfide or thiosulfate, but formed ATP during oxidation of sulfite (37 nmol per 100 nmol sulfite). This, and the effects of AMP, pyrophosphate and molybdate on sulfite oxidation, suggested that sulfate is formed via the (reversed) sulfate activation pathway (involving APS reductase and ATP sulfurylase). Thiosulfate oxidation with O₂ probably required a reductive first step, since it was obtained only with energized intact cells.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - APS adenosine phosphosulfate or adenylyl sulfate     

Growth of Desulfovibrio in Lactate or Ethanol Media Low in Sulfate in Association with H2-Utilizing Methanogenic Bacteria

In the analysis of an ethanol-CO₂ enrichment of bacteria from an anaerobic sewage digestor, a strain tentatively identified as Desulfovibrio vulgaris and an H₂-utilizing methanogen resembling Methanobacterium formicicum were isolated, and they were shown to represent a synergistic association of two bacterial species similar to that previously found between S organism and Methanobacterium strain MOH isolated from Methanobacillus omelianskii. In lowsulfate media, the desulfovibrio produced acetate and H₂ from ethanol and acetate, H₂, and, presumably, CO₂ from lactate; but growth was slight and little of the energy source was catabolized unless the organism was combined with an H₂-utilizing methanogenic bacterium. The type strains of D. vulgaris and Desulfovibrio desulfuricans carried out the same type of synergistic growth with methanogens. In mixtures of desulfovibrio and strain MOH growing on ethanol, lactate, or pyruvate, diminution of methane produced was stoichiometric with the moles of sulfate added, and the desulfovibrios grew better with sulfate addition. The energetics of the synergistic associations and of the competition between the methanogenic system and sulfate-reducing system as sinks for electrons generated in the oxidation of organic materials such as ethanol, lactate, and acetate are discussed. It is suggested that lack of availability of H₂ for growth of methanogens is a major factor in suppression of methanogenesis by sulfate in natural ecosystems. The results with these known mixtures of bacteria suggest that hydrogenase-forming, sulfate-reducing bacteria could be active in some methanogenic ecosystems that are low in sulfate.