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Sulfidogenesis is a promising technology for the selective recovery of chalcophile bulk metals (e.g. Cu, Zn, and Co) from metal-contaminated waters such as acid mine drainage (AMD) and metallurgy waste streams. The use of elemental sulfur (S 0 ) instead of sulfate (SO 4 2- ) as electron acceptor reduces electron donor requirements four-fold, lowering process costs, and expanding the range of operating conditions to a more acidic pH. We previously reported autotrophic S 0 reduction using an industrial mesophilic granular sludge as inoculum under thermoacidophilic conditions. Here, we examined the effect of pH on the S 0 reduction performance of the same inoculum, in a gas-lift reactor run at 30°C under neutral (pH 6.9) and acidic (pH 3.8) conditions, continuously fed with mineral media and H 2 and CO 2 . Steady-state volumetric sulfide production rates (VSPR) dropped 2.5-fold upon transition to acidic pH, from 1.79 ± 0.18 g S 2- ·L -1 ·d -1 to 0.71 ± 0.07 g S 2- ·L -1 ·d -1 . Microbial community composition was analyzed using 16S rRNA gene amplicon sequencing. At neutral pH (6.9), the high relative abundance of the S 0 -reducing genus Sulfurospirillum, previously known only for heterotrophic members, combined with the presence of Acetobacterium and detection of acetate, suggests an important role for heterotrophic S 0 reduction facilitated by acetogenesis. Conversely, at acidic pH (3.9), S 0 reduction appeared autotrophic, as indicated by the high relative abundance of Desulfurella., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Biological desulfurization under haloalkaline conditions has been applied worldwide to remove hydrogen sulfide (H 2 S) from sour gas steams. The process relies on sulfide-oxidizing bacteria (SOB) to oxidize H 2 S to elemental sulfur (S 8 ), which can then be recovered and reused. Recently, a dual-reactor biological desulfurization system was implemented where an anaerobic (sulfidic) bioreactor was incorporated as an addition to a micro-oxic bioreactor, allowing for higher S 8 selectivity by limiting by-product formation. The highly sulfidic bioreactor environment enabled the SOB to remove (poly)sulfides (S x 2- ) in the absence of oxygen, with S x 2- speculated as a main substrate in the removal pathway, thus making it vital to understand its role in the process. The SOB are influenced by the oxidation-reduction potential (ORP) set-point of the micro-oxic bioreactor as it is used to control the product of oxidation (S 8 vs. SO 4 2- ), while the uptake of S x 2- by SOB has been qualitatively linked to pH. Therefore, to quantify these effects, this work determined the concentration and speciation of S x 2- in the biological desulfurization process under various pH values and ORP set-points. The total S x 2- concentrations in the sulfidic zone increased at elevated pH (8.9) compared to low pH (< 8.0), with on average 3.3 ± 1.0 mM-S more S x 2- . Chain lengths varied, with S 7 2- only doubling in concentration while S 5 2- increased 9 fold, which is in contrast with observations from abiotic systems. Changes to the ORP set-point of the micro-oxic reactor did not produce substantial changes in S x 2- concentration in the sulfidic zone. This illustrates that the reduction degree of the SOB in the micro-oxic bioreactor does not enhance their ability to interact with S x 2- in the sulfidic bioreactor. This increased understanding of how both pH and ORP affect changes in S x 2- concentration and chain length can lead to improved efficiency and design of the dual-reactor biological desulfurization process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Thermophilic and hyper-thermophilic anaerobic digestion (AD) are promising techniques for the treatment of concentrated black water (toilet fraction of domestic wastewater collected by low flush volume toilets; BW), recovery of nutrients and simultaneous pathogen removal for safe recovery and reuse of those nutrients. This study showed that thermophilic AD (55 °C) of concentrated BW reaches the same methanisation and COD removal as mesophilic anaerobic treatment of BW (conventional vacuum toilets) and kitchen waste while applying a higher loading rate (OLR) (2.5-4.0 kgCOD/m 3 /day). With a retention time of 8.7 days, and an OLR of >3 kgCOD/m 3 /day, COD removal of 70% and a methanisation of 62% (based on COD t ) was achieved during thermophilic AD. Hyper-thermophilic (70 °C) reached lower levels of methanisation (38%). Start-up time of thermophilic AD was 12 days. And during thermophilic AD, a shift from acetoclastic methanogenesis towards syntrophic acetate oxidation was observed., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Sulfur reduction at hyperthermoacidophilic conditions represents a promising opportunity for metal sulfide precipitation from hot acidic metallurgical streams, avoiding costly cooling down. The suitability of mesophilic anaerobic sludges as the inoculum for sulfur-reducing bioreactors operated at high temperature and low pH was explored. We examined sludges from full-scale anaerobic reactors for sulfur-reducing activity at pH 2.0-3.5 and 70 or 80 °C, with H 2 as an electron donor. At pH 3.5 in batch experiments, sulfidogenesis started within 4 days, reaching up to 100-200 mg·L -1 of dissolved sulfide produced after 19-24 days, depending on the origin of the sludge. Sulfidogenesis resumed after removing H 2 S by flushing with nitrogen gas, indicating that sulfide was limiting the conversion. The best performing sludge was used to inoculate a 4 L gas-lift reactor fed with H 2 as the electron donor, CO 2 as the carbon source, and elemental sulfur as the electron acceptor. The reactor was operated in semibatch mode at a pH 3.5 and 80 °C, and stable sulfide production rates of 60-80 mg·L -1 ·d -1 were achieved for a period of 24 days, without formation of methane or acetate. Our results reveal the potential of mesophilic anaerobic sludges as seed material for sulfur-reducing bioprocesses operated at hyperthermoacidophilic conditions. The process needs further optimization of the volumetric sulfide production rate to gain relevance for practice.

In the biotechnological desulfurization process under haloalkaline conditions, dihydrogen sulfide (H 2 S) is removed from sour gas and oxidized to elemental sulfur (S 8 ) by sulfide-oxidizing bacteria. Besides S 8 , the byproducts sulfate (SO 4 2- ) and thiosulfate (S 2 O 3 by a new process line-up for biological gas desulfurization, applying two bioreactors with different substrate conditions (i.e., sulfidic and microaerophilic), instead of one (i.e., microaerophilic). A 111-day continuous test, mimicking full scale operation, demonstrated that S 2- ) are formed, which consume caustic and form a waste stream. The aim of this study was to increase selectivity toward S 8 by a new process line-up for biological gas desulfurization, applying two bioreactors with different substrate conditions (i.e., sulfidic and microaerophilic), instead of one (i.e., microaerophilic). A 111-day continuous test, mimicking full scale operation, demonstrated that S 8 formation was 96.6% on a molar H 2 S supply basis; selectivity for SO 4 2- and S 2 O 3 2- were 1.4 and 2.0% respectively. The selectivity for S 8 formation in a control experiment with the conventional 1-bioreactor line-up was 75.6 mol %. At start-up, the new process line-up immediately achieved lower SO 4 2- and S 2 O 3 2- formations compared to the 1-bioreactor line-up. When the microbial community adapted over time, it was observed that SO 4 2- formation further decreased. In addition, chemical formation of S 2 O 3 2- was reduced due to biologically mediated removal of sulfide from the process solution in the anaerobic bioreactor. The increased selectivity for S 8 formation will result in 90% reduction in caustic consumption and waste stream formation compared to the 1-bioreactor line-up.

Simultaneous recovery of calcium phosphate granules (CaP granules) and methane in anaerobic treatment of source separated black water (BW) has been previously demonstrated. The exact mechanism behind the accumulation of calcium phosphate (Ca x (PO 4 ) y ) in CaP granules during black water treatment was investigated in this study by examination of the interface between the outer anaerobic biofilm and the core of CaP granules. A key factor in this process is the pH profile in CaP granules, which increases from the edge (7.4) to the center (7.9). The pH increase enhances supersaturation for Ca x (PO 4 ) y phases, creating internal conditions preferable for Ca x (PO 4 ) y precipitation. The pH profile can be explained by measured bioconversion of acetate and H 2 , HCO 3 - and H + into CH 4 in the outer biofilm and eventual stripping of CO 2 and CH 4 (biogas) from the granule. Phosphorus content and Ca x (PO 4 ) y crystal mass quantity in the granules positively correlated with the granule size, in the reactor without Ca 2+ addition, indicating that the phosphorus rich core matures with the granule growth. Adding Ca 2+ increased the overall phosphorus content in granules >0.4 mm diameter, but not in fine particles (<0.4 mm). Additionally, H + released from aqueous phosphate species during Ca x (PO 4 ) y crystallization were buffered by internal hydrogenotrophic methanogenesis and stripping of biogas from the granule. These insights into the formation and growth of CaP granules are important for process optimization, enabling simultaneous Ca x (PO 4 ) y and CH 4 recovery in a single reactor. Moreover, the biological induction of Ca x (PO 4 ) y crystallization resulting from biological increase of pH is relevant for stimulation and control of (bio)crystallization and (bio)mineralization in real environmental conditions.

Chain elongation is an open-culture biotechnological process which converts volatile fatty acids (VFAs) into medium chain fatty acids (MCFAs) using ethanol and other reduced substrates. The objective of this study was to investigate the quantitative effect of CO 2 loading rate on ethanol usages in a chain elongation process. We supplied different rates of CO 2 to a continuously stirred anaerobic reactor, fed with ethanol and propionate. Ethanol was used to upgrade ethanol itself into caproate and to upgrade the supplied VFA (propionate) into heptanoate. A high CO 2 loading rate (2.5 L CO2 ·L -1 ·d -1 ) stimulated excessive ethanol oxidation (EEO; up to 29%) which resulted in a high caproate production (10.8 g·L -1 ·d -1 ). A low CO 2 loading rate (0.5 L CO2 ·L -1 ·d -1 ) reduced EEO (16%) and caproate production (2.9 g·L -1 ·d -1 ). Heptanoate production by VFA upgrading remained constant (∼1.8 g·L -1 ·d -1 ) at CO 2 loading rates higher than or equal to 1 L CO2 ·L -1 ·d -1 . CO 2 was likely essential for growth of chain elongating microorganisms while it also stimulated syntrophic ethanol oxidation. A high CO 2 loading rate must be selected to upgrade ethanol (e.g., from lignocellulosic bioethanol) into MCFAs whereas lower CO 2 loading rates must be selected to upgrade VFAs (e.g., from acidified organic residues) into MCFAs while minimizing use of costly ethanol.

In microbial electrosynthesis (MES), CO 2 can be reduced preferably to multi-carbon chemicals by a biocathode-based process which uses electrochemically active bacteria as catalysts. A mixed anaerobic consortium from biological origin typically produces methane from CO 2 reduction which circumvents production of multi-carbon compounds. This study aimed to develop a stable and robust CO 2 reducing biocathode from a mixed culture inoculum avoiding the methane generation. An effective approach was demonstrated based on (i) an enrichment procedure involving inoculum pre-treatment and several culture transfers in H 2 :CO 2 media, (ii) a transfer from heterotrophic to autotrophic growth and (iii) a sequential batch operation. Biomass growth and gradual acclimation to CO 2 electro-reduction accomplished a maximum acetate production rate of 400mgL catholyte -1 d -1 at -1V (vs. Ag/AgCl). Methane was never detected in more than 300days of operation. Accumulation of acetate up to 7-10gL -1 was repeatedly attained by supplying (80:20) CO 2 :N 2 mixture at -0.9 to -1V (vs. Ag/AgCl). In addition, ethanol and butyrate were also produced from CO 2 reduction. Thus, a robust CO 2 reducing biocathode can be developed from a mixed culture avoiding methane generation by adopting the specific culture enrichment and operation procedures without the direct addition of chemical inhibitor., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Bioelectrochemical power-to-gas (BEP2G) is considered a potentially convenient way of storing renewable surplus electricity in the form of methane. In methane-producing bioelectrochemical systems (BESs), carbon dioxide and electrical energy are converted into methane, using electrodes that supply either electrons or hydrogen to methanogenic archaea. This review summarizes the performance of methane-producing BESs in relation to cathode potential, electrode materials, operational strategies, and inoculum. Analysis and estimation of energy input and production rates show that BEP2G may become an attractive alternative for thermochemical methanation, and biochemical methanogenesis. To determine if BEP2G can become a future power-to-gas technology, challenges relating to cathodic energy losses, choice of a suitable electron donor, efficient reactor design/operation, and experience with large reactors need to be overcome., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Anaerobic digestion at low temperature is an attractive technology especially in moderate climates, however, low temperature results in low microbial activity and low rates of methane formation. This study investigated if bioelectrochemical systems (BESs) can enhance methane production from organic matter in low-temperature anaerobic digestion (AD). A bioelectrochemical reactor was operated with granular activated carbon as electrodes at 10 °C. Our results showed that bioelectrochemical systems can enhance CH4 yield, accelerate CH4 production rate and increase acetate removal efficiency at 10 °C. The highest CH4 yield of 31 mg CH4-COD/g VSS was achieved in the combined BES-AD system at a cathode potential of -0.90 V (Ag/AgCl), which was 5.3-6.6 times higher than that in the AD reactor at 10 °C. CH4 production rate achieved in the combined BES-AD system at 10 °C was only slightly lower than that in the AD reactor at 30 °C. The presence of an external circuit between the acetate-oxidizing bioanode and methane-producing cathode provided an alternative pathway from acetate via electrons to methane, potentially via hydrogen. This alternative pathway seems to result in higher CH4 production rates at low temperature compared with traditional methanogenesis from acetate. Integration of BES with AD could therefore be an attractive alternative strategy to enhance the performance of anaerobic digestion in cold areas., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Chain elongation into medium-chain carboxylates, such as n-caproate and n-caprylate, with ethanol as an electron donor and with open cultures of microbial consortia (i.e., reactor microbiomes) under anaerobic conditions is being developed as a biotechnological production platform. The goal is to use the high thermodynamic efficiency of anaerobic fermentation to convert organic biomass or organic wastes into valuable biochemicals that can be extracted. Several liter-scale studies have been completed and a first pilot-plant study is underway. However, the underlying microbial pathways are not always well understood. In addition, an interdisciplinary approach with knowledge from fields ranging from microbiology and chemical separations to biochemistry and environmental engineering is required. To bring together research from different fields, we reviewed the literature starting with the microbiology and ending with the bioprocess engineering studies that already have been performed. Because understanding the microbial pathways is so important to predict and steer performance, we delved into a stoichiometric and thermodynamic model that sheds light on the effect of substrate ratios and environmental conditions on product formation. Finally, we ended with an outlook.

The use of methane and acetate as electron donors for biological reduction of thiosulphate in a 5-L laboratory membrane bioreactor was studied and compared to disproportionation of thiosulphate as competing biological reaction. The reactor was operated for 454 days in semi-batch mode; 30 % of its liquid phase was removed and periodically replenished (days 77, 119, 166, 258, 312 and 385). Although the reactor was operated under conditions favourable to promote thiosulphate reduction coupled to methane oxidation, thiosulphate disproportionation was the dominant microbial process. Pyrosequencing analysis showed that the most abundant microorganisms in the bioreactor were phototrophic green sulphur bacteria (GSB) belonging to the family Chlorobiaceae and thiosulphate-disproportionating bacteria belonging to the genus Desulfocapsa. Even though the reactor system was surrounded with opaque plastic capable of filtering most of the light, the GSB used it to oxidize the hydrogen sulphide produced from thiosulphate disproportionation to elemental sulphur. Interrupting methane and acetate supply did not have any effect on the microbial processes taking place. The ultimate goal of our research was to develop a process that could be applied for thiosulphate and sulphate removal and biogenic sulphide formation for metal precipitation. Even though the system achieved in this study did not accomplish the targeted conversion using methane as electron donor, it does perform microbial conversions which allow to directly obtain elemental sulphur from thiosulphate.

Simultaneous nitrogen and methane removal by the slow growing denitrifying methanotrophic bacterium 'Candidatus Methylomirabilis oxyfera' offers opportunities for a new approach to wastewater treatment. However, volumetric nitrite consumption rates should be increased by an order of magnitude before application in wastewater treatment becomes possible. A maximum volumetric nitrite consumption rate of 36 mg NO2(-)-N/L d was achieved in a membrane bioreactor inoculated with wastewater sludge and operated at 20°C. This rate is similar to maximum rates reported in literature, though it was thought that by strict biomass retention using membranes, higher rates would be achieved. In experiments lasting several years, growth was not stable: every experiment showed a decrease in activity after 1-2 years. The cause remains unknown. Rates increased after addition of copper and operating a membrane bioreactor at shorter hydraulic retention times. Further research should focus on long-term effects of copper addition and operation at hydraulic retention times in the order of hours using membrane bioreactors., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A methane-producing biocathode that converts CO(2) into methane was studied electrochemically and microbiologically. The biocathode produced methane at a maximum rate of 5.1 L CH(4)/m(2) projected cathode per day (1.6 A/m(2)) at -0.7 V versus NHE cathode potential and 3.0 L CH(4)/m(2) projected cathode per day (0.9 A/m(2)) at -0.6 V versus NHE cathode potential. The microbial community at the biocathode was dominated by three phylotypes of Archaea and six phylotypes of bacteria. The Archaeal phylotypes were most closely related to Methanobacterium palustre and Methanobacterium aarhusense. Besides methanogenic Archaea, bacteria seemed to be associated with methane production, producing hydrogen as an intermediate. Biomass density varied greatly with part of the carbon electrode covered with a dense biofilm, while only clusters of cells were found on other parts. Based on our results, we discuss how inoculum enrichment and changing operational conditions may help to increase biomass density and to select for microorganisms that produce methane.

Despite many advantages of anaerobic sewage treatment over conventional activated sludge treatment, it has not yet been applied in temperate zones. This is especially because effluent from low-temperature anaerobic treatment contains nitrogen and dissolved methane. The presence of nitrogen and methane offers the opportunity to develop a reactor in which methane is used as electron donor for denitrification. Such a reactor could be used in a new concept for low-temperature anaerobic sewage treatment, consisting of a UASB-digester system, a reactor for denitrification coupled to anaerobic methane oxidation, and a nitritation reactor. In the present study denitrifying methanotrophic bacteria similar to 'Candidatus Methylomirabilis oxyfera' were enriched. Maximum volumetric nitrite consumption rates were 33.5 mg NO(2)(-)-N/Ld (using synthetic medium) and 37.8 mg NO(2)(-)-N/Ld (using medium containing effluent from a sewage treatment plant), which are similar to the maximum rate reported so far. Though the goal was to increase the rates, in both reactors, after reaching these maximum rates, volumetric nitrite consumption rates decreased in time. Results indicate biomass washout may have significantly decelerated enrichment. Therefore, to obtain higher volumetric consumption rates, further research should focus on systems with complete biomass retention., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Physicochemical processes, such as the Lo-cat and Amine-Claus process, are commonly used to remove hydrogen sulfide from hydrocarbon gas streams such as landfill gas, natural gas, and synthesis gas. Biodesulfurization offers environmental advantages, but still requires optimization and more insight in the reaction pathways and kinetics. We carried out experiments with gas lift bioreactors inoculated with haloalkaliphilic sulfide-oxidizing bacteria. At oxygen-limiting levels, that is, below an O(2)/H(2)S mole ratio of 1, sulfide was oxidized to elemental sulfur and sulfate. We propose that the bacteria reduce NAD(+) without direct transfer of electrons to oxygen and that this is most likely the main route for oxidizing sulfide to elemental sulfur which is subsequently oxidized to sulfate in oxygen-limited bioreactors. We call this pathway the limited oxygen route (LOR). Biomass growth under these conditions is significantly lower than at higher oxygen levels. These findings emphasize the importance of accurate process control. This work also identifies a need for studies exploring similar pathways in other sulfide oxidizers such as Thiobacillus bacteria.

The anaerobic treatment of low strength domestic sewage at low temperature is an attractive and important topic at present. The upflow anaerobic sludge bed (UASB)-digester system is one of the anaerobic systems to challenge low temperature and concentrations. The effect of sludge recirculation rate on a UASB-digester system treating domestic sewage at 15 °C was studied in this research. A sludge recirculation rate of 0.9, 2.6 and 12.5% of the influent flow rate was investigated. The results showed that the total chemical oxygen demand (COD) removal efficiency rose with increasing sludge recirculation rate. A sludge recirculation rate of 0.9% of the influent flow rate led to organic solids accumulation in the UASB reactor. After the sludge recirculation rate increased from 0.9 to 2.6%, the stability of the UASB sludge was substantially improved from 0.37 to 0.15 g CH₄-COD/g COD, and the bio-gas production in the digester went up from 2.9 to 7.4 L/d. The stability of the UASB sludge and bio-gas production in the digester were not significantly further improved by increasing sludge recirculation rate to 12.5% of the influent flow rate, but the biogas production in the UASB increased from 0.37 to 1.2 L/d. It is recommended to apply a maximum sludge recirculation rate of 2-2.5% of the influent flow rate in a UASB-digester system, as this still allows energy self-sufficiency of the system.

Anaerobic oxidation of methane (AOM) in marine sediments is coupled to sulfate reduction (SR). AOM is mediated by distinct groups of archaea, called anaerobic methanotrophs (ANME). ANME co-exist with sulfate-reducing bacteria, which are also involved in AOM coupled SR. The microorganisms involved in AOM coupled to SR are extremely difficult to grow in vitro. Here, a novel well-mixed submerged-membrane bioreactor system is used to grow and enrich the microorganisms mediating AOM coupled to SR. Four reactors were inoculated with sediment sampled in the Eckernförde Bay (Baltic Sea) and operated at a methane and sulfate loading rate of 4.8 L L(-1) day(-1) (196 mmol L(-1) day(-1)) and 3.0 mmol L(-1) day(-1). Two bioreactors were controlled at 15 degrees C and two at 30 degrees C, one reactor at 30 degrees C contained also anaerobic granular sludge. At 15 degrees C, the volumetric AOM and SR rates doubled approximately every 3.8 months. After 884 days, an enrichment culture was obtained with an AOM and SR rate of 1.0 mmol g(volatile suspended solids) (-1) day(-1) (286 micromol g(dry weight) (-1) day(-1)). No increase in AOM and SR was observed in the two bioreactors operated at 30 degrees C. The microbial community of one of the 15 degrees C reactors was analyzed. ANME-2a became the dominant archaea. This study showed that sulfate reduction with methane as electron donor is possible in well-mixed bioreactors and that the submerged-membrane bioreactor system is an excellent system to enrich slow-growing microorganisms, like methanotrophic archaea.

High rate sulfate reduction under acidic conditions opens possibilities for new process flow sheets that allow the selective recovery of metals from mining and metallurgical waste and process water. However, knowledge about high-rate sulfate reduction under acidic conditions is limited. This paper investigates sulfate reduction in a membrane bioreactor at a controlled pH of 5. Sulfate and formate were dosed using a pH-auxostat system while formate was converted into hydrogen, which was used for sulfate reduction. Sulfide was removed from the gas phase to prevent sulfide inhibition. This study shows a high-rate sulfate-reducing bioreactor system for the first time at pH 5, with a volumetric activity of 188mmol SO(4)(2-)/I/d and a specific activity of 81mmol SO(4)(2-) volatile suspended. The microbial community at the end of the reactor run consisted of a diverse mixed population including sulfate-reducing bacteria.

Process streams with high concentrations of metals and sulfate are characteristic for the mining and metallurgical industries. This study aims to selectively recover nickel from a nickel-iron-containing solution at pH 5.0 using a single stage bioreactor that simultaneously combines low pH sulfate reduction and metal-sulfide formation. The results show that nickel was selectively precipitated in the bioreactor at pH 5.0 and the precipitates consisted of >or=83% of the nickel content. The nickel-iron precipitates were partly crystalline and had a metal/sulfur ratio of 1, suggesting these precipitates were NiS and FeS. Experiments focusing on nickel recovery at pH 5.0 and 5.5 reached a recovery of >99.9%, resulting in a nickel effluent concentration<0.05 microM. The mixed microbial population included known sulfate reducers and acetogens. This study shows that selective metal precipitation in a single stage sulfate reducing bioreactor operated at low pH has the potential to produce metal-sulfides that can be used by the metallurgical industry as a resource for metal production.

Biotechnological treatment of sulfate- and metal-ionscontaining acidic wastewaters from mining and metallurgical activities utilizes sulfate-reducing bacteria to produce sulfide that can subsequently precipitate metal ions. Reducing sulfate at a low pH has several advantages above neutrophilic sulfate reduction. This study describes the effect of sulfide removal on the reactor performance and microbial community in a high-rate sulfidogenic gas-lift bioreactor fed with hydrogen at a controlled internal pH of 5. Under sulfide removal conditions, 99% of the sulfate was converted at a hydraulic retention time of 24 h, reaching a volumetric activity as high as 51 mmol sulfate/l/d. Under nonsulfide removal conditions, <25% of the sulfate was converted at a hydraulic retention time of 24 h reaching volumetric activities of <13mmol sulfate/l/d. The absence of sulfide removal at a hydraulic retention time of 24 h resulted in an average H2S concentration of 18.2 mM (584 mg S/l). The incomplete sulfate removal was probably due to sulfide inhibition. Molecular phylogenetic analysis identified 11 separate 16S rRNA bands under sulfide stripping conditions, whereas under nonsulfide removal conditions only 4 separate 16S rRNA bands were found. This shows that a less diverse population was found in the presence of a high sulfide concentration.

Bioelectrochemical systems (BESs), such as microbial fuel cells (MFCs) and microbial electrolysis cells (MECs), are generally regarded as a promising future technology for the production of energy from organic material present in wastewaters. The current densities that can be generated with laboratory BESs now approach levels that come close to the requirements for practical applications. However, full-scale implementation of bioelectrochemical wastewater treatment is not straightforward because certain microbiological, technological and economic challenges need to be resolved that have not previously been encountered in any other wastewater treatment system. Here, we identify these challenges, provide an overview of their implications for the feasibility of bioelectrochemical wastewater treatment and explore the opportunities for future BESs.

Many industrial waste and process waters contain high concentrations of sulfate, which can be removed by sulfate-reducing bacteria (SRB). This paper reports on mesophilic (30 degrees C) sulfate reduction at pH 6 with formate as electron donor in a membrane bioreactor with a pH-auxostat dosing system. A mixed microbial community from full-scale industrial wastewater treatment bioreactors operated at pH 7 was used as inoculum. The pH-auxostat enabled the bacteria to convert sulfate at a volumetric activity of 302 mmol sulfate reduced per liter per day and a specific activity of 110 mmol sulfate reduced per gram volatile suspended solids per day. Biomass grew in 15 days from 0.2 to 4 g volatile suspended solids per liter. This study shows that it is possible to reduce sulfate at pH 6 with formate as electron donor at a high volumetric and specific activity with inocula from full-scale industrial wastewater treatment bioreactors operated at neutral pH. The combination of a membrane bioreactor and a pH-auxostat is a useful research tool to study processes with unknown growth rates at maximum activities.

A biotechnological process is described to remove hydrogen sulfide (H(2)S) from high-pressure natural gas and sour gases produced in the petrochemical industry. The process operates at halo-alkaline conditions and combines an aerobic sulfide-oxidizing reactor with an anaerobic sulfate (SO(4) (2-)) and thiosulfate (S(2)O(3) (2-)) reducing reactor. The feasibility of biological H(2)S oxidation at pH around 10 and total sodium concentration of 2 mol L(-1) was studied in gas-lift bioreactors, using halo-alkaliphilic sulfur-oxidizing bacteria (HA-SOB). Reactor operation at different oxygen to sulfide (O(2):H(2)S) supply ratios resulted in a stable low redox potential that was directly related with the polysulfide (S(x) (2-)) and total sulfide concentration in the bioreactor. Selectivity for SO(4) (2-) formation decreased with increasing S(x) (2-) and total sulfide concentrations. At total sulfide concentrations above 0.25 mmol L(-1), selectivity for SO(4) (2-) formation approached zero and the end products of H(2)S oxidation were elemental sulfur (S(0)) and S(2)O(3) (2-). Maximum selectivity for S(0) formation (83.3+/-0.7%) during stable reactor operation was obtained at a molar O(2):H(2)S supply ratio of 0.65. Under these conditions, intermediary S(x) (2-) plays a major role in the process. Instead of dissolved sulfide (HS(-)), S(x) (2-) seemed to be the most important electron donor for HA-SOB under S(0) producing conditions. In addition, abiotic oxidation of S(x) (2-) was the main cause of undesirable formation of S(2)O(3) (2-). The observed biomass growth yield under SO(4) (2-) producing conditions was 0.86 g N mol(-1) H(2)S. When selectivity for SO(4) (2-) formation was below 5%, almost no biomass growth was observed., ((c) 2007 Wiley Periodicals, Inc.)

ABSTRACT BACKGROUND The treatment of source separated urine allows for a more sustainable approach to nutrients recovery in actual wastewater treatment plants. Struvite precipitation from urine yields a slow-release fertilizer (struvite) with a high marketable value for agricultural use. Extensive research on struvite precipitation has been carried out at laboratory scale, but there is still a lack of specific knowledge about the operation and key parameters at larger scale and over the long term. The effectiveness of the struvite reactor was assessed, as well as the quality of the harvested struvite in terms of size and composition. RESULTS The efficiency of recovery of P was maintained at 85-99% for 1 year of continuous operation. Phosphorus was recovered in the form of struvite with purity over 90%. Struvite was harvested as round pellets between 0.3 and 6 mm in diameter, with heavy metals concentration well below the legal limits for fertilizers in The Netherlands. The post-treatment of the reactor effluent yielded almost complete removal (>95%) of calcium and magnesium. A loss of 26 ± 5% of the influent chemical oxygen demand and 16 ± 1% of the NH4+-N occurred during the magnesium ammonium phosphate ( MAP) precipitation and the post-treatment. CONCLUSION This work shows how the treatment of source-separated urine by struvite precipitation in a fluidized bed reactor allows for efficient recovery of phosphorus in the form of a potential fertilizer. © 2016 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Acidic industrial process and wastewaters often contain high sulfate and metal concentrations and their direct biological treatment is thus far not possible as biological processes at pH < 5 have been neglected. Sulfate-reducing bacteria convert sulfate to sulfide that can subsequently be used to recover metals as metal-sulfides precipitate. This study reports on high-rate sulfate reduction with a mixed microbial community at pH 4.0 and 4.5 with hydrogen and/or formate as electron donors. The maximum sulfate reducing activity at pH 4.0 was sustained for over 40 days with a specific activity 500-fold greater than previously reported values: 151 mmol sulfate reduced/L reactor liquid per day with a maximum specific activity of 84 mmol sulfate per gram of volatile suspended solids per day. The biomass yield gradually decreased from 38 to 0.4 g volatile suspended solids per kilogram of sulfate when decreasing the reactor pH from pH 6 to 4. The microorganisms had a high maintenance requirement probably due maintaining pH homeostasis and the toxicity of sulfide at low pH. The microbial community diversity in the pH 4.0 membrane bioreactor decreased over time, while the diversity of the sulfate reducing community increased. Thus, a specialized microbial community containing a lower proportion of microorganisms capable of activity at pH 4 developed in the reactor compared with those present at the start of the experiment. The 16S rRNA genes identified from the pH 4.0 grown mixed culture were most similar to those of Desulfovibrio species and Desulfosporosinus sp. M1. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [ABSTRACT FROM AUTHOR]

Background: Chain elongation forms a new platform technology for the circular production of biobased chemicals from renewable carbon and energy sources. This study aimed to develop a continuous methanol-based chain elongation process for the open-culture production of a new-generation biofuel precursor and potential platform chemical: n-valerate. Propionate was used as a substrate for chain elongation to n-valerate in an anaerobic open-culture bioreactor. In addition, the co-production of n- and iso-butyrate in addition to n-valerate via, respectively, acetate and propionate elongation was investigated. Results: n-Valerate was produced during batch and continuous experiments with a pH in the range 5.5–5.8 and a hydraulic retention time of 95 h. Decreasing the pH from 5.8 to 5.5 caused an increase of the selectivity for n-valerate formation (from 58 up to 70 wt%) during methanol-based propionate elongation. n-Valerate and both n- and iso-butyrate were produced during simultaneous methanol-based elongation of propionate and acetate. Propionate was within the open-culture preferred over acetate as a substrate with 10–30% more consumption. Increasing the methanol concentration in the influent (from 250 to 400 mM) resulted in a higher productivity (from 45 to 58 mmol C/L/day), but a lower relative product selectivity (from 49 to 43 wt%) of n-valerate. The addition of acetate as a substrate did not change the average n-valerate productivities. Within the continuous bioreactor experiments, 6 to 17 wt% of formed products was methane. The microbial community during all steady-states in both methanol-based elongation bioreactors was dominated by species related to Clostridium luticellarii and Candidatus Methanogranum. C. luticellarii is the main candidate for n-valerate formation from methanol and propionate. Conclusions: n-Valerate was for the first time proven to be produced from propionate and methanol by an open-culture bioreactor. Methanogenic activity can be inhibited by decreasing the pH, and the n-valerate productivity can be improved by increasing the methanol concentration. The developed process can be integrated with various biorefinery processes from thermochemical, (bio)electrochemical, photovoltaic and microbial technologies. The findings from this study form a useful tool to steer the process of biological production of chemicals from biomass and other carbon and energy sources. [ABSTRACT FROM AUTHOR]

Abstract: Dissolved zinc is present in natural waters and process streams generated by the mining and metallurgical industry. These streams usually have a low pH. By using sulfate reducing bacteria, sulfide can be produced that precipitates with zinc as zinc sulfide (sphalerite), which can be easily separated from the wastewater and even reused as zinc concentrate. In this study, a sulfate reducing gas-lift bioreactor was operated at pH 5.5 using hydrogen as electron donor for sulfate reduction. We demonstrate effective zinc removal (7.2mmolL−1 d−1) with low zinc effluent concentrations (0.65–8.8μM) in a system combining sulfide generation by sulfate reducing bacteria (7.2–10.6mmolSO4 2− L−1 d−1) at low pH (5.5) with the bio-precipitation of crystalline sphalerite. To investigate the effect of the sulfide excess on the settling properties of the sphalerite precipitates, the sulfide excess concentration was varied about two orders of magnitude (0.008–2.2mM). The results show that crystalline sphalerite was formed in all cases, but larger particles with better settling properties were formed at lower sulfide concentrations. [Copyright &y& Elsevier]

In a biotechnological process for hydrogen sulfide (H2S) removal from gas streams, operating at natronophilic conditions, formation of thiosulfate (S2O32-) is unfavorable, as it leads to a reduced sulfur production. Thiosulfate formation was studied in gas-lift bioreactors, using natronophilic biomass at [Na+]+[K+] = 2 mol L-1. The results show that at sulfur producing conditions, selectivity for S2O32- formation mainly depends on the equilibrium between free sulfide (HS-) and polysulfide (Sx2-), which can be controlled via the pH. At pH 8.6, 21% of the total dissolved sulfide is present as Sx2- and selectivity for S2O32- formation is 3.9-5.5%. At pH 10, 87% of the total dissolved sulfide is present as Sx2- and 20-22% of the supplied H2S is converted to S2O32-, independent of the H2S loading rate. Based on results of bioreactor experiments and biomass activity tests, a mechanistic model is proposed to describe the relation between S2O32- formation and pH. [ABSTRACT FROM AUTHOR]

Chain elongation is an open-culture biotechnological process which converts short chain fatty acids and an electron donor to medium chain fatty acids (MCFAs). With this letter we present the first observation of granular sludge formation in a chain elongation process. This discovery was made in a continuously stirred anaerobic reactor producing caproate (10.8 g L −1 d −1 ) and heptanoate (1.8 g L −1 d −1 ) as main MCFAs. Concurrently granular and suspended sludge were shaped and attributed to 85% and 15% respectively of the total sludge. Both sludge types showed equal product distributions and contributed similarly to MCFA production. Granules had irregular shapes, diameters up to ∼1.5 mm, settling velocities of 4–36 m h −1 and contained micro-organisms with various shapes. The in-situ settler retained sludge in the bioreactor resulting in a SRT of 4.7 days at an HRT of 17 h. Granular sludge based chain elongation can be optimised as a high rate biotechnological process. [ABSTRACT FROM AUTHOR]

Although literature suggests that aquatic worms can help to enhance the methane production from excess activated sludge, clear evidence for this is missing. Therefore, anaerobic digestion tests were performed at 20 and at 30 °C with sludge from a high-loaded membrane bioreactor, the aquatic worm Lumbriculus variegatus , feces from these worms and with mixtures of these substrates. A significant synergistic effect of the worms or their feces on methane production from the high-loaded sludge or on its digestion rate was not observed. However, a positive effect on low-loaded activated sludge, which generally has a lower anaerobic biodegradability, cannot be excluded. The results furthermore showed that the high-loaded sludge provides an excellent feed for L. variegatus , which is promising for concepts where worm biomass is considered a resource for technical grade products such as coatings and glues. [ABSTRACT FROM AUTHOR]

Abstract: Removal of the toxic selenium compounds selenite and selenate from waste water before discharge is becoming increasingly imperative in industrialized countries. Bacteria can reduce selenate to selenite, but also further to elemental selenium, selenide or organic selenium. In this paper, we aim to exclusively bio-reduce selenate to selenite in an open high-rate bioreactor. This conversion could be part of a two-stage process in which the selenite is subsequently reduced by chemical means under optimal conditions to produce a biomass-free selenium product. In the process, yield and reduction rate of biological selenate to selenite should be maximized, while formation of elemental selenium, selenide and organic selenium compounds should be avoided. Fed-batch experiments with a liquid volume of 0.25–0.75 L at different temperatures 20–30–40–50 °C, pH settings 6–7–8–9, initial biomass concentration of 1 or 5 g wet weight granular Eerbeek sludge and various lactic acid concentrations were performed to determine their effect on the biological conversion of selenate to selenite. Furthermore, the effect on selenite losses by further biological reduction or, if present, chemical reduction was investigated as well. Optimal selenate reduction to selenite was found at 30 °C and pH 6 or 7 or 8 with 25 mM selenate and 13.75 mM lactic acid in the influent, with a selenite yield of 79–95%. In all the other conditions, less selenate was reduced to selenite. Also a 5 times higher electron donor concentration resulted in less selenite production, with only 22% of the selenate converted to selenite. The high yield and the high biological reduction rate of at least 741 mg Se/g initial VSS/day detected in the 1 g initial biomass experiment implicate that biological selenate conversion to selenite is a feasible process. [Copyright &y& Elsevier]

Abstract: Direct anaerobic treatment of municipal waste waters allows for energy recovery in the form of biogas. A further decrease in the energy requirement for waste water treatment can be achieved by removing the ammonium in the anaerobic effluent with an autotrophic process, such as anammox. Until now, anammox has mainly been used for treating warm (>30 °C) and concentrated (>500 mg N/L) waste streams. Application in the water line of municipal waste water treatment poses the challenges of a lower nitrogen concentration (<100 mg N/L) and a lower temperature (≤20 °C). Good biomass retention and a short HRT are required to achieve a sufficiently high nitrogen loading rate. For this purpose a 4.5 L gaslift reactor was inoculated with a small amount of anammox granules and operated for 253 days at 20 °C. The synthetic influent contained (69 ± 5) mg (NH4 + + )/L and 20 vol.% of anaerobically stabilised effluent. Results showed a clear increase in nitrogen loading rate (NLR) up to 0.31 g (NH4 + NO2)-N/(L × d) at a hydraulic retention time (HRT) of 5.3 h. A low effluent concentration of 0.03–0.17 mg ()-N/L could be achieved. Anammox biomass was retained as granules and as a biofilm on the reactor walls, which contributed 54 and 46%, respectively, towards total activity. The biomass was further characterised by an estimated net growth rate of 0.040 d−1 and an apparent activation energy of 72 kJ/mol. The results presented in this paper showed that anammox bacteria can be applied for autotrophic nitrogen removal from the water line at a municipal waste water treatment plant. Combining direct anaerobic treatment with autotrophic nitrogen removal opens opportunities for energy-efficient treatment of municipal waste waters. [Copyright &y& Elsevier]

Abstract: Treatment of domestic waste water results in the production of waste sludge, which requires costly further processing. A biological method to reduce the amount of waste sludge and its volume is treatment in an aquatic worm reactor. The potential of such a worm reactor with the oligochaete Lumbriculus variegatus has been shown at small scale. For scaling up purposes, a new configuration of the reactor was designed, in which the worms were positioned horizontally in the carrier material. This was tested in a continuous experiment of 8 weeks where it treated all the waste sludge from a lab-scale activated sludge process. The results showed a higher worm growth rate compared to previous experiments with the old configuration, whilst nutrient release was similar. The new configuration has a low footprint and allows for easy aeration and faeces collection, thereby making it suitable for full scale application. [Copyright &y& Elsevier]

Abstract: To liberate gold from refractory pyrite, oxidative destruction techniques that consume lots of energy and generate acidic waste streams are custom. As an alternative the “bio-reduction” of pyrite is proposed and investigated in this study. Bio-reduction is an anaerobic process based on sulfate/sulfur reducing bacteria which are thought to be able to use pyrite-sulfur as a possible electron acceptor. The conversion of pyrite-sulfur into hydrogen sulfide is advantageous because energy is saved and the generation of an acidic waste stream is prevented. In addition, the generated H2S can be used to produce elemental sulfur, or even gold lixiviants such as thiosulfate or bisulfide. Batch experiments under anaerobic conditions showed that two effects can inhibit bio-reduction; methane formation and sulfide accumulation. In a gas lift loop reactor operated at pH 5, temperature of 35°C, and with continuous sulfide removal no evidence of pyrite bio-reduction was found. Though the sulfate reducing bacteria survived, they did not utilize pyrite-sulfur as an electron acceptor under the chosen conditions. [Copyright &y& Elsevier]