Mixed-culture Reactors Research Articles

Enhanced elemental sulfur recovery and nitrogen removal through coupling of sulfide-dependent denitrification and anammox processes during ammonium- and sulfide-laden waste stream treatment

Influence of anammox process on autotrophic denitrification was investigated in this study, during simultaneous ammonium, nitrate and sulfide treatment. Elemental sulfur production and nitrogen removal by sulfide-dependent denitrifier (S-den) and mixed culture (S-den-anammox) were compared. Accumulation of nitrite (100%) and elemental sulfur (46.3%) was achieved at pH 7.0 and 30 °C with S2O32−/NO3− ratio of 1:1 by autotrophic denitrifier. Enhanced elemental sulfur production (56.5%–99.5%) by autotrophic denitrifier was obtained after coupling with anammox process. Total nitrogen removal increased to 80.5% in mixed culture reactor, which was 72.0% in anammox reactor. N2O emission in anammox reactor was not affected (<0.5% gaseous nitrogen) after coupling with autotrophic denitrification. Therefore, this study demonstrated enhanced elemental sulfur recovery and nitrogen removal through coupling of sulfide-dependent denitrification and anammox processes, which might be a potential technology for water eutrophication and corrosion control, and atmospheric acid rain, PM2.5 and global warming control.

Immobilized mixed-culture reactor (IMcR) for hydrogen and methane production from glucose

Immobilized cell technology is a new technique to produce biogas. In the present study, an immobilized mixed-culture reactor (IMcR) in batch-mode operation was used for the production of hydrogen and methane simultaneously from glucose. Several factors, such as glucose concentration, temperature and fermentation time, were evaluated to determine the optimal conditions for hydrogen and methane production. Gas chromatography with a thermal conductivity detector (GC-TCD) and high-performance liquid chromatography (HPLC) were used to analyse the gas and effluent. The morphologies of the immobilized cells were characterized using scanning electron microscopy (SEM). The optimal conditions for hydrogen and methane production were obtained using a substrate with 5.0 g/L glucose at 60 °C for fermentation times of 48.0 h (hydrogen) and 72.0 h (methane). The maximum yields of hydrogen and methane at these optimal conditions were 37.0 ± 0.0 (×10−3) mol/mol glu and 39.0 ± 0.0 (×10−3) mol/mol glu, respectively. The chemical oxygen demand (COD) and pH gradually decreased with increasing fermentation time and temperature. However, the performance of the IMcR decreased over time due to cell damage and microorganism detachment from the cell. In conclusion, the IMcR system is a potential system for the simultaneous production of hydrogen and methane.

In-situ biogas upgrading with pulse H2 additions: The relevance of methanogen adaption and inorganic carbon level

Surplus electricity from fluctuating renewable power sources may be converted to CH4 via biomethanisation in anaerobic digesters. The reactor performance and response of methanogen population of mixed-culture reactors was assessed during pulsed H2 injections. Initial H2 uptake rates increased immediately and linearly during consecutive pulse H2 injections for all tested injection rates (0.3 to 1.7LH2/Lsludge/d), while novel high throughput mcrA sequencing revealed an increased abundance of specific hydrogenotrophic methanogens. These findings illustrate the adaptability of the methanogen population to H2 injections and positively affects the implementation of biomethanisation. Acetate accumulated by a 10-fold following injections exceeding a 4:1 H2:CO2 ratio and may act as temporary storage prior to biomethanisation. Daily methane production decreased for headspace CO2 concentrations below 12% and may indicate a high sensitivity of hydrogenotrophic methanogens to CO2 limitation. This may ultimately decide the biogas upgrading potential which can be achieved by biomethanisation.

Quantitative Polymerase Chain Reaction for Microbial Growth Kinetics of Mixed Culture System.

Microbial growth kinetics is often used to optimize environmental processes owing to its relation to the breakdown of substrate (contaminants). However, the quantification of bacterial populations in the environment is difficult owing to the challenges of monitoring a specific bacterial population within a diverse microbial community. Conventional methods are unable to detect and quantify the growth of individual strains separately in the mixed culture reactor. This work describes a novel quantitative PCR (qPCR)-based genomic approach to quantify each species in mixed culture and interpret its growth kinetics in the mixed system. Batch experiments were performed for both single and dual cultures of Pseudomonas putida and Escherichia coli K12 to obtain Monod kinetic parameters (μmax and Ks). The growth curves and kinetics obtained by conventional methods (i.e., dry weight measurement and absorbance reading) were compared with that obtained by qPCR assay. We anticipate that the adoption of this qPCR-based genomic assay can contribute significantly to traditional microbial kinetics, modeling practice, and the operation of bioreactors, where handling of complex mixed cultures is required.

Carbon dioxide reduction by mixed and pure cultures in microbial electrosynthesis using an assembly of graphite felt and stainless steel as a cathode

Carbon dioxide (CO2) reduction to multi-carbon compounds at the cathode using chemolithoautotrophs is an emerging application of microbial electrosynthesis (MES). In this study, CO2 reduction in MES was investigated at hydrogen evolving potentials, separately by a mixed culture and Clostridium ljungdahlii, using a graphite felt and stainless steel assembly as cathode. The mixed culture reactor produced acetate at the maximum rate of 1.3mMd−1, along with methane and hydrogen at −1.1V/Ag/AgCl. Over 160days of run-time in four fed-batches, 26% of bicarbonate was converted to acetate between day 28 and 41, whereas in the late batches, methane production prevailed. Out of 45days of run-time in the C. ljungdahlii reactor, 2.4mMd−1 acetate production was achieved at −0.9V/Ag/AgCl in Batch 1. Simultaneous product degradation occurred when the mixed culture was not selectively enriched. Hydrogen evolution is potentially the rapid way of transferring electrons to the biocatalysts for higher bioproduction rates.

Alamethicin suppresses methanogenesis and promotes acetogenesis in bioelectrochemical systems.

Microbial electrosynthesis (MES) systems with mixed cultures often generate a variety of gaseous and soluble chemicals. Methane is the primary end product in mixed-culture MES because it is the thermodynamically most favorable reduction product of CO2. Here, we show that the peptaibol alamethicin selectively suppressed the growth of methanogens in mixed-culture MES systems, resulting in a shift of the solution and cathode communities to an acetate-producing system dominated by Sporomusa, a known acetogenic genus in MES systems. Archaea in the methane-producing control were dominated by Methanobrevibacter species, but no Archaea were detected in the alamethicin-treated reactors. No methane was detected in the mixed-culture reactors treated with alamethicin over 10 cycles (∼ 3 days each). Instead, acetate was produced at an average rate of 115 nmol ml(-1) day(-1), similar to the rate reported previously for pure cultures of Sporomusa ovata on biocathodes. Mixed-culture control reactors without alamethicin generated methane at nearly 100% coulombic recovery, and no acetate was detected. These results show that alamethicin is effective for the suppression of methanogen growth in MES systems and that its use enables the production of industrially relevant organic compounds by the inhibition of methanogenesis.

Hydrogen Sulfide Removal from Biogas Using Pure and Mixed Cultures of Sulfide-Oxidizing Bacteria Biofiltration

A novel biofiltration system using pure and mixed cultures of sulfide-oxidizing bacteria immobilized on granular activated carbon (GAC) to purify synthetic and real biogas from a sulfate reaction reactor of concentrated latex industry, contaminated by H2S, was studied. Complete H2S removal from synthetic biogas was found in pure and mixed culture reactors at 200 ppm initial H2S concentration, 35 l h−1 of biogas flow rate and 5.83 l h−1 of airflow rate. H2S removal using biofiltration had little effect on the methane content of synthetic biogas. Pure and mixed culture reactors at airflow rates of 0.75–5.83 l h−1 also showed complete H2S removal from real biogas. However, H2S removal efficiency at the airflow rate of 0.53 l h−1 decreased to 99%. Thus, H2S removal using biofiltration had little effect on the methane content of real biogas. The results of this study suggest the feasibility of developing using mixed culture of sulfide-oxidizing bacteria from concentrated latex wastewater for industrial application to remove H2S from biogas through biofiltration.

Hydrogen-based, hollow-fiber membrane biofilm reactor for reduction of perchlorate and other oxidized contaminants

Many oxidized pollutants, such as nitrate, perchlorate, bromate, and chlorinated solvents, can be microbially reduced to less toxic or less soluble forms. For drinking water treatment, an electron donor must be added. Hydrogen is an ideal electron donor, as it is non-toxic, inexpensive, and sparsely soluble. We tested a hydrogen-based, hollow-fiber membrane biofilm reactor (MBfR) for reduction of perchlorate, bromate, chlorate, chlorite, chromate, selenate, selenite, and dichloromethane. The influent included 5 mg/L nitrate or 8 mg/L oxygen as a primary electron accepting substrate, plus 1 mg/L of the contaminant. The mixed-culture reactor was operated at a pH of 7 and with a 25 minute hydraulic detention time. High recirculation rates provided completely mixed conditions. The objective was to screen for the reduction of each contaminant. The tests were short-term, without allowing time for the reactor to adapt to the contaminants. Nitrate and oxygen were reduced by over 99 percent for all tests. Removals for the contaminants ranged from a minimum of 29% for chlorate to over 95% for bromate. Results show that the tested contaminants can be removed as secondary substrates in an MBfR, and that the MBfR may be suitable for treating these and other oxidized contaminants in drinking water.

Effect of carbon to nitrogen ratio on the physical and chemical properties of activated sludge

The effect of carbon to nitrogen ratio (C/N) of the feed on the physical and surface chemical properties of activated sludge is investigated. Semi‐continuous reactors with 2 liters volume were operated at a mean cell residence time of 8 days. These mixed culture reactors were operated at 3 different carbon to nitrogen ratio. The operation of conventional activated sludge plants treating municipal wastewater is represented with reactors having a C/N ratio of 21 (in terms of the ratio of COD to TKN). Carbon and nitrogen limited activated sludge systems are represented by reactors with a C/N ratio of 9 and 43, respectively. The results show that C/N ratio has a profound effect on the ultimate physical and surface chemical properties of activated sludge. Both the steady state microorganism concentration and the amount of microbial extracellular polymers produced increase with increasing C/N ratio. The sludge becomes much harder to dewater and settle and it becomes more viscous as the C/N ratio increases. A decrease in surface hydrophobicity and an increase in surface charge of the sludge accompany these physical changes.

Impact of the addition of pure cultures on the performance of mixed culture reactors

Exploitation of the metabolic capabilities of specialized microorganisms requires availability of specific process control methods. The specialists must be enabled to compete favorably in multi-species microbial systems. Bench scale sequencing batch reactor (SBR) experiments were conducted to study the effectiveness of selected process strategies with respect to maintenance of the degradative capabilities of added microorganisms. 3-Chlorobenzoate (3-CB) was applied as a model substance for chloroorganic pollutants, mixed with sodium acetate and peptone as model substances for readily biodegradable pollutants. Pseudomonas putida PRS 2015 pAC 27 served as a model specialist. The bacteria were added to activated sludge from a municipal wastewater treatment plant. The SBRs were operated at a fast fill strategy (0.5 h), frequent cycle; repetition (8 h cycle time) and a volumetric exchange ratio of 20%. On the long run, no significant differences between the supplemented and non-supplemented system was observed. In comparison with the reactor which was not bio-supplemented the start-up period was significantly reduced, however, and the special metabolic capabilities remained resident, even when 3-CB was not fed over several days (up to 7 days).