Anaerobic Bioprocesses Research Articles

Assessing the potential of olive mill solid waste as feedstock for methane and volatile fatty acids production via anaerobic bioprocesses

The extensive production of olive mill solid waste (OMSW) from olive oil industry in the Mediterranean basin claims effective treatments and valorization strategies. This study aims to elucidate the potential of anaerobic digestion (AD) and anaerobic fermentation (AF) to convert pre-treated OMSW into biogas (CH4) and volatile fatty acids (VFA), respectively. The two thermal treatment conditions (65 °C and 180 °C) that are being implemented in the industry that manages the OMSW were tested. Comparing the two treatments aims to demonstrate the influence on the AD process of the degree of solubilization and degradation of the metabolites produced from the same substrate. AD of OMSW treated at low-temperature (65 °C) exhibited similar methane yields (195 ± 8 mL CH4/g volatile solid (VS)) to raw OMSW. AD of the solid phase (SP) after high-temperature treatment with acid addition at 180 °C resulted in methane yields comparable to raw OMSW while the liquid phase (LP) exhibited low methane yields (85 ± 10 mL CH4/g VS). Nevertheless, LP/180 °C exhibited the highest VFA bioconversion at 27.6 %, compared to less than 10 % for SP/180 ºC, SP/65 °C, and raw OMSW. The VFA profile showed notable variations with thermal treatment temperatures. Propionic acid dominated at SP/65 °C, while acetic acid became the primary VFA at 180 °C. Furthermore, significant degradation rates of phenolic compounds and furans were observed during the final day of both anaerobic processes. Overall, these findings suggest that AD is more suitable for raw OMSW, treated at low temperature and SP at high temperature, while AF offers a promising alternative for high-temperature-treated LP.

Advancement in constructed wetland microbial fuel cell process for wastewater treatment and electricity generation: a review.

The constructed wetland coupled with a microbial fuel cell (CW-MFC) is a wastewater treatment process that combines contaminant removal with electricity production, making it an environmentally friendly option. This hybrid system primarily relies on anaerobic bioprocesses for wastewater treatment, although other processes such as aerobic bioprocesses, plant uptake, and chemical oxidation also contribute to the removal of organic matter and nutrients. CW-MFCs have been successfully used to treat various types of wastewater, including urban, pharmaceutical, paper and pulp industry, metal-contaminated, and swine wastewater. In CW-MFC, macrophytes such as rice plants, Spartina angalica, Canna indica, and Phragmites australis are used. The treatment process can achieve a chemical oxygen demand removal rate of between 80 and 100%. Initially, research focused on enhancing power generation from CW-MFC, but recent studies have shifted towards resource recovery from wastewater. This review paper provides an overview of the development of constructed wetland microbial fuel cell technology, from its early stages to its current applications. The paper also highlights research gaps and potential directions for future research.

Effect and mechanism of Mn2+ on urease activity during anaerobic biological treatment of landfill leachate.

As a crucial hydrolytic enzyme, urease plays a vital role in anaerobic biological treatment. It is well-known that manganese ions are abundant in landfill leachate, but their concentration fluctuates significantly. However, few studies have investigated the effect and mechanism of different concentrations of Mn2+ on urease activity during anaerobic biological treatment of landfill leachate. This paper aimed to investigate the effects and mechanisms of different concentrations of Mn2+ on urease activity. The results showed that an appropriate amount of Mn2+ could significantly enhance urease activity, while a high concentration of Mn2+ could inhibit it. Insight into the mechanisms behind this phenomenon, various methods such as Zeta potential, particle size, ultraviolet spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy, and statistical analysis were employed in our study. Research suggested that, on one hand, Mn2+ may form hydrogen bonds with the side chain amino or carboxyl groups of urease amino acid residues, affecting the structure of urease through hydrogen bonding. Additionally, Mn2+ also binds to urease through hydrophobic interactions. On the other hand, the C-OH and C-N functional groups in urease have a strong affinity for Mn2+, and changes in these functional groups can greatly enhance the activity of urease. Furthermore, under the action of high concentrations of Mn2+, while the structure of urease becomes more stable, there is also a steric hindrance phenomenon that affects the substrate from entering the catalytic center. Therefore, studying the mechanism of Mn2+ affecting urease activity has significant biological significance and provides a new perspective for exploring the impact of metals on anaerobic bioprocessing of landfill leachate.

Methylotrophic methanogenesis induced by ammonia nitrogen in an anaerobic digestion system

This lab-scale study aimed to investigate the effect of total ammonia nitrogen (TAN) stress on the methanogenic activity and the taxonomic and functional profiles of the microbial community of anaerobic sludge (AS) from a full-scale bioreactor. The AS was subjected to a stepwise increase in TAN every 14 days at concentrations of 1, 2, 2.5, 3, 3.5, and 4g TAN/L (Acclimated-AS or AAS). This acclimation stage was followed by an ammonia stress stage (4g/L). A blank-AS (BAS) was maintained without TAN during the acclimation stage. In the second stress stage (ST), the BAS was divided into two new treatments: a control (BAS') and one that received a shock load of TAN of 4g/L (SBAS'). Methane production was measured, and a metagenomic analysis was conducted to describe the microbial community. A decrease in the relative abundance of Methanothrix soehngenii of 16% was related to a decrease of 23% in the methanogenic capacity of AAS when comparing with the final stage of BAS. However, recovery was observed at 3.5g TAN/L, and a shift to methylotrophic metabolism occurred, indicated by a 4-fold increase in abundance of Methanosarcina mazei. The functional analysis of sludge metagenomes indicated that no statistical differences (p>0.05, RM ANOVA) were found in the relative abundance of methanogenic genes that initiate acetoclastic and hydrogenotrophic pathways (acetyl-CoA synthetase, ACSS; acetate kinase, ackA; phosphate acetyltransferase, pta; and formylmethanofuran dehydrogenase subunit A, fwdA) into the BAS and AAS during the acclimation phase. The same was observed between groups of genes associated with methanogenesis from methylated compounds. In contrast, statistical differences (p<0.05, one-way ANOVA) in the relative abundance of these genes were recorded during ST. The functional profiles of the genes involved in acetoclastic, hydrogenotrophic, and methylotrophic methanogenic pathways were brought to light for acclimatation and stress experimental stages. TAN inhibited methanogenic activity and acetoclastic metabolism. The gradual acclimatization to TAN leads to metabolic and taxonomic changes that allow for the subsequent recovery of methanogenic functionality. The study highlights the importance of adequate management of anaerobic bioprocesses with high nitrogen loads to maintain the methanogenic functionality of the microbial community.

Unveiling the Mechanism Underlying the Effects of Ammonia on n-Caproate Production: Influenced Pathways, Key Enzymes, and Microbiota Functions

n-Caproate, which is produced via chain elongation (CE) using waste biomass, can supply various fossil-derived products, thus advancing the realization of carbon neutrality. Ammonia released from the degradation of nitrogen-rich waste biomass can act as a nutrient or an inhibitor in anaerobic bioprocesses, including CE, with the distinction being primarily dependent on its concentration. Currently, the optimal concentration of ammonia and the threshold of toxicity for open-culture n-caproate production using ethanol as an electron donor, along with the underlying mechanisms, remain unclear. This study revealed that the optimal concentration of ammonia for n-caproate production was 2.0 g∙L−1, whereas concentrations exceeding this threshold markedly suppressed the CE performance. Exploration of the mechanism revealed the involvement of two forms of ammonia (i.e., ammonium ions and free ammonia) in this inhibitory behavior. High ammonia levels (5.0 g∙L−1) induced excessive ethanol oxidation and suppressed the reverse β-oxidation (RBO) process, directly leading to the enhanced activities of enzymes (phosphotransacetylase and acetate kinase) responsible for acetate formation and diminished activities of butyryl-coenzyme A (CoA):acetyl-CoA transferase, caproyl-CoA:butyryl-CoA transferase, and caproyl-CoA:acetyl-CoA transferase that are involved in the syntheses of n-butyrate and n-caproate. Furthermore, the composition of the microbial community shifted from Paraclostridium dominance (at 0.1 g∙L−1 ammonia) to a co-dominance of Fermentimonas, Clostridium sensu stricto 12, and Clostridium sensu stricto 15 at 2.0 g∙L−1 ammonia. However, these CE-functional bacteria were mostly absent in the presence of excessive ammonia (5.0 g∙L−1 ammonia). Metagenomic analysis revealed the upregulation of functions such as RBO, fatty acid synthesis, K+ efflux, adenosine triphosphatase (ATPase) metabolism, and metal cation export in the presence of 2.0 g∙L−1 ammonia, collectively contributing to enhanced n-caproate production. Conversely, the aforementioned functions (excluding metal cation export) and K+ influx were suppressed by excessive ammonia, undermining both ammonia detoxification and n-caproate biosynthesis. The comprehensive elucidation of ammonia-driven mechanisms influencing n-caproate production, as provided in this study, is expected to inspire researchers to devise effective strategies to alleviate ammonia-induced inhibition.

Development of a Kinetics Based Onsite Treatment System for Faecal Sludge Treatment and Resource Recovery

The management of faecal sludge in cities without a sewerage system is a challenging problem that requires immediate attention. The high organic and nutrient content, pathogen load, and pharmaceutical residues in faecal sludge necessitate special attention for its treatment to ensure environmental and public health safety. Onsite treatment of faecal sludge with a focus on resource recovery is a sustainable approach to its management. However, studies in these areas are limited, particularly in the Indian context. In this study, the aim is to generate primary data on the current status of faecal sludge management in a model region without a sewerage network. Additionally, kinetic parameters were examined to design an anaerobic bioprocess and an engineered system for resource-oriented faecal sludge treatment. One of the wards in Thiruvananthapuram Corporation, Kerala was selected for the study. Fresh faecal sludge collected from an apartment complex was used for the experimental studies. Anaerobic treatment of fresh faecal sludge was validated in bench-scale experimental bioreactors using different combinations: with mixing without inoculum, with mixing with inoculum, without mixing without inoculum, and without mixing with inoculum. The primary focus was on monitoring the organic removal efficiency as Chemical Oxygen Demand (COD) of these combinations. Among the four combinations tested the units "without mixing with inoculum" and "with mixing with inoculum" demonstrated COD removal efficiencies of 20.89% and 18.91%, respectively, with a first-order rate constant of 0.0008 hr-1 for both combinations. The other combinations exhibited comparatively lower COD removal efficiency and rate constants. Since the combinations with mixing and inoculum showed higher COD removal efficiency, either of them can be adopted for design purposes. Based on these results, a combined anaerobic-aerobic treatment unit is proposed for the treatment of faecal sludge.

New insights into enhanced biodegradation of 4-bromphenol in a nitrate-reducing system: Process performance and mechanism

Due to the recalcitrant nature of halogenated phenol, conventional anaerobic bioprocess is often limited by low removal efficiency and poor process stability. At the presence of electron acceptors such as nitrate, 4-bromophenol (4-BP) removal efficiency is significantly higher than that in the anaerobic control system, but the mechanism involved is still unclear. Therefore, an up-flow nitrate-reducing bioreactor (NRBR) was designed and consecutively performed for 215 days to explore the synergistic mechanism for BPs biodegradation and nitrate reduction. Complete 4-BP biodegradation could be obtained in NRBR at HRT and 4-BP loading rate of 24 h and 0.29 mol m − 3d − 1, while the TOC removal and nitrate reduction efficiencies were as high as 91.33±2.11% and 98.31±1.33%, respectively. Population evolution analyses revealed that the microorganisms involved in 4-BP debromination and biodegradation (Candidatus Peregrinibacteria, Denitratisoma, Anaerolineaceae and Ignavibacterium) as well as nitrate reduction (Denitratisoma, Anaerolineaceae, Limnobacter and Ignavibacterium) were significantly enriched in NRBR. Major intermediates during 4-BP biodegradation, including 4-bromocatechol, 4‑bromo-6-oxo-hexanoic acid and succinic acid were identified, while a distinct 4-BP biodegradation pathway via hydration, aromatic-ring cleavage, hydrolysis debromination and oxidation was expounded. Metagenomic analysis indicated that oxidation (had, pht4, boh, butA), hydrolysis debromination ((S)-2-haloacid dehalogenase) and bio-mineralization (gabD, sdhA) of 4-BP were largely enhanced in NRBR. Moreover, carbon, nitrogen, energy and amino acid metabolisms were significantly facilitated with the injection of nitrate in order to provide energy and electron, thus enhanced microbial activities and enzymatic reactions in NRBR. The proposed mechanism provides new insights into our mechanistic understanding of halogenated phenol biodegradation and the development of sustainable bioremediation strategies.

Boosting butyrate and hydrogen production in acidogenic fermentation of food waste and sewage sludge mixture: a pilot scale demonstration

Within the urban scenario, the application of a biorefinery technology value chain can foster the conversion of different organic substrates into marketable and added-value products. In this work, a pilot-scale dark fermentation (DF) process has been carried out as a key step for volatile fatty acids (VFA) and hydrogen production from the liquid fraction of sewage sludge and food waste mixture. Six operating conditions have been monitored in terms of yield and process stability, by changing the hydraulic retention time (HRT) from 4 to 6 days and applying a short-term hyper-thermophilic hydrolysis (70 °C, 8 h) on the same feedstock mixture. A tubular centrifugation was utilized to remove part of the biosolids (driven to biogas production) before the DF step, which was applied on the soluble and/or colloidal organic matter only. The hydrolysis step favored the following acidification process, in which a fermentation yield up to 0.42 g CODVFA/g VS0 was achieved at 5 days as HRT. Hydrogen production (up to 34.4% v/v and 0.046 m3 H2/kg VS0) was positively affected by the hydrolysis application and by the decrease of the HRT, highlighting the possibility to produce biohythane in the modeled two-phases anaerobic bioprocess. On the other hand, without the application of the hydrolysis, a selective production of butyric acid (up to 75% COD basin) was achieved, furnishing a different valorization route for the chosen urban organic feedstock. Changes in operating conditions and performances were also reflected by the adaptation of the microbial community, whose characterization highlighted the occurrence of several fermentative microorganisms (e.g., Clostridiaceae, Ruminococcaceae).

Metabolic engineering of Mannheimia succiniciproducens for malic acid production using dimethylsulfoxide as an electron acceptor.

Microbial production of various TCA intermediates and related chemicals through the reductive TCA cycle has been of great interest. However, rumen bacteria that naturally possess strong reductive TCA cycle have been rarely studied to produce these chemicals, except for succinic acid, due to their dependence on fumarate reduction to transport electrons for ATP synthesis. In this study, malic acid (MA), a dicarboxylic acid of industrial importance, was selected as a target chemical for mass production using Mannheimia succiniciproducens, a rumen bacterium possessing a strong reductive branch of the TCA cycle. The metabolic pathway was reconstructed by eliminating fumarase to prevent MA conversion to fumarate. The respiration system of M. succiniciproducens was reconstructed by introducing the Actinobacillus succinogenes dimethylsulfoxide (DMSO) reductase to improve cell growth using DMSO as an electron acceptor. Also, the cell membrane was engineered by employing Pseudomonas aeruginosa cis-trans isomerase to enhance MA tolerance. High inoculum fed-batch fermentation of the final engineered strain produced 61 g/L of MA with an overall productivity of 2.27 g/L/h, which is the highest MA productivity reported to date. The systems metabolic engineering strategies reported in this study will be useful for developing anaerobic bioprocesses for the production of various industrially important chemicals.

Managing the Effluents of Anaerobic Fermentations by Bioprocess Schemes Involving Membrane Bioreactors and Bio-Electrochemical Systems: A Mini-Review

Anaerobic bioprocesses, such as anaerobic digestion and dark fermentation, provide energy carriers in the form of methane and hydrogen gases, respectively. However, their wastewater-type residues, that is, the fermentation effluents, must be treated carefully due to the incomplete and non-selective conversion of organic matter fed to the actual system. For these reasons, the effluents contain various secondary metabolites and unutilized substrate, in most cases. Only a fraction of anaerobic effluents can be directly applied for fertilization under a moderate climate. Conventional wastewater treatment technologies may be used to clean the remainder, but that approach leads to a net loss of energy and of potentially useful agricultural input materials (organic carbon and NPK fertilizer substitutes). The rationale of this paper is to provide an overview of promising new research results in anaerobic effluent management strategies as a part of technological downstream that could fit the concept of new-generation biorefinery schemes aiming towards zero-waste discharge, while keeping in mind environmental protection, as well as economical perspectives. According to the literature, the effluents of the two above processes can be treated and valorized relying either on membrane bioreactors (in case of anaerobic digestion) or bio-electrochemical apparatus (for dark fermentation). In this work, relevant findings in the literature will be reviewed and analyzed to demonstrate the possibilities, challenges, and useful technical suggestions for realizing enhanced anaerobic effluent management. Both membrane technology and bio-electrochemical systems have the potential to improve the quality of anaerobic effluents, either separately or in combination as an integrated system.

Role of microparticles in membrane fouling from acidogenesis to methanogenesis phases in an anaerobic baffled reactor

Microparticles (0.45–10 μm) have been recognized as key foulants in anaerobic membrane bioreactors (AnMBRs). However, their characteristics and fouling behaviors are often understood in single-stage and completely mixed reactors, failing to elucidate the occurrence of microparticles in the multi-stage anaerobic bioprocess. Here, a lab-scale anaerobic baffled reactor with four compartments (C1-C4) was employed to explore the composition and fouling potential of microparticles in different compartments. Photometric analysis showed that the microparticles had an increasing percentage in the total organics of the top supernatant but a decreasing concentration from C1 to C4. Long-term filtration and dead-end filtration tests revealed that the top supernatant in C1 had much higher fouling potential than those in C2-C4. The supernatant microparticles significantly accumulated in the cake layers for each compartment (68–95% of the total organics), particularly the fraction of 1–5 μm, and the fouling rate was positively correlated with the biomass accumulation rate. Based on reactor performance and 16S rRNA gene sequences, a significant bio-phase separation occurred between C1 (acidogenesis) and C2-C4 (methanogenesis). And hydrolytic and fermentative bacteria in the family Veillonellaceae, Streptococcaceae, and Enterobacteriaceae were dominant in the supernatant microparticles, particularly in C1, which had a positive correlation with the fouling rate and biomass accumulation rate. These above results all revealed that the microparticles in the acidogenesis phase had higher fouling potential. In summary, our results suggest that the tactic of pre-hydrolysis and acidification with feedstocks and constructing AnMBRs by coupling with multi-phase anaerobic bioprocesses and membrane units could be beneficial to fouling control.

Progress and prospects of applying carbon-based materials (and nanomaterials) to accelerate anaerobic bioprocesses for the removal of micropollutants.

SummaryCarbon‐based materials (CBM), including activated carbon (AC), activated fibres (ACF), biochar (BC), nanotubes (CNT), carbon xenogels (CX) and graphene nanosheets (GNS), possess unique properties such as high surface area, sorption and catalytic characteristics, making them very versatile for many applications in environmental remediation. They are powerful redox mediators (RM) in anaerobic processes, accelerating the rates and extending the level of the reduction of pollutants and, consequently, affecting positively the global efficiency of their partial or total removal. The extraordinary conductive properties of CBM, and the possibility of tailoring their surface to address specific pollutants, make them promising as catalysts in the treatment of effluents containing diverse pollutants. CBM can be combined with magnetic nanoparticles (MNM) assembling catalytic and magnetic properties in a single composite (C@MNM), allowing their recovery and reuse after the treatment process. Furthermore, these composites have demonstrated extraordinary catalytic properties. Evaluation of the toxicological and environmental impact of direct and indirect exposure to nanomaterials is an important issue that must be considered when nanomaterials are applied. Though the chemical composition, size and physical characteristics may contribute to toxicological effects, the potential toxic impact of using CBM is not completely clear and is not always assessed. This review gives an overview of the current research on the application of CBM and C@MNM in bioremediation and on the possible environmental impact and toxicity.

Rationale for the Combined Use of Biological Processes and AOPs in Wastewater Treatment Tasks

This paper aims to form a unified concept of the integrated use of different wastewater treatment methods to form a resistant biological treatment stage of technological systems under the influence of such toxic factors as antibiotics and surfactants. The processes of mechanical treatment, ozonation, UV irradiation, and electrolytic anodic oxidation were implemented in an electrotechnological wastewater treatment facility. Wastewater treatment quality was determined by the concentration of nitrogen compounds in aqueous solutions according to the method of Lurie. Biodiagnostics of the investigated activated sludge via surfactant action was carried out at polyethylene oxide concentrations of 10, 30, and 50 mg/dm3. As a result of experiments on wastewater treatment after aquaculture, an improvement in the reduction of pollutants only by the indicator “nitrate concentration” was determined: by 20% after anodic oxidation, and by 15% after photolysis. At almost all surfactant concentrations studied, the activated sludge was not completely recovered, which was expressed in a decrease in its quantity and in the inability to aggregate flakes of activated sludge. The diameter of the growth retardation of the standard disk with antibiotic (amoxiclav) by the accumulative culture of activated sludge was 17.3 ± 2 mm at a concentration of 4 mg/dm3 and 31.3 ± 3 mm at a concentration of 6 mg/dm3. In the process of studying the state of the activated sludge’s biocenosis under the influence of such toxicants, several regularities were revealed. The directions of using combined approaches of water treatment and wastewater treatment were defined. The structural model of treatment facilities using aerobic and anaerobic bioprocesses together with advanced oxidative technologies was substantiated.

Physiological characteristics of Corynebacterium glutamicum as a cell factory under anaerobic conditions.

Corynebacterium glutamicum, a gram-positive and facultative anaerobic bacterium, is widely used for the industrial production of amino acids, such as L-glutamate and L-lysine. C. glutamicum grows and produces amino acids under aerobic conditions. When restricted under anaerobic conditions, it produces organic acids, such as L-lactate and succinate, through metabolic shift. With the increasing threat of global warming, these organic acids have drawn considerable attention as bio-based plastic monomers. In addition to the organic acids, the anaerobic bioprocess is also used to produce other value-added compounds, including isobutanol, ethanol, 3-methyl-1-butanol, 2,3-butanediol, L-alanine, and L-valine. Therefore, C. glutamicum is now a versatile cell factory for producing a wide variety of useful chemicals under both aerobic and anaerobic conditions. The growth and metabolism of the bacterium depend on the oxygen levels, which modulate the rearrangement of the carbon flux by reprogramming gene expression patterns and intracellular redox states. Anaerobic cell growth and L-lysine production as well as aerobic succinate production have been demonstrated by engineering the metabolic pathways or supplying a terminal electron acceptor instead of oxygen. In this review, we discuss the physiological and metabolic changes in C. glutamicum associated with its application as a cell factory under different oxygen states. Physiological switching in bacteria is initiated with the sensing of oxygen availability. While such a sensor has not been identified in C. glutamicum yet, the molecular mechanism for oxygen sensing in related bacteria is also discussed. KEY POINTS: • C. glutamicum produces a wide variety of useful compounds under anaerobic conditions. • C. glutamicum is a versatile cell factory under both aerobic and anaerobic conditions. • Metabolic fate can be overcome by engineering metabolic pathways.

Tuning microbial community in non-conventional two-stage anaerobic bioprocess for microalgae biomass valorization into targeted bioproducts

Unspecific microorganisms consortia are normally used in anaerobic biodegradation of solid wastes. However, these consortia can be tuned to optimally obtain determined bioproducts. In this study, high value-added products and biogas were obtained via an innovative two-stage anaerobic bioprocess from microalgae biomass. The anaerobic fermentation (AF) entailed the production of short-chain fatty acids (SCFAs) and subsequently, only the solid spent of AF effluent was valorized for methane production via conventional anaerobic digestion (AD). Applied conditions in AF (25 °C, HRT 8 days) favored Firmicutes predominance (64%) enabling a conversion efficiency of 32.1% g SCFAs-COD/g CODin. Opposite, a wider microbial biodiversity was determined in the AD reactor (35 °C, HRT 20 days), being mainly composed by Firmicutes (28.6%), Euryarchaeota (17.7%) and Proteobacteria (15.3%). AD of the AF-solid spent reached 168.9 mL CH4 /g CODin. Strikingly, operational conditions imposed mediated a microbial specialization that maximized product output.

Detoxification of Ciprofloxacin in an Anaerobic Bioprocess Supplemented with Magnetic Carbon Nanotubes: Contribution of Adsorption and Biodegradation Mechanisms.

In anaerobic bioreactors, the electrons produced during the oxidation of organic matter can potentially be used for the biological reduction of pharmaceuticals in wastewaters. Common electron transfer limitations benefit from the acceleration of reactions through utilization of redox mediators (RM). This work explores the potential of carbon nanomaterials (CNM) as RM on the anaerobic removal of ciprofloxacin (CIP). Pristine and tailored carbon nanotubes (CNT) were first tested for chemical reduction of CIP, and pristine CNT was found as the best material, so it was further utilized in biological anaerobic assays with anaerobic granular sludge (GS). In addition, magnetic CNT were prepared and also tested in biological assays, as they are easier to be recovered and reused. In biological tests with CNM, approximately 99% CIP removal was achieved, and the reaction rates increased ≈1.5-fold relatively to the control without CNM. In these experiments, CIP adsorption onto GS and CNM was above 90%. Despite, after applying three successive cycles of CIP addition, the catalytic properties of magnetic CNT were maintained while adsorption decreased to 29 ± 3.2%, as the result of CNM overload by CIP. The results suggest the combined occurrence of different mechanisms for CIP removal: adsorption on GS and/or CNM, and biological reduction or oxidation, which can be accelerated by the presence of CNM. After biological treatment with CNM, toxicity towards Vibrio fischeri was evaluated, resulting in ≈ 46% detoxification of CIP solution, showing the advantages of combining biological treatment with CNM for CIP removal.

Direct Medium-Chain Carboxylic Acid Oil Separation from a Bioreactor by an Electrodialysis/Phase Separation Cell.

Medium-chain carboxylic acids (MCCAs) are valuable platform chemicals and can be produced from waste biomass sources or syngas fermentation effluent through microbial chain elongation. We have previously demonstrated successful approaches to separate >90% purity oil with different MCCAs (MCCA oil) by integrating the anaerobic bioprocess with membrane-based liquid-liquid extraction (pertraction) and membrane electrolysis. However, two-compartment membrane electrolysis unit without pertraction was not able to separate MCCA oil. Therefore, we developed a five-compartment electrodialysis/phase separation cell (ED/PS). First, we tested an ED/PS cell in series with pertraction and achieved a maximum MCCA-oil flux of 1.7 × 103 g d-1 per projected area (m2) (19 mL oil d-1) and MCCA-oil transfer efficiency [100% × moles MCCA-oil moles electrons-1] of 74% at 15 A m-2. This extraction system at 15 A m-2 demonstrated a ∼10 times lower electric-power consumption (1.1 kWh kg-1 MCCA oil) than membrane electrolysis in series with pertraction (9.9 kWh kg-1 MCCA oil). Second, we evaluated our ED/PS as a stand-alone unit when integrated with the anaerobic bioprocess and demonstrated that we can selectively extract and separate MCCA oil directly from chain-elongating bioreactor broth with just an abiotic electrochemical cell. However, the electric-power consumption increased considerably due to the lower MCCA concentrations in the bioreactor broth compared to the pertraction broth.

Effects of cellulolytic lactic acid bacteria on the lignocellulose degradation, sugar profile and lactic acid fermentation of high-moisture alfalfa ensiled in low-temperature seasons

The objective of this study was to investigate the effects of cellulolytic lactic acid bacteria on the lignocellulose degradation, sugar profile and lactic acid fermentation of high-moisture alfalfa during anaerobic bioprocessing in low-temperature seasons. Last cut and low dry matter (DM) alfalfa was ensiled with five additives, commercial cellulase (EN), a combination of Lactobacillus plantarum and commercial cellulase (LPEN), a combination of engineered Lactococcus lactis strains (HT2) containing cellulase gene, Enterococcus faecium Y83 (EF83), or Enterococcus faecalis JF85 (EF85), and without additives, as the control, with four replicates each. After anaerobic preservation in a silo from late fall through winter (3 to 20 °C) for 140 days, the ensiled alfalfa was sampled and analysed. Compared with the control, EF85 increased the relative contents of neutral detergent fibre (NDF), acid detergent fibre (ADF), and cellulose and decreased contents of water-soluble carbohydrates (WSCs) and lactic acid, and the conversion of WSCs into lactic acid. Consequently, lactic acid fermentation was decreased in EF85. Compared with the control, supplementation with EN, EF83, HT2, and LPEN decreased contents of NDF, ADF, cellulose, and hemicellulose and increased the contents of WSCs, disaccharide, xylose, and arabinose; interestingly, compared with EN, supplementation with EF83, HT2 and LPEN increased the lactic acid content and the ratio of lactic acid to acetic acid as indicated by strong homo-lactic acid fermentation. Furthermore, HT2 increased the lactic acid concentration to a maximum of 72.2 g/kg DM and had a higher conversion of WSCs into lactic acid than EF83 (36.3%) and LPEN (17.9%), indicating that HT2 had a greater ability to ferment lignocellulose of alfalfa into lactic acid. Therefore, HT2 could enhance the lactic acid production of alfalfa during ensiling in low-temperature seasons.

Enhancement of tribromophenol removal in a sequencing batch reactor via submicron magnetite.

Conductive magnetite (Fe3O4) has been applied into some anaerobic bioprocesses to accelerate direct interspecies electron transfer (DIET), however, Fe3O4 is usually dissolved by iron-reducing bacteria under anaerobic conditions, resulting in the loss of magnetite. Therefore, submicron magnetite particles were added to the sequencing batch reactor (SBR) to build a Fe3O4/SBR system, which could alleviate magnetite dissolution and simultaneously remove tribromophenol (TBP) effectively. The average removal efficiencies of chemical oxygen demand (COD) and TBP in Fe3O4/SBR system were 81% and 91%, respectively, which were 51% and 18% higher than those of the control group without Fe3O4 (SBR system). The enhanced TBP biodegradation was likely related to potential DIET, which was supported by the scanning electron microscopy (SEM) analysis, the increase of dehydrogenase and heme c (fivefold and 1.7-fold), and the enrichment of iron-redoxing bacteria (Geobacter and Thiobacillus). Furthermore, magnetite mainly remained intact in structure as indicated by X-ray diffraction (XRD), which might be ascribed to in situ iron redox cycle and magnetite biosynthesis via Magnetospirillum. Notably, the content of hydrogen peroxide (H2O2) and hydroxyl radical (⋅OH) in Fe3O4/SBR system was 4-5 times higher than that of SBR system. These findings could provide insights into the development of cost-effective strategy for the removal of refractory organic pollutants.

Online measurement of CO2 and total gas production in parallel anaerobic shake flask cultivations

Online measurements of off-gas streams are often crucial for studying bioconversion processes. However, for anaerobic processes, options for online off-gas analysis are typically restricted to lab-scale bioreactors or larger systems, while gas measurements at smaller scales typically do not discriminate between different gases. In this work, a method for online measurement of CO2 and total gas production in anaerobic fermentations at the shake flask scale is described, extending capabilities of a previously reported device developed for aerobic processes to anaerobic bioprocesses. The novel design allows anaerobic fermentations to be performed in multiple parallel vessels, all of which collect online gas signals. The online gas signals are used to calculate the transfer rates, allowing near real-time visualization of the progress of eight fermentations operating in parallel. Conditions such as carbon source depletion, inhibition of growth, and exhaustion of a single carbon source in a dual-substrate fermentation can all be clearly distinguished. The combination of online signals and offline analysis allowed for carbon balances to be performed with high degrees of closure. The new design allows for higher throughput screening of anaerobic bioprocesses, an area lacking in small-scale options with off-gas analysis capabilities.