Enhancing Electricity Generation Using a Laccase-Based Microbial Fuel Cell with Yeast Galactomyces reessii on the Cathode.

The fungi associated with termites secrete enzymes such as laccase (multi-copper oxidase) that can degrade extracellular wood matrix. Laccase uses molecular oxygen as an electron acceptor to catalyze the degradation of organic compounds. Owing to its ability to transfer electrons from the cathodic electrode to molecular oxygen, laccase has the potential to be a biocatalyst on the surface of the cathodic electrode of a microbial fuel cell (MFC). In this study, a two-chamber MFC using the laccase-producing fungus Galactomyces reessii was investigated. The fungus cultured on coconut coir was placed in the cathode chamber, while an anaerobic microbial community was maintained in the anode chamber fed by industrial rubber wastewater and supplemented by sulfate and a pH buffer. The laccase-based biocathode MFC (lbMFC) produced the maximum open circuit voltage of 250 mV, output voltage of 145 mV (with a 1,000 Ω resistor), power density of 59 mW/m2, and current density of 278 mA/m2, and a 70% increase in half-cell potential. This study demonstrated the capability of laccase-producing yeast Galactomyces reessii as a biocatalyst on the cathode of the two-chamber lbMFC.

Assessment of molecular detection of anaerobic ammonium-oxidizing (anammox) bacteria in different environmental samples using PCR primers based on 16S rRNA and functional genes.

Eleven published PCR primer sets for detecting genes encoding 16S ribosomal RNA (rRNA), hydrazine oxidoreductase (HZO), cytochrome cd 1-containing nitrite reductase (NirS), and hydrazine synthase subunit A (HzsA) of anaerobic ammonium-oxidizing (anammox) bacteria were assessed for the diversity and abundance of anammox bacteria in samples of three environments: wastewater treatment plant (WWTP), wetland of Mai Po Nature Reserve (MP), and the South China Sea (SCS). Consistent phylogenetic results of three biomarkers (16S rRNA, hzo, and hzsA) of anammox bacteria were obtained from all samples. WWTP had the lowest diversity with Candidatus Kuenenia dominating while the SCS was dominated by Candidatus Scalindua. MP showed the highest diversity of anammox bacteria including C. Scalindua, C. Kuenenia, and Candidatus Brocadia. Comparing different primer sets, no significant differences in specificity for 16S rRNA gene could be distinguished. Primer set CL1 showed relatively high efficiency in detecting the anammox bacterium hzo gene from all samples, while CL2 showed greater selectivity for WWTP samples. The recently reported primer sets of the hzsA gene resulted in high efficiencies in detecting anammox bacteria while nirS primer sets were more selective for specific samples. Results collectively indicate that the distribution of anammox bacteria is niche-specific within different ecosystems and primer specificity may cause biases on the diversity detected.

Identifiability of large-scale non-linear dynamic network models applied to the ADM1-case study.

In this work, both the structural and practical identifiability of the Anaerobic Digestion Model no. 1 (ADM1) is investigated, which serves as a relevant case study of large non-linear dynamic network models. The structural identifiability is investigated using the probabilistic algorithm, adapted to deal with the specifics of the case study (i.e., a large-scale non-linear dynamic system of differential and algebraic equations). The practical identifiability is analyzed using a Monte Carlo parameter estimation procedure for a 'non-informative' and 'informative' experiment, which are heuristically designed. The model structure of ADM1 has been modified by replacing parameters by parameter combinations, to provide a generally locally structurally identifiable version of ADM1. This means that in an idealized theoretical situation, the parameters can be estimated accurately. Furthermore, the generally positive structural identifiability results can be explained from the large number of interconnections between the states in the network structure. This interconnectivity, however, is also observed in the parameter estimates, making uncorrelated parameter estimations in practice difficult.

Assessment of anaerobic bacterial diversity and its effects on anaerobic system stability and the occurrence of antibiotic resistance genes.

This study evaluated the link between anaerobic bacterial diversity and, the biodegradation of antibiotic combinations and assessed how amending antibiotic combination and increasing concentration of antibiotics in a stepwise fashion influences the development of resistance genes in anaerobic reactors. The biodegradation, sorption and occurrence of the known antibiotic resistance genes (ARGs) of erythromycin and tetracycline were investigated using the processes of UV-HPLC and qPCR analysis respectively. Ion Torrent sequencing was used to detect microbial community changes in response to the addition of antibiotics. The overall results indicated that changes in the structure of a microbial community lead to changes in biodegradation capacity, sorption of antibiotics combinations and occurrence of ARGs. The enhanced biodegradation efficiency appeared to generate variations in the structure of the bacterial community. The results suggested that controlling the ultimate Gram-negative bacterial community, especially Acinetobacter-related populations, may promote the successful biodegradation of antibiotic combinations and reduce the occurrence of ARGs.

Simultaneous Removal of Phenol and Dissolved Solids from Wastewater Using Multichambered Microbial Desalination Cell.

Microbial desalination cell (MDC) has great potential toward direct electricity generation from wastewater and concurrent desalination through potential difference developed due to microbial activity. Degradation of phenol by isolate Pseudomonas aeruginosa in anodic chamber and simultaneous desalination of water in middle desalination chamber of multichamber MDC is demonstrated in this study. Performance of the MDCs with different anodic inoculum conditions, namely pure culture of P. aeruginosa (MDC-1), 50 % v/v mixture of P. aeruginosa and anaerobic mixed consortia (MDC-2) and anaerobic mixed consortia (MDC-3), was evaluated to compare the phenol degradation in anodic chamber, bioelectricity generation, and simultaneous total dissolved solids (TDS) removal from saline water in desalination chamber. Synergistic effect between P. aeruginosa and mixed anaerobic consortia as inoculum was evident in MDC-2 demonstrating phenol degradation of 90 %, TDS removal of 75 % in 72 h of reaction time along with higher power generation of 27.5 mW/m(2) as compared to MDC-1 (95 %, 64 %, 12.8 mW/m(2), respectively) and MDC-3 (58 %, 52 %, 4.8 mW/m(2), respectively). The results illustrate that the multichamber MDC-2 is effective for simultaneous removal of phenol and dissolved solids contained in industrial wastewaters.

Protein costs do not explain evolution of metabolic strategies and regulation of ribosomal content: does protein investment explain an anaerobic bacterial Crabtree effect?

Protein investment costs are considered a major driver for the choice of alternative metabolic strategies. We tested this premise in Lactococcus lactis, a bacterium that exhibits a distinct, anaerobic version of the bacterial Crabtree/Warburg effect; with increasing growth rates it shifts from a high yield metabolic mode [mixed-acid fermentation; 3 adenosine triphosphate (ATP) per glucose] to a low yield metabolic mode (homolactic fermentation; 2 ATP per glucose). We studied growth rate-dependent relative transcription and protein ratios, enzyme activities, and fluxes of L. lactis in glucose-limited chemostats, providing a high-quality and comprehensive data set. A three- to fourfold higher growth rate rerouted metabolism from acetate to lactate as the main fermentation product. However, we observed hardly any changes in transcription, protein levels and enzyme activities. Even levels of ribosomal proteins, constituting a major investment in cellular machinery, changed only slightly. Thus, contrary to the original hypothesis, central metabolism in this organism appears to be hardly regulated at the level of gene expression, but rather at the metabolic level. We conclude that L. lactis is either poorly adapted to growth at low and constant glucose concentrations, or that protein costs play a less important role in fitness than hitherto assumed.

Establishment and assessment of an integrated citric acid-methane production process.

To solve the problem of extraction wastewater in citric acid industrial production, an improved integrated citric acid-methane production process was established in this study. Extraction wastewater was treated by anaerobic digestion and then the anaerobic digestion effluent (ADE) was stripped by air to remove ammonia. Followed by solid-liquid separation to remove metal ion precipitation, the supernatant was recycled for the next batch of citric acid fermentation, thus eliminating wastewater discharge and reducing water consumption. 130U/g glucoamylase was added to medium after inoculation and the recycling process performed for 10 batches. Fermentation time decreased by 20% in recycling and the average citric acid production (2nd-10th) was 145.9±3.4g/L, only 2.5% lower than that with tap water (149.6g/L). The average methane production was 292.3±25.1mL/g CODremoved and stable in operation. Excessive Na(+) concentration in ADE was confirmed to be the major challenge for the proposed process.

Enhancing the functional and economical efficiency of a novel combined thermo chemical disperser disintegration of waste activated sludge for biogas production.

In this investigation, an effort was made to pretreat surplus waste activated sludge (WAS) inexpensively by a novel combined process involving thermo chemical disperser pretreatment. This pretreatment was found to be efficient at a specific energy (SE) consumption of 3360.94 kJ/kg TS, with the chemical oxygen demand (COD) solubilization of 20%. This was comparatively higher than thermo chemically treated sludge where the solubilization was found to be 15.5% at a specific energy consumption of 10,330 kJ/kg TS respectively. Higher production of volatile fatty acids (VFA) (675 mg/L) in anaerobic fermentation of pretreated WAS indicates better hydrolysis performance. The biogas production potential of sludge pretreated through this combined technique was found to be 0.455 (L/gVS) and comparatively higher than thermo chemically pretreated sludge. Economic investigation provides 90% net energy savings in this combined pretreatment. Therefore, this combined process was considered to be potentially effective and economical in sludge disintegration.

A bench-scale assessment of ozone pre-treatments for landfill leachates.

Leachate from stabilized landfill can pose unique challenges to conventional biological wastewater treatment. Ozone-based advanced oxidation processes have garnered recent consideration as an option to reduce the organic strength and recalcitrance of aged landfill leachate. With a bench-scale investigation, the reported work examines the potential for leachate conditioning for further biological treatment by treatment with low-mg/L doses of ozone (0-7.5 mg/L 03). While not sufficient for significant organics mineralization, the tested ozone doses could potentially produce both selective and non-selective oxidation of recalcitrant leachate organic compounds leaving bio-available products in the pre-treated leachate. Leachate conditioning by 03 or 03/H202 was assessed via monitoring of three anthropogenic organic leachate contaminants(tris-(2-chloroethyl) phosphate, tris-(butoxyethyl)-phosphate and 17beta-estradiol (E2)) with ozonation, and ozonation followed by anaerobic incubation. In addition, chemical oxygen demand (COD) and BOD5 analysis of the ozonated leachate, and methane and total gas formation during the anaerobic incubation were used to assess the degree of leachate conditioning. When treated with O3 alone, 58% removal of E2 was observed with an ozone dose of 4.5-5.4mg/L. Direct oxidation of the three leachate contaminants was limited with O3/H202 pre-treatment. However, this pre-treatment was observed to have significantly improved degradation of E2 during anaerobic incubation of ozonated leachates (removal rate of E2 was 53.7% with 15 days of incubation), indicating the potential for ozone synthesized co-metabolism. However, overall anaerobic microbial activity was not significantly impacted by the applied ozone pre-treatments, as measured by methane formation, total gas formation, and COD removal during incubation.

[Methods for the assessment of organic compounds biodegradability: A review].

There are plenty of methods used for the assessment of organic compounds biodegradability, but lack of comprehensive literatures in systematically introducing the assessment methods and guiding the choice of the methods. This paper briefly introduced the indicative parameters in assessing organic compounds biodegradability and the assessment methods of aerobic and anaerobic biodegradation, with the focus on the principles and criteria in the choice of the assessment methods, and analyzed in detail the reasons for the differences in the biodegradation rates of organic compounds when different test methods were adopted. Taking the standards established by the Organization for Economic Co-operation and Development (OECD) as an example, the process of choosing and applying the assessment methods was introduced, the systematic method-choosing route was integrated, and the advantages of the OECD standard methods were pointed out.

Life cycle assessment of biohydrogen and biomethane production and utilisation as a vehicle fuel.

Environmental burdens for the production and utilisation of biomethane vehicle fuel or a biohydrogen/biomethane blend produced from food waste or wheat feed, based on data from two different laboratory experiments, have been compared. For food waste treated by batch processes the two stage system gave high hydrogen yields (84.2l H2kg(-1) VS added) but a lower overall energy output than the single stage system. Reduction in environmental burdens compared with diesel was achieved, supported by the diversion of waste from landfill. For wheat feed, the semi continuously fed two stage process gave low hydrogen yields (7.5l H2kg(-1) VS added) but higher overall energy output. The process delivers reduction in fossil fuel burdens, and improvements in process efficiencies will lead to reduction in CO2 burdens compared with diesel. The study highlights the importance of understanding and optimising biofuel production parameters according to the feedstock utilised.

Performance and energy economics of mesophilic and thermophilic digestion in anaerobic hybrid reactor treating coal wastewater.

Two anaerobic hybrid AHRs (AHR), mesophilic (35 °C) and thermophilic (55 °C) were operated with coal wastewater at different hydraulic retention times (HRT) ranging from 3-0.5 to 3.12-0.6d with organic loading rates (OLR) of 1.12-6.72 g L(-1) d(-1). Synthetic coal wastewater with an average chemical oxygen demand (COD) of 2240 mg L(-1) and phenolics concentration of 752 mg L(-1) was used as substrate. At each HRT, the thermophilic AHR gave a better performance, measured in terms of phenolics/COD removal and gas production. The specific methane yield was also higher for thermophilic AHR at each HRT compared to mesophilic one. The volatile fatty acid concentration in the effluent increased with the lowering of HRT. The Stover-Kincannon model was applicable at both temperatures and showed higher substrate utilization in thermophilic AHR. Energy economic study of the AHRs revealed that 11,938 MJ d(-1) more energy can be generated using thermophilic AHR than mesophilic.

Life cycle assessment of two emerging sewage sludge-to-energy systems: evaluating energy and greenhouse gas emissions implications.

A "cradle-to-grave" life cycle assessment was conducted to examine the energy and greenhouse gas (GHG) emission footprints of two emerging sludge-to-energy systems. One system employs a combination of anaerobic digestion (AD) and fast pyrolysis for bioenergy conversion, while the other excludes AD. Each system was divided into five process phases: plant construction, sludge pretreatment, sludge-to-bioenergy conversion, bioenergy utilizations and biochar management. Both systems achieved energy and GHG emission benefits, and the AD-involving system performed better than the AD-excluding system (5.30 vs. 0.63 GJ/t sludge in net energy gain and 0.63 vs. 0.47 t CO(2)eq/t sludge in emission credit for base case). Detailed contribution and sensitivity analyses were conducted to identify how and to what degree the different life-cycle phases are responsible for the energy and emission impacts. The energy and emission performances were significantly affected by variations in bioenergy production, energy requirement for sludge drying and end use of bioenergy.

The environmental sustainability of anaerobic digestion as a biomass valorization technology.

This paper studies the environmental sustainability of anaerobic digestion from three perspectives. First, reference electricity is compared to electricity production from domestic organic waste and energy crop digestion. Second, different digester feed possibilities in an agricultural context are studied. Third, the influence of applying digestate as fertilizer is investigated. Results highlight that biomass is converted at a rational exergy (energy) efficiency ranging from 15.3% (22.6) to 33.3% (36.0). From a life cycle perspective, a saving of over 90% resources is achieved in most categories when comparing biobased electricity to conventional electricity. However, operation without heat valorization results in 32% loss of this performance while using organic waste (domestic and agricultural residues) as feedstock avoids land resources. The use of digestate as a fertilizer is beneficial from a resource perspective, but causes increased nitrogen and methane emissions, which can be reduced by 50%, making anaerobic digestion an environmentally competitive bioenergy technology.

Assessment of the anaerobic acidogenesis of wet olive cake from a two-phase olive oil mill.

A study of the anaerobic acidogenesis of wet olive cake or olive mill solid waste (OMSW) from the two-phase olive oil mill industry was carried out. Eight different hydraulic retention times (HRTs) ranging from 50.0-10.7 days were studied. An increase of 935.7 % in total volatile fatty acids (VFA) over the initial acidic concentration in the OMSW (1.4 g L(-1) expressed as acetic acid) was achieved. The results showed a maximum total VFA generation rate of 5.05 g COD L(-1)d(-1), this rate being achieved at the same hydraulic retention time as the maximum acetic acid production (8.2 g L(-1)) and as the maximum acidification degree (34.4 %).

Continuous thermal hydrolysis and anaerobic digestion of sludge. Energy integration study.

Experimental data obtained from the operation in a pilot plant are used to perform mass and energy balances to a global process combining units of thermal hydrolysis (TH) of secondary sludge, anaerobic digestion (AD) of hydrolysed secondary sludge together with fresh primary sludge, and cogeneration from biogas by using a gas engine in which the biogas produces electricity and heat from the exhaust gases. Three scenarios were compared, corresponding to the three digesters operated: C (conventional AD, 17 days residence time), B (combined TH + AD, same time), and A (TH + AD at half residence time). The biogas production of digesters B and A was 33 and 24% better, respectively when compared with C. In the case of the combined TH + AD process (scenarios A and B), the key factors in the energy balance were the recovery of heat from hot streams, and the concentration of sludge. The results of the balances showed that for 8% DS concentration of the secondary sludge tested in the pilot plant, the process can be energetically self-sufficient, but a fraction of the biogas must by-pass the gas engine to be directly burned. From an economic point of view, scenario B is more profitable in terms of green energy and higher waste removal, while scenario A reduces the digester volume required by a half. Considering a population of 100,000 inhabitants, the economic benefit is 87,600 €/yr for scenario A and 132,373 €/yr for B. This value can be increased to 223,867 €/yr by increasing the sludge concentration of the feeding to the TH unit to a minimum value that allows use of all the biogas to produce green energy. This concentration is 13% DS, which is still possible from a practical point of view. Additional benefits gained with the combined TH + AD process are the enhancement of the digesters rheology and the possibility of getting Class A biosolids. The integration study presented here set the basis for the scale-up to a demonstration plant.

Removal of oxytetracycline (OTC) in a synthetic pharmaceutical wastewater by a sequential anaerobic multichamber bed reactor (AMCBR)/completely stirred tank reactor (CSTR) system: biodegradation and inhibition kinetics.

An anaerobic multichamber bed reactor (AMCBR) was effective in removing both molasses-chemical oxygen demand (COD), and the antibiotic oxytetracycline (OTC). The maximum COD and OTC removals were 99% in sequential AMCBR/completely stirred tank reactor (CSTR) at an OTC concentration of 300 mg L(-1). 51%, 29% and 9% of the total volatile fatty acid (TVFA) was composed of acetic, propionic acid and butyric acids, respectively. The OTC loading rates at between 22.22 and 133.33 g OTC m(-3) d(-1) improved the hydrolysis of molasses-COD (k), the maximum specific utilization of molasses-COD (k(mh)) and the maximum specific utilization rate of TVFA (k(TVFA)). The direct effect of high OTC loadings (155.56 and -177.78 g OTC m(-3) d(-1)) on acidogens and methanogens were evaluated with Haldane inhibition kinetic. A significant decrease of the Haldane inhibition constant was indicative of increases in toxicity at increasing loading rates.

Novel anaerobic digestion process with sludge ozonation for economically feasible power production from biogas.

A novel process scheme was developed to achieve economically feasible energy recovery from anaerobic digestion. The new process scheme employs a hybrid configuration of mesophilic and thermophilic anaerobic digestion with sludge ozonation: the ozonated sludge is first degraded in a thermophilic digester and then further degraded in a mesophilic digester. In small-scale pilot experiments of the new process scheme, degradation of VSS improved by 3.5% over the control (mesophilic-only configuration) with 20% less ozone consumption. Moreover, biogas conversion also improved by 7.1% over the control. Selective enrichment of inorganic compounds during centrifugation produced a dewatered sludge cake with very low water content (59.4%). This low water content in the sludge cake improved its auto-thermal combustion potential during incineration and added to the overall energy savings. We conducted a case study to evaluate power generation from biogas for a municipal wastewater treatment plant with an average dry weather flow of 43,000 m3/d. Electricity production cost was 5.2 ¢/kWh for the advanced process with power generation, which is lower than the current market price of 7.2 ¢/kWh. The new anaerobic digestion scheme with power generation may reduce greenhouse gas emissions by about 1,000 t-CO(2)/year compared with the conventional process without power generation.

The effect of acid pretreatment on the anaerobic digestion and dewatering of waste activated sludge.

Waste activated sludge (WAS) is difficult to degrade in anaerobic digestion systems and pretreatments have been shown to speed up the hydrolysis stage. Here the effects of acid pretreatment (pH 6-1) using HCl on subsequent digestion and dewatering of WAS have been investigated. Optimisation of acid dosing was performed considering digestibility benefits and level of acid required. Pretreatment to pH 2 was concluded to be the most effective. In batch digestion this yielded the same biogas after 13 days as compared to untreated WAS at 21 days digestion. In semi-continuous digestion experiments (12 day hydraulic retention time at 35°C) it resulted in a 14.3% increase in methane yield compared to untreated WAS, also Salmonella was eradicated in the digestate. Dewatering investigations suggested that the acid pretreated WAS required 40% less cationic polymer addition to achieve the same cake solid content. A cost analysis was also carried out.

Assessment of optimum dilution ratio for biohydrogen production by anaerobic co-digestion of press mud with sewage and water.

Anaerobic co-digestion of press mud with water or sewage at ratios of 1:7.5, 1:10 and 1:12.5 were performed in continuously fed UASB reactors for hydrogen production. At a constant hydraulic retention time of 30 h, the specific hydrogen production rate was 187 mL/g volatile solids (VS) reduced during maximum biohydrogen production of 7960 mL/day at a 1:10 ratio of press mud to sewage. Chemical oxygen demand (COD) and VS reductions of 61% and 59% were noted on peak biohydrogen yield. A pH range of 5-6 was suitable at ambient temperature for entire process; a lower pH was inhibitory. Co-digestion of acidic press mud with sewage controlled pH for fermentation. Hence press mud can be exploited for biohydrogen production.