For the anaerobic biological treatment of saline wastewater, Anaerobic Digestion (AD) is currently a possibility, even though elevated salt concentrations can be a major obstacle. Anaerobic consortia and especially methanogenic archaea are very sensitive to fluctuations in salinity. When working with Upflow Sludge Blanket Reactor (UASB) technology, in which the microorganisms are aggregated and retained in the system as a granular biofilm, high sodium concentration negatively affects aggregation and consequently process performances. In this research, we analysed the structure of the biofilm and granules formed during the anaerobic treatment of high salinity (at 10 and 20 g/L of sodium) synthetic wastewater at lab scale. The acclimated inoculum was able to accomplish high rates of organics removal at all the salinity levels tested. 16S rRNA gene clonal analysis and Fluorescence In Situ Hybridization (FISH) analyses identified the acetoclastic Methanosaeta harundinacea as the key player involved acetate degradation and microbial attachment/granulation. When additional calcium (1 g/L) was added to overcome the negative effect of sodium on microbial aggregation, during the biofilm formation process microbial attachment and acetate degradation decreased. The same result was observed on granules formation: while calcium had a positive effect on granules strength when added to UASB reactors, Methanosaeta filaments were not present and the degradation of the partially acidified substrate was negatively influenced. This research demonstrated the possibility to get granulation at high salinity, bringing to the forefront the importance of a selection towards Methanosaeta cells growing in filamentous form to obtain strong and healthy granules.
Coastal eutrophication is one of the pivotal factors driving occurrence of harmful algal blooms (HABs), whose underlying mechanism remained unclear. To better understand the nutrient regime triggering HABs and their formation process, the phytoplankton composition and its response to varying nitrogen (N) and phosphorus (P), physio-chemical parameters in water and sediment in Johor Strait in March 2019 were analyzed. Surface and sub-surface HABs were observed with the main causative species of Skeletonema, Chaetoceros and Karlodinium. The ecophysiological responses of Skeletonema to the low ambient N/P ratio such as secreting alkaline phosphatase, regulating cell morphology (volume; surface area/volume ratio) might play an important role in dominating the community. Anaerobic sediment iron-bound P release and simultaneous N removal by denitrification and anammox, shaped the stoichiometry of N and P in water column. The decrease of N/P ratio might shift the phytoplankton community into the dominance of HABs causative diatoms and dinoflagellates.
Safe disposal of effluent from palm oil production poses an environmental concern. The highly polluting effluent is customarily treated by unsustainable open ponds with low efficiency, direct emissions, and massive land use. This study looks into an application of integrated anaerobic/oxic/oxic scheme for treatment of high strength palm oil mill effluent. The anaerobic reactors functioned as a prime degrader that removed up to 97.5% of the chemical oxygen demand (COD), while the aerobic reactors played a role of an effluent polisher that further reduced the COD. Their complementing roles resulted in a remarkable removal of 99.7%. Assessment of emission mitigation and biogas energy revealed that yearly energy of 53.2 TJ, emissions reduction of 239,237 tCO2 and revenue of USD 1.40 millions can be generated out of electricity generation and heating. The integrated scheme provides a viable and sustainable treatment of the high strength palm oil mill effluent.
Mixed culture anaerobic fermentation generates a wide range of products from simple sugars, and is potentially an effective process for producing renewable commodity chemicals. However it is difficult to predict product spectrum, and to control the process. One of the key control handles is pH, but the response is commonly dependent on culture history. In this work, we assess the impact of pH regulation mode on the product spectrum. Two regulation modes were applied: in the first, pH was adjusted from 4.5 to 8.5 in progressive steps of 0.5 and in the second, covered the same pH range, but the pH was reset to 5.5 before each change. Acetate, butyrate, and ethanol were produced throughout all pH ranges, but there was a shift from butyrate at pH < 6.5 to ethanol at pH > 6.5, as well as a strong and consistent shift from hydrogen to formate as pH increased. Microbial analysis indicated that progressive pH resulted in dominance by Klebsiella, while reset pH resulted in a bias towards Clostridium spp., particularly at low pH, with higher variance in community between different pH levels. Reset pH was more responsive to changes in pH, and analysis of Gibbs free energy indicated that the reset pH experiments operated closer to thermodynamic equilibrium, particularly with respect to the formate/hydrogen balance. This may indicate that periodically resetting pH conforms better to thermodynamic expectations.
The biggest agricultural sector that contributes to the Malaysian economy is the oil palm industry. The effluent generated during the production of crude palm oil known as palm oil mill effluent (POME). POME undergoes anaerobic treatment that requires long retention time and produces large amount of methane that consequently contributes to global warming. In this study, an isolated bacteria was selected based on its ability to degrade kraft lignin (KL) and identified as Ochrobactrum sp. The bacteria were able to treat POME (from anaerobic pond) under the aerobic condition without addition of nutrient, resulting in a significant chemical oxygen demand (COD) removal of 71 %, removal rate of 1385 mg/l/day, and 12.3 times higher than that of the ponding system. It has also resulted in 60 % removal of ammoniacal nitrogen and 55 % of total polyphenolic after 6-day treatment period with the detection of lignocellulolytic enzymes.
Pseudogenes in the Escherichia coli genome are assumed to be non-functional. In this study, Keio collection BW25113∆yqiG and YqiG-producing strain (BW25113/pCA24N-YqiG) were used to evaluate the importance of pseudogene yqiG in hydrogen metabolism. Our results show pseudogene protein YqiG was identified as an essential protein in the production of biohydrogen from glucose. The mutant yqiG decreased biohydrogen production from 37 µmol mg-1 protein to 6 µmol mg-1 protein compared to the wild-type strain, and glucose consumption was reduced by 80%. Through transcriptional analysis, we found that the yqiG mutation represses pflB transcription tenfold; pflB encodes pyruvate-formate lyase, one of the key enzymes in the anaerobic metabolism of E. coli. Moreover, production of YqiG stimulated glycolysis and increased biohydrogen productivity 1.5-fold compared to that of the wild-type strain. Thus, YqiG is important for the central glycolysis reaction and is able to influence hydrogen metabolism activity in E. coli.
The present study intends to evaluate the potential of co-digestion for utilizing Organic fraction of Municipal Solid Waste (OFMSW) and sewage sludge (SS) for enhanced biogas production. Metagenomic analysis was performed to identify the dominant bacteria, archaea and fungi, changes in their communities with time and their functional roles during the course of anaerobic digestion (AD). The cumulative biogas yield of 586.2 mL biogas/gVS with the highest methane concentration of 69.5% was observed under an optimum ratio of OFMSW:SS (40:60 w/w). Bacteria and fungi were found to be majorly involved in hydrolysis and initial stages of AD. Probably, the most common archaea Methanosarsina sp. primarily followed the acetoclastic pathway. The hydrogenotrophic pathway was less followed as indicated by the reduction in abundance of syntrophic acetate oxidizers. An adequate understanding of microbial communities is important to manipulate and inoculate the specific microbial consortia to maximize CH4 production through AD.
Anammox bacteria can easily undergo starvation due to fluctuations in feed flowrate and concentration in wastewater treatment plants. In this study, we analyzed the effects of different types of storage conditions (presence of ammonium (Ra), nitrite (Rn), hydrazine (Rh), and no substrate (Rc)) in aiding the viability of anammox bacteria during starvation and recovery. After starvation, the bacteria were subjected to a 15-week recovery period. Anammox bacteria showed better results during starvation and recovery in Rh as compared to other conditions. Decay rate values obtained after starvation in Ra, Rn, Rh, and Rc were 0.032/day, 0.042/day, 0.019/day, and 0.037/day, respectively. Meanwhile, µmax values obtained in Rh, Ra, Rn, and Rc on the 15th week of recovery were 0.092, 0.075, 0.011, and 0.067 d-1, respectively. This indicated that the availability of hydrazine helps to reduce the mortality rate of anammox bacteria during starvation and enhances the recovery of anammox process.
The sustainable application of an up-flow anaerobic baffled reactor (UABR) to treat real paper and cardboard industrial effluent (PCIE) containing bronopol (2-bromo-2-nitropropan-1, 3-diol) was investigated. At a hydraulic retention time (HRT) of 11.7 h and a bronopol concentration of 7.0 mg L-1, the removal efficiencies of total chemical oxygen demand (CODtotal), CODsoluble, CODparticulate, total suspended solids (TSS), volatile suspended solids (VSS), carbohydrates, and proteins were 55.3 ± 5.2%, 26.8 ± 2.3%, 94.4 ± 4.6%, 89.4 ± 2.6%, 84.5 ± 3.2%, 72.1 ± 1.8%, and 22.4 ± 1.8%, respectively. The conversion of complex organics (e.g., carbohydrates and proteins) into bio-methane (CH4) was assisted via enzyme activities of, in U (100 mL)-1, α-amylase (224-270), α-xylanase (171-188), carboxymethyl cellulase (CM-cellulase) (146-187), polygalacturonase (56-126), and protease (67,000-75300). The acidogenic condition was dominant at a short HRT of 2.9 h, where methane yield dropped by 32.5%. Under this condition, the growth of methanogenic bacteria could be inhibited by volatile fatty acids (VFA) accumulation. The analysis of Fourier-transform infrared (FTIR) spectra detected peaks relevant to methylene and nitro groups in the sludge samples, suggesting that entrapment/adsorption by the sludge bed could be a major mechanism for removing bronopol. The economic feasibility of UABR, as proposed to receive 100 m3 d-1 of PCIE, showed a payback period (profits from environmental benefits, biogas recovery, and carbon credit) of 7.6 yr. The study outcomes showed a high connection to the environmental-, economic-, and social-related sustainable development goals (SDGs).
An anaerobic membrane bioreactor (AnMBR) was employed as primary treatment unit for anaerobic treatment of simulated wastewater to produce high effluent quality. A lab scale hollow fiber membrane was used to scrutinize the performance of AnMBR as a potential treatment system for simulated milk wastewater and analyze its energy recovery potential. The 15 L bioreactor was operated continuously at mesophilic conditions (35 °C) with a pH constant of 7.0. The membrane flux was in the range of 9.6-12.6 L/m2. h. The different organic loading rates (OLRs) of 1.61, 3.28, 5.01, and 8.38 g-COD/L/d, of simulated milk wastewater, were fed to the reactor and the biogas production rate was analyzed, respectively. The results revealed that the COD removal efficiencies of 99.54 ± 0.001% were achieved at the OLR of 5.01 gCOD/L/d. The highest methane yield was found to be at OLR of 1.61 gCOD/L/d at HRT of 30 d with the value of 0.33 ± 0.01 L-CH4/gCOD. Moreover, based on the analysis of energy balance in the AnMBR system, it was found that energy is positive at all the given HRTs. The net energy production (NEP) ranged from 2.594 to 3.268 kJ/gCOD, with a maximum NEP value of 3.268 kJ/gCOD at HRT 10 d HRT. Bioenergy recovery with the maximum energy ratio, of 4.237, was achieved with an HRT of 5 d. The study suggests a sizable energy saving with the anaerobic membrane process.
Recycled paper mill effluent (RPME) contains high levels of organic and solid compounds, causing operational problems for anaerobic biological treatment. In this study, a unique modified anaerobic inclining-baffled reactor (MAI-BR) has been developed to treat RPME at various initial chemical oxygen demand (COD) concentrations (1000-4000 mg/L) and hydraulic retention times (HRTs) (3 and 1 day). The COD removal efficiency was decreased from 96 to 83% when the organic loading rate (OLR) was increased from 0.33 to 4 g/L day. Throughout the study, a maximum methane yield of 0.25 L CH4/g COD was obtained, while the pH fluctuated in the range of 5.8 to 7.8. The reactor performance was influenced by the development and distribution of the microbial communities. Based on the next-generation sequencing (NGS) analysis, the microbial community represented a variety of bacterial phyla with significant homology to Euryarchaeota (43.06%), Planctomycetes (24.68%), Proteobacteria (21.58%), Acidobacteria (4.12%), Chloroflexi (3.14%), Firmicutes (1.12%), Bacteroidetes (1.02%), and others (1.28%). The NGS analysis showed that the microbial community was dominated by Methanosaeta concilii and Candidatus Kuenenia stuttgartiensis. This can be supported by the presence of filamentous and spherical microbes of different sizes. Additionally, methanogenic and anaerobic ammonium oxidation (ANAMMOX) microorganisms coexisted in all compartments, and these contributed to the overall degradation of substances in the RPME. Graphical abstract ᅟ.
This study aimed at investigating the biohydrogen and biomethane potential of co-digestion from palm oil mill effluent (POME) and concentrated latex wastewater (CLW) in a two-stage anaerobic digestion (AD) process under thermophilic (55 ± 3 °C) and at an ambient temperature (30 ± 3 °C) conditions, respectively. The batch experiments of POME:CLW mixing ratios of 100:0, 70:30, 50:50, 30:70, and 0:100 was investigated with the initial loadings at 10 g-VS/L. The highest hydrogen yield of 115.57 mLH2/g-VS was obtained from the POME: CLW mixing ratio of 100:0 with 29.0 of C/N ratio. While, the highest subsequent methane production yield of 558.01 mLCH4/g-VS was achieved from hydrogen effluent from POME:CLW mixing ratio of 70:30 0 with 21.8 of C/N ratio. This mixing ratio revealed the highest synergisms of about 9.21% and received maximum total energy of 19.70 kJ/g-VS. Additionally, continuous hydrogen and methane production were subsequently performed in a series of continuous stirred tank reactor (CSTR) and up-flow anaerobic sludge blanket reactor (UASB) to treat the co-substate. The results indicated that the highest hydrogen yield of POME:CLW mixing ratio at 70:30 of 95.45 mL-H2/g-VS was generated at 7-day HRT, while methane production was obtained from HRT 15 days with a yield of 204.52 mL-CH4/g-VS. Thus, the study indicated that biogas production yield of CLW could be enhanced by co-digesting with POME. In addition, the two-stage AD model under anaerobic digestion model no. 1 (ADM-1) framework was established, 9.10% and 2.43% of error fitting of hydrogen and methane gas between model simulation data and experimental data were found. Hence, this research work presents a novel approach for optimization and feasibility for co-digestion of POME with CLW to generate mixed gaseous biofuel potentially.
This critical review for anaerobic degradation of complex organic compounds like butyrate using reactors has been enormously applied for biogas production. Biogas production rate has a great impact on: reactor granulation methanogenesis, nutrient content, shear velocity, organic loading and loss of nutrients taking place in the reactor continuously. Various technologies have been applied to closed anaerobic reactors to improve biogas production and treatment efficiency. Recent reviews showed that the application of closed anaerobic reactors can accelerate the degradation of organics like volatile fatty acid-butyrate and affect microbial biofilm formation by increasing the number of methanogens and increase methane production 16.5 L-1 CH4 L-1 POME-1. The closed anaerobic reactors with stable microbial biofilm and established organic load were responsible for the improvement of the reactor and methane production. The technology mentioned in this review can be used to monitor biogas concentration, which directly correlates to organic concentrations. This review attempts to evaluate interactions among the: degradation of organics, closed anaerobic reactors system, and microbial granules. This article provides a useful picture for the improvement of the degradation of organic butyrate for COD removal, biogas and methane production in an anaerobic closed reactor.
The augmentation of biogas production can be achieved by incorporating metallic nanoparticles as additives within anaerobic digestion. The objective of this current study is to examine the synthesis of Fe-Ni-Zn and Fe-Co-Zn trimetallic nanoparticles using the co-precipitation technique and assess its impact on anaerobic digestion using palm oil mill effluent (POME) as carbon source. The structural morphology and size of the synthesised trimetallic nanoparticles were analysed using a range of characterization techniques, such as X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), and Energy-dispersive X-ray spectroscopy (EDX) . The average size of Fe-Ni-Zn and Fe-Co-Zn were 19-25.5 nm and 19.1-30.5 nm respectively. Further, investigation focused on examining the diverse concentrations of trimetallic nanoparticles, ranging from 0 to 50 mgL-1. The biogas production increased by 55.55% and 60.11% with Fe-Ni-Zn and Fe-Co-Zn trimetallic nanoparticles at 40 mgL-1 and 20 mgL-1, respectively. Moreover, the lowest biogas of 11.11% and 38.11% were found with 10 mgL-1 of Fe-Ni-Zn and Fe-Co-Zn trimetallic nanoparticles. The findings of this study indicated that the trimetallic nanoparticles exhibited interactions with anaerobes, thereby enhancing the degradation process of palm oil mill effluent (POME) and biogas production. The study underscores the potential efficacy of trimetallic nanoparticles as a viable supplement for the promotion of sustainable biogas generation.
A two-stage fermentation process consisting of dark and photo-fermentation periods was carried out in a batch reactor. In the first stage, glucose was fermented in the dark stage using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564; CSN1-4) to produce acetate, CO2 and H2. The acetate produced in the first stage is fermented to H2 and CO2 by Rhodobacter sphaeroides NCIMB 8253 for further hydrogen production in the second, illuminated stage. The yield of hydrogen in the first stage was about 3.10 mol H2 (mol glucose)(-1) at a glucose concentration of 10 g L(-1), pH 6 +/- 0.2 and 37 degrees C and the second stage yield was about 1.10-1.25 mol H2 (mol acetic acid)(-1) at pH 6.8 +/- 0.2 and 32 degrees C, without removal of the Clostridium CSN1-4. The overall yield of hydrogen in the two-stage process, with glucose as the main substrate was higher than single-stage fermentation.
Effects of antibiotic residues on methane production in anaerobic digestion are commonly studied using the following two antibiotic addition methods: (1) adding manure from animals that consume a diet containing antibiotics, and (2) adding antibiotic-free animal manure spiked with antibiotics. This study used chlortetracycline (CTC) as a model antibiotic to examine the effects of the antibiotic addition method on methane production in anaerobic digestion under two different swine wastewater concentrations (0.55 and 0.22mg CTC/g dry manure). The results showed that CTC degradation rate in which manure was directly added at 0.55mg CTC/g (HSPIKE treatment) was lower than the control values and the rest of the treatment groups. Methane production from the HSPIKE treatment was reduced (p<0.05) by 12% during the whole experimental period and 15% during the first 7days. The treatments had no significant effect on the pH and chemical oxygen demand value of the digesters, and the total nitrogen of the 0.55mg CTC/kg manure collected from mediated swine was significantly higher than the other values. Therefore, different methane production under different antibiotic addition methods might be explained by the microbial activity and the concentrations of antibiotic intermediate products and metabolites. Because the primary entry route of veterinary antibiotics into an anaerobic digester is by contaminated animal manure, the most appropriate method for studying antibiotic residue effects on methane production may be using manure from animals that are given a particular antibiotic, rather than adding the antibiotic directly to the anaerobic digester.
The composting of lignocellulosic oil palm empty fruit bunch (OPEFB) with continuous addition of palm oil mill (POME) anaerobic sludge which contained nutrients and indigenous microbes was studied. In comparison to the conventional OPEFB composting which took 60-90 days, the rapid composting in this study can be completed in 40 days with final C/N ratio of 12.4 and nitrogen (2.5%), phosphorus (1.4%), and potassium (2.8%), respectively. Twenty-seven cellulolytic bacterial strains of which 23 strains were closely related to Bacillus subtilis, Bacillus firmus, Thermobifida fusca, Thermomonospora spp., Cellulomonas sp., Ureibacillus thermosphaericus, Paenibacillus barengoltzii, Paenibacillus campinasensis, Geobacillus thermodenitrificans, Pseudoxanthomonas byssovorax which were known as lignocellulose degrading bacteria and commonly involved in lignocellulose degradation. Four isolated strains related to Exiguobacterium acetylicum and Rhizobium sp., with cellulolytic and hemicellulolytic activities. The rapid composting period achieved in this study can thus be attributed to the naturally occurring cellulolytic and hemicellulolytic strains identified.
Tylosin has been considered inhibiting COD removal in anaerobic digestion. In this study it is proven that this is not always the case. Accordingly, elevated concentrations of Tylosin (100-800mgL-1) could be tolerated by the anaerobic system. The influence of Tylosin concentrations on an up-flow anaerobic stage reactor (UASR) was assessed using additions of Tylosin phosphate concentrate. Results showed high efficiency for COD removal (average 93%) when Tylosin was present at concentrations ranging from 0 to 400 mg L-1. However, at Tylosin concentrations of 600 and 800 mg L-1 treatment efficiency declined to 85% and 75% removal respectively. The impact of Tylosin concentrations on archaeal activity were investigated and the analysis revealed that archaeal cells dominated the reactor, confirming that there was no detectable inhibition of the methanogens at Tylosin levels between 100 and 400mg L-1. Nevertheless, the investigation showed a slight reduction in the number of methanogens at Tylosin levels of 600 and 800 mg L-1. These results demonstrated that the methanogens were well adapted to Tylosin. It would not be expected that the process performance of the UASR would be affected, not even at a level well in excess of those appearing in real wastewater from a Tylosin production site.
Periphyton dominated by the cellulose-rich filamentous green alga Cladophora forms conspicuous growths along rocky marine and freshwater shorelines worldwide, providing habitat for diverse epibionts. Bacterial epibionts have been inferred to display diverse functions of biogeochemical significance: N-fixation and other redox reactions, phosphorus accumulation, and organic degradation. Here, we report taxonomic diversity of eukaryotic and prokaryotic epibionts and diversity of genes associated with materials cycling in a Cladophora metagenome sampled from Lake Mendota, Dane Co., WI, USA, during the growing season of 2012. A total of 1,060 distinct 16S, 173 18S, and 351 28S rRNA operational taxonomic units, from which >220 genera or species of bacteria (~60), protists (~80), fungi (6), and microscopic metazoa (~80), were distinguished with the use of reference databases. We inferred the presence of several algal taxa generally associated with marine systems and detected Jaoa, a freshwater periphytic ulvophyte previously thought endemic to China. We identified six distinct nifH gene sequences marking nitrogen fixation, >25 bacterial and eukaryotic cellulases relevant to sedimentary C-cycling and technological applications, and genes encoding enzymes in aerobic and anaerobic pathways for vitamin B12 biosynthesis. These results emphasize the importance of Cladophora in providing habitat for microscopic metazoa, fungi, protists, and bacteria that are often inconspicuous, yet play important roles in ecosystem biogeochemistry.
This paper critically reviews the impacts of supplementing trace elements on the anaerobic digestion performance. The in-depth knowledge of trace elements as micronutrients and metalloenzyme components justifies trace element supplementation into the anaerobic digestion system. Most of the earlier studies reported that trace elements addition at (sub)optimum dosages had positive impacts mainly longer term on digester stability with greater organic matter degradation, low volatile fatty acids (VFA) concentration and higher biogas production. However, these positive impacts and element requirements are not fully understood, they are explained on a case to case basis because of the great variance of the anaerobic digestion operation. Iron (Fe), nickel (Ni) and cobalt (Co) are the most studied and desirable elements. The right combination of multi-elements supplementation can have greater positive impact. This measure is highly recommended, especially for the mono-digestion of micronutrient-deficient substrates. The future research should consider the aspect of trace element bioavailability.