Archaeal community composition affects the function of anaerobic co-digesters in response to organic overload

Microbial community diversity in two thermophilic laboratory-scale and three full-scale anaerobic co-digesters was analysed by genetic profiling based on PCR-amplified partial 16S rRNA genes. In parallel operated laboratory reactors a stepwise increase of the organic loading rate (OLR) resulted in a decrease of methane production and an accumulation of volatile fatty acids (VFAs). However, almost threefold different OLRs were necessary to inhibit the gas production in the reactors. During stable reactor performance, no significant differences in the bacterial community structures were detected, except for in the archaeal communities. Sequencing of archaeal PCR products revealed a dominance of the acetoclastic methanogen Methanosarcina thermophila, while hydrogenotrophic methanogens were of minor importance and differed additionally in their abundance between reactors. As a consequence of the perturbation, changes in bacterial and archaeal populations were observed. After organic overload, hydrogenotrophic methanogens (Methanospirillum hungatei and Methanoculleus receptaculi) became more dominant, especially in the reactor attributed by a higher OLR capacity. In addition, aggregates composed of mineral and organic layers formed during organic overload and indicated tight spatial relationships between minerals and microbial processes that may support de-acidification processes in over-acidified sludge.Comparative analyses of mesophilic stationary phase full-scale reactors additionally indicated a correlation between the diversity of methanogens and the VFA concentration combined with the methane yield. This study demonstrates that the coexistence of two types of methanogens, i.e. hydrogenotrophic and acetoclastic methanogens is necessary to respond successfully to perturbation and leads to stable process performance.     

Application of an early warning indicator and CaO to maximize the time–space-yield of an completely mixed waste digester using rape seed oil as co-substrate

In order to increase the organic loading rate (OLR) and hereby the performance of biogas plants an early warning indicator (EWI-VFA/Ca) was applied in a laboratory-scale biogas digester to control process stability and to steer additive dosing. As soon as the EWI-VFA/Ca indicated the change from stable to instable process conditions, calcium oxide was charged as a countermeasure to raise the pH and to bind long-chain fatty acids (LCFAs) by formation of aggregates. An interval of eight days between two increases of the OLR, which corresponded to 38% of the hydraulic residence time (HRT), was sufficient for process adaptation. An OLR increase by a factor of three within six weeks was successfully used for biogas production. The OLR was increased to 9.5 kg volatile solids (VS) m^?3 d^?1 with up to 87% of fat. The high loading rates affected neither the microbial community negatively nor the biogas production process. Despite the increase of the organic load to high rates, methane production yielded almost its optimum, amounting to 0.9 m³ (kg VS)^?1. Beneath several uncharacterized members of the phylum Firmicutes mostly belonging to the family Clostridiaceae, a Syntrophomonas-like organism was identified that is known to live in a syntrophic relationship to methanogenic archaea. Within the methanogenic group, microorganisms affiliated to Methanosarcina, Methanoculleus and Methanobacterium dominated the community.     

Microbial oxidation of methane from old landfills in biofilters

Landfill gas emissions are among the largest sources of the greenhouse gas methane. For this reason, the possibilities of microbial methane degradation in biofilters were investigated. Different filter materials were tested in two experimental plants, a bench-scale plant (total filter volume 51 l) and a pilot plant (total filter volume 4 m3). Three months after the beginning of the experiment, very high degradation rates of up to 63 g CH4/(m3h) were observed in the bench-scale plant at mean methane concentrations of 2.5% v/v and with fine-grained compost as biofilter material. However, the degradation rates of the compost biofilter decreased in the fifth month of the experiment, probably due to the accumulation of exopolymeric substances formed by the microorganisms. A mixture of compost, peat, and wood fibers showed stable and satisfactory degradation rates around 20 g/(m3h) at mean concentrations of 3% v/v over a period of one year. In this material, the wood fibers served as a structural material and prevented clogging of the biofilter. Extrapolation of the experimental data indicates that biofilters for methane oxidation have to be at least 100 times the volume of biofilters for odor control to obtain the same cleaning efficiency per unit volume flow of feed gas.     

Feasibility assessment of thermophilic anaerobic digestion process of food waste

In this study, a lab-scale thermophilic anaerobic digestion of food waste collected from G-district in Seoul was performed to assess its feasibility and applicability in field-scale biogas plants. Monitoring parameters included biogas production, methane composition, pH, alkalinity, and volatile fatty acid (VFA) concentrations. Accumulation of VFA caused successive depression in pH, which inhibited microbial activity of methane-forming microorganisms. Signals of biological instability and inhibition of methanogenesis suggest possible process failure, as indicated by reduction in methane production. Results revealed that modifications in certain conditions, such as decreased organic loading rate (OLR) or additional insertion of alkalinity, must be made for its application in industrial-scale biogas plants, and that thermophilic anaerobic digestion of food waste may not be feasible without any modification.     

Effects of various fatty acid amendments on a microbial digester community in batch culture

Since biogas production is becoming increasingly important the understanding of anaerobic digestion processes is fundamental. However, large-scale digesters often lack online sensor equipment to monitor key parameters. Furthermore the possibility to selectively change fermenting parameter settings in order to investigate methane output or microbial changes is limited. In the present study we examined the possibility to investigate the microbial community of a large scale (750,000 L) digester within a laboratory small-scale approach. We studied the short-term response of the downscaled communities on various fatty acids and its effects on gas production and compared it with data from the original digester sludge. Even high loads of formic acid led to distinct methane formation, whereas high concentrations of other acids (acetic, butyric, propionic acid) caused a marked inhibition of methanogenesis coupled with an increase in hydrogen concentration. Molecular microbial techniques (DGGE/quantitative real-time-PCR) were used to monitor the microbial community changes which were related to data from GC and HPLC analysis. DGGE band patterns showed that the same microorganisms which were already dominant in the original digester re-established again in the lab-scale experiment. Very few microorganisms dominated the whole fermenting process and species diversity was not easily influenced by moderate varying fatty acid amendments - Methanoculleus thermophilus being the most abundant species throughout the variants. MCR-copy number determined via quantitative real-time-PCR - turned out to be a reliable parameter for quantification of methanogens, even in a very complex matrix like fermenter sludge. Generally the downscaled batch approach was shown to be appropriate to investigate microbial communities from large-scale digesters.     

Effect of substrate feeding frequencies on the methane production and microbial communities of laboratory-scale anaerobic digestion reactors

Even though full-scale digesters have been designed based on laboratory-scale tests, the substrate feeding modes of laboratory-scale tests might be different from those of full-scale digesters. The effect of substrate feeding frequencies on the performance and microbial community of laboratory-scale anaerobic digestion reactors was investigated. Feeding frequencies of twice a day, once a day, and every two days were tested in three 2-L reactors with an organic loading rate of 0.5 g-glucose/L/day under mesophilic condition. According to the results of this study, all the reactors showed similar methane production rates and SCOD removal efficiencies after sufficient time of acclimation, but frequently feeding promoted more stable digestion. Although there was no significant difference in microbial diversities from pyrosequencing analyses, the changes of archaeal community composition were observed. The decrease in feeding frequency appeared to cause shifts from acetoclastic methanogens affiliated with Methanosaeta to H₂-utilizing methanogens. The increase of Methanosaeta at a frequently feeding might contribute to the stability of reactor operation. Since this study uses glucose as the substrate, there is still possibility of different results for more complex substrates, such as sludge, food waste, etc.     

Anaerobic digestion of the liquid fraction of dairy manure in pilot plant for biogas production: residual methane yield of digestate

The performance of the only dairy manure biogas plant in Cantabria (Northern coast of Spain) was evaluated in terms of liquid-solid separation and anaerobic digestion of the liquid fraction. Screened liquid fraction was satisfactorily treated in a CSTR digester at HRTs from 20 to 10 days with organic loading rates ranging from 2.0 to 4.5 kg VS/(m(3)d). Stable biogas productions from 0.66 to 1.47 m(3)/(m(3)d) were achieved. Four anaerobic effluents collected from the digester at different HRTs were analyzed to measure their residual methane potentials, which ranged from 12.7 to 102.4 L/gVS. These methane potentials were highly influenced by the feed quality and HRT of the previous CSTR anaerobic digestion process. Biomethanization of the screened liquid fraction of dairy manure from intensive farming has the potential to provide up to 2% of total electrical power in the region of Cantabria.     

Increasing biogas production from sewage sludge anaerobic co-digestion process by adding crude glycerol from biodiesel industry

In an effort to convert waste streams to energy in a green process, glycerol from biodiesel manufacturing has been used to increase the gas production and methane content of biogas within a mesophilic anaerobic co-digestion process using primary sewage sludge. Glycerol was systematically added to the primary digester from 0% to 60% of the organic loading rate (OLR). The optimum glycerol loading range was from 25% to 60% OLR. This resulted in an 82–280% improvement in specific gas production. Following the feeding schedule described, the digesters remained balanced and healthy until inhibition was achieved at 70% glycerol OLR. This suggests that high glycerol loadings are possible if slow additions are upheld in order to allow the bacterial community to adjust properly. Waste water treatment plant operators with anaerobic digesters can use the data to increase loadings and boost biogas production to enhance energy conversion. This process provides a safe, environmentally friendly method to convert a typical waste stream to an energy stream of biogas.     

Wet explosion of wheat straw and codigestion with swine manure: Effect on the methane productivity

The continuously increasing demand for renewable energy sources renders anaerobic digestion to one of the most promising technologies for renewable energy production. Twenty-two (22) large-scale biogas plants are currently under operation in Denmark. Most of these plants use manure as the primary feedstock but their economical profitable operation relies on the addition of other biomass products with a high biogas yield. Wheat straw is the major crop residue in Europe and the second largest agricultural residue in the world. So far it has been used in several applications, i.e. pulp and paper making, production of regenerated cellulose fibers as an alternative to wood for cellulose-based materials and ethanol production. The advantage of exploiting wheat straw for various applications is that it is available in considerable quantity and at low-cost. In the present study, the codigestion of swine manure with wheat straw in a continuous operated system was investigated, as a method to increase the efficiency of biogas plants that are based on anaerobic digestion of swine manure. Also, the pretreatment of wheat straw with the wet explosion method was studied and the efficiency of the wet explosion process was evaluated based on (a) the sugars release and (b) the methane potential of the pretreated wheat straw compared to that of the raw biomass. It was found that, although a high release of soluble sugars was observed after wet explosion, the methane obtained from the wet-exploded wheat straw was slightly lower compared to that from the raw biomas s. On the other hand, the results from the codigestion of raw (non-pretreated) wheat straw with swine manure were very promising, suggesting that 4.6 kg of straw added to 1 t of manure increase the methane production by 10%. Thus, wheat straw can be considered as a promising, low-cost biomass for increasing the methane productivity of biogas plants that are based mainly on swine manure.     

Changes in soil chemical and microbiological properties during 4 years of application of various organic residues

A 4-year field trial was established in eastern Sweden to evaluate the effects of organic waste on soil chemical and microbiological variables. A simple crop rotation with barley and oats was treated with either compost from household waste, biogas residue from household waste, anaerobically treated sewage sludge, pig manure, cow manure or mineral fertilizer. All fertilizers were amended in rates corresponding to 100kgNha(-1)year(-1). The effects of the different types of organic waste were evaluated by subjecting soil samples, taken each autumn 4 weeks after harvest, to an extensive set of soil chemical (pH, Org-C, Tot-N, Tot-P, Tot-S, P-AL, P-Olsen, K-AL, and some metals) and microbiological (B-resp, SIR, microSIR active and dormant microorganisms, PDA, microPDA, PAO, Alk-P and N-min) analyses. Results show that compost increased pH, and that compost as well as sewage sludge increased plant available phosphorus; however, the chemical analysis showed few clear trends over the 4 years and few clear relations to plant yield or soil quality. Biogas residues increased substrate induced respiration (SIR) and, compared to the untreated control amendment of biogas residues as well as compost, led to a higher proportion of active microorganisms. In addition, biogas residues increased potential ammonia oxidation rate (PAO), nitrogen mineralization capacity (N-min) as well as the specific growth rate constant of denitrifiers (microPDA). Despite rather large concentrations of heavy metals in some of the waste products, no negative effects could be seen on either chemical or microbiological soil properties. Changes in soil microbial properties appeared to occur more rapidly than most chemical properties. This suggests that soil microbial processes can function as more sensitive indicators of short-term changes in soil properties due to amendment of organic wastes.     

Ultrasound assisted biogas production from landfill leachate

The aim of this study is to increase biogas production and methane yield from landfill leachate in anaerobic batch reactors by using low frequency ultrasound as a pretreatment step. In the first part of the study, optimum conditions for solubilization of organic matter in leachate samples were investigated using various sonication durations at an ultrasound frequency of 20 kHz. The level of organic matter solubilization during ultrasonic pretreatment experiments was determined by calculating the ratio of soluble chemical oxygen demand (sCOD) to total chemical oxygen demand (tCOD). The sCOD/tCOD ratio was increased from 47% in raw leachate to 63% after 45 min sonication at 600 W/l. Non-parametric Friedman’s test indicated that ultrasonic pretreatment has a significant effect on sCOD parameter for leachate (p < 0.05). In the second part of the study, anaerobic batch reactors were operated for both ultrasonically pretreated and untreated landfill leachate samples in order to assess the effect of sonication on biogas and methane production rate. In anaerobic batch reactor feed with ultrasonically pretreated leachate, 40% more biogas was obtained compared to the control reactor. For statistical analysis, Mann–Whitney U test was performed to compare biogas and methane production rates for raw and pretreated leachate samples and it has been found that ultrasonic pretreatment significantly enhanced biogas and methane production rates from leachate (p < 0.05) in anaerobic batch reactors. The overall results showed that low frequency ultrasound pretreatment can be potentially used for wastewater management especially with integration of anaerobic processes.     

Methane generation from corncobs treated with xylanolytic consortia

Consortia were developed for the treatment of corncobs for use as a feedstock in a biogas fermentor. The treatment of corncobs with xylanolytic consortia enhanced the production of methane and biogas. All five consortia developed produced the maximum biogas and methane at a 6% loading rate and 20 days hydraulic retention time (HRT). The maximum biogas yield of 0.59m³/kg volatile solids (VS) with a methane content of 62% was produced with the KK-10 consortium. This was apparently due to a maximum hemicellulose degradation of 88%.     

Analysis on biochemical methane potential of agricultural byproducts with different types of silage storage

The aim of this work is to estimate biogas production by anaerobic digestion of agricultural byproduct silage at the low carbon green village in South Korea. The composition of agricultural byproduct from hot pepper farms was analyzed and it was found to be favorable with anaerobic digestion. In the cases of silage materials, the theoretical methane potentials of all of the ensiled materials were increased with silage storage having an increased range from 103 to 120 % compared to that without ensiling. The biochemical methane potential (BMP) tests showed that the ultimate methane potential of ensiled material was measured to be higher than that of raw material without silage storage, while the first order hydrolysis constant was lower. All of the silage materials containing microbial additives used in this study showed higher ultimate methane potentials and first order hydrolysis constants than raw material and silage material without additives. The change of ultimate methane potential was analyzed over time, and all of the test materials, except Day 2, showed higher ultimate methane potential than raw material, Day 0, and the highest was found on Day 40.     

Benefits of supplementing an industrial waste anaerobic digester with energy crops for increased biogas production

Currently, there is increasing competition for waste as feedstock for the growing number of biogas plants. This has led to fluctuation in feedstock supply and biogas plants being operated below maximum capacity. The feasibility of supplementing a protein/lipid-rich industrial waste (pig manure, slaughterhouse waste, food processing and poultry waste) mesophilic anaerobic digester with carbohydrate-rich energy crops (hemp, maize and triticale) was therefore studied in laboratory scale batch and continuous stirred tank reactors (CSTR) with a view to scale-up to a commercial biogas process. Co-digesting industrial waste and crops led to significant improvement in methane yield per ton of feedstock and carbon-to-nitrogen ratio as compared to digestion of the industrial waste alone. Biogas production from crops in combination with industrial waste also avoids the need for micronutrients normally required in crop digestion. The batch co-digestion methane yields were used to predict co-digestion methane yield in full scale operation. This was done based on the ratio of methane yields observed for laboratory batch and CSTR experiments compared to full scale CSTR digestion of industrial waste. The economy of crop-based biogas production is limited under Swedish conditions; therefore, adding crops to existing industrial waste digestion could be a viable alternative to ensure a constant/reliable supply of feedstock to the anaerobic digester.     

Anaerobic digestion of food waste: comparing leachate exchange rates in sequential batch systems digesting food waste and biosolids

Over 35,000 tonnes of food waste are generated from high concentration point sources (i.e., restaurants, hospitals and markets) in metropolitan Adelaide (Australia) each year. Anaerobic digestion is a preferred method of treatment to degrade highly putrescible waste streams such as food waste due its high methane potential. To maximise methane yield, a sequential batch anaerobic system was chosen as the most appropriate system. Two sets of sequential batch systems consisting of mature and start-up reactors in triplicate exchanged leachate. One set exchanged twice as much leachate by volume as the other set to determine the effects of different leachate exchange volumes. Results show that by increasing the leachate volume between mature and start-up reactors, the time to degrade feedstock decreases, but total methane generation yields did not markedly differ, being 229LCH4 kg(-1) VSadded and 214LCH4 kg(-1) VSadded. Process parameters used to determine when to cease leachate exchange in start-up reactors were a pH of 6.5 and methane generation rate of >0.5LCH4 kg(-1) VSadded day(-1). Changes in carbon dioxide and methane biogas composition in the mature reactors reflected process stress caused by the addition of leachate with high VFA concentrations from the start-up reactors and indicate there may be limits to leachate exchange rates and subsequent loading of mature reactors.     

Stabilization of sewage sludge in the presence of nanoscale zero-valent iron (nZVI): abatement of odor and improvement of biogas production

The potential benefits of nanoscale zero-valent iron (nZVI) on sludge stabilization, either the abatement of odor or the improvement of biogas production, were investigated in this study. Two commercial-grade microscale iron powders were also utilized for comparison. Adding 0.10 wt% of nZVI in sludge during anaerobic incubation significantly reduced the concentration of H₂S in biogas by 98.0 % (96.2–98.9 %), probably attributed by reactions between sulfides and the neo-formed hydrous Fe(II)/Fe(III) oxides layer at the surface of ZVI nanoparticles. Meanwhile, the percentage of P in bioavailable fractions decreased from 76.8 to 52.5 %, possibly due to the formation of vivianite [Fe₃(PO₄)₂]. Furthermore, 0.10 wt% of nZVI in anaerobic digestion for 17 days enhanced the concentration of CH₄ in biogas by 5.1–13.2 % and improved the production of biogas and methane by 30.4 and 40.4 %, respectively. The amendment of iron nanoparticles during anaerobic digestion can not only effectively reduce H₂S in biogas, but also potentially boost methane production significantly.     

Comparison of different liquid anaerobic digestion effluents as inocula and nitrogen sources for solid-state batch anaerobic digestion of corn stover

Effluents from three liquid anaerobic digesters, fed with municipal sewage sludge, food waste, or dairy waste, were evaluated as inocula and nitrogen sources for solid-state batch anaerobic digestion of corn stover in mesophilic reactors. Three feedstock-to-effluent (F/E) ratios (i.e., 2, 4, and 6) were tested for each effluent. At an F/E ratio of 2, the reactor inoculated by dairy waste effluent achieved the highest methane yield of 238.5 L/kgVS_feed, while at an F/E ratio of 4, the reactor inoculated by food waste effluent achieved the highest methane yield of 199.6 L/kgVS_feed. The microbial population and chemical composition of the three effluents were substantially different. Food waste effluent had the largest population of acetoclastic methanogens, while dairy waste effluent had the largest populations of cellulolytic and xylanolytic bacteria. Dairy waste also had the highest C/N ratio of 8.5 and the highest alkalinity of 19.3 g CaCO₃/kg. The performance of solid-state batch anaerobic digestion reactors was closely related to the microbial status in the liquid anaerobic digestion effluents.     

Anaerobic microbial degradation of poly (3-hydroxyalkanoates) with various terminal electron acceptors

The microbial degradation of poly (3-hydroxyalkanoates) (PHAs) under anaerobic conditions with various terminal electron acceptors was examined. Nitrate-reducing consortia were established using activated sludge, and PHAs were shown to be biodegradable under these conditions. A positive correlation between carbon dioxide production and nitrate reduction was demonstrated. Nitrous oxide accumulated as the main N-containing product of nitrate reduction. The amount of PHAs in activated sludge cultures decreased approximately 20% within 40 days of incubation. Attempts were made to establish iron- and sulfate-reducing consortia from spring water, yet it could not be demonstrated that the mixed cultures were capable of degrading PHAs. Pure cultures of iron- and sulfate-reducing bacteria could not utilize PHAs as sole carbon sources. Methanogenic environments sampled included pond sediment and rumen fluid. PHAs were fermented to methane and carbon dioxide after 10 weeks by a sediment consortium, with 43 to 57% of the substrate carbon transformed to methane. Although it could not be demonstrated that PHAs were biodegraded by a rumen fluid consortium, a facultative anaerobic bacterium, identified as aStaphylococcus sp., that could grow on PHAs was isolated from rumen fluid.     

Effects of a gradually increased load of fish waste silage in co-digestion with cow manure on methane production

This study examined the effects of an increased load of nitrogen-rich organic material on anaerobic digestion and methane production. Co-digestion of fish waste silage (FWS) and cow manure (CM) was studied in two parallel laboratory-scale (8 L effective volume) semi-continuous stirred tank reactors (designated R1 and R2). A reactor fed with CM only (R0) was used as control. The reactors were operated in the mesophilic range (37 °C) with a hydraulic retention time of 30 days, and the entire experiment lasted for 450 days. The rate of organic loading was raised by increasing the content of FWS in the feed stock. During the experiment, the amount (volume%) of FWS was increased stepwise in the following order: 3% – 6% – 13% – 16%, and 19%. Measurements of methane production, and analysis of volatile fatty acids, ammonium and pH in the effluents were carried out. The highest methane production from co-digestion of FWS and CM was 0.400 L CH4 gVS^?1, obtained during the period with loading of 16% FWS in R2. Compared to anaerobic digestion of CM only, the methane production was increased by 100% at most, when FWS was added to the feed stock. The biogas processes failed in R1 and R2 during the periods, with loadings of 16% and 19% FWS, respectively. In both reactors, the biogas processes failed due to overloading and accumulation of ammonia and volatile fatty acids.     

Anaerobic co-digestion of municipal solid organic waste with melon residues to enhance biodegradability and biogas production

Management of solid organic waste has become a major challenge in developing countries. Raw solid organic waste can be converted into biogas through anaerobic digestion; however, the efficiency of the process is influenced by various factors including the composition of the substrate. The present study was designed with the objective of enhancing the biodegradability of the organic fraction of municipal solid waste (OFMSW) and biogas production through co-digestion of the substrate with melon residues. The study was conducted in batch mode in four phases. The results revealed that an addition of melon waste at the rate of 300?g?kg^?1 OFMSW substantially increased the biodegradation rate and biogas production compared to OFMSW alone. The removal of up to 57.2?% volatile solids and a carbon to nitrogen (C/N) ratio of 15.9 was achieved at a 60?% water level when the digestion mixture was treated with inocula collected from partially-degraded food waste. The findings of this study reveal that melon residues could be used as a potential co-substrate to enhance the biodegradability of OFMSW and biogas production.