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.     

A dynamic simulation of methane fermentation process receiving heterogeneous food wastes and modelling acidic failure

Microbial response on volatile fatty acids (VFAs) is a key for methane fermentation processes since accumulation of VFAs often causes an acidic failure, especially treating such organics as food wastes composed of mostly readily biodegradable materials. To evaluate the impact of VFA accumulation, a lab-scale continuous experiment was performed for 110 days with sequential feeding of heterogeneous food wastes. When the volumetric loading rate was increased from 6 to 8 kg-COD/m³/day, a sudden decrease of methane production was observed with an accumulation of acetate and propionate in the fermenter. After discontinuation of feeding for 10 days, the digestate in the fermenter was centrifuged and washed with tap water to reduce the VFAs to be acceptable concentration below 1000 mg-COD/L. Nevertheless, no recovery of methane production was observed and VFA concentrations consistently increased. To model the event, a modification of ADM1 was made assuming the methanogens in the fermenter were irreversibly inactivated under very high VFA. Also considering the different nature of the fed food wastes over 11 samples, decomposition kinetics of individual food wastes were manipulated. The modified ADM1 with methanogenic activity decay reasonably reproduced the responses for soluble material concentrations and methane gas production rate over the experimental period.     

Recovery of energy from Taro (Colocasia esculenta) with solid-feed anaerobic digesters (SOFADs)

We present studies on solid-feed anaerobic digesters (SOFADs) in which chopped Colocasia esculenta was fed without any other pretreatment, in an attempt to develop an efficient means of utilizing the semi-aquatic weed that is otherwise an environmental nuisance. Two types of SOFADs were studied. The first type had a single vessel with two compartments. The lower portion of the digester, 25% of the total volume, was separated from the upper by a perforated PVC disk. The weed was charged from the top and inoculated with anaerobically digested cow dung-water slurry. The fermentation of the weed in the digester led to the formation of volatile fatty acids (VFAs) plus some biogas. The bioleachate, rich in the VFAs, passed through the perforated PVC disk and was collected in the lower compartment of the digester. The other type of digesters, referred to as anaerobic multi-phase high-solids digesters (AMHDs), had the same type of compartmentalized digester unit as the first type and an additional methaniser unit. Up-flow anaerobic filters (UAFs) were used as methaniser units, which converted the bioleachate into combustible biogas consisting of approximately 60% methane. All SOFADs developed a consistent performance in terms of biogas yield within 20 weeks from the start. Among the two types of digesters studied, the AMHDs were found to perform better with a twofold increase in biogas yield compared to the first type of digesters.     

Volatile fatty acid evolution in biomass mixture composts prepared in open and closed bioreactors

In this study we observed the production of volatile fatty acids (VFAs) during the composting process of compost heaps in two different bioreactors (open and closed) at three different depths (0, 40 and 80 cm). The compost was prepared as a mixture of bio-waste, horse manure, grass and sawdust to ensure sufficient pH conditions in compost heaps. VFA contents in the composting materials were analysed weekly over 14–119 d. The degradation process was monitored, along with temperature, pH, total organic carbon, oxidizable carbon and mono- and oligosaccharides. VFA contents were evaluated with regard to the depth of the sample site in the compost heap and to conditions in the bioreactors. The maximum VFA occurrence was observed during the first 35 d; acetic and propionic acids in particular were determined to occur in each sample. Considerable variations in their formation and elimination were observed in the two bioreactors as well as at the various depths in the compost heaps. Significant correlations were found between individual VFAs, as well as between VFA concentrations and organic carbon contents.     

Anaerobic co-digestion of tannery waste water and tannery solid waste using two-stage anaerobic sequencing batch reactor: focus on performances of methanogenic step

In this study, anaerobic co-digestion of the tannery waste water (TWW) and tannery solid waste (TSW) with four TWW to TSW mixing ratios (100:0, 75:25, 50:50 and 25:75) was carried out using semi-continuous two-phase anaerobic sequencing batch reactor system under mesophilic temperature (38?±?2 °C). During the experimental study, effluents resulted from previously optimized acidogenic reactors were used to feed subsequent methanogenic reactors and then operated at hydraulic retention time (HRT) of 20, 15 and 10 days and equivalent organic loading rate. The findings revealed that methanogenic reactor of 50:50 (TWW:TSW) treating the effluent from previously optimized acidogenic step exhibits best process performances in terms of daily biogas (415 ml/day), methane production (251 ml/day), methane content (60.5%) and COD removal efficiency (75%) when operated at HRT of 20 days. Process stability of methanogenic step also evaluated and the obtained results showed suitable pH (6.8), no VFA accumulation, i.e., VFA/Alkalinity (0.305), alkalinity (3210 mgCaCO₃/l) and ammonia (246 mg/l with in optimum operating range). In general, improved process stability as well as performance was achieved during anaerobic co-digestion of TWW with TSW compared to mono-digestion of TWW.     

Microbiological characterization and specific methanogenic activity of anaerobe sludges used in urban solid waste treatment

This study presents the microbiological characterization of the anaerobic sludge used in a two-stage anaerobic reactor for the treatment of organic fraction of urban solid waste (OFUSW). This treatment is one alternative for reducing solid waste in landfills at the same time producing a biogas (CH(4) and CO(2)) and an effluent that can be used as biofertilizer. The system was inoculated with sludge from a wastewater treatment plant (WWTP) (Río Frío Plant in Bucaramanga-Colombia) and a methanogenic anaerobic digester for the treatment of pig manure (Mesa de los Santos in Santander). Bacterial populations were evaluated by counting groups related to oxygen sensitivity, while metabolic groups were determined by most probable number (MPN) technique. Specific methanogenic activity (SMA) for acetate, formate, methanol and ethanol substrates was also determined. In the acidogenic reactor (R1), volatile fatty acids (VFA) reached values of 25,000 mg L(-1) and a concentration of CO(2) of 90%. In this reactor, the fermentative population was predominant (10(5)-10(6)MPN mL(-1)). The acetogenic population was (10(5)MPN mL(-1)) and the sulphate-reducing population was (10(4)-10(5)MPN mL(-1)). In the methanogenic reactor (R2), levels of CH(4) (70%) were higher than CO(2) (25%), whereas the VFA values were lower than 4000 mg L(-1). Substrate competition between sulphate-reducing (10(4)-10(5)MPN mL(-1)) and methanogenic bacteria (10(5)MPN mL(-1)) was not detected. From the SMA results obtained, acetoclastic (2.39 g COD-CH(4)g(-1)VSS(-1)day(-1)) and hydrogenophilic (0.94 g COD-CH(4)g(-1)VSS(-1)day(-1)) transformations as possible metabolic pathways used by methanogenic bacteria is suggested from the SMA results obtained. Methanotrix sp., Methanosarcina sp., Methanoccocus sp. and Methanobacterium sp. were identified.     

Effects of lipid concentration on anaerobic co-digestion of municipal biomass wastes

The influence of the lipid concentration on the anaerobic co-digestion of municipal biomass waste and waste-activated sludge was assessed by biochemical methane potential (BMP) tests and by bench-scale tests in a mesophilic semi-continuous stirred tank reactor. The effect of increasing the volatile solid (VS) concentration of lipid from 0% to 75% was investigated. BMP tests showed that lipids in municipal biomass waste could enhance the methane production. The results of bench-scale tests showed that a lipids concentration of 65% of total VS was the inhibition concentration. Methane yields increased with increasing lipid concentration when lipid concentrations were below 60%, but when lipid concentration was set as 65% or higher, methane yields decreased sharply. When lipid concentrations were below 60%, the pH values were in the optimum range for the growth of methanogenic bacteria and the ratios of volatile fatty acid (VFA)/alkalinity were in the range of 0.2–0.6. When lipid concentrations exceeded 65%, the pH values were below 5.2, the reactor was acidized and the values of VFA/alkalinity rose to 2.0. The amount of Brevibacterium decreased with increasing lipid content. Long chain fatty acids stacked on the methanogenic bacteria and blocked the mass transfer process, thereby inhibiting anaerobic digestion.     

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.     

Dry anaerobic digestion in batch mode: Design and operation of a laboratory-scale,completely mixed reactor

A laboratory-scale (40 l) reactor was designed to investigate dry anaerobic digestion. The reactor is equipped with an intermittent paddle mixer, enabling complete mixing in the reactor. Three consecutive batch dry digestion tests of municipal solid waste were performed under mesophilic conditions and compared to operation results obtained on a pilot-scale (21 m³) with the same feedstock. Biogas and methane production at the end of the tests were similar (around 200 m³ CH₄STP/tVS), and the dynamics of methane production and VFA accumulation concurred. However, the maximal levels of VFA transitory accumulation varied between reactors and between runs in a same reactor. Ammonia levels were similar in both reactors. These results show that the new reactor accurately imitates the conditions found in larger ones. Adaptation of micro-organisms to the waste and operating conditions was also pointed out along the consecutive batches.     

Anaerobic co-digestion of food waste and landfill leachate in single-phase batch reactors

In order to investigate the effect of raw leachate on anaerobic digestion of food waste, co-digestions of food waste with raw leachate were carried out. A series of single-phase batch mesophilic (35 ± 1 °C) anaerobic digestions were performed at a food waste concentration of 41.8 g VS/L. The results showed that inhibition of biogas production by volatile fatty acids (VFA) occurred without raw leachate addition. A certain amount of raw leachate in the reactors effectively relieved acidic inhibition caused by VFA accumulation, and the system maintained stable with methane yield of 369–466 mL/g VS. Total ammonia nitrogen introduced into the digestion systems with initial 2000–3000 mgNH₄–N/L not only replenished nitrogen for bacterial growth, but also formed a buffer system with VFA to maintain a delicate biochemical balance between the acidogenic and methanogenic microorganisms. UV spectroscopy and fluorescence excitation–emission matrix spectroscopy data showed that food waste was completely degraded.We concluded that using raw leachate for supplement water addition and pH modifier on anaerobic digestion of food waste was effective. An appropriate fraction of leachate could stimulate methanogenic activity and enhance biogas production.     

Degradation of lignocellulosic materials under sulfidogenic and methanogenic conditions

Biochemical methane potential (BMP) assays, typically used to assess anaerobic biodegradability of liquid wastes with added nutrients and bacteria, were adapted to compare hydrolysis of lignocellulosic material under sulfidogenic and methanogenic environments. A method based on selective inhibition of microorganism activity, by 3% toluene, was used to measure the hydrolysis rate of lignocellulosic material and the accumulation of sugar. The neutral sugars, galactose, glucose, and xylose, which were released from lignocellulosic material such as office paper and newspaper in the presence 3% toluene, clearly accumulated over time under sulfidogenic conditions. The accumulation rates of sugars, glucose, and xylose, were higher in the sulfidogenic condition than in the methanogenic condition, indicating a faster degradation of lignocellulosic materials under the sulfidogenic condition.     

Differences in volatile methyl siloxane (VMS) profiles in biogas from landfills and anaerobic digesters and energetics of VMS transformations

The objectives of this study were to compare the types and levels of volatile methyl siloxanes (VMS) present in biogas generated in the anaerobic digesters and landfills, evaluate the energetics of siloxane transformations under anaerobic conditions, compare the conditions in anaerobic digesters and municipal solid waste (MSW) landfills which result in differences in siloxane compositions. Biogas samples were collected at the South District Wastewater Treatment Plant and South Dade Landfill in Miami, Florida. In the digester gas, D4 and D5 comprised the bulk of total siloxanes (62% and 27%, respectively) whereas in the landfill gas, the bulk of siloxanes were trimethylsilanol (TMSOH) (58%) followed by D4 (17%). Presence of high levels of TMSOH in the landfill gas indicates that methane utilization may be a possible reaction mechanism for TMSOH formation. The free energy change for transformation of D5 and D4 to TMSOH either by hydrogen or methane utilization are thermodynamically favorable. Either hydrogen or methane should be present at relatively high concentrations for TMSOH formation which explains the high levels present in the landfill gas. The high bond energy and bond distance of the Si–O bond, in view of the atomic sizes of Si and O atoms, indicate that Si atoms can provide a barrier, making it difficult to break the Si–O bonds especially for molecules with specific geometric configurations such as D4 and D5 where oxygen atoms are positioned inside the frame formed by the large Si atoms which are surrounded by the methyl groups.     

Transformation of phthalates in young landfill cells

Phthalic acid diesters are additives in a variety of materials that can end up in landfills. Leachates from a series of full-scale young landfill cells were analysed over time for dimethyl, diethyl, dibutyl, butylbenzyl, and di(2-ethylhexyl) phthalate (respectively designated DMP, DEP, DBP, BBP, and DEHP), and their corresponding monoesters monomethyl, monoethyl, monobutyl, monobenzyl, and mono(2-ethylhexyl) phthalate (MMP, MEP, MbutP, MbenzP, and MEHP, respectively), as well as o-phthalic acid (PA). One landfill cell was created in each of three consecutive years by deposition of the same type of waste in July and August. The pH, volatile fatty acids (VFAs), and total organic carbon (TOC) were measured to characterise development of the degradation phases in three landfill cells, which revealed early acidogenic to initial methanogenic stages. Analysis of the phthalate compounds showed that observed concentrations of the degradation products were below the detection limit in the acidogenic leachates but exceeded concentrations of their corresponding diesters in leachates from cells in the initial methanogenic phase. Maximum and average concentrations of phthalic acid were 50 and 23 mg/l, respectively, and the corresponding values for the other phthalates were 430 and 27 microg/l. The concentrations of all phthalates decreased during the establishment of stable methanogenic conditions.     

Pilot project of biogas production from pig manure and urine mixture at ambient temperature in Ventanilla (Lima, Peru)

Parque Porcino de Ventanilla has an extension of 840ha with 2200 farmers dedicated to pig production. There is a lack of services in the area (i.e., water supply, electricity, or waste collection). Anaerobic treatment of pig manure would replace current dumping and incineration, reducing environmental pollution and hazards to public health, as well as providing an organic fertilizer and biogas. The objective of the present work was to study the viability of ambient temperature anaerobic digestion of pig manure diluted in urine, by means of on-site pilot scale reactors. The final goal was to establish design parameters for anaerobic digesters to be implemented; since it was part of a project to improve life conditions for the farmers through the incorporation of better management techniques. Experiments were carried out in a low-cost pilot plant, which consists of three anaerobic digesters (225L total volume), without heating or agitation, placed in a greenhouse. The start-up of the digestion process was performed with a mixture of temperature adapted pig manure-sludge and fresh rumen, and showed a good performance regardless of the dilution of pig manure with water or urine, which is a key parameter due to the scarcity of water in the area under study.     

Report: anaerobic digestion of a sulphate-rich high-strength landfill leachate: the effect of differential dosing with FeCl3.

The effect of different dose concentrations of FeCl3 on the performance of a mesophilic anaerobic digester treating a highly alkaline, high-strength and sulphate-rich landfill leachate was tested. The results indicated that sulphate reduction was the predominant reaction and methanogenic processes were entirely inhibited in the reactor without FeCl3 addition. Adding FeCl3 into the reactor restored some methanogenic activity and also improved the rates of sulphate reduction. A combination of sulphate reduction and methanogenic activity resulted in up to 75% chemical oxygen demand (COD) removal and 85% sulphate removal. Sulphate reduction remained the principle mechanism by which COD removal took place with a methane yield of only between 0.14 and 0.18 m3 CH4 kg(-1) COD removed being achieved. The process was, however, stable and offered advantages for the further treatment or conveyance of the anaerobically treated leachate.     

Two-phase anaerobic digestion within a solid waste/wastewater integrated management system

A two-phase, wet anaerobic digestion process was tested at laboratory scale using mechanically pre-treated municipal solid waste (MSW) as the substrate. The proposed process scheme differs from others due to the integration of the MSW and wastewater treatment cycles, which makes it possible to avoid the recirculation of process effluent. The results obtained show that the supplying of facultative biomass, drawn from the wastewater aeration tank, to the solid waste acidogenic reactor allows an improvement of the performance of the first phase of the process which is positively reflected on the second one. The proposed process performed successfully, adopting mesophilic conditions and a relatively short hydraulic retention time in the methanogenic reactor, as well as high values of organic loading rate. Significant VS removal efficiency and biogas production were achieved. Moreover, the methanogenic reactor quickly reached optimal conditions for a stable methanogenic phase. Studies conducted elsewhere also confirm the feasibility of integrating the treatment of the organic fraction of MSW with that of wastewater.     

Thickness of the buffer layer in the 'reaction front' model of solid-state digestion

Conventional models of anaerobic digestion assume dispersed reaction, in the aqueous phase and at solid-liquid interfaces. However, a second, localized mechanism might operate in a bed of nutrient-rich wastes, such as occurs in many landfills and some waste digesters. A 'reaction front' (comprising two distinct reaction zones separated by a buffer zone) might develop around 'seed bodies' (local concentrations of methanogenic inoculum) then propagate through the waste. The initial conditions, notably the mode of seeding, would determine whether which mechanism is dominant. This paper presents a method of predicting the thickness of such a front and indicates a thickness of the order of 3.5 cm. This would suggest that the seed material must be present in the form of 'seed bodies' of over 7 cm diameter or layers over 7 cm thick. The probability of the formation of a reaction front is discussed, as are the implications for operational practice.     

Regulating the hydrolysis of organic wastes by micro-aeration and effluent recirculation

In this study, the effects of micro-aeration and liquid recirculation on the hydrolysis of vegetable and flower wastes during two-phase solid–liquid anaerobic digestion were assessed. To accomplish this, we evaluated the hydrolysis of five batches of waste that were treated under the following conditions: anaerobic, insufficient micro-aeration (aeration for 5 min every 24 h), and sufficient micro-aeration (aeration for 5 min every 12, 4 and 1 h). Hydrolysis was found to depend on the level of micro-aeration. Specifically, insufficient micro-aeration led to unstable and decreased performance. Conversely, sufficient micro-aeration promoted the hydrolysis of easily biodegradable carbohydrates and proteins, but the microbial activity was later impaired by liquid recirculation using methanogenic effluent. The hydrolysis efficiency under anaerobic conditions was comparable to the efficiency observed under sufficient micro-aeration, while the cumulative TOC of the anaerobic batch was 1.4–2.4 times higher than that of the micro-aerated batches. In addition, liquid recirculation did not have a negative effect on the development of microbial activity under anaerobic conditions, which resulted in the lignocelluloses having a higher hydrolysis efficiency.     

Methane production potential of leachate generated from Korean food waste recycling facilities: a lab-scale study

This paper examines the applicability of food waste leachate (FWL) in bioreactor landfills or anaerobic digesters to produce methane as a sustainable solution to the persisting leachate management problem in Korea. Taking into account the climatic conditions in Korea and FWL characteristics, the effect of key parameters, viz., temperature, alkalinity and salinity on methane yield was investigated. The monthly average moisture content and the ratio of volatile solids to total solids of the FWL were found to be 84% and 91%, respectively. The biochemical methane potential experiment under standard digestion conditions showed the methane yield of FWL to be 358 and 478 ml/g VS after 10 and 28 days of digestion, respectively, with an average methane content of 70%. Elemental analysis showed the chemical composition of FWL to be C(13.02)H(23.01)O(5.93)N(1). The highest methane yield of 403 ml/g VS was obtained at 35 degrees C due to the adaptation of seed microorganisms to mesophilic atmosphere, while methane yields at 25, 45 and 55 degrees C were 370, 351 and 275 ml/g VS, respectively, at the end of 20 days. Addition of alkalinity had a favorable effect on the methane yield. Dilution of FWL with salinity of 2g/l NaCl resulted in 561 ml CH(4)/g VS at the end of 30 days. Considering its high biodegradability (82.6%) and methane production potential, anaerobic digestion of FWL in bioreactor landfills or anaerobic digesters with a preferred control of alkalinity and salinity can be considered as a sustainable solution to the present emergent problem.