Effects of organic loading rate and hydraulic retention time on bioaugmentation performance to tackle ammonia inhibition in anaerobic digestion.

Three continuously stirred-tank reactors fed with manure operating under high ammonia levels (5.0 g NH3-N L-1) and with increased organic loading rate (OLR), (2.09 R1, 3.02 R2 and 4.0 R3 g VS L-1 d-1), achieved through glucose amendment in R2 and R3, were inoculated with an ammonia-acclimatized microbial culture. Successful bioaugmentation was endured only in R2 and R3, both reactors characterized by high OLR, resulting in 19.6 and 24.5% increase in methane production, respectively. The high OLRs in these reactors favored the co-occurrence of the hydrogenotrophic (Methanobacteriaceae), methylotrophic (Methanomethylophilaceae) and aceticlastic methanogenic pathways. The latter was supported by the successful establishment of ammonium-tolerant Methanosarcina, prevailing in the inoculum. Oppositely in R1, the low OLR prevented the establishment of Methanosarcina, leading to an exclusive hydrogenotrophic methanogenesis and reduced methane production. HRT shortening resulted in limited effect on biomethane performance, indicating a well establishment of the introduced bioaugmentation culture in the reactors.

Ammonia-induced inhibition of manure-based continuous biomethanation process under different organic loading rates and associated microbial community dynamics.

Three Continuously Stirred Tank Reactors (CTSRs) were operating at steady state conditions with Organic Loading Rates (OLR) of 2.09, 3.024 and 4.0 g VS L-1 d-1. Glucose was used as the sole factor for increasing the OLR, linking the increase of the OLR with the C/N ratio increase. The reactors were stressed by increasing the ammonia concentration to 5 g L-1 from 1.862 g L-1. The results showed elevating inhibition of the anaerobic process by increasing the C/N ratio just by increasing the OLR, under the high ammonia concentration. A different response of the bacterial and archaeal community under ammonia stressed conditions was also observed. Under the high ammonia concentration, hydrogen-depended methylotrophic was the dominant methanogenesis route at OLR of 2.09 g VS L-1d-1, while the hydrogenotrophic route was the dominant at the high OLR of 4 g VS L-1d-1, which coincided with high acetate and propionate concentrations.

Biological CO2 fixation in up-flow reactors via exogenous H2 addition.

Gas fermentation for the production of building block molecules and biofuels is lately gaining attention as a means to eliminate the greenhouse gases emissions. Especially CO2 capture and recycling are in focus. Thus, the biological coupling of CO2 and H2 is of high interest. Therefore, the focus of the present work was to evaluate the performances of two up-flow reactors for CO2 and H2 assimilation. Process monitoring showed that the gas-liquid H2 transfer was highly affected by reactor design. A reactor filled with Raschig rings could lift up gases utilization leading to a CH4 content of 81% at 6 h gas retention time and 8.8 L/LR.h gas recirculation rate. In contrast, limited biomethanation was achieved in the absence of Raschig rings highlighting the positive role of packing material to the performance of up-flow-reactors. Additionally, high-throughput 16S rRNA sequencing revealed that the microbial community was ultimately resided by Methanothermobacter methanogens.

Two-year microbial adaptation during hydrogen-mediated biogas upgrading process in a serial reactor configuration.

Microbial dynamics in an upgrading biogas reactor system undergoing a more than two years-period at stable operating conditions were explored. The carbon dioxide generated during biomass degradation in the first reactor of the system was converted to methane into the secondary reactor by addition of external hydrogen. Considering the overall efficiency, the long-term operation period resulted in an improved biogas upgrading performance (99% methane content). However, a remarkable accumulation of acetate was revealed, indicating the enhancement of homoacetogenic activity. For this reason, a shift in the anaerobic digestion microbiome was expected and evaluated by 16S rRNA amplicon analysis. Results demonstrated that the most abundant archaeal species identified in the first time point, Candidatus Methanoculleus thermohydrogenotrophicum, was replaced by Methanothermobacter thermautotrophicus, becoming dominant after the community adaptation. The most interesting taxonomic units were clustered by relative abundance and six main long-term adaptation trends were found, characterizing functionally related microbes (e.g. homoacetogens).

Bioaugmentation with hydrolytic microbes to improve the anaerobic biodegradability of lignocellulosic agricultural residues.

Bioaugmentation with hydrolytic microbes was applied to improve the methane yield of bioreactors fed with agricultural wastes. The efficiency of Clostridium thermocellum and Melioribacter roseus to degrade lignocellulosic matter was evaluated in batch and continuously stirred tank reactors (CSTRs). Results from batch assays showed that C. thermocellum enhanced the methane yield by 34%. A similar increase was recorded in CSTR during the bioaugmentation period; however, at steady-state the effect was noticeably lower (7.5%). In contrast, the bioaugmentation with M. roseus did not promote markedly the anaerobic biodegradability, as the methane yield was increased up to 10% in batch and no effect was shown in CSTR. High-throughput 16S rRNA amplicon sequencing was used to assess the effect of bioaugmentation strategies on bacterial and archaeal populations. The microbial analysis revealed that both strains were not markedly resided into biogas microbiome. Additionally, the applied strategies did not alter significantly the microbial communities.

Improving the energy balance of grass-based anaerobic digestion through combined harvesting and pretreatment.

An important challenge that has to be addressed to achieve sustainable anaerobic digestion of lignocellulosic substrates is the development of energy and cost efficient pretreatment methods. Technologies orientated to simultaneously harvest and mechanically pretreat the biomass at the field could meet these criteria as they can potentially reduce the energy losses. The objective of this study was to elucidate the effect of two full-scale harvesting machines to enhance the biogas production and subsequently, improve energy balance. The performances of Disc-mower and Excoriator were assessed on meadow and cultivated grass silages. The results showed that relatively high methane production can be achieved from meadow and cultivated grass harvested in different seasons. The findings indicated that the bioenergy production can be improved based on the selection of the appropriate harvesting technology. More specifically, Excoriator, which cuts and subsequently applies shearing forces on harvested biomass, enhanced the methane production up to 10% and the overall energy budget was improved proportionally to the driving speed increase.

Effect of micro-aeration and inoculum type on the biodegradation of lignocellulosic substrate.

The effect of various micro-aeration strategies on the anaerobic digestion (AD) of wheat straw was thoroughly examined using a mixture of inocula, containing compost and well digested sludge from biogas plant. The aim was to determine the most efficient oxygen load, pulse repetition and treatment duration, resulting in the highest methane production. The oxygen load had the largest impact on the biodegradability of straw, among the examined variables. More specifically, a micro-aeration intensity of 10mLO2/gVS was identified as the critical threshold above which the AD performance was more susceptible to instability. The highest enhancement in biogas production was achieved by injecting 5mLO2/gVS for a consecutive 3-day treatment period, presenting a 7.2% increase compared to the untreated wheat straw. Nevertheless, the results from optimisation case study indicated a higher increase of 9% by injecting 7.3mLO2/gVS, distributed in 2 pulses during a slightly shorter treatment period (i.e. 47h).

Optimising the anaerobic co-digestion of urban organic waste using dynamic bioconversion mathematical modelling.

Mathematical anaerobic bioconversion models are often used as a convenient way to simulate the conversion of organic materials to biogas. The aim of the study was to apply a mathematical model for simulating the anaerobic co-digestion of various types of urban organic waste, in order to develop strategies for controlling and optimising the co-digestion process. The model parameters were maintained in the same way as the original dynamic bioconversion model, albeit with minor adjustments, to simulate the co-digestion of food and garden waste with mixed sludge from a wastewater treatment plant in a continuously stirred tank reactor. The model's outputs were validated with experimental results obtained in thermophilic conditions, with mixed sludge as a single substrate and urban organic waste as a co-substrate at hydraulic retention times of 30, 20, 15 and 10 days. The predicted performance parameter (methane productivity and yield) and operational parameter (concentration of ammonia and volatile fatty acid) values were reasonable and displayed good correlation and accuracy. The model was later applied to identify optimal scenarios for an urban organic waste co-digestion process. The simulation scenario analysis demonstrated that increasing the amount of mixed sludge in the co-substrate had a marginal effect on the reactor performance. In contrast, increasing the amount of food waste and garden waste resulted in improved performance.

Improving methane production from digested manure biofibers by mechanical and thermal alkaline pretreatment.

Animal manure digestion is associated with limited methane production, due to the high content in fibers, which are hardly degradable lignocellulosic compounds. In this study, different mechanical and thermal alkaline pretreatment methods were applied to partially degradable fibers, separated from the effluent stream of biogas reactors. Batch and continuous experiments were conducted to evaluate the efficiency of these pretreatments. In batch experiments, the mechanical pretreatment improved the degradability up to 45%. Even higher efficiency was shown by applying thermal alkaline pretreatments, enhancing fibers degradability by more than 4-fold. In continuous experiments, the thermal alkaline pretreatment, using 6% NaOH at 55°C was proven to be the most efficient pretreatment method as the methane production was increased by 26%. The findings demonstrated that the methane production of the biogas plants can be increased by further exploiting the fraction of the digested manure fibers which are discarded in the post-storage tank.

Co-digestion of food and garden waste with mixed sludge from wastewater treatment in continuously stirred tank reactors.

Co-digestions of urban organic waste were conducted to investigate the effect of the mixing ratio between sludge, food waste, grass clippings and green waste at different hydraulic retention times (HRTs). Compared to the digestion of 100% sludge, the methane yield increased by 48% and 35%, when co-digesting sludge with food waste, grass clippings and garden waste with a corresponding %VS of 10:67.5:15.75:6.75 (R1) and 10:45:31.5:13.5 (R2), respectively. The methane yield remained constant at around 425 and 385 NmL CH4/g VS in R1 and R2, respectively, when the reactors were operated at HRTs of 15, 20 and 30 days. However, the methane yield dropped significantly to 356 (R1) and 315 (R2) NmL CH4/g VS when reducing the HRT to 10 days, indicating that the process was stressed. Since the methane production rate improved significantly with decreasing HRT, the trade-off between yield and productivity was obtained at 15 days HRT.

Counteracting foaming caused by lipids or proteins in biogas reactors using rapeseed oil or oleic acid as antifoaming agents.

Foaming is one of the major operational problems in biogas plants, and dealing with foaming incidents is still based on empirical practices. Various types of antifoams are used arbitrarily to combat foaming in biogas plants, but without any scientific support this action can lead to serious deterioration of the methanogenic process. Many commercial antifoams are derivatives of fatty acids or oils. However, it is well known that lipids can induce foaming in manure based biogas plants. This study aimed to elucidate the effect of rapeseed oil and oleic acid on foam reduction and process performance in biogas reactors fed with protein or lipid rich substrates. The results showed that both antifoams efficiently suppressed foaming. Moreover rapeseed oil resulted in stimulation of the biogas production. Finally, it was reckoned that the chemical structure of lipids, and more specifically their carboxylic ends, is responsible for their foam promoting or foam counteracting behaviour. Thus, it was concluded that the fatty acids and oils could suppress foaming, while salt of fatty acids could generate foam.

Biogas production from ensiled meadow grass; effect of mechanical pretreatments and rapid determination of substrate biodegradability via physicochemical methods.

As the biogas sector is rapidly expanding, there is an increasing need in finding new alternative feedstock to biogas plants. Meadow grass can be a suitable co-substrate and if ensiled it can be supplied to biogas plants continuously throughout the year. Nevertheless, this substrate is quite recalcitrant and therefore efficient pretreatment is needed to permit easy access of microbes to the degradable components. In this study, different mechanical pretreatment methods were applied on ensiled meadow grass to investigate their effect on biomass biodegradability. All the tested pretreatments increased the methane productivity and the increase ranged from 8% to 25%. The best mechanical pretreatment was the usage of two coarse mesh grating plates. Additionally, simple analytical methods were conducted to investigate the possibility of rapidly determining the methane yield of meadow grass. Among the methods, electrical conductivity test showed the most promising calibration statistics (R(2)=0.68).

Anaerobic digestion foaming in full-scale biogas plants: a survey on causes and solutions.

Anaerobic digestion foaming is a common operation problem in biogas plants with negative impacts on the biogas plants economy and environment. A survey of 16 Danish full-scale biogas plants on foaming problems revealed that most of them had experienced foaming in their processes up to three times per year. Foaming incidents often lasted from one day to three weeks, causing 20-50% biogas production loss. One foaming case at Lemvig biogas plant has been investigated and the results indicated that the combination of feedstock composition and mixing pattern of the reactor was the main cause of foaming in this case. Moreover, no difference in bacterial communities between the foaming and non-foaming reactors was observed, showing that filamentous bacteria were not the main reason for foaming in this case.

Foam suppression in overloaded manure-based biogas reactors using antifoaming agents.

Foam control is an imperative need in biogas plants, as foaming is a major operational problem. In the present study, the effect of oils (rapeseed oil, oleic acid, and octanoic acid) and tributylphosphate on foam reduction and process performance in batch and continuous manure-based biogas reactors was investigated. The compounds were tested in dosages of 0.05%, 0.1% and 0.5% v/vfeed. The results showed that rapeseed oil was most efficient to suppress foam at the dosage of 0.05% and 0.1% v/vfeed, while octanoic acid was most efficient to suppress foam at dosage of 0.5% v/vfeed. Moreover, the addition of rapeseed oil also increased methane yield. In contrast, tributylphosphate, which was very efficient antifoam, was found to be inhibitory to the biogas process.

Antifoaming effect of chemical compounds in manure biogas reactors.

A precise and efficient antifoaming control strategy in bioprocesses is a challenging task as foaming is a very complex phenomenon. Nevertheless, foam control is necessary, as foam is a major operational problem in biogas reactors. In the present study, the effect of 14 chemical compounds on foam reduction was evaluated at concentration of 0.05%, 0.1% and 0.5% v/v(sample), in raw and digested manure. Moreover, two antifoam injection methods were compared for foam reduction efficiency. Natural oils (rapeseed and sunflower oil), fatty acids (oleic, octanoic and derivative of natural fatty acids), siloxanes (polydimethylsiloxane) and ester (tributylphosphate) were found to be the most efficient compounds to suppress foam. The efficiency of antifoamers was dependant on their physicochemical properties and greatly correlated to their chemical characteristics for dissolving foam. The antifoamers were more efficient in reducing foam when added directly into the liquid phase rather than added in the headspace of the reactor.

Effect of organic loading rate and feedstock composition on foaming in manure-based biogas reactors.

Foaming is one of the major problems that occasionally occur in biogas plants, affecting negatively the overall digestion process. In the present study, the effect of organic loading rate (OLR) and feedstock composition on foaming was elucidated in continuous reactor experiments. By stepwise increasing the OLR and the concentration of proteins or lipids in the substrate, foaming in biogas reactors was investigated. No foam formation was observed at the OLR of 3.5 g volatile solids/(L-reactor·day). Organic loading was the main factor affecting foam formation in manure digester, while the organic composition, such as content of proteins or lipids were factors that in combination with the organic loading were triggering foaming. More specifically, gelatine could initiate foam formation at a lower OLR than sodium oleate. Moreover, the volume of foam produced by gelatine was relatively stable and was not increased when further increasing either OLR or gelatine concentration in the feed.

Pilot-scale application of an online VFA sensor for monitoring and control of a manure digester.

A volatile fatty acids (VFA) sensor based on headspace chromatography was tested for online monitoring and control of a pilot-scale manure digester. The sensor showed satisfying results in terms of sensitivity and reliability for monitoring of the digester. The online VFA and biogas production data were used for automatic control of the digester based on feed flow manipulation. The control approach was based on optimization of biogas production while using VFA concentration as the alarm threshold. A rule-based supervisory system with a cascade controller was used to optimize the biogas production from the digester. The alarm state was set at 40 mM total VFA and 10 mM propionate concentration. The control algorithms could successfully maximize the biogas production without overloading the process. However, as the algorithm was based on a fixed biogas yield parameter and only used the biogas parameter for optimization, it could not distinguish between the decreases of biogas production from inhibition and from lower organic content in the substrate, which resulted in undesired decreasing of the control gas setpoint when the substrate was diluted. It was necessary to adjust the yield parameter in order to get this control approach to function properly, which is not suitable for the full-scale biogas plant where the organic content of waste streams can vary. An alternative approach could be a modified rule-based algorithm that includes VFA parameters to help distinguish between different process scenarios.

Effect of substrates and intermediate compounds on foaming in manure digestion systems.

Manure contains several compounds that can potentially cause foaming during anaerobic digestion. Understanding the effect of substrates and intermediate compounds on foaming tendency and stability could facilitate strategies for foaming prevention and recovery of the process. In this study, the effect of physicochemical properties of substrates and intermediate compounds on liquid properties such as surface tension, surfactant property, and hydrophobicity were investigated and compared with the effect on foaming tendency and foam stability. The results showed that there was no consistent correlation between foaming potential and hydrophobicity, oil displacement area (ODA) or surface tension of the tested solutions, and the best way to determine the foaming property of the solution was to directly measure foaming tendency and foam stability. Na-oleate and acetic acid showed the highest potential to create foam in a manure digester. Moreover, high organic loading of lipids and protein, and high concentrations of acetic and butyric acids also showed a strong tendency to create foaming during anaerobic digestion. Due to their great ability to stabilize foam, high organic loadings of Na-oleate or gelatine were considered to be the main potential foaming problem.

Long-chain fatty acids inhibition and adaptation process in anaerobic thermophilic digestion: batch tests, microbial community structure and mathematical modelling.

Biomass samples taken during the continuous operation of thermophilic anaerobic digestors fed with manure and exposed to successive inhibitory pulses of long-chain fatty acids (LCFA) were characterized in terms of specific metabolic activities and 16S rDNA DGGE profiling of the microbial community structure. Improvement of hydrogenotrophic and acidogenic (beta-oxidation) activity rates was detected upon successive LCFA pulses, while different inhibition effects over specific anaerobic trophic groups were observed. Bioreactor recovery capacity and biomass adaptation to LCFA inhibition were verified. Population profiles of eubacterial and archaeal 16S rDNA genes revealed that no significant shift on microbial community composition took place upon biomass exposure to LCFA. DNA sequencing of predominant DGGE bands showed close phylogenetic affinity to ribotypes characteristic from specific beta-oxidation bacterial genera (Syntrophomonas and Clostridium), while a single predominant syntrophic archaeae was related with the genus Methanosarcina. The hypothesis that biomass adaptation was fundamentally of physiological nature was tested using mathematical modelling, taking the IWA ADM1 as general model. New kinetics considering the relation between LCFA inhibitory substrate concentration and specific biomass content, as an approximation to the adsorption process, improved the model fitting and provided a better insight on the physical nature of the LCFA inhibition process.