A strict anaerobic extreme thermophilic hydrogen-producing culture enriched from digested household waste

Aims:  The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H₂) producers from digested household solid wastes.
Methods and Results:  A strict anaerobic extreme thermophilic H₂ producing bacterial culture was enriched from a lab-scale digester treating household wastes at 70°C. The enriched mixed culture consisted of two rod-shaped bacterial members growing at an optimal temperature of 80°C and an optimal pH 8·1. The culture was able to utilize glucose, galactose, mannose, xylose, arabinose, maltose, sucrose, pyruvate and glycerol as carbon sources. Growth on glucose produced acetate, H₂ and carbon dioxide. Maximal H₂ production rate on glucose was 1·1 mmol l⁻¹ h⁻¹ with a maximum H₂ yield of 1·9 mole H₂ per mole glucose. 16S ribosomal DNA clone library analyses showed that the culture members were phylogenetically affiliated to the genera Bacillus and Clostridium. Relative abundance of the culture members, assessed by fluorescence in situ hybridization, were 87 ± 5% and 13 ± 5% for Bacillus and Clostridium , respectively.
Conclusions:  An extreme thermophilic, strict anaerobic, mixed microbial culture with H₂-producing potential was enriched from digested household wastes.
Significance and Impact of the Study:  This study provided a culture with a potential to be applied in reactor systems for extreme thermophilic H₂ production from complex organic wastes.     

A novel process configuration for anaerobic digestion of source-sorted household waste using hyper-thermophilic post-treatment

A novel reactor configuration was investigated for anaerobic digestion (AD) of the organic fraction of municipal solid waste (OFMSW). An anaerobic hyper-thermophilic (68 degrees C) reactor R68 was implemented as a post-treatment step for the effluent of a thermophilic reactor R1 (55 degrees C) in order to enhance hydrolysis of recalcitrant organic matter, improve sanitation and ease the stripping of ammonia from the reactor. The efficiency of the combined system was studied in terms of methane yield, volatile solids (VS) reduction, and volatile fatty acid (VFA) production at different hydraulic retention times (HRT). A single-stage thermophilic (55 degrees C) reactor R2 was used as control. VS reduction and biogas yield of the combined system was 78-89% and 640-790 mL/g VS, respectively. While the VS reduction in the combined system was up to 7% higher than in the single-stage treatment, no increase in methane yield was observed. Shifting the HRT of the hyper-thermophilic reactor from 5 days to 3 days resulted in a drop in the methanogenic activity in the hydrolysis reactor to a minimum. Operation of R68 at HRTs of 24-48 h was sufficient to achieve high VS conversion into VFAs. Removal of pathogens was enhanced by the hyper-thermophilic post-treatment. 7% of the ammonia load was removed in the hyper-thermophilic reactor with a flow of headspace gas through the reactor equivalent to four times the biogas flow produced in reactor R1.     

Limitations of thermophilic anaerobic wastewater treatment and the consequences for process design

Thermophilic anaerobic digestion offers an attractive alternative for the treatment of medium- and high-strength wastewaters. However, literature reports reveal that thermophilic wastewater treatment systems are often more sensitive to environmental changes than the well-defined high-rate reactors at the mesophilic temperature range. Also, in many cases a poorer effluent quality is experienced while the carry over of suspended solids in the effluent is relatively high. In this paper recent achievements are discussed regarding the process stability of thermophilic anaerobic wastewater treatment systems. Laboratory experiments reveal a relatively low sensitivity to temperature changes if high-rate reactors with immobilized biomass are used. Other results show that if a staged process is applied, thermophilic reactors can be operated for prolonged periods of time under extreme loading conditions (80–100 kg chemical oxygen demand.m^-3.day^-1), while the concentrations of volatile fatty acids in the effluent remain at a low level.     

Thermophilic anaerobic digestion of source-sorted household solid waste: the effects of enzyme additions

Thermophilic (55° C) methanation of source-sorted household solid waste (HSW) was studied in batch and in continuous experiments. Furthermore, the effects of additions of xylanase, lipase, protease and a mixture of these on the methanation were tested. In the batch studies, comparative assays with active and inactive enzymes were used to elucidate the role of the added enzymes. The results showed that the HSW was readily digestible, up to 400–590 mlCH₄·g^–1 volatile solids (VS) was produced. Only with protease added, at a concentration of 1.1 Anson protease units·kg^–1 VS was a higher specific methanogenic activity found with active enzymes compared to inactive (autoclaved) enzymes or without enzyme addition. The methane yield by conversion of the HSW in the batch assays and in the reactor studies was not increased by enzyme additions (enzyme mixture). Correspondence to: B. K. Ahring     

Super blue box recycling (SUBBOR) enhanced two-stage anaerobic digestion process for recycling municipal solid waste: laboratory pilot studies

The super blue box recycling (SUBBOR) process is an enhanced, multi-stage anaerobic digestion process for mixed municipal solid waste (MSW) and other biomass feedstock materials. The technology centers on enhanced high solids, thermophilic digestion after steam-pressure disruption of the ligno-cellulosic fiber components that are recalcitrant to conventional anaerobic digestion. Mixed MSW, rich in organic components but also containing inorganic materials, such as glass, aluminum and steel, as well as non-digestible plastic materials, has been laboratory pilot tested with a fully integrated process train designed to treat and recycle all of the MSW components. Methane yields from the MSW were 0.36 m3 CH4/kg volatile solids (VS) representing a 40% increase over the yield obtained from conventional single stage digestion. The secondary digestion step after steam pressure disruption also provided a 40% improvement in total solids and VS reduction. The residual organic fraction following two-stage digestion was fine in texture and was recovered as a clean peat fraction with reduced contents of heavy metal and other fugitive non-digested contaminants. Mass and energy balance determinations indicated a high degree of MSW diversion from landfill disposal (>80%) was achievable by the SUBBOR process as well as substantial net electrical and thermal energy production. Continuous long-term trials of the SUBBOR process at 25,000 tonnes/year are underway.     

Treatment of leachate from a solid waste landfill site using a two-stage anaerobic filter

Raw leachate was treated using a two-stage upflow anaerobic filter process. Leachate from a solid waste landfill site, which received both municipal and industrial wastes, contained high organic matter (17-21 g/L COD, 13-14 g/L BOD, and 3.5-4.6 g/L volatile acids), and low metal (Zn and Fe) concentrations. Depending on sampling time, leachate composition and characteristics varied considerably. At an organic loading up to 4 g COD/day(2) media area, the BOD and COD removal percentages were 98 and 91%, respectively. The biofilters were also effective for metal removal. However, the filter effluent contained a high concentration of ammonia. System overloading was characterized by the accumulation of large quantities of volatile acids and by a now ratio of alkalinity/volatile acids, resulting in low COD removal and reduced gas production. Once the first filter was upset, the second stage could only partially respond to the volatile acids accumulated in the effluent of first filter.     

Co-production of hydrogen and methane from potato waste using a two-stage anaerobic digestion process

Hydrogen and methane co-production from potato waste was examined using a two-stage process of anaerobic digestion. The hydrogen stage was operated in continuous flow under a pH of 5.5 and a HRT of 6h. The methane stage was operated in both continuous and semi-continuous flows under HRTs of 30 h and 90 h, respectively, with pH controlled at 7. A maximum gas production rate of 270 ml/h and an average of 119 ml/h were obtained from the hydrogen stage during the operation over 110 days. The hydrogen concentration contained in the gas was 45% (v/v), on average. The maximum and average gas production rates observed from methane reactor during the 74 days of semi-continuous flow operation were 187 and 141 ml/h, respectively, with an average methane concentration of 76%. Overall, 70% of VS, 64% of total COD in the feedstock were removed. The hydrogen and methane yields from the potato waste were 30 l/kg TS (with a maximum of 68 l/kg) and 183 l/kg TS (with a maximum of 225 l/kg), respectively. The total energy yield obtained was 2.14 kW h/kg TS, with a maximum of 2.74 kW h/kg TS.     

Effect of pre-aeration and inoculum on the start-up of batch thermophilic anaerobic digestion of municipal solid waste

In this study, a short pre-aeration step was investigated as pre-treatment for thermophilic anaerobic digestion of the organic fraction of municipal solid waste (OFMSW). It was found that pre-aeration of 48 h generated enough biological heat to increase the temperature of bulk OFMSW to 60 °C. This was sufficient self-heating of the bulk OFMSW for the start-up of thermophilic anaerobic digestion without the need for an external heat source. Pre-aeration also reduced excess easily degradable organic compounds in OFMSW, which were the common cause of acidification during the start-up of the batch system. Careful consideration however must be taken to avoid over aeration as this consumes substrate, which would otherwise be available to methanogens to produce biogas. To accelerate methane production and volatile solids destruction, the anaerobic digestion in this study was operated as a wet process with the anaerobic liquid recycled through the OFMSW. Appropriate anaerobic liquid inoculum was found to be particularly beneficial. It provided high buffer capacity as well as suitable microbial inoculum. As a result, acidification during start-up was kept to a minimum. With volatile fatty acids (VFAs-acetate in particular) and H₂ accumulation typical of hydrolysis and fermentation of the easily degradable substrates during start-up, inoculum with high numbers of hydrogenotrophic methanogens was critical to not only maximise CH₄ production but also reduce H₂ partial pressure in the system to allow VFAs degradation. In a lab-scale bioreactor, the combined pre-aeration and wet thermophilic anaerobic digestion was able to stabilise the OFMSW within a period of only 12 days. The stabilised inert residual material can be used as a soil amendment product.     

Use of respirometer in evaluation of process and toxicity of thermophilic anaerobic digestion for treating kitchen waste

A thermophilic anaerobic digestion (TAnD, 55 degrees C) system was adopted to hydrolyze the kitchen waste for 3 days, which was then fermented for a hydraulic retention time (HRT) of 10 days. The TAnD system performed much better than a similar system without thermal pre-treatment. A bubble respirometer was employed to study the effects of thermal pre-treatment, which showed that pre-treatment at 60 degrees C yielded the highest Total COD (TCOD) removal efficiency (79.2%) after 300h reaction. Respirometer results also indicated that oil and grease (O and G) began to inhibit the TAnD system at a concentration of approximately 1000mg/L and the gas production was inhibited by 50% at a concentration of approximately 7500mg/L of sodium.     

Semi-dry thermophilic anaerobic digestion of the organic fraction of municipal solid waste: focusing on the start-up phase

The paper concerns the results of a pilot-scale study of the simulation of the start-up phase of the thermophilic semi-dry anaerobic digestion of the organic fraction of municipal solid wastes. The aim of the study was to aid and shorten the start-up phase of the full-scale plant (500 t/d) in Verona--Ca' del Bue, where the semi-dry anaerobic digestion process is being used. The substrate used in the experimentation was the mechanically sorted organic fraction of municipal solid waste (MS-OFMSW) enriched with the putrescent fraction from the source sorted OFMSW in order to simulate the substrate which is dealt with in the Verona plant. The results of the pilot scale study agreed with literature data and previous work of the authors: it showed a specific gas production of 0.23 m3/kg TVSfeed and a gas production rate of 2.1 m3/m3 d when operating at a specific organic loading rate of 0.135 kgTVSfeed/kgTVSreacter d. No problems regarding process stability were encountered in the gradual acclimation of the biomass. The design organic loading rate of 9 kg TVSfeed/m3reactor d was reached in about 30 days, during which the total solids content in the feedwas increased. Only a partial comparison with the full scale start-up, which is now in progress, is possible: this shows an initial general concordance with the results found in previous work.     

Batch anaerobic co-digestion of cow manure and waste milk in two-stage process for hydrogen and methane productions

Anaerobic co-digestion of cow manure (CM) and waste milk (WM), produced by sick cows during treatment with antibiotics, was evaluated in two-stage process under thermophilic condition (55 °C) to determine the effect of WM addition on hydrogen (H₂) and methane (CH₄) production potentials, volatile solids (VS) removal, and energy recovery. Six CM to WM VS ratios of 100:0, 90:10, 70:30, 50:50, 30:70, and 10:90 were examined using 1-L batch digesters. The WM VS ratio of 30 % was found to be the minimum limit for significant increases in specific H₂ and CH₄ yields, and VS removal as compared to digestion of manure alone (P < 0.05). The highest specific H₂ and CH₄ yields, VS removal and energy yield were 38.2 mL/g VS, 627.6 mL/g VS, 78.4 % and 25,459.8 kJ/kg VS, respectively, in CM:WM 30:70. Lag phases to H₂ and CH₄ productions were observed in CM–WM mixtures, increased with increasing the amount of WM in the feedstock and were greater than 72 h in CM:WM 50:50 and 30:70. The digestion system failed in CM:WM 10:90. The results suggest that CM:WM 30:70 was optimum, however, due to limited amount of WM usually generated and long lag phase at this ratio which may make the process uneconomical, CM:WM 70:30 is recommended in practice.     

A half-submerged integrated two-phase anaerobic reactor for agricultural solid waste codigestion

Anaerobic digestion is widely used in bioenergy recovery from waste. In this study, a half-submerged, integrated, two-phase anaerobic reactor consisting of a top roller acting as an acidogenic unit and a recycling bottom reactor acting as a methanogenic unit was developed for the codigestion of wheat straw (WS) and fruit/vegetable waste (FVW). The reactor was operated for 21 batches (nearly 300 d). Anaerobic granular sludge was inoculated into the methanogenic unit. The residence time for the mixed waste was maintained as 10 d when the operation stabilized, and the temperature was kept at 35 °C. The highest organic loading rate was 1.37 kg VS/(m³ d), and the maximum daily biogas production was 328 L/d. Volatile solid removal efficiencies exceeded 85%. WS digestion could be confirmed, and efficiency was affected by both the ratio of WS to FVW and the loading rate. The dominant bacteria were Bacteroides-like species, which are involved in glycan and cellulose decomposition. Methanogenic community structures, pH levels, and volatile fatty acid concentrations in the acidogenic and methanogenic units differed, indicating successful phase separation. This novel reactor can improve the mass transfer and microbial cooperation between acidogenic and methanogenic units and can efficiently and steady codigest solid waste.     

Design of acidogenic reactors for the anaerobic treatment of the organic fraction of solid food waste

Volatile Fatty Acids (VFA) production by anaerobic fermentation of organic solid wastes was studied at laboratory scale. The influence of initial substrate concentration was evaluated on VFA production. Completely mixed reactors (0.9?l) were used at mesophilic temperature (35?°C). Food wastes had 43.8% Total Solids content. Three dilutions of substrate (1/25, 1/10 and 1/5) corresponding to 1.75%, 4.38% and 8.76% of Total Solids and five values of Organic Loading Rates: 2, 5, 10, 12.5 and 25?kg COD/m³?d were studied. It was found that substrate 1/10 led to 14?g VFA/l at a loading rate of 12.5?kg COD/m³?d and an hydraulic retention time of 3.7 d. The main VFA produced were especially acetate and butyrate. Substrate diluted 1/5 led to 26.1?g VFA/l at a loading of 5?kg COD/m³?d and an hydraulic retention time of 15.1 d, but biomass production was not optimal. In a second study, a cascade of three reactors was used. An effluent with 42?g VFA/l was obtained at steady-state conditions at a loading of 12.5?kg of COD/m³?d and an hydraulic retention time of 12.5?d. The distribution of VFA was the following: 36% of propionate, 34% of acetate and 22.5% of butyrate.     

Screening of thermophilic anaerobic bacteria for solid substrate cultivation on lignocellulosic substrates

Interest in solid substrate cultivation (SSC) techniques is gaining for biochemical production from renewable resources; however, heat and mass transfer problems may limit application of this technique. The use of anaerobic thermophiles in SSC offers a unique solution to overcoming these challenges. The production potential of nine thermophilic anaerobic bacteria was examined on corn stover, sugar cane bagasse, paper pulp sludge, and wheat bran in submerged liquid cultivation (SmC) and SSC. Production of acetate, ethanol, and lactate was measured over a 10 day period, and total product concentrations were used to compare the performance of different organism-substrate combinations using the two cultivation methods. Overall microbial activity in SmC and SSC was dependent on the organism and growth substrate. Clostridium thermocellum strains JW20, LQRI, and 27405 performed significantly better in SSC when grown on sugar cane bagasse and paper pulp sludge, producing at least 70 and 170 mM of total products, respectively. Growth of C. thermocellum strains in SSC on paper pulp sludge proved to be most favorable, generating at least twice the concentration of total products produced in SmC (p-value < 0.05). Clostridium thermolacticum TC21 demonstrated growth on all substrates producing 30-80 and 60-116 mM of total product in SmC and SSC, respectively. Bacterial species with optimal growth temperatures of 70 degrees C grew best on wheat bran in SmC, producing total product concentrations of 45-75 mM. For some of the organism-substrate combinations total end product concentrations in SSC exceeded those in SmC, indicating that SSC may be a promising alternative for microbial activity and value-added biochemical production.     

Thermophilic anaerobic digestion of solid waste for fuel gas production.

Anaerobic digestion offers a potential means of converting organic solid waste into fuel gas and thereby provide a supplemental and readily utilizable source of energy. We are particularly interested in the use of thermophilic digestion over a mesophilic operation for it can achieve higher rates of digestion, greater conversion of waste organics to gas, faster solid-liquid separation, and minimization of bacterial and viral pathogen accumulation. Our results comparing mesophilic (37 degree C) and thermophilic (65 degree C) anaerobic digestion of domestic solid waste confirm the increased rate and conversion of waste to methane. In addition, utilizing radioactive labeling of glucose and acetic acid, we have measured the volumetric rates of volatile acid production and disappearance under both mesophilic and thermophilic conditions.     

Optimization of the anaerobic treatment of a waste stream from an enhanced oil recovery process

The aim of this work was to optimize the anaerobic treatment of a waste stream from an enhanced oil recovery (EOR) process. The treatment of a simulated waste water containing about 150 mg chemical oxygen demand (COD)/L of total petroleum hydrocarbons (TPH) and the saturation level of CO₂ was evaluated. A two-step anaerobic system was undertaken in the mesophilic temperature range (30-40 °C). The method of evolutionary operation EVOP factorial design was used to optimize pH, temperature and organic loading rate with the target parameters of CO₂ reduction and CH₄ production in the first reactor and TPH removal in the second reactor. The results showed 98% methanogenic removal of CO₂ and CH₄ yield of 0.38 L/gCOD in the first reactor and 83% TPH removal in the second reactor. In addition to enhancing CO₂ and TPH removal and CH₄ production, application of this method showed the degree of importance of the operational variables and their interactive effects for the two reactors in series.     

Thermophilic anaerobic digestion: the best option for waste treatment

After introducing thermophilic anaerobic digestion (AD), characteristics of thermophilic methanogens are provided. Accordingly, (a) site of occurrence, (b) morphological characteristics (shape and motility), (c) biochemical characteristics (Gram character and % G+C profile), (d) nutritional characteristics (NaCl requirement and substrate specificity), and (e) growth characteristics (pH and temperature) of thermophilic methanogens are described. Some studies of the thermophilic AD are cited with their operational management problems. Subsequently, strategies to maximize net energy production are given, including mode of heating the bioreactors, role of agitation to promote AD performance and mode/intensity of mixing. Finally, advantages as well as drawbacks of AD under thermophilic conditions are given, concluding with its applications.     

Plastics from household waste as a source of heavy metal pollution

An inventory study to the levels of cadmium in the plastic component of household waste was carried out utilizing INAA as the analytical technique. In a 2-h irradiation, 2-d decay, and 1-h measurement, protocol adequate sensitivities could be obtained for Cd, but also for a group of other metals: Cr, Co, Ni, Cu, Sr, Zn, As, Se, Mo, Sn, Sb, Ba, and Hg. Red-, orange-, and yellow-colored plastics either contain Cd at high levels (over 1000 mg/kg) or have relatively low Cd concentrations (<50 mg/kg). High concentrations were also occasionally found for Sr, Se, Ba, Sb, and Hg. INAA appeared very well to be routinely usable for such analysis because of the absence of a destruction step, adequate sensitivity, high accuracy, and multielement results.