Vermistabilization and nutrient enhancement of anaerobic digestate through earthworm species Perionyx excavatus and Perionyx sansibaricus

The aim of the study was to examine safe reuse and recycling of organic waste digestate obtained from a biogas plant (5 % total solid) with enhanced nutrient value through vermitechnology. Two indigenous epigeic earthworm species Perionyx excavates and Perionyx sansibaricus were tried individually for this purpose. The results clearly show a significant increase in pH values from 6.5–7.4, electrical conductivity (21.3–21.7 %), total N (84.5–94.6 %), total P (35.9–47.1 %), total K (49.8–52.6 %), Ca (41.9–41.9 %) and a significant decrease in total organic C (17.1–22.4 %), C/N ratio (7.2–6.9), C/P ratio (20.3–20.6), COD (51.9–55.7 %), BOD (85.5–91.2 %). Similarly, indicator organisms (fecal coliforms and fecal streptococci) showed decrement at the end of the composting period (60 days). Fecal Coliforms reduce to nil, while in fecal streptococci a 6 log reduction was observed. Oxygen uptake rate dropped to 67.4–70 % for vermireactors. Overall, the aforementioned findings highlighted that the indigenous earthworm species could enhance the nutrient value of the anaerobic digestate, which could be utilized as an efficient soil conditioner.     

Anaerobic co-digestion of aquatic flora and quinoa with manures from Bolivian Altiplano

Quinoa stalk (Chenopodium quinoa Willd.) from agricultural crop residue, totora (Schoenoplectus tatora) and o-macrophytes (aquatic flora) from Lake Titicaca (on the Bolivian Altiplano) were studied in a wet anaerobic co-digestion process together with manure from llama, cow and sheep. Anaerobic semi-continuous experiments were performed in (10) 2-l reactors at a temperature of 25 degrees C with 30 days of hydraulic retention time (HRT) and an organic loading rate (OLR) of 1.8kgVSm(-3)d(-1). Totora was found to be the best co-substrate. In mixture ratios of 1:1 (VS basis), it increased the biogas productivity by 130% for llama manure, 60% for cow manure, and 40% for sheep manure. It was possible to use up to 58% (VS basis) of totora in the substrate. Higher concentrations (including pure totora) could not be digested, as that caused acidification problems similar to those caused by other lignocellulosic materials. When quinoa and o-macrophytes were used as co-substrates, the increase in biogas productivity was slightly less. However, these co-substrates did not cause any operational problems. An additional advantage of quinoa and o-macrophytes was that they could be used in any proportion (even in pure form) without causing any destabilization problems in the anaerobic digestion process.     

Evaluation of heavy metal content in digestate from batch anaerobic co-digestion of sunflower hulls and poultry manure

In this experimental study, the biogas digestate from mesophilic batch anaerobic co-digestion of poultry manure and an agricultural residue, sunflower hulls, was characterized, particularly in terms of heavy metal content, in order to evaluate whether the biogas digestate was suitable for land applications. Ni, Zn, Cu, Pb, Cr, Cd, and Hg were detected in the biogas digestate in each trial, however, their concentrations were always lower than the limit values stated in Turkish regulations. The main source of heavy metals in the biogas digestate seemed to be the poultry manure, not the agricultural residue. The commercial feedstuffs that are frequently supplemented with various essential elements to promote optimum nutrient supply and optimum growth rates may have contributed to heavy metals presence in the biogas digestate. The results indicated that the biogas digestate from anaerobic co-digestion of manure and agricultural residue could be utilized as fertilizer in agricultural applications.     

Tertiary recycling of PVC-containing plastic waste by copyrolysis with cattle manure

The corrosion from pyrolysis of PVC in plastic waste was reduced by copyrolysis of PVC with cattle manure. The optimization of pyrolysis conditions between PVC and cattle manure was studied via a statistical method, the Box-Behnken model. The pyrolysis reaction was operated in a tubular reactor. Heating rate, reaction temperature and the PVC:cattle manure ratio were optimized in the range of 1-5 degrees C/min, 250-450 degrees C and the ratio of 1:1-1:5, respectively. The suitable conditions which provided the highest HCl reduction efficiency were the lowest heating rate of 1 degrees C/min, the highest reaction temperature of 450 degrees C, and the PVC:cattle manure ratio of 1:5, with reliability of more than 90%. The copyrolysis of the mixture of PVC-containing plastic and cattle manure was operated at optimized conditions and the synergistic effect was studied on product yields. The presence of manure decreased the oil yield by about 17%. The distillation fractions of oil at various boiling points from both the presence and absence of manure were comparable. The BTX concentration decreased rapidly when manure was present and the chlorinated hydrocarbon was reduced by 45%. However, the octane number of the gasoline fraction was not affected by manure and was in the range of 99-100.     

Survival of weed seeds and animal parasites as affected by anaerobic digestion at meso- and thermophilic conditions

Anaerobic digestion of residual materials from animals and crops offers an opportunity to simultaneously produce bioenergy and plant fertilizers at single farms and in farm communities where input substrate materials and resulting digested residues are shared among member farms. A surplus benefit from this practice may be the suppressing of propagules from harmful biological pests like weeds and animal pathogens (e.g. parasites). In the present work, batch experiments were performed, where survival of seeds of seven species of weeds and non-embryonated eggs of the large roundworm of pigs, Ascaris suum, was assessed under conditions similar to biogas plants managed at meso- (37 °C) and thermophilic (55 °C) conditions. Cattle manure was used as digestion substrate and experimental units were sampled destructively over time. Regarding weed seeds, the effect of thermophilic conditions (55 °C) was very clear as complete mortality, irrespective of weed species, was reached after less than 2 days. At mesophilic conditions, seeds of Avena fatua, Sinapsis arvensis, Solidago canadensis had completely lost germination ability, while Brassica napus, Fallopia convolvulus and Amzinckia micrantha still maintained low levels (～1%) of germination ability after 1 week. Chenopodium album was the only weed species which survived 1 week at substantial levels (7%) although after 11 d germination ability was totally lost. Similarly, at 55 °C, no Ascaris eggs survived more than 3 h of incubation. Incubation at 37 °C did not affect egg survival during the first 48 h and it took up to 10 days before total elimination was reached. In general, anaerobic digestion in biogas plants seems an efficient way (thermophilic more efficient than mesophilic) to treat organic farm wastes in a way that suppresses animal parasites and weeds so that the digestates can be applied without risking spread of these pests.     

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.     

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.     

Co-digestion of cattle manure with food waste and sludge to increase biogas production

Anaerobic co-digestion strategies are needed to enhance biogas production, especially when treating certain residues such as cattle/pig manure. This paper presents a study of co-digestion of cattle manure with food waste and sewage sludge. With the aim of maximising biogas yields, a series of experiments were carried out under mesophilic and thermophilic conditions using continuously stirred-tank reactors, operating at different hydraulic residence times. Pretreatment with ultrasound was also applied to compare the results with those obtained with non-pretreated waste. Specific methane production decreases when increasing the OLR and decreasing HRT. The maximum value obtained was 603 LCH(4)/kg VS(feed) for the co-digestion of a mixture of 70% manure, 20% food waste and 10% sewage sludge (total solid concentration around 4%) at 36°C, for an OLR of 1.2g VS/Lday. Increasing the OLR to 1.5g VS/Lday led to a decrease of around 20-28% in SMP. Lower methane yields were obtained when operating at 55°C. The increase in methane production when applying ultrasound to the feed mixtures does not compensate for the energy spent in this pretreatment.     

Improved biogas production from rice straw by co-digestion with kitchen waste and pig manure

In order to investigate the effect of feedstock ratios in biogas production, anaerobic co-digestions of rice straw with kitchen waste and pig manure were carried out. A series of single-stage batch mesophilic (37 ± 1 °C) anaerobic digestions were performed at a substrate concentration of 54 g/L based on volatile solids (VS). The results showed that the optimal ratio of kitchen waste, pig manure, and rice straw was 0.4:1.6:1, for which the C/N ratio was 21.7. The methane content was 45.9–70.0% and rate of VS reduction was 55.8%. The biogas yield of 674.4 L/kg VS was higher than that of the digestion of rice straw or pig manure alone by 71.67% and 10.41%, respectively. Inhibition of biogas production by volatile fatty acids (VFA) occurred when the addition of kitchen waste was greater than 26%. The VFA analysis showed that, in the reactors that successfully produced biogas, the dominant intermediate metabolites were propionate and acetate, while they were lactic acid, acetate, and propionate in the others.     

Comparative fertilizer properties of digestates from mesophilic and thermophilic anaerobic digestion of dairy manure: focusing on plant growth promoting bacteria (PGPB) and environmental risk

The fertilizer properties of anaerobic digestate depend on the feedstock and operating conditions of digestion. In this study, the comparative fertilizer properties of mesophilic and thermophilic digestates from dairy manure were evaluated for plant nutrient contents, and special attention was paid to plant growth promoting bacteria (PGPB). Two digestates contained similar plant nutrient contents, while the thermophilic digestate contained higher contents of NH₄⁺–N. The quantity of Bacillus and Pseudomonas in the mesophilic digestate was significantly higher than in the thermophilic digestate. Furthermore, Bacillus showed siderophore production and antifungal activity (43.5–75.3%), and Pseudomonas showed siderophore and phytohormone production (4.2–75.2 µg ml^?1). One phosphate solubilizing isolate was also detected in the mesophilic digestate. These results indicated that two digestates showed different fertilizer properties with respect to nutrient contents and PGPB, and digestates had the potential to increase the availability of phosphorus and iron in the soil, both to provide phytohormones to plant roots and protect plants from fungal phytopathogens. The contents of indicator bacteria and heavy metals were analyzed to determine their environmental risk, and the results showed a high reduction in indicator bacteria and lower levels of heavy metals than in other feedstocks.     

Microbiological assessments of compost toilets: in situ measurements and laboratory studies on the survival of fecal microbial indicators using sentinel chambers

Compost toilet systems were assessed for their ability to reduce microbial indicators and pathogens. Bacterial pathogens were not detected in any samples indicating a low survival rate in composting feces and/or an initial low occurrence. Indicator bacteria showed large variations with no clear trend of lower bacterial numbers after longer storage. In controlled composting experiments, thermophilic conditions were only reached when amendments were made (grass and a sugar solution). Even then it was impossible to ensure a homogenous temperature in the composting fecal material and therefore difficult to achieve a uniform reduction and killing of indicator organisms. Presumptive thermotolerant coliforms, Salmonella typhimurium Phage 28 B and eggs of Ascaridia galli, proved useful as indicators. However, regrowth was detected for enterococci and total numbers of bacteria grown at 36 degrees C. These indicator parameters may therefore overestimate the level of other (pathogenic) bacteria present in the material and can not be recommended for use as reliable indicator organisms in composting toilet systems. The addition of indicator bacteria to fecal material contained in semi-permeable capsules proved to be a useful technique to ensure that microorganisms were contained in a small test volume.     

Hygienic quality of faeces treated in urine diverting vermicomposting toilets

On-site sanitation solutions have gained much interest in recent years. One such solution is the urine diverting vermicomposting toilet (UDVT). This study evaluated the hygienic quality of the composted material in six UDVTs in operation in France. Samples were taken from three sampling positions in each toilet, with increasing distance from the fresh material. The concentration of Salmonella spp., Enterococcus spp., thermotolarent coliforms and coliphages were analysed and plotted against a number of variables. The variables found to have the greatest impact was the pH (for Enterococcus spp. and thermotolarent coliforms (TTC)) and time since last maintenance (coliphages). The pH was found to correlate with the material maturity. The current practise of maintenance can cause recontamination of the stabilised material and increase the risk of regrowth of pathogenic microorganisms. A modification in the maintenance procedure, in which a fourth maturation point is introduced, would eliminate this risk. UDVTs were found to be a good on-site sanitation option as the maintenance requirement is small and the system effectively reduced odour and concentration of pathogen and indicator organisms in human waste while keeping the accumulation of material down to a minimum. If the vermicompost is to be used for crops consumed raw, an additional sanitisation step is recommended.     

Anaerobic biogasification of undiluted dairy manure in leaching bed reactors

Dry anaerobic digestion of high solids animal manure is of increasing importance since conventional slurry digestion is not an effective system for these manures. The investment costs for large-size reactors, costs for heating these reactors, handling, dewatering, and the disposal of the digested residue decrease the benefits of conventional slurry anaerobic digestion for high solids animal manure. Even though leaching bed reactors (LBR) constitute a promising option for dry anaerobic biogasification of animal manure, no study is cited in the literature for animal manure, excluding a single study on cattle waste which utilized a similar concept in a different experimental set-up, namely a packed bed digester. Therefore, this work was undertaken to investigate the anaerobic biogasification of undiluted dairy manure in LBRs. To this purpose anaerobic leaching bed reactors (ALBR) packed with a mixture of dairy manure, anaerobic seed and wood powder/chips were operated. The ALBRs were fed with water, and the leachate that was collected from their effluents was subjected to biochemical methane potential (BMP) experiments to determine the biogas production. The results revealed that LBRs can successfully be applied to anaerobic digestion of undiluted dairy manure with around 25% improvement in biogas production relative to conventional (slurry) anaerobic digesters.     

Mechanical properties of dewatered sewage sludge

The mechanical properties of dewatered, anaerobically digested sewage sludge were determined from soil laboratory tests. The sludge material is largely composed of organic clay sized-particles, a sizable fraction of which is in an active state of biological digestion which can continue over many years under field conditions. Moderately digested sludge material was found to have a typical specific gravity of solids value of 1.55, and loss on ignition (LOI) value of 70% dry mass. Strongly digested sludge, produced by digesting the liquid sludge further at 35 degrees C in the laboratory, was found to have a lower LOI value of 55% dry mass, and a higher specific gravity of solids value of about 1.72. The maximum dry density of 0.56 tonne/m3 for the dried sludge material was produced using standard Proctor compaction at roughly 85% moisture content (54% solids content). Air-dried, compacted sludge material was tested in quick-undrained triaxial compression and vane shear. Undrained shear strength-moisture content plots are presented. Shear strength values measured in triaxial compression and vane shear were consistent. The effective angle of shearing resistance (phi') was determined from consolidated-undrained, triaxial compression tests on pasteurized, normally consolidated samples of the sludge material. The mechanical properties of the sludge material changed with the level of sludge digestion. The phi' value increased from 32 degrees for moderately digested sludge, to 37 degrees for strongly digested sludge. The effective cohesion of the sludge material remained zero throughout. The shrinkage, swelling and adhesion properties of the sludge material were also studied. Significant shrinkage occurred as the compacted material dried. The sludge material lost its adhesion below about 95% moisture content (51% solids content). Re-hydration of the dry material caused the bulk volume to double.     

Appraisal of methane production and anaerobic fermentation kinetics of livestock manures using artificial neural networks and sinusoidal growth functions

This study aimed to perform a comparative analysis of the performance of five models (Gompertz, logistic, Richards, the first-order, artificial neural networks) in predicting methane production rate from anaerobic digestion of livestock manures. The input variables were fermentation time, digestion temperature, biogas temperature, ambient temperature, pH, and specific biogas production rate. The physicochemical compositions of cow manure and sheep manure showed that volatile solid (VS) contents were close to each other in manure compositions (77.6% and 64.7%, respectively), while the potential of methane production from cow manure (673.44 mL CH₄/g VS) was greater than that from sheep manure (320.32 mL CH₄/g VS). The determination coefficients (R²) for logistic function, Gompertz, Richards, the first-order, and ANN models were obtained as 0.968, 0.967, 0.975, 0.825, and 0.995 for the cow manure, respectively. In case of the sheep manure, the R² values obtained from these models were 0.976, 0.979, 0.981, 0.968 and 0.991, respectively. Although the determination coefficients of all models were in satisfactory agreement with the experimental data, the ANN model showed competitive lower RMSE values of 0.111 and 0.164 for cow and sheep manure data sets, respectively, indicating its superior performance than other models.

     

Temperature and deactivation of microbial faecal indicators during small scale co-composting of faecal matter

Small scale co-composting of faecal matter from dry toilet systems with shredded plant material and food waste was investigated in respect to heat development and deactivation of faecal indicators under tropical semiarid conditions. Open (uncovered) co-composting of faecal matter with shredded plant material alone did not generate temperatures high enough (<55 °C) to reduce the indicators sufficiently. The addition of food waste and confinement in chambers, built of concrete bricks and wooden boards, improved the composting process significantly. Under these conditions peak temperatures of up to 70 °C were achieved and temperatures above 55 °C were maintained over 2 weeks. This temperature and time is sufficient to comply with international composting regulations. The reduction of Escherichia coli, Enterococcus faecalis and Salmonella senftenberg in test containment systems placed in the core of the compost piles was very efficient, exceeding 5 log₁₀-units in all cases, but recolonisation from the cooler outer layers appeared to interfere with the sanitisation efficiency of the substrate itself. The addition of a stabilisation period by extending the composting process to over 4 months ensured that the load of E. coli was reduced to less than 10³ cfu^?g and salmonella were undetectable.     

Effects of municipal solid waste compost application on the microbial biomass of cultivated and non-cultivated soil in a semi-arid zone.

The aim of this study was to assess whether soil microbial biomass could be used as an indicator of environmental changes following the application of organic residue (compost of municipal solid waste and farmyard manure) or mineral fertilizers (N and P) into cultivated or uncultivated loam-clayey soil, for three consecutive years. The carbon and nitrogen of the microbial biomass (B(C) and B(N) were studied using the fumigation-extraction method. For the two cultivated and uncultivated plots, B(N) and B(C) were more important in the superficial profile (0-20 cm) than in the deep one (20-40 cm). In the uncultivated soil, we observed a good linear relation between B(C) and B(N) at the level of upper soil horizon during the wet season with r coefficients of 0.95, 0.71 and 0.80 for the consecutive years 2000, 2001 and 2002, respectively. Microbial biomasses C and N increased during the rainy season and decreased during the dry season. Microbial biomass C and N showed the higher content with compost and farmyard manure at 40 tonnes ha(-1). Moreover, the results showed that at the beginning of the experiment, the microbial biomass was higher in the ploughed wheat-cultivated plot than in the uncultivated one. Microbial biomass C and N in the cultivated plot amended with compost at 40 tonnes ha(-1) were significantly different in comparison with the soil microbial biomass amended with farmyard manure. The combining of chemical fertilizer with the organic fertilizer, such as compost at 40 or 80 tonnes ha(-1) and farmyard manure, increased the microbial biomasses C and N after 1 and 2 years. In the cultivated or uncultivated plots the results revealed that the best application rate of the compost was 40 tonnes ha(-1) and when the compost rate was increased from 40 to 80 tonnes ha(-1) both B(C) and B(N) decreased significantly.     

Kinetics and dynamic modelling of batch anaerobic digestion of municipal solid waste in a stirred reactor

A series of batch, slurry anaerobic digestion experiments were performed where the soluble and insoluble fractions, and unwashed MSW were separately digested in a 200l stirred stainless steel vessel at a pH of 7.2 and a temperature of 38 degrees C. It was found that 7% of the total MSW COD was readily soluble, of which 80% was converted to biogas; 50% of the insoluble fraction was solubilised, of this only 80% was converted to biogas. The rate of digesting the insoluble fraction was about four times slower than the rate of digesting the soluble fraction; 48% of the total COD was converted to biogas and 40% of the total nitrogen was converted to ammonia. Soluble and insoluble fractions were broken down simultaneously. The minimum time to convert 95% of the degradable fraction to biogas was 20 days. The lag phase for the degradation of insoluble fraction of MSW can be overcome by acclimatising the culture with the soluble fraction. The rate of digestion and the methane yield was not affected by particle size (within the range of 2-50mm). A dynamic model was developed to describe batch digestion of MSW. The parameters of the model were estimated using data from the separate digestion of soluble and insoluble fractions and validated against data from the digestion of unwashed MSW. Trends in the specific aceticlastic and formate-utilising methanogenic activity were used to estimate initial methanogenic biomass concentration and bacterial death rate coefficient. The kinetics of hydrolysis of insoluble fraction could be adequately described by a Contois equation and the kinetics of acidogenesis, and aceticlastic and hydrogen utilising methanogenesis by Monod equations.     

Sanitary aspect of using partially treated landfill leachate as a water source in green waste composting

Shredded green wastes were composted in windrows, at the Harewood Whin landfill, near the city of York, in West Yorkshire, UK. Landfill leachate were added twice during the second and fourth week of the process in two piles. One pile was turned once every week for eight weeks and the other was turned twice, during the same period. Each time approximately, 2 m3 of leachate was added, into each pile. The two piles each contained about 45 m3 of shredded green waste. The effect of adding leachate on the sanitisation of the green waste during composting, was evaluated based on the changes in the levels of faecal coliforms and faecal streptococci. The results suggested that using leachate as the moisture source had no significant effect (tested with two factors ANOVA test) on the sanitisation process when compared with two similar piles, used as the control, for which tap water was used for moisture addition. In all four piles sanitisation was almost complete and below the acceptable levels. Additionally, the results indicated that there was no significant effect on the sanitisation process of the turning frequency.     

Aerobic thermophilic bacteria enhance biogas production

The enhancing effect of aerobic thermophilic (AT) bacteria on the production of biogas from anaerobically digested sewage sludge (methanogenic sludge) was investigated. Sewage sludge (5%, w/w) was incubated at 65°C with shaking for a few months to prepare the AT seed sludge. AT sludge was prepared by incubation of the AT seed sludge (5%, v/v) and sewage sludge (5%, w/w) at 65°C with shaking. The addition of this AT sludge (1.2% ± 0.5% of total volatile solids) to methanogenic sludge enhanced the production of biogas. The optimum volume of the addition and the pretreatment temperature of the AT sludge for optimum biogas production were 5% (v/v) and 65°C. Batch-fed anaerobic digestion was covered with the addition of various AT sludges. The AT sludge prepared with the AT seed sludge improved the biogas production by 2.2 times relative to that from the sewage sludge addition. The addition of sludge without AT seed sludge weakly enhanced biogas production. An aerobic thermophilic bacterium (strain AT1) was isolated from the AT seed sludge. Strain AT1 grew well in a synthetic medium. The production of biogas from the anaerobic digestion of sewage sludge was improved by the addition of 5% (v/v) AT1 bacterial culture compared with that from the sewage sludge addition. The addition of AT1 culture reduced the volatile solids by 21%, which was higher than the 12.6% achieved with the sewage sludge addition. The AT1 bacterial culture enhanced the biogas production more than the AT seed sludge. The phylogenetic analysis of the 16S rRNA gene revealed that strain AT1 is closely related to Geobacillus thermodenitrificans (100% sequence similarity). The improvement in the production of biogas with the AT sludge could be caused by thermophilic bacterial activity in the AT sludge.