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.     

Investigating the effects of anaerobic and aerobic post-treatment on quality and stability of organic fraction of municipal solid waste as soil amendment

The use of OFMSW for biogas and compost production is considered as a sustainable strategy in saving valuable landfill space while producing valuable product for soil application. This study examines the effects of anaerobic and aerobic post-treatment of OFMSW on the stability of anaerobic digestate and compost and soil quality using seed germination tests. Anaerobic digestion of OFMSW was carried out for fifteen days after which the residual anaerobic digestate was subjected to aerobic post-treatment for seventy days. Seed germination tests showed that fresh feedstock and digestates collected during anaerobic digestion and during the early stages of aerobic post-treatment were phytotoxic. However, phytotoxic effects were not observed in soils amended with the fully stabilised anaerobic digestate compost, ADC. It was also found that seed germination increases with dilution and incubation time, suggesting that lower soil application rates and longer lag periods between soil application of ADC and planting can reduce the amount of biodegradable organics in the ADC, thus enhancing the benefits of ADC as soil amendment.     

Physical degradation of wheat straw by the in-vessel and windrow methods of mushroom compost production

Mushroom compost manufacturers in Ireland are moving away from the traditional outdoor phase I windrow method, favouring in-vessel production. Composters and growers have reported better quality compost with faster spawn run and higher yields produced by this process. In the present study, physical examination of samples highlighted differences when comparing the windrow and in-vessel methods of compost production. Observations using scanning electron microscopy suggest that the cuticle of wheat straw from in-vessel production is damaged during phase I, peeling away from the surface in fragments, and exposing the epidermis. Changes in silicon levels on the straw surface acted as a marker for cuticle damage when comparing both composting systems. Cuticle damage may be important during composting and afterwards, as substrate colonisation is faster, and consequently spawn run is shorter. The phase I compost microbial community is altered by the in-vessel technique, producing a predominantly thermophilic bacterial flora in contrast to the mesophilic and thermophilic bacteria and fungi found in windrow phase I compost. These differences may be significant in mushroom compost production.     

Biological nutrients removal from the supernatant originating from the anaerobic digestion of the organic fraction of municipal solid waste

Abstract

This study critically evaluates the biological processes and techniques applied to remove nitrogen and phosphorus from the anaerobic supernatant produced from the treatment of the organic fraction of municipal solid waste (OFMSW) and from its co-digestion with other biodegradable organic waste (BOW) streams. The wide application of anaerobic digestion for the treatment of several organic waste streams results in the production of high quantities of anaerobic effluents. Such effluents are characterized by high nutrient content, because organic and particulate nitrogen and phosphorus are hydrolyzed in the anaerobic digestion process. Consequently, adequate post-treatment is required in order to comply with the existing land application and discharge legislation in the European Union countries. This may include physicochemical and biological processes, with the latter being more advantageous due to their lower cost. Nitrogen removal is accomplished through the conventional nitrification/denitrification, nitritation/denitritation and the complete autotrophic nitrogen removal process; the latter is accomplished by nitritation coupled with the anoxic ammonium oxidation process. As anaerobic digestion effluents are characterized by low COD/TKN ratio, conventional denitrification/nitrification is not an attractive option; short-cut nitrogen removal processes are more promising. Both suspended and attached growth processes have been employed to treat the anaerobic supernatant. Specifically, the sequencing batch reactor, the membrane bioreactor, the conventional activated sludge and the moving bed biofilm reactor processes have been investigated. Physicochemical phosphorus removal via struvite precipitation has been extensively examined. Enhanced biological phosphorus removal from the anaerobic supernatant can take place through the sequencing anaerobic/aerobic process. More recently, denitrifying phosphorus removal via nitrite or nitrate has been explored. The removal of phosphorus from the anaerobic supernatant of OFMSW is an interesting research topic that has not yet been explored. At the moment, standardization in the design of facilities that treat anaerobic supernatant produced from the treatment of OFMSW is still under development. To move toward this direction, it is first necessary to assess the performance of alternative treatment options. It study concentrates existing data regarding the characteristics of the anaerobic supernatant produced from the treatment of OFMSW and from their co-digestion with other BOW. This provides data documenting the effect of the anaerobic digestion operating conditions on the supernatant quality and critically evaluates alternative options for the post-treatment of the liquid fraction produced from the anaerobic digestion process.     

Enhancement of the thermophilic stage in cattle waste composting by addition of tofu residue

The microbial degradation and temperature rise during the composting of a cattle waste and rice straw mixture blended with tofu (soybean curd) residue was investigated using an insulated and unheated in-vessel composter (effective volume, 12 1) and a static pile with passive aeration. The addition of 11% (dry weight basis) of tofu residue shortened the time required for temperature to reach the thermophilic phase and increased the duration of the temperatures above 55 degrees C significantly, but the maximum temperature was not affected by the additive level. As shown by the change in BOD, most of the easily biodegradable matter in the tofu residue was consumed during 12 days of composting. The same results were observed in the temperature profile of the static pile with passive aeration. Tofu residue addition yielded a higher maximum temperature and a nearly two times longer duration of temperatures above 55 degrees C in almost all locations of the pile. The use of tofu residue as a co-composting material would promote thermophilic degradation throughout the entire composting mass.     

Bioremediation of PAHs-contaminated soil through composting: Influence of bioaugmentation and biostimulation on contaminant biodegradation

The degradation of several polycyclic aromatic hydrocarbons (PAHs) in soil through composting was investigated. The selected PAHs included: fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo(a)anthracene, and chrysene, with concentrations simulating a real creosote sample. The degradation of PAHs (initial concentration 1 g of total PAHs kg⁻¹ dry soil) was assessed applying bioaugmentation with the white-rot fungi Trametes versicolor and biostimulation using compost of the source-selected organic fraction of municipal solid waste (OFMSW) and rabbit food as organic co-substrates. The process performance during 30 days of incubation was evaluated through different analyses including: dynamic respiration index (DRI), cumulative oxygen consumption during 5 days (AT₅), enzymatic activity, and fungal biomass. These analyses demonstrated that the introduced T. versicolor did not significantly enhance the degradation of PAHs. However, biostimulation was able to improve the PAHs degradation: 89% of the total PAHs were degraded by the end of the composting period (30 days) compared to the only 29.5% that was achieved by the soil indigenous microorganisms without any co-substrate (control, not amended). Indeed, the results showed that stable compost from the OFMSW has a greater potential to enhance the degradation of PAHs compared to non-stable co-substrates such as rabbit food.     

Ammonia inhibition in anaerobic digestion: A review

Even though ammonia is an essential nutrient for bacterial growth, it may inhibit methanogenesis during anaerobic digestion process if it is available at high concentrations. Therefore, ammonia is regarded as a potential inhibitor during anaerobic digestion, particularly when dealing with complex type of substrates such as manure or the organic fraction of municipal solid waste (OFMSW). Ammonia is produced through biological degradation of nitrogenous matter. Ammonium ion (NH₄⁺) and free ammonia (NH₃) are the two principal forms of inorganic ammonia nitrogen. Both forms can directly and indirectly cause inhibition in an anaerobic digestion system. Particularly, free ammonia (FAN) is a powerful inhibitor in an anaerobic digester above threshold concentrations. Process inhibition is related to the particular characteristics of the substrate to be anaerobically digested, pH, process temperature (mesophilic or thermophilic), type of the seed sludge (inoculum), the reactor configuration and to the concentrations of ammonium and ammonia. In this paper, ammonia inhibition in anaerobic digestion systems and the recovery efforts after inhibition are discussed. Furthermore, the impacts of ammonia inhibition on the microbial population available in anaerobic digesters, namely bacteria and Archaea, are also evaluated in detail.     

Environmental impact of two aerobic composting technologies using life cycle assessment

Background, aim, and scope  Composting is a viable technology to treat the organic fraction of municipal solid waste (OFMSW) because it stabilizes biodegradable organic matter and contributes to reduce the quantity of municipal solid waste to be incinerated or land-filled. However, the composting process generates environmental impacts such as atmospheric emissions and resources consumption that should be studied. This work presents the inventory data and the study of the environmental impact of two real composting plants using different technologies, tunnels (CT) and confined windrows (CCW). Materials and methods  Inventory data of the two composting facilities studied were obtained from field measurements and from plant managers. Next, life cycle assessment (LCA) methodology was used to calculate the environmental impacts. Composting facilities were located in Catalonia (Spain) and were evaluated during 2007. Both studied plants treat source separated organic fraction of municipal solid waste. In both installations the analysis includes environmental impact from fuel, water, and electricity consumption and the main gaseous emissions from the composting process itself (ammonia and volatile organic compounds). Results and discussion  Inventory analysis permitted the calculation of different ratios corresponding to resources consumption or plant performance and process yield with respect to 1 t of OFMSW. Among them, it can be highlighted that in both studied plants total energy consumption necessary to treat the OFMSW and transform it into compost was between 130 and 160 kWh/t OFMSW. Environmental impact was evaluated in terms of global warming potential (around 60 kg CO₂/t OFMSW for both plants), acidification potential (7.13 and 3.69 kg SO₂ eq/t OFMSW for CT and CCW plant respectively), photochemical oxidation potential (0.1 and 3.11 kg C₂H₄ eq/t OFMSW for CT and CCW plant, respectively), eutrophication (1.51 and 0.77 kg /t OFMSW for CT and CCW plant, respectively), human toxicity (around 15 kg 1,4-DB eq/t OFMSW for both plants) and ozone layer depletion (1.66 × 10⁻⁵ and 2.77 × 10⁻⁵ kg CFC⁻¹¹ eq/t OFMSW for CT and CCW plant, respectively). Conclusions  This work reflects that the life cycle perspective is a useful tool to analyze a composting process since it permits the comparison among different technologies. According to our results total energy consumption required for composting OFMSW is dependent on the technology used (ranging from 130 to 160 kWh/t OFMSW) as water consumption is (from 0.02 to 0.33 m³ of water/t OFMSW). Gaseous emissions from the composting process represent the main contribution to eutrophication, acidification and photochemical oxidation potentials, while those contributions related to energy consumption are the principal responsible for global warming. Recommendations and perspectives  This work provides the evaluation of environmental impacts of two composting technologies that can be useful for its application to composting plants with similar characteristics. In addition, this study can also be part of future works to compare composting with other OFMSW treatments from a LCA perspective. Likewise, the results can be used for the elaboration of a greenhouse gasses emissions inventory in Catalonia and Spain.     

Community structure and population dynamics of ammonia oxidizers in composting processes of ammonia-rich livestock waste

This study investigated the relationship between the population dynamics of ammonia-oxidizing bacteria (AOB) and archaea (AOA), and changes in the concentrations of nitrogenous compounds during ammonia-rich livestock waste-composting processes. The data showed that ammonia in beef and dairy cow livestock waste-composting piles was slowly oxidized to nitrite and nitrate after approximately 21–35 days under thermophilic or moderately thermophilic and mesophilic conditions. Real-time quantitative PCR (qPCR) assays showed a relative abundance of betaproteobacterial AOB during ammonia oxidation but did not detect AOA in any composting stage. Furthermore, real-time qPCR and terminal-restriction fragment length polymorphism (T-RFLP) analyses for the AOB in two composting processes (beef and dairy cow livestock waste) out of the three studied found that thermophilic or moderately thermophilic uncultured betaproteobacterial AOB from the “compost AOB cluster” contributed to ammonia oxidation during hot composting stages. Non-metric multidimensional scaling analyses of the data from T-RFLP showed that only a few analogous species predominated during composting of beef, dairy cow and pig livestock wastes, and thus, the AOB community structures in the three composting piles operating under different conditions were similar. AOB-targeted clone library analyses revealed that uncultured members of the “compost AOB cluster”, which could be clearly distinguished from the authentic species of the genus Nitrosomonas, were the major constituents of the AOB populations. These results suggested that a limited and unique species of AOB played a role in ammonia oxidation during the composting of ammonia-rich livestock waste.     

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.     

Monitoring the biological activity of the composting process: Oxygen uptake rate (OUR), respirometric index (RI), and respiratory quotient (RQ)

Composting of several organic wastes of different chemical composition (source-separated organic fraction of municipal solid waste, dewatered raw sludge, dewatered anaerobically digested sludge and paper sludge) was carried out under controlled conditions to study the suitability of different biological indexes (oxygen uptake rate, respirometric index, and respiratory quotient) to monitor the biological activity of the composting process. Among the indexes tested, oxygen uptake rate (also referred to as dynamic respirometric index) provided the most reliable values of microbial activity in a compost environment. On the other hand, values of the static respirometric index measured at process temperature, especially in the early stages of the composting process, were significantly lower than those of the dynamic index, which was probably due to oxygen diffusion limitations present in static systems. Both static and dynamic indexes were similar during the maturation phase. Static respirometric index measured at 37 degrees C should not be used with samples obtained during the thermophilic phase, since it resulted in an underestimation of the respiration values. Respiratory quotient presented only slight variations when changing the process temperature or the waste considered, and its use should be restricted to ensure aerobic conditions in the composting matrix.     

Anaerobic digestion of municipal solid waste and agricultural waste and the effect of co-digestion with dairy cow manure

Anaerobic digestion of dairy cow manure (CM), the organic fraction of municipal solid waste (OFMSW), and cotton gin waste (CGW) was investigated with a two-phase pilot-scale anaerobic digestion (AD) system. The OFMSW and CM were digested as single wastes and as combined wastes. The single waste digestion of CM resulted in 62m(3) methane/ton of CM on dry weight basis. The single waste digestion of OFMSW produced 37m(3) methane/ton of dry waste. Co-digestion of OFMSW and CM resulted in 172m(3) methane/ton of dry waste. Co-digestion of CGW and CM produced 87m(3) methane/ton of dry waste. Comparing the single waste digestions with co-digestion of combined wastes, it was shown that co-digestion resulted in higher methane gas yields. In addition, co-digestion of OFMSW and CM promotes synergistic effects resulting in higher mass conversion and lower weight and volume of digested residual.     

Eradication of Polymyxa betae by Thermal and Anaerobic Conditions and in the Presence of Compost Leachate

The abiotic conditions required for eradication of Polymyxa betae, the vector of Beet necrotic yellow vein virus in sugar beet, were investigated. Survival of resting spores of P. betae was determined under aerobic (30 min, 4 days and 21 days) and anaerobic (4 days) conditions under several temperature regimes in a water suspension and in leachate extracted from an aerobic compost heap. In water under aerobic conditions the lethal temperature was 60, 55 and 40°C for exposure times of 30 min, 4 days and 21 days, respectively. The effect of compost leachate and/or anaerobic conditions on survival of P. betae depended on temperature. After incubation for 4 days at 20°C, no significant effects of anaerobic conditions or leachate on the survival of P. betae were found. However, at 40°C for 4 days under anaerobic conditions, survival of P. betae was significantly lower than survival under aerobic conditions in water as well as in leachate. In leachate taken from an aerobic compost heap, aerobically incubated at 40°C for 4 days, survival of P. betae was significantly lower than survival in water at the same temperature. As anaerobic spots are prevalent in aerobic compost heaps, especially during the thermophilic phase, actual inactivation temperatures under composting conditions are likely to be lower than the temperatures we found for eradication in water under aerobic conditions.     

Activity and composition of ammonia oxidizing bacterial communities and emission dynamics of NH3 and N2O in a compost reactor treating organic household waste

Aims:  To monitor emissions of NH₃ and N₂O during composting and link these to ammonia oxidation rates and the community structure of ammonia oxidizing bacteria (AOB).
Methods and Results:  A laboratory-scale compost reactor treating organic household waste was run for 2 months. NH₃ emissions peaked when pH started to increase. Small amounts of N₂O and CH₄ were also produced. In total, 16% and less than 1% of the initial N was lost as NH₃-N and N₂O-N respectively. The potential ammonia oxidation rate, determined by a chlorate inhibition assay, increased fourfold during the first 9 days and then remained high. Initially, both Nitrosospira and Nitrosomonas populations were detected using DGGE analysis of AOB specific 16S rRNA fragments. Only Nitrosomonas europaea was detected under thermophilic conditions, but Nitrosospira populations re-established during the cooling phase.
Conclusions:  Thermophilic conditions favoured high potential ammonia oxidation rates, suggesting that ammonia oxidation contributed to reduced NH₃ emissions. Small but significant amounts of N₂O were emitted during the thermophilic phase. The significance of different AOBs detected in the compost for ammonia oxidation is not clear.
Significance and Impact of Study:  This study shows that ammonia oxidation occurs at high temperature composting and therefore most likely reduces NH₃ emissions.     

Assessing amendment properties of digestate by studying the organic matter composition and the degree of biological stability during the anaerobic digestion of the organic fraction of MSW

The transformation of organic matter during anaerobic digestion of mixtures of energetic crops, cow slurry, agro-industrial waste and organic fraction of municipal solid waste (OFMSW) was studied by analysing different samples at diverse points during the anaerobic digestion process in a full-scale plant. Both chemical (fiber analysis) and spectroscopic approaches (¹³C CPMAS NMR) indicated the anaerobic digestion process proceeded by degradation of more labile fraction (e.g. carbohydrate-like molecules) and concentration of more recalcitrant molecules (lignin and non-hydrolysable lipids). These modifications determined a higher degree of biological stability of digestate with respect to the starting mixture, as suggested, also, by the good correlations found between the cumulative oxygen uptake (OD₂₀), and the sum of (cellulose + hemicellulose + cell soluble) contents of biomasses detected by fiber analysis (r = 0.99; P < 0.05), and both O–alkyl-C (r = 0.98; P < 0.05) and alkyl-C (r = −0.99; P < 0.05) measured by ¹³C CPMAS NMR.     

Composting of bio-waste, aerobic and anaerobic sludges--effect of feedstock on the process and quality of compost

In-vessel composting of three stocks with originally different degree of organic matter degradation was conducted for: (1) kitchen source-separated bio-waste (BW), (2) aerobic (AS) as well as (3) anaerobic sludges (AnS) from municipal wastewater treatment plant. Composting experiment lasted over a year. The highest activity of the process was in the BW compost. It was implied by the highest temperature, CO(2) release, ammonification and nitrification, intensive accumulation and removal of low-weight carboxylic acids (water- and NaOH-extractable). Between the sludges higher mineralization and CO2 release was in AnS, while ammonification and nitrification were higher in AS compost; no significant difference between sludge composts was noticed for dynamics of pH, conductivity, concentrations of LWCA, and some nutrient compounds and heavy metals. Nitrogen content of the final compost increased in BW, but decreased in AS and AnS. Phytotoxicity of Lepidium sativum was eliminated faster in sludge composts compared to BW compost.     

Rapid start-up of thermophilic anaerobic digestion with the turf fraction of MSW as inoculum

This study aims to determine suitable start-up conditions and inoculum sources for thermophilic anaerobic digestion. Within days of incubation MSW at 55 °C, methane was produced at a high rate. In an attempt to narrow down which components of typical MSW contained the thermophilic methanogens, vacuum cleaner dust, banana peel, kitchen waste, and garden waste were tested as inoculum for thermophilic methanogenesis with acetate as the substrate. Results singled out grass turf as the key source of thermophilic acetate degrading methanogenic consortia. Within 4 days of anaerobic incubation (55 °C), anaerobically incubated grass turf samples produced methane accompanied by acetate degradation enabling successful start-up of thermophilic anaerobic digestion. Other essential start-up conditions are specified. Stirring of the culture was not conducive for successful start-up as it resulted specifically in propionate accumulation.     

Evolution of organic matter in a full-scale composting plant for the treatment of sewage sludge and biowaste by respiration techniques and pyrolysis-GC/MS

A full-scale composting plant treating in two parallel lines sewage sludge and the source-selected organic fraction of municipal solid waste (OFMSW or biowaste) has been completely monitored. Chemical routine analysis proved not to be suitable for an adequate plant monitoring in terms of stabilization and characterization of the process and final compost properties. However, the dynamic respiration index demonstrated to be the most feasible tool to determine the progression of the degradation and stabilization of organic matter for both sewage sludge and OFMSW lines. Both lines exhibited an important degree of stabilization of organic matter using rapid and cumulative respiration indices. Pyrolysis-GC/MS was applied to the most important inputs, outputs, and intermediate points of the plant. It proved to be a powerful tool for the qualitative characterization of molecular composition of organic matter present in solid samples. A full characterization of the samples considered is also presented.     

Production and characterization of β-glucosidase from Aspergillus niger fermentation: Application for organic fraction of municipal solid waste hydrolysis and methane enhancement

The anaerobic digestion of the organic fraction of municipal solid waste (OFMSW) is currently an attractive treatment process with energy production in the form of biogas. Hydrolysis is the rate-limiting step for the anaerobic digestion of solid wastes. Thus, in the present study fungal enzymatic pretreatment of OFMSW was applied to enhance biogas production. Two enzyme cocktails rich on β-glucosidase were produced from submerged fermentation of Aspergillus niger on basal medium using OFMSW as carbon source and urea (Urea cocktail) and Ulva rigida as nitrogen source (Ulva cocktail). Ulva cocktail displayed an important effect on OFMSW solubilization. Therefore, an increase of reducing sugar concentration about 60% was obtained which was in correlation with chemical oxygen demand (COD) increase. The performance of enzymatic pretreatment on anaerobic digestion of OFMSW was studied by conducting biochemical methane potential tests. Results showed that the enzymatic pretreatment improved methane yield of OFMSW even at high solid concentration. High methane yield about 500 ml/g total volatile solid was obtained, which corresponds up to 68% enhancement over the control.     

Microbial Community Growth and Utilization of Carbon Constituents During Thermophilic Composting at Different Oxygen Levels

Composting is characterized by dramatic changes in microbial community structure, to a high extent driven by changes in temperature and in the composition of the organic substrate. This study focuses on the interrelationships between decomposition of major classes in the organic material and dynamics in microbial populations during thermophilic composting of source-separated organic household waste. Experiments were performed in a 200-L laboratory reactor at 16, 2.5, and 1% O₂ in the compost atmosphere. Major classes of carbon constituents were analyzed by chemical methods, and the microbial biomass and community structure determined by fatty acid analyses with phospholipid fatty acids (PLFA) and total ester-linked fatty acids (EL) methods. At all three O₂ levels, the process was characterized by a rapid increase in microbial activity and biomass in the early thermophilic phase, although this period was delayed at the lower O₂ concentrations. Starch and fat were the main substrates utilized at all three O₂ levels during this period. The depletion of the starch fraction coincided with the beginning of a microbial biomass decrease, suggesting thatstarch is an important carbon substrate for the growth of thermophilic microorganisms during composting. Growth yields in the microbial community based on consumption of major carbon constituent classes in the high-activity period fell between 22 and 28%. Multivariate statistical analysis of changes in fatty acid composition revealed small, but statistically significant differences in the microbial community succession. At 16% O₂, 10Me fatty acids from Actinomycetes and cyclopropyl fatty acids (from Gram-negative bacteria) became more important with time, whereas 18:1ω7t was characteristic at 2.5 and 1% O₂, indicating a more stressed bacterial community at the lower O₂ concentrations. Although adequate composting was achieved at O₂ levels as low as 2.5 and 1%, it is not recommended to compost at such low levels in large-scale systems, because the heterogeneous gas transport through the material in these systems might lead to anaerobic conditions and inefficient composting.