W. P. Clarke

Methane as a resource: can the methanotrophs add value?

Methane is an abundant gas used in energy recovery systems, heating, and transport. Methanotrophs are bacteria capable of using methane as their sole carbon source. Although intensively researched, the myriad of potential biotechnological applications of methanotrophic bacteria has not been comprehensively discussed in a single review. Methanotrophs can generate single-cell protein, biopolymers, components for nanotechnology applications (surface layers), soluble metabolites (methanol, formaldehyde, organic acids, and ectoine), lipids (biodiesel and health supplements), growth media, and vitamin B12 using methane as their carbon source. They may be genetically engineered to produce new compounds such as carotenoids or farnesene. Some enzymes (dehydrogenases, oxidase, and catalase) are valuable products with high conversion efficiencies and can generate methanol or sequester CO2 as formic acid ex vivo. Live cultures can be used for bioremediation, chemical transformation (propene to propylene oxide), wastewater denitrification, as components of biosensors, or possibly for directly generating electricity. This review demonstrates the potential for methanotrophs and their consortia to generate value while using methane as a carbon source. While there are notable challenges using a low solubility gas as a carbon source, the massive methane resource, and the potential cost savings while sequestering a greenhouse gas, keeps interest piqued in these unique bacteria.

Identification, Detection, and Spatial Resolution of Clostridium Populations Responsible for Cellulose Degradation in a Methanogenic Landfill Leachate Bioreactor

ABSTRACT An anaerobic landfill leachate bioreactor was operated with crystalline cellulose and sterile landfill leachate until a steady state was reached. Cellulose hydrolysis, acidogenesis, and methanogenesis were measured. Microorganisms attached to the cellulose surfaces were hypothesized to be the cellulose hydrolyzers. 16S rRNA gene clone libraries were prepared from this attached fraction and also from the mixed fraction (biomass associated with cellulose particles and in the planktonic phase). Both clone libraries were dominated by Firmicutes phylum sequences (100% of the attached library and 90% of the mixed library), and the majority fell into one of five lineages of the clostridia. Clone group 1 (most closely related to Clostridium stercorarium), clone group 2 (most closely related to Clostridium thermocellum), and clone group 5 (most closely related to Bacteroides cellulosolvens) comprised sequences in Clostridium group III. Clone group 3 sequences were in Clostridium group XIVa (most closely related to Clostridium sp. strain XB90). Clone group 4 sequences were affiliated with a deeply branching clostridial lineage peripherally associated with Clostridium group VI. This monophyletic group comprises a new Clostridium cluster, designated cluster VIa. Specific fluorescence in situ hybridization (FISH) probes for the five groups were designed and synthesized, and it was demonstrated in FISH experiments that bacteria targeted by the probes for clone groups 1, 2, 4, and 5 were very abundant on the surfaces of the cellulose particles and likely the key cellulolytic microorganisms in the landfill bioreactor. The FISH probe for clone group 3 targeted cells in the planktonic phase, and these organisms were hypothesized to be glucose fermenters.

Effect of recirculated leachate volume on MSW degradation

Landfilling represents the most economical method for the disposal of municipal solid waste (MSW). After depletion of the limited volume of air available in void spaces of a waste bed, decomposition in a landfill takes place under anaerobic conditions. Anaerobic digestion requires moisture and a lack of water is generally responsible for retarding degradation of MSW in conventional landfills. Furthermore, the moisture that may be present is seldom uniformly distributed. Bioreac tor landfills are being researched as a means to provide a rapid and predictable stabilisation of waste and landfill gas produc tion. This is achieved primarily by control and management of the liquid flow within the landfill.

Degradation of unsorted municipal solid waste by a leach-bed process

In current landfills breakdown of municipal solid waste (MSW) occurs slowly and the landfill leaves a legacy of care, management, monitoring and potential catastrophic failure over several generations. Social concern over these long term issues, with their legislative and economic implementation, increasingly favour practices which promote short stabilisation times and minimise environmental impact. This paper describes experiments carried out on mixed and unsorted municipal solid waste (MSW) in which 75% of the rapidly biodegradable fraction was degraded in about 2 months with an average yield of 0.18 m(3) CH4/kg volatile solids at s.t.p. The experiments served to demonstrate that with proper leachate management very rapid decomposition of waste can be accomplished by taking the waste through a series of controlled degradation stages

The anaerobic degradability of thermoplastic starch: polyvinyl alcohol blends: potential biodegradable food packaging materials.

A systematic study on the anaerobic degradability of a series of starch:polyvinyl alcohol (TPS:PVOH) blends was performed to determine their fate upon disposal in either anaerobic digesters or bioreactor landfills. The aims of the study were to measure the rate and extent of solubilisation of the plastics. The extent of substrate solubilisation on a COD basis reached 60% for a 90:10 (w/w) blend of TPS:PVOH, 40% for 75:25, 30% for 50:50 and 15% for PVOH only. The rate of substrate solubilisation was most rapid for the 90:10 blend (0.041 h(-1)) and decreased with the amount of starch in the blend in the following order 0.034 h(-1)(75:25); 0.023 h(-1)(50:50). The total solids that remained after 900 h were 10 wt.% (90:10); 23 wt.% (75:25); 55 wt.% (50:50); 90 wt.% (0:100). Starch containing substrates produced a higher concentration of volatile fatty acids (VFAs) and biogas, compared to the 0:100 substrate. The major outcome was that PVOH inhibited the degradation of the starch from the blend.

Removal of sulfate from high-strength wastewater by crystallisation

Sulfate causes considerable problems in anaerobic digesters, related to generation of sulfides, loss of electrons (and hence methane), and contamination of gas streams. Removal of sulfides is generally expensive, and still results in methane losses. In this paper, we evaluate the use of precipitation for low-cost sulfate removal, in highly contaminated streams (>1 gS L(-1)). The main precipitate assessed is calcium sulfate (gypsum), though the formation of complex precipitates such as jarosite and ettringite to remove residual sulfate is also evaluated. The four main concerns in contaminated wastewater are:- high solubility, caused by high ion activity and ion pairing; slow kinetics; inhibition of nucleation; and poisoning of crystals by impurities, rendering product unsuitable for reuse as seed. These concerns were addressed through batch experiments on a landfill wastewater with a similar composition to other sulfate rich industrial wastewaters (high levels of organic and inorganic contaminants). Crystallisation rates were rapid and comparable to what is observed by others for pure solutions (2-5 h). The kinetics of crystallisation showed a 2nd order dependence on supersaturation, which have implications for crystalliser design, as discussed in the paper. No spontaneous nucleation was observed (seed was required). Seed poisoning did not occur, and product crystals were as effective as pure seed. Solubility was increased by an order of magnitude compared to a pure solution (2.6x10(-3) M2 vs. 0.22x10(-3) M2). As evaluated using equilibrium modelling, this was caused equally by non-specific ion activity, and specific ion pairing. Jarosite and ettringite could not be formed at reasonable pH and temperature levels. Given the lack of complex precipitates, and relatively high solubility, gypsum crystallisation cannot practically be used to remove sulfate to very low levels, and gas-sulfide treatment will likely still be required. It can however, be used for low-cost bulk removal of sulfate.

Evaluation of methanogenic activities during anaerobic digestion of municipal solid waste

Numerous researchers have demonstrated that the rate and extent of the degradation of municipal solid waste (MSW) can be enhanced beyond that observed in a conventional landfill by adding moisture, buffering agents and sources of microorganisms such as anaerobically digested sludge. One method of achieving the addition of these agents is by directing leachate that has trickled through a bed of anaerobically stabilised waste to beds of fresh MSW. Proper operational strategies need to be developed for successful implementation of this process on a large scale or in a landfill. Operational parameters of primary importance are the point of time at which a stabilised waste bed can be used for sequencing, the period of sequencing and the minimal amount of leachate that has to be recirculated to rapidly attain balanced microbial activity in a fresh waste bed. Assays that measure a substrate-specific methanogenic activity of an anaerobic microbial consortia have been previously developed by researchers. These assays were employed in this study to evaluate the microbial activity of the leachate for utilisation of substrates like cellulose, acetate and formate. Activity in leachate samples, taken from a batch of MSW at various times during the degradation process, was measured in terms of the amount of methane produced in 4 h after spiking the sample with one of the selected substrates. Activity resulting from the utilisation of formate and cellulose showed considerable promise as indicators for optimising operational strategies. It was observed that the formate degradation activity followed the methane production rate with both reaching a maximum at the same time and that this could be used as an indicator for determining the period of sequencing. Cellulase activity in fresh waste beds responded to flushes of mature leachate and peaked a few days after sequencing was terminated.

Cellulolytic activity in leachate during leach-bed anaerobic digestion of municipal solid waste

The degradation of municipal solid waste (MSW) under mesophilic conditions can be enhanced by exchanging leachate between fresh waste and stabilised waste. The optimum point in time when leachate from an anaerobically digesting waste bed can be used to initiate degradation of another waste bed might occur when the leachate of the digesting waste bed is highly active with cellulolytic and methanogenic bacteria. In this study, the cellulolytic activity of the leachate was measured using the cellulose-azure assay. As products of hydrolysis are soluble compounds, the rate of generation of these compounds was estimated based on a soluble chemical oxygen demand (SCOD) balance around the fresh waste bed. It was found that once the readily soluble material present in MSW was washed out there was very little generation of SCOD without the production of methane, indicating that flushing leachate from a stabilised waste bed resulted in a balanced inoculation of the fresh waste bed. With the onset of sustained methanogenesis, the rate of SCOD generation equalled the SCOD released from the digester as methane. The experimental findings also showed that cellulolytic activities of the leachate samples closely followed the trend of SCOD generation. reserved.

A methanotroph-based biorefinery: potential scenarios for generating multiple products from a single fermentation

Methane, a carbon source for methanotrophic bacteria, is the principal component of natural gas and is produced during anaerobic digestion of organic matter (biogas). Methanotrophs are a viable source of single cell protein (feed supplement) and can produce various products, since they accumulate osmolytes (e.g. ectoine, sucrose), phospholipids (potential biofuels) and biopolymers (polyhydroxybutyrate, glycogen), among others. Other cell components, such as surface layers, metal chelating proteins (methanobactin), enzymes (methane monooxygenase) or heterologous proteins hold promise as future products. Here, scenarios are presented where ectoine, polyhydroxybutyrate or protein G are synthesised as the primary product, in conjunction with a variety of ancillary products that could enhance process viability. Single or dual-stage processes and volumetric requirements for bioreactors are discussed, in terms of an annual biomass output of 1000 tonnesyear(-1). Product yields are discussed in relation to methane and oxygen consumption and organic waste generation.

Composting of waste algae: A review

Although composting has been successfully used at pilot scale to manage waste algae removed from eutrophied water environments and the compost product applied as a fertiliser, clear guidelines are not available for full scale algae composting. The review reports on the application of composting to stabilize waste algae, which to date has mainly been macro-algae, and identifies the peculiarities of algae as a composting feedstock, these being: relatively low carbon to nitrogen (C/N) ratio, which can result in nitrogen loss as NH3 and even N2O; high moisture content and low porosity, which together make aeration challenging; potentially high salinity, which can have adverse consequence for composting; and potentially have high metals and toxin content, which can affect application of the product as a fertiliser. To overcome the challenges that these peculiarities impose co-compost materials can be employed.