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Coal seam gas (CSG) is being touted as a transition fuel as the world moves towards low-carbon economies. However, the development of CSG reserves will generate enormous volumes of saline water. In this work, we investigate the potential of using this saline water to support mass algae production. Water and brine from a CSG water treatment facility (1.6 and 11.6 g total dissolved solids per litre (TDS L(-1)) respectively) were inoculated with algal biomass from freshwater and seawater environments and supplemented with nutrients in open, fed-batch reactors. Significant algal growth was recorded, with maximum specific growth rates in CSG water and CSG brine of 0.20 +/- 0.05 d(-1) and 0.26 +/- 0.04 d(-1) respectively. These maximum specific growth rates were equal to or greater than specific growth rates in deionized water and seawater diluted to the same salinity. However, algal growth lag time in CSG brine was between 7 and 9 times longer than in other waters. Microscopy and terminal-restriction fragment length polymorphism (T-RFLP) were used to monitor community structure in the reactors. The same few algal species dominated all of the reactors, except for the CSG brine reactor at day 15. This result indicates that conditions in CSG brine select for different species of algae compared to seawater of the same salinity and other waters tested. The findings suggest that mass algae production in CSG water is feasible but algae community composition may be a function of CSG water chemistry. This has implications for the downstream use of algae.

Microalgal lipid recovery for biodiesel production is currently considered suboptimal, but pre-treatment of algal biomass, the use of solvent mixtures and the positioning of transesterification can lead to increased yields. Here, the effect of various reportedly successful pre-treatments and solvent mixtures were directly compared to each other and combined with direct and indirect transesterification methods using the oleaginous microalga Tetraselmis sp. M8. Microwave and thermal pre-treatments were applied and the total lipid and fatty acid methyl ester (FAME) recoveries were investigated. The application of pre-treatments increased FAME recovery through indirect transesterification when a Soxhlet system was used but they had no significant effect for direct transesterification. Gravimetric analyses of total lipids revealed that lipid recovery was highest when utilizing the chloroform-based Bligh and Dyer extraction method; however FAME yield was the highest when applying a Soxhlet system utilizing a solvent mixture of hexane-ethanol (3:1). Total lipid recovery did not necessarily correlate with the recovery of FAMEs. The highest FAME recovery was achieved from thermal or microwave pre-treated biomass followed by indirect transesterification through Soxhlet extraction. FAME recovery could be more than doubled (increase of up to 171%) under these conditions. We conclude that a simple thermal pre-treatment (80°C for 10 min) in combination with solvent mixture extraction through indirect transesterification may present a cost-effective and scalable option for large-scale lipid extraction from microalgae.

Acidogenic fermentation is an anaerobic process of double purpose, while treating organic residues it produces chemical compounds, such as hydrogen, ethanol and organic acids. Therefore, acidogenic fermentation arises as an attractive biotechnology process towards the biorefinery concept. Moreover, this process does not need sterile operating conditions and works under a wide range of pH. Changes of operating conditions produce metabolic shifts, inducing variability on acidogenic product yield. To induce those changes, experiments, based on reactor headspace N(2)-flushing (gas phase), were designed. A major result was the hydrogen yield increase from 1 to 3.25±0.4 ( [Formula: see text] ) at pH 4.5 and N(2)-flushing of 58.4 (L·d(-1)). This yield is close to the theoretical acidogenic value (4 [Formula: see text] ). The mechanisms that explain this increase on hydrogen yield shifts are related to the thermodynamics of three metabolic reactions: lactate hydrogenase, NADH hydrogenase and homoacetogenesis, which are affected by the low hydrogen partial pressures.

Phosphorus removal in waste stabilization ponds (WSP) is highly variable, but the reasons for this are not well understood. Luxury uptake of phosphorus by microalgae has been studied in natural systems such as lakes but not under the conditions found in WSP. This work reports on the effects of phosphate concentration, light intensity, and temperature on luxury uptake of phosphorus by WSP microalgae in continuous culture bioreactors. Increasing temperature had a statistically significant "positive effect" on intracellular acid-insoluble polyphosphate concentration. It is likely that elevated temperature increased the rate of polyphosphate accumulation, but because the biomass was not starved of phosphate, the stored acid-insoluble polyphosphate was not utilized. Increasing light intensity had no effect on acid-insoluble polyphosphate but had a "negative effect" on the acid-soluble polyphosphate. A possible explanation for this is that the faster growth rate at high light intensity results in this form of polyphosphate being utilized by the cells for synthesis of cellular constituents at a rate that exceeds replenishment. The variability in the phosphorus content of the microalgal biomass shows that with this new understanding ofthe luxury uptake mechanism there is the potential to optimize WSP for biological phosphorus removal.

The use of mixed microbial cultures for the production of polyhydroxyalkanoates (PHAs) is emerging as a viable technology. In this study, 16S rRNA gene amplicon pyrosequencing was used to analyse fluctuations in populations over a 63-day period within a PHA-storing mixed microbial community enriched on fermented whey permeate. This community was dominated by the genera Flavisolibacter and Zoogloea as well as an unidentified organism belonging to the phylum Bacteroidetes. The population was observed to cycle through an increase in Zoogloea followed by a return to a community composition similar to the initial one (highly enriched in Flavisolibacter). It was found that the PHA accumulation capacity of the community was robust to population flux during enrichment and even PHA accumulation, with final polymer composition dependent on the overall proportion of acetic to propionic acids in the feed. This community adaptation suggests that mixed culture PHA production is a robust process.

In this study, the production of polyhydroxyalkanoates (PHAs) from waste activated sludge (WAS) was evaluated. PHAs were produced from fermented WAS pretreated via high-pressure thermal hydrolysis, a stream characterised by high levels of nutrients (approximately 3.5 g N L(-1) and 0.5 g P L(-1)) and soluble organics. PHA-storing organisms were successfully enriched at high organic loading rates (6 g COD(sol) L(-1) d(-1)) under aerobic dynamic feeding in sequencing batch reactors at a sludge retention time of 6 d with a short feast length less than 20% of the cycle, and a maximum substrate concentration during feast of 1 g COD(VFA) L(-1). The biomass enrichment, characterised by a decrease in species evenness based on Lorenz curves, provided a biomass that accumulated 25% PHA on a dry-biomass basis with yields on VFA of 0.4 Cmol Cmol(-1) in batch tests. The PHA consisted of ∼70 mol% 3-hydroxybutyrate and ∼30 mol% 3-hydroxyvalerate, and presented high thermal stability (T(d) = 283-287 °C) and a molecular mass ranging from 0.7 to 1.0 × 10(6) g mol(-1). Overall PHA storage was comparable to that achieved with other complex substrates; however, lower PHA storage rates (0.04-0.05 Cmol PHA(-1) Cmol X(-1) h(-1)) and productivities (3-4 Cmol PHA L(-1) h(-1)) were probably associated with a biomass-growth and high-respiration response induced by high levels of non-VFA organics (40-50% of COD(sol) in feed) and nutrients. PHA production is feasible from pretreated WAS, but the enrichment and accumulation process require further optimisation. A milder WAS pretreatment yielding lower levels of non-VFA organics and readily available nutrients may be more amenable for improved performance.

AbstractA method for detailed investigation of aerobic carbon degradation processes by microorganisms is presented. The method relies on an integrated use of the respirometric, titrimetric, and off‐gas CO2 measurements. The oxygen uptake rate (OUR), hydrogen ion production rate (HPR), and the carbon dioxide transfer rate (CTR) resulting from the biological as well as physicochemical processes, coupled with a metabolic model characterizing both the growth and carbon storage processes, enables the comprehensive study of the carbon degradation processes. The method allows the formation of carbon storage products and the biomass growth rates to be estimated without requiring any off‐line biomass or liquid‐phase measurements, although the practical identifiability of the system could be improved with additional measurements. Furthermore, the combined yield for biomass growth and carbon storage is identifiable, along with the affinity constant with respect to the carbon substrate. However, the individual yields for growth and carbon storage are not identifiable without further knowledge about the metabolic pathways employed by the microorganisms in the carbon conversion. This is true even when more process variables are measured. The method is applied to the aerobic carbon substrate degradation by a full‐scale sludge using acetate as an example carbon source. The sludge was able to quickly take up the substrate and store it as poly‐β‐hydroxybutyrate (PHB). The PHB formation rate was a few times faster than the biomass growth rate, which was confirmed by off‐line liquid‐ and solid‐phase analysis. The estimated combined yield for biomass growth and carbon storage compared closely to that determined from the theoretical yields reported in literature based on thermodynamics. This suggests that the theoretical yields may be used as default parameters for modeling purposes. © 2004 Wiley Periodicals, Inc.