Hybrid microalgae-activated sludge system for carbon-efficient wastewater treatment.

Engineered microalgae-bacteria systems can play a key role in the realisation of energy-efficient carbon-neutral wastewater treatment technologies. An attempt was made to develop a hybrid microalgae-activated sludge (HMAS) system coupling carbon capture with domestic wastewater treatment. Photobioreactors internally illuminated with red light-emitting diodes (LEDs), and inoculated with mixed microbial culture, resulted in substantial savings in operational cost. System performance was evaluated at about 600 µmol/m2 s LED irradiance while treating synthetic municipal wastewater in a chemostat for about 2 months, containing about 250 mg/L soluble chemical oxygen demand (SCOD), 90 mg/L NH3-N and 10 mg/L orthophosphate. Carbon dioxide was supplied into the HMAS at 25 mL/min, 25% v/v. SCOD was efficiently removed from the wastewater (up to 70%) and bacterial oxygen requirement of >2 mg/L was met through microalgal photosynthesis. The system demonstrated its potential in achieving carbon-efficient wastewater treatment.

Evaluating an anaerobic digestion (AD) feedstock derived from a novel non-source segregated municipal solid waste (MSW) product.

In many nations industrial scale AD of non-agricultural waste materials (such as MSW) has not yet reached its full potential, often constrained by the lack of secure, inexpensive, high quality AD feedstocks, and markets for the resulting digestate material. We tested the output material of a high throughput novel industrial process to define its potential as an AD feedstock (based on quality and consistency). This process, designed to circumvent the constraints of source segregation while still generating segregated waste streams, resulted in the production of a temporally homogenous fibrous material with: an average moisture content of 44.2 (±2.33)%; C:N ratio of ∼32.9:1 (±3.46:1), C:P ratio of ∼228:1 and gross calorific value of 17.4 (±0.29)MJ/kg(DM). This material provided a CH4 yield of between 201 and 297m3 CH4/tonne(DM) (271-401m3CH4/tonne(vs)) comparable to commonly used AD feedstocks. Material contaminant levels were temporally consistent (P>0.05), (average values being Cd 0.63 (±0.19), Cu 56.3 (±7.45), Crtot 51.4 (±4.41), Hg<0.3, Ni 28.9 (±5.17), Pb 79.2 (±23.71), Zn 202 (±44.5), total polyaromatic hydrocarbons (PAH) 2.2 (±0.3), and total polychlorinated biphenyls (PCB) (<0.2)mg/kg(DM)). Calculated digestate contaminant levels were below the median contaminant threshold limits for anaerobic digestates of all countries within the European Union i.e. of Cd 3.35, Cu 535, Crtot 535, Hg 8.15, Ni 185, Pb 397.5, Zn 2100mg/kg(DM). We suggest that novel high throughput processes that produce high quality AD feedstocks, may have a place in further diversion of waste from landfill.

Energy-efficient stirred-tank photobioreactors for simultaneous carbon capture and municipal wastewater treatment.

Algal based wastewater treatment (WWT) technologies are attracting renewed attention because they couple energy-efficient sustainable treatment with carbon capture, and reduce the carbon footprint of the process. A low-cost energy-efficient mixed microalgal culture-based pilot WWT system, coupled with carbon dioxide (CO2) sequestration, was investigated. The 21 L stirred-tank photobioreactors (STPBR) used light-emitting diodes as the light source, resulting in substantially reduced operational costs. The STPBR were operated at average optimal light intensity of 582.7 µmol.s(-1).m(-2), treating synthetic municipal wastewater containing approximately 250, 90 and 10 mg.L(-1) of soluble chemical oxygen demand (SCOD), ammonium (NH4-N), and phosphate, respectively. The STPBR were maintained for 64 days without oxygen supplementation, but had a supply of CO2 (25 mL.min(-1), 25% v/v in N2). Relatively high SCOD removal efficiency (>70%) was achieved in all STPBR. Low operational cost was achieved by eliminating the need for mechanical aeration, with microalgal photosynthesis providing all oxygenation. The STPBR achieved an energy saving of up to 95%, compared to the conventional AS system. This study demonstrates that microalgal photobioreactors can provide effective WWT and carbon capture, simultaneously, in a system with potential for scaling-up to municipal WWT plants.

Tolerance of the antibiotic tylosin on treatment performance of an up-flow anaerobic stage reactor (UASR).

Tylosin has been considered inhibiting COD removal in anaerobic digestion. In this study it is proven that this is not always the case. Accordingly, elevated concentrations of Tylosin (100-800mgL-1) could be tolerated by the anaerobic system. The influence of Tylosin concentrations on an up-flow anaerobic stage reactor (UASR) was assessed using additions of Tylosin phosphate concentrate. Results showed high efficiency for COD removal (average 93%) when Tylosin was present at concentrations ranging from 0 to 400 mg L-1. However, at Tylosin concentrations of 600 and 800 mg L-1 treatment efficiency declined to 85% and 75% removal respectively. The impact of Tylosin concentrations on archaeal activity were investigated and the analysis revealed that archaeal cells dominated the reactor, confirming that there was no detectable inhibition of the methanogens at Tylosin levels between 100 and 400mg L-1. Nevertheless, the investigation showed a slight reduction in the number of methanogens at Tylosin levels of 600 and 800 mg L-1. These results demonstrated that the methanogens were well adapted to Tylosin. It would not be expected that the process performance of the UASR would be affected, not even at a level well in excess of those appearing in real wastewater from a Tylosin production site.

Granule development in a split-feed anaerobic baffled reactor.

Operating anaerobic reactors at high organic loading rates during start-up can lead to instability, accumulation of volatile fatty acids and low pH, such problems being exacerbated in reactors that exhibit plug-flow characteristics. Moreover, plug-flow conditions increase the exposure of biomass to any toxic components in the feed. To overcome these limitations, an anaerobic baffled reactor (ABR), a reactor exhibiting partial plug-flow characteristics, was modified by splitting the feed between the individual compartments to produce the split-feed ABR (SFABR). Consequently, more favourable conditions were created in the initial compartments, such as lower, longer hydraulic retention time and longer cell retention time; conditions in the final compartments were also improved by the increased food availability for microorganisms. Other benefits included better gas mixing characteristics as a result of the more balanced gas production across the reactor. Granule development was compared in SFABR and normally fed ABR by analysing sludge samples, taken during start-up and continuous operation, using scanning electron microscopy. Photomicrographs allowed tentative conclusions to be made concerning the effect of split-feeding on the distribution of bacterial populations within the granule architecture and the role of extracellular polymers on granule formation.

Improved split feed anaerobic baffled reactor (SFABR) for shorter start-up period and higher process performance.

In this paper, the Split Feed Anaerobic Baffled Reactor (SFABR) is introduced and investigations into the use of modified seed material are described. It was shown that shorter and reliable start-up times could be achieved with SFABR when using improved seed material, even for the treatment of particularly problematic wastewater, i.e. ice-cream wastewater. Scanning electron photo-micrographs (SEM) revealed that granulation process occurs relatively rapidly in the SFABR compared with other reactor configurations, and that the reactor contained a highly mixed population of methanogens in all compartments. The use of polymer-conditioned anaerobic sludge and granular sludge as seed proved advantageous over the use of suspended growth anaerobic sludge, and the "improved" SFABR consequently performed more efficiently and also showed greater stability than the conventional ABR.

The effect of polymer addition on granulation in an anaerobic baffled reactor (ABR). Part I: process performance.

The stability and performance of an anaerobic baffled reactor (ABR) treating an ice-cream wastewater at several organic loading rates have been investigated. Specifically, it was determined whether an ABR would promote phase separation and if a polymer additive was capable of enhancing granule formation in an ABR. In order to achieve these goals, two ABRs, having identical dimensions and configurations, were used to study the above objectives using a synthetic ice-cream wastewater. The ABR proved to be an efficient reactor configuration for the treatment of a high-strength synthetic ice-cream wastewater. An organic loading rate of around 15 kg CODm(-3) d(-1) was treated with a 99% COD removal efficiency. From the jar test and inhibition assay, it was concluded that Kymene SLX-2 was the most effective and least inhibitory polymer tested. The methane yield was higher in the polymer-amended reactor compared to the control reactor. In addition, polymer addition resulted in a considerably higher degree of biomass retention and lower solids washout from the ABR. Consequently, it demonstrated that there was a considerable potential for sludge conditioning in ABRs by facilitating better biomass retention within the reactor which in turn led to better process performance. Granulation was achieved in both ABRs within 3 months. However, the granules from the polymer-amended reactor appeared earlier and were generally larger and more compact, although this was not quantified in detail during the present study. The main advantage of using an ABR comes from its compartmentalised structure. The first compartment of an ABR may act as a buffer zone to all toxic and inhibitory material in the feed thus allowing the later compartments to be loaded with a relatively harmless, balanced and mostly acidified influent. In this respect, the latter compartments would be more likely to support active populations of the relatively sensitive methanogenic bacteria and partly explains why the best granules and the highest methane yield were obtained in Compartment 2. It is unlikely that a complete separation of phases (acidogenic and methanogenic) occurred within the ABRs since methane production was observed in all compartments, although this was low (approximately 40% of all gas composition) in Compartment 1, becoming higher (approximately 70%) in the following compartments.

The effect of polymer addition on granulation in an anaerobic baffled reactor (ABR). Part II: compartmentalization of bacterial populations.

The microbial ecology of wastewater treatment plants remains one of the least understood aspects in both aerobic and anaerobic systems, despite the fact that both processes are ultimately dependent on an active biomass for operational efficiency. Ultimately, future developments in anaerobic treatment processes will require a much greater understanding of the fundamental relationships between bacterial populations within the biomass if optimum process efficiency is to be fully realised. This study assesses the influence of polymer addition on granule formation within an ABR and compares the ecology of the biomass in each compartment of two ABRs treating ice-cream wastewater. To our knowledge, this is the first reported characterisation of the microbiology of acidogenic and methanogenic bacteria in the individual compartments of an ABR. The polymer-amended reactor contained sludge that had a greater density of anaerobic bacteria and larger and denser granules than the control reactor, indicating that polymer addition possibly contributed to the retention of active biomass within the ABR. The average fraction of autofluorescent methanogens was lower, with 1.5% being in the initial compartments of the ABRs, compared to the last compartment which had 15%, showing that each compartment of an ABR had a unique microbial composition. Partial spatial separation of anaerobic bacteria appeared to have taken place with acidogenic bacteria predominating in the initial compartments and methanogenic bacteria predominating in the final compartments. Scanning electron micrographs have revealed that the dominant bacteria in the initial compartments of the ABR (Compartments 1 and 2) were those which could consume H2/CO2 and formate as substrate, i.e. Methanobrevibacter, Methanococcus, with populations shifting to acetate utilisers, i.e. Methanosaeta, Methanosarcina, in the final compartments (Compartments 3 and 4). In addition, there appeared to be a stratified structure to the bacterial genera present within the granules.

Biochemical characterization of a haloalcohol dehalogenase from Arthrobacter erithii H10a.

Arthrobacter erithii H10a possesses two enzymes capable of catalyzing the dehalogenation of vicinal halohydrins which have been designated as dehalogenases DehA and DehC. The DehA dehalogenase demonstrated greater activity toward 1,3-dichloro-2-propanol (1,3-DCP) while the DehC dehalogenase showed higher activity toward 3-chloro-1,2-propanediol (3-CPD) and brominated alcohols. The DehA dehalogenase was composed of two non-identical subunits (relative molecular mass of 31.5 and 34 kDa) which probably associate with other proteins to form a large catalytically active protein of 200 kDa. The two subunits were purified and the amino acid sequence of their tryptic digests determined. The DehA enzyme catalyzed the conversion of vicinal halohydrins to epoxides and the reverse reaction in the presence of an excess of halogen. This enzyme had maximum activity at 50 degrees C and a broad pH optimum over the range 8.5-10.5. The apparent K(m) and Vmax values for dehalogenation of 1,3-DCP and 3-CPD were 0.105 mM and 223 mumol min-1 mg-1; and 2.366 mM and 1.742 mumol min-1 mg-1, respectively. The enzyme was inhibited by 2-chloroacetic acid (MCA) and 2,2-dichloroacetic acid (DCA). The inhibition pattern suggested a mixed type inhibition which was predominantly uncompetitive. Amino acid modification experiments demonstrated that one or more cysteine and arginine residues are likely to be involved in catalysis or play an important role in the maintenance of the enzyme structure. The characteristics of the DehA enzyme are compared to those of previously reported haloalcohol dehalogenases and discussed in terms of diversity of this type of dehalogenase.

Dehalogenation of haloalkanes by Rhodococcus erythropolis Y2. The presence of an oxygenase-type dehalogenase activity complements that of an halidohydrolase activity.

Rhodococcus erythropolis Y2 produced two types of dehalogenase: a hydrolytic enzyme, that is an halidohydrolase, which was induced by C3 to C6 1-haloalkane substrates, and at least one oxygenase-type dehalogenase induced by C7 to C16 1-haloalkanes and n-alkanes. The oxygenase-type activity dehalogenated C4 to C18 1-chloroalkanes with an optimum activity towards 1-chlorotetradecane. The halidohydrolase catalysed the dehalogenation of a wide range of 1- and alpha,omega-disubstituted haloalkanes and alpha,omega-substituted haloalcohols. In resting cell suspensions of hexadecane-grown R. erythropolis Y2 the oxygenase-type dehalogenase had a specific activity of 12.9 mU (mg protein)-1 towards 1-chlorotetradecane (3.67 mU mg-1 towards 1-chlorobutane) whereas the halidohydrolase in 1-chlorobutane-grown batch cultures had a specific activity of 44 mU (mg protein)-1 towards 1-chlorobutane. The significance of the two dehalogenase systems in a single bacterial strain is discussed in terms of their contribution to the overall catabolic potential of the organism.

Isolation and characterization of a haloalkane halidohydrolase from Rhodococcus erythropolis Y2.

Rhodococcus erythropolis strain Y2, isolated from soil by enrichment culture using 1-chlorobutane, was able to utilize a range of halogenated aliphatic compounds as sole sources of carbon and energy. The ability to utilize 1-chlorobutane was conferred by a single halidohydrolase-type haloalkane dehalogenase. The presence of the single enzyme in cell-free extracts was demonstrated by activity strain polyacrylamide gel electrophoresis. The purified enzyme was a monomeric protein with a relative molecular mass of 34 kDa and demonstrated activity against a broad range of haloalkanes, haloalcohols and haloethers. The highest activity was found towards alpha, omega disubstituted chloro- and bromo- C2-C6 alkanes and 4-chlorobutanol. The Km value of the enzyme for 1-chlorobutane was 0.26 mM. A comparison of the R. erythropolis Y2 haloalkane halidohydrolase with other haloalkane dehalogenases is discussed on the basis of biochemical properties and N-terminal amino acid sequence data.

The characterization of amidohydrolases in a freshwater lake sediment.

The properties of three amidohydrolases, i.e., urease (I) EC 3.5.1.5, L-asparaginase (II) EC 3.5.1.1, and L-glutaminase (III) EC 3.5.1.2, were studied in sediment samples taken from a shallow eutrophic freshwater lake.Sediment samples were air dried (ADS) and stored for at least 3 months before being enzymically characterized. The pH optimum of I, II, and III were pH 7.0, 8.4, and 6.5-7.0, respectively, while III in soluble extracts from ADS was most active between pH 8.0 and 9.0. The temperature response of the three enzymes in ADS gave Ea values of 38.9, 41.6, and 35.9 kJmol(-1) for I, II, and III, respectively. Km and Vmax values for ADS I, II, and III were 1.2 mM and 1.9µmol NH3 g(-1)h(-1); 0.8 mM and 4.1µmol NH3 g(-1)h(-1); and 1.25 mM and 17.4µmol NH3 g(-1)h(-1). Km values for all three enzymes in ADS extracts were at least an order of magnitude greater than those of the ADS. The susceptability of each enzyme to proteolysis was followed in ADS and fresh wet sediment and compared with that of III in an ADS extract. All sediment enzymes were found to be more resistant than the commercial preparation of bacterial L-glutaminase subjected to the same treatment. These results suggested that I, II, and III all exist to some extent as colloid-immobilized enzyme fractions in freshwater sediments and are analogous to the stable enzyme fractions in soils.