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Third generation biofuels, e.g. biofuels production from algal biomass, have gained attention due to increased interest on global renewable energy. However, crop-based biofuels compete with food production and should be avoided. Microalgal cultivation for biofuel production offers an alternative to crops and can become economically viable when combined with the use of used water resources. Besides nutrients and water, harvesting microalgal biomass represents one of the major costs related to biofuel production and thus efficient and cheap solutions are needed. In bacterial-algal systems, there is the potential to produce energy by co-digesting the two types of biomass. We present an innovative approach to recover microalgal biomass via a two-step flocculation using bacterial biomass after the destabilisation of microalgae with conventional cationic polymer. A short solids retention time (SRT) enhanced biological phosphorus removal (EBPR) system was combined with microalgal cultivation. Two different bacterial biomass removal strategies were assessed whereby bacterial biomass was collected from the solid-liquid separation after the anaerobic phase and after the aerobic phase. Microalgal recovery was tested by jar tests where three different chemical coagulants in coagulation-flocculation tests (AlCl3, PDADMAC and Greenfloc 120) were assessed. Furthermore, jar tests were conducted to assess the microalgal biomass recovery by a two-step flocculation method, involving chemical coagulants in the first step and bacterial biomass used in the second step to enhance the flocculation. Up to 97% of the microalgal biomass was recovered using 16 mg polymer/g algae and 0.1 g algae/g bacterial biomass. Moreover, the energy recovery by the short-SRT EBPR system combined with microalgal cultivation was assessed via biomethane potential tests. Up to 560 ± 24 mL CH4/gVS methane yield was obtained by co-digesting bacterial biomass collected after the anaerobic phase and microalgal biomass. The energy recovery in terms of methane production obtained in the short-SRT EBPR system is about 40% of the influent chemical energy.

A sorptive slurry bioscrubber adds powdered activated carbon (PAC) to a conventional suspended-growth bioscrubber. The activated carbon increases pollutant removal from the gas phase due to adsorption on carbon. The carbon is bioregenerated in the oxidation reactor and recycled to the scrubbing column. A three-stage, conventional bioscrubber was tested with and without carbon. The experiments showed that the PAC improved the removal efficiency of the system and that bioregeneration occurred. At an inlet gas-phase acetone concentration of 50 ppmv, the steady-state removal increased from 88 to 95% when activated carbon was added to the biological slurry.

Kinetic characterization of biological processes via batch respirometry requires an accurate estimate of the biomass yield coefficient because it provides the stoichiometric link between biomass synthesis, substrate consumption and oxygen uptake. Expressions for biomass yield coefficients describing autotrophic ammonia and nitrite oxidation were derived from a mechanistically based electron balanced equation. We demonstrate that applying the conventional expression used to calculate the heterotrophic biomass yield results in erroneous estimates for the autotrophic biomass yield. Yield coefficients for autotrophic NH4(+)-N to NO2(-)-N oxidation and NH4(+)-N to NO3(-)-N oxidation were overestimated by 27 to 36%. Due to correlation between the maximum specific growth rate and the biomass yield, the error in yield values propagated in 30 to 40% overestimates of the maximum specific growth rate coefficient for NH4(+)-N oxidation determined from batch respirograms. Therefore, it is essential to employ the correct expression to estimate the autotrophic biomass yield coefficient from batch respirograms due its inadvertent impact on subsequent parameter estimation.

Anaerobic ammonium oxidation (anammox) is a cost-effective process to treat high-strength nitrogenous wastewater. Even without organic carbon input, the effluent contains bioproducts from autotrophic and heterotrophic bacteria. In this work, excitation-emission matrix (EEM) fluorescence spectroscopy was used to characterize the effluent dissolved organic matter (EfOM) from an anammox reactor treating synthetic wastewater. Two dominant EEM components were identified as humic acid-like (component 1) and protein-like (component 2) substances with excitation/emission peaks at <240, 355, 420/464 nm and <240, 280, 330/346 nm, respectively. The presence of both compounds in the effluent was tracked during an activity recovery period (nitrogen load increased from 0.2 to 1.3 kg Nm(-3)d(-1)). The effluent concentration of both components increased during this period, indicating correlation between production and bacterial activity. The dynamics of these bioproducts during both substrate consumption and starvation phases was analyzed in batch experiments. Component 1 was only formed during substrate consumption in a rate proportional to ammonium removal and was considered an up-take associated product characteristic of anammox activity. The results show that the composition of the EfOM was qualitatively and quantitatively influenced by process performance. Monitoring the EfOM could, therefore, offer a useful approach to assess anammox process performance and must be further explored.

A comprehensive and global sensitivity analysis was conducted under a range of operating conditions. The relative importance of mass transfer resistance versus kinetic parameters was studied and found to depend on the operating regime as follows: Operating under the optimal loading ratio of 1.90(gO(2)/m(3)/d)/(gN/m(3)/d), the system was influenced by mass transfer (10% impact on nitrogen removal) and performance was limited by AOB activity (75% impact on nitrogen removal), while operating above, AnAOB activity was limiting (68% impact on nitrogen removal). The negative effect of oxygen mass transfer had an impact of 15% on nitrogen removal. Summarizing such quantitative analyses led to formulation of an optimal operation window, which serves a valuable tool for diagnosis of performance problems and identification of optimal solutions in nitritation/anammox applications.

One-stage autotrophic nitrogen (N) removal, requiring the simultaneous activity of aerobic and anaerobic ammonium oxidizing bacteria (AOB and AnAOB), can be obtained in spatially redox-stratified biofilms. However, previous experience with Membrane-Aerated Biofilm Reactors (MABRs) has revealed a difficulty in reducing the abundance and activity of nitrite oxidizing bacteria (NOB), which drastically lowers process efficiency. Here we show how sequential aeration is an effective strategy to attain autotrophic N removal in MABRs: Two separate MABRs, which displayed limited or no N removal under continuous aeration, could remove more than 5.5 g N/m(2)/day (at loads up to 8 g N/m(2)/day) by controlled variation of sequential aeration regimes. Daily averaged ratios of the surficial loads of O(2) (oxygen) to NH(4)(+) (ammonium) (L(O(2))/L(NH(4))) were close to 1.73 at this optimum. Real-time quantitative PCR based on 16S rRNA gene confirmed that sequential aeration, even at elevated average O(2) loads, stimulated the abundance of AnAOB and AOB and prevented the increase in NOB. Nitrous oxide (N(2)O) emissions were 100-fold lower compared to other anaerobic ammonium oxidation (Anammox)-nitritation systems. Hence, by applying periodic aeration to MABRs, one-stage autotrophic N removal biofilm reactors can be easily obtained, displaying very competitive removal rates, and negligible N(2)O emissions.

The use of respirometric for from the evaluation of intrinsic biodegradation kinetic parameters for single organic compounds is discussed. Emphasis is placed on the preliminary assessment of the data set to determine whether it is suitable for kinetic parameter estimation. Careful preliminary examination of the data avoids attempting parameter estimation with unacceptable data. Furthermore, the use of unbiased respirometric data helps ensure that the estimated parameters truly reflect the intrinsic kinetics for biodegradation of a single substrate by the culture tested. Both experimental and theoretical oxygen uptake curves are used to illustrate how various conditions can limit the utility of a given data set. The effect of substrate inhibition, dual or multiple substrate limited growth, inaccuracies in the initial conditions assumed for curve fitting, and the use of poorly acclimated cultures are discussed. Techniques are presented which allow identification of whether a data set is unsuitable and should not be used for parameter estimation. In addition, experimental procedures which can help avoid the collection of aberrant data are discussed.

Extracellular polymeric substances (EPS) have a presumed determinant role in the structure, architecture, strength, filterability, and settling behaviour of microbial solids in biological wastewater treatment processes. Consequently, numerous EPS extraction protocols have recently been published that aim to optimize the trade off between high EPS recovery and low cell lysis. Despite extensive efforts, the obtained results are often contradictory, even when analysing similar biomass samples and using similar experimental conditions, which greatly complicates the selection of an extraction protocol. This study presents a rigorous and critical assessment of existing physical and chemical EPS extraction methods applied to mixed-culture biomass samples (nitrifying, nitritation-anammox, and activated sludge biomass). A novel fluorescence-based method was developed and calibrated to quantify the lysis potential of different EPS extraction protocols. We concluded that commonly used methods to assess cell lysis (DNA concentrations or G6PDH activities in EPS extracts) do not correlate with cell viability. Furthermore, we discovered that the presence of certain chemicals in EPS extracts results in severe underestimation of protein and carbohydrate concentrations by using standard analytical methods. Keeping both maximum EPS extraction yields and minimal biomass lysis as criteria, it was identified a sonication-based extraction method as the best to determine and compare tightly-bound EPS fractions in different biomass samples. Protein was consistently the main EPS component in all analysed samples. However, EPS from nitrifying enrichments was richer in DNA, the activated sludge EPS had a higher content in humic acids and carbohydrates, and the nitritation-anammox EPS, while similar in composition to the nitrifier EPS, had a lower fraction of hydrophobic biopolymers. In general, the easily-extractable EPS fraction was more abundant in carbohydrates and humic substances, while DNA could only be found in tightly bound EPS fractions. In conclusion, the methodology presented herein supports the rational selection of analytical tools and EPS extraction protocols in further EPS characterization studies.