Mark C.M. van Loosdrecht

Microbiology and application of the anaerobic ammonium oxidation ('anammox') process.

Ten years ago, an anaerobic ammonium oxidation (anammox) process was discovered in a denitrifying pilot plant reactor. From this system, a highly enriched microbial community was obtained, dominated by a single deep-branching planctomycete, Candidatus Brocadia anammoxidans. Phylogenetic inventories of different wastewater treatment plants with anammox activity have suggested that at least two genera in Planctomycetales can catalyse the anammox process. Electron microscopy of the ultrastructure of B. anammoxidans has shown that several membrane-bounded compartments are present inside the cytoplasm. Hydroxylamine oxidoreductase, a key anammox enzyme, is found exclusively inside one of these compartments, tentatively named the anammoxosome.

Modeling the PAO–GAO competition: Effects of carbon source, pH and temperature

The influence of different carbon sources (acetate to propionate ratios), temperature and pH levels on the competition between polyphosphate- and glycogen-accumulating organisms (PAO and GAO, respectively) was evaluated using a metabolic model that incorporated the carbon source, temperature and pH dependences of these microorganisms. The model satisfactorily described the bacterial activity of PAO (Accumulibacter) and GAO (Competibacter and Alphaproteobacteria-GAO) laboratory-enriched cultures cultivated on propionate (HPr) and acetate (HAc) at standard conditions (20 degrees C and pH 7.0). Using the calibrated model, the effects of different influent HAc to HPr ratios (100-0, 75-25, 50-50 and 0-100%), temperatures (10, 20 and 30 degrees C) and pH levels (6.0, 7.0 and 7.5) on the competition among Accumulibacter, Competibacter and Alphaproteobacteria-GAO were evaluated. The main aim was to assess which conditions were favorable for the existence of PAO and, therefore, beneficial for the biological phosphorus removal process in sewage treatment plants. At low temperature (10 degrees C), PAO were the dominant microorganisms regardless of the used influent carbon source or pH. At moderate temperature (20 degrees C), PAO dominated the competition when HAc and HPr were simultaneously supplied (75-25 and 50-50% HAc to HPr ratios). However, the use of either HAc or HPr as sole carbon source at 20 degrees C was not favorable for PAO unless a high pH was used (7.5). Meanwhile, at higher temperature (30 degrees C), GAO tended to be the dominant microorganisms. Nevertheless, the combined presence of acetate and propionate in the influent (75-25 and 50-50% HAc to HPr ratios) as well as a high pH (7.5) appear to be potential factors to favor the metabolism of PAO over GAO at higher sewage temperature (30 degrees C).

Improved nitrogen removal by application of new nitrogen-cycle bacteria

In order to meet increasingly stringentEuropean discharge standards, new applicationsand control strategies for the sustainableremoval of ammonia from wastewater have to beimplemented. In this paper we discuss anitrogen removal system based on the processesof partial nitrification and anoxic ammoniaoxidation (anammox). The anammox process offersgreat opportunities to remove ammonia in fullyautotrophic systems with biomass retention. Noorganic carbon is needed in such nitrogenremoval system, since ammonia is used aselectron donor for nitrite reduction. Thenitrite can be produced from ammonia inoxygen-limited biofilm systems or in continuousprocesses without biomass retention. Forsuccessful implementation of the combinedprocesses, accurate biosensors for measuringammonia and nitrite concentrations, insight inthe complex microbial communities involved, andnew control strategies have to be developed andevaluated.

Aerobic sludge granulation: A tale of two polysaccharides?

Aerobic sludge granules are suspended biofilms with the potential to reduce the cost and footprint of secondary wastewater treatment. Attempts to answer how and why they form leads to a consideration of the role of their extracellular polymeric substances (EPS) in determining their physical and microbiological properties. The exopolysaccharide components of this matrix, in particular, have received attention as putative structural, gel-forming agents. Two quite different exopolysaccharides have been proposed as the gel-forming constituents, with their gel properties clearly different from those of activated sludge EPS. This review aims to address the question of whether more than one gel-forming exopolysaccharide exist in granules. Based on the available structural data, it seems likely that they are different gel-forming polymers and their differences are not artifacts of the analytical methods used. Nonetheless, both proposed structural gel polymers are extracted and purified based on procedures selecting for anionic polar polysaccharides soluble at high pH, and both contain hexuronic acids. Granulation does not result from EPS synthesis by any single microbial population, nor from production of a single exopolysaccharide. Future studies using solvents suitable for recalcitrant polysaccharides are likely to reveal important structural roles for other polysaccharides. It is hoped that this article will serve as a guide for subsequent studies into understanding the roles of exopolysaccharides in aerobic granular sludge.

Evaluating sludge minimization caused by predation and viral infection based on the extended activated sludge model No. 2d

The Activated Sludge Model No. 2d (ASM2d) was extended to incorporate the processes of both predation and viral infection. The extended model was used to evaluate the contributions of predation and viral infection to sludge minimization in a sequencing batch reactor (SBR) system enriching polyphosphate-accumulating organisms (PAOs). Three individual decay processes formulated according to the general model rules were used in the extended model. The model was firstly calibrated and validated by different experimental results. It was used to evaluate the potential extent of predation and viral infection on sludge minimization. Simulations indicate that predation contributes roughly two times more to sludge minimization than viral infection in the SBR system enriching PAOs. The sensitivity analyses of the selected key parameters reveal that there are thresholds on both predation and viral infection rates, if they are too large a minimal sludge retention time is obtained and the effluent quality is deteriorating. Due to the thresholds, the contributions of predation and viral infection to sludge minimization are limited to a maximal extent of about 21% and 9%, respectively. However, it should be noted that the parameters concerning predation and viral infection were not calibrated separately by independent experiment in our study due to the lack of an effective method, especially for the parameters regarding viral infection. Therefore, it is essential to better evaluate these parameters in the future.

A mathematical model for electrochemically active filamentous sulfide-oxidising bacteria

Oxygen and sulfide in ocean sediments can be consumed biologically over long spatial distances by way of filamentous bacteria in electron-conducting sheaths. To analyse observations, a mathematical model of these filamentous sulfur-oxidising bacteria was developed, including electrical conduction between reactive zones. Mechanisms include Nernst-Planck diffusion and migration of ions coupled with Ohms law for conduction along filaments, and metabolic activity throughout the filaments. Simulations predict outward biomass growth toward the boundaries of the sediment floor and top surface, resulting in two distinct zones with anode (sulfide consumption) and cathode (oxygen consumption) reactions enabled by electron conduction. Results show inward fluxes of 4.6 mmol O2/m(2)/d and 2.5 mmol S/m(2)/d, with consumption increasing with growth to final fluxes of 8.2 mmol O2/m(2)/d and 4.34 mmol S/m(2)/d. Qualitatively, the effect of varying cell conductivity and substrate affinity is evaluated. Controlling mechanisms are identified to shift from biomass limitation, to substrate limitation, and to conductivity limitations as the lengths of the filaments increase. While most observed data are reflected in the simulation results, a key discrepancy is the lower growth rates, which are largely fixed by thermodynamics, indicating that microbes may utilise secondary substrates or an alternative metabolism.