Growth media in anaerobic fermentative processes: The underestimated potential of thermophilic fermentation and anaerobic digestion.

Fermentation and anaerobic digestion of organic waste and wastewater is broadly studied and applied. Despite widely available results and data for these processes, comparison of the generated results in literature is difficult. Not only due to the used variety of process conditions, but also because of the many different growth media that are used. Composition of growth media can influence biogas production (rates) and lead to process instability during anaerobic digestion. To be able to compare results of the different studies reported, and to ensure nutrient limitation is not influencing observations ascribed to process dynamics and/or reaction kinetics, a standard protocol for creating a defined growth medium for anaerobic digestion and mixed culture fermentation is proposed. This paper explains the role(s) of the different macro- and micronutrients, as well as the choices for a growth medium formulation strategy. In addition, the differences in nutrient requirements between mesophilic and thermophilic systems are discussed as well as the importance of specific trace metals regarding specific conversion routes and the possible supplementary requirement of vitamins. The paper will also give some insight into the bio-availability and toxicity of trace metals. A remarkable finding is that mesophilic and thermophilic enzymes are quite comparable at their optimum temperatures. This has consequences for the trace metal requirements of thermophiles under certain conditions. Under non-limiting conditions, the trace metal requirement of thermophilic systems is about 3 times higher than for mesophilic systems.

Influence of chemically enhanced primary treatment on anaerobic digestion and dewaterability of waste sludge.

To lower energy consumption at a sewage treatment plant (STP), primary settling could be enhanced to direct more chemical oxygen demand (COD) to anaerobic digestion (AD) for increased biogas production and decreased aeration. Primary settling can be chemically enhanced by applying flocculation aids (FAs). FAs are refractory compounds that may affect all sludge treatment facilities. In this study the consequences are investigated of the application of FAs for chemically enhanced primary treatment (CEPT) on AD and subsequent dewatering of digested sludge in a conventional STP. It was found that FAs maintain their effect throughout all sludge processing facilities. With CEPT, more readily degradable solids were removed, resulting in a higher bio methane potential of the primary sludge. In AD, FAs lowered the viscosity; meanwhile an increased hydrolysis rate was observed. But FAs also partially irreversibly bound substrate in such way that it is not available for biological degradation anymore. In subsequent dewatering of digested sludge, a higher dry solids concentration was observed with CEPT. A computer simulation showed that in a conventional STP, CEPT would not be economically feasible. However, several benefits were discussed that can make CEPT an interesting option for future low COD/N-tolerant STPs with, for example, Anammox processes for N removal.

Full scale performance of the aerobic granular sludge process for sewage treatment.

Recently, aerobic granular sludge technology has been scaled-up and implemented for industrial and municipal wastewater treatment under the trade name Nereda(®). With full-scale references for industrial treatment application since 2006 and domestic sewage since 2009 only limited operating data have been presented in scientific literature so far. In this study performance, granulation and design considerations of an aerobic granular sludge plant on domestic wastewater at the WWTP Garmerwolde, the Netherlands were analysed. After a start-up period of approximately 5 months, a robust and stable granule bed (>8 g L(-1)) was formed and could be maintained thereafter, with a sludge volume index after 5 min settling of 45 mL g(-1). The granular sludge consisted for more than 80% of granules larger than 0.2 mm and more than 60% larger than 1 mm. Effluent requirements (7 mg N L(-1) and 1 mg P L(-1)) were easily met during summer and winter. Maximum volumetric conversion rates for nitrogen and phosphorus were respectively 0.17 and 0.24 kg (m(3) d)(-1). The energy usage was 13.9 kWh (PE150·year)(-1) which is 58-63 % lower than the average conventional activated sludge treatment plant in the Netherlands. Finally, this study demonstrated that aerobic granular sludge technology can effectively be implemented for the treatment of domestic wastewater.

Pilot-scale evaluation of anammox-based mainstream nitrogen removal from municipal wastewater.

Autotrophic nitrogen removal in the mainstream wastewater treatment process is suggested to be a prerequisite of energy autarkic wastewater treatment plants (WWTP). Whilst the application of anammox-related technologies in the side-stream is at present state of the art, the feasibility of this energy-efficient process at mainstream conditions is still under development. Lower operating temperature and ammonium concentration, together with required high nitrogen removal efficiency, represent the main challenges to face in order to reach this appealing new frontier of the wastewater treatment field. In this study, we report the evaluation of the process in a plug-flow granular sludge-based pilot-scale reactor (4 m3) continuously fed with the actual effluent of the A-stage of the WWTP of Dokhaven, Rotterdam. The one-stage partial nitritation-anammox system was operated for more than 10 months at 19±1°C. Observed average N-removal and ammonium conversion rates were comparable or higher than those of conventional N-removal systems, with 182±46 and 315±33 mg-N L(-1) d(-1), respectively. Biochemical oxygen demand was also oxidized in the system with an average removal efficiency of 90%. Heterotrophic biomass grew preferentially in flocs and was efficiently washed out of the system. Throughout the experimentation, the main bottleneck was the nitritation process that resulted in nitrite-limiting conditions for the anammox conversion. Anammox bacteria were able to grow under mainstream WWTP conditions and new granules were formed and efficiently retained in the system.

Quantification of continual anthropogenic pollutants released in swimming pools.

Disinfection in swimming pools is often performed by chlorination, However, anthropogenic pollutants from swimmers will react with chlorine and form disinfection by-products (DBPs). DBPs are unwanted from a health point of view, because some are irritating, while others might be carcinogenic. The reduction of anthropogenic pollutants will lead to a reduction in DBPs. This paper investigates the continual release of anthropogenic pollutants by means of controlled sweat experiments in a pool tank during laboratory time-series experiments (LTS experiments) and also during on-site experiments (OS experiments) in a swimming pool. The sweat released during the OS and LTS experiments was very similar. The sweat rate found was 0.1-0.2 L/m(2)/h at water temperatures below 29 °C and increased linearly with increasing water temperatures to 0.8 L/m(2)/h at 35 °C. The continual anthropogenic pollutant release (CAPR) not only consisted of sweat, particles (mainly skin fragments and hair) and micro-organisms, but also sebum (skin lipids) has to be considered. The release of most components can be explained by the composition of sweat. The average release during 30 min of exercise is 250 mg/bather non-purgeable organic carbon (NPOC), 77.3 mg/bather total nitrogen (TN), 37.1 mg/bather urea and 10.1 mg/bather ammonium. The release of NPOC cannot be explained by the composition of sweat and is most probably a result of sebum release. The average release of other components was 1.31 × 10(9) # particles/bather (2-50 µm), 5.2 µg/bather intracellular adenosine triphosphate (cATP) and 9.3 × 10(6) intact cell count/bather (iCC). The pool water temperature was the main parameter to restrain the CAPR. This study showed that a significant amount of the total anthropogenic pollutants release is due to unhygienic behaviour of bathers.

Evaluating the main and side effects of high salinity on aerobic granular sludge.

Salinity can adversely affect the performance of most biological processes involved in wastewater treatment. The effect of salt on the main conversion processes in an aerobic granular sludge (AGS) process accomplishing simultaneous organic matter, nitrogen, and phosphate removal was evaluated in this work. Hereto, an AGS sequencing batch reactor was subjected to different salt concentrations (0.2 to 20 g Cl(-) l(-1)). Granular structure was stable throughout the whole experimental period, although granule size decreased and a significant effluent turbidity was observed at the highest salinity tested. A weaker gel structure at higher salt concentrations was hypothesised to be the cause of such turbidity. Ammonium oxidation was not affected at any of the salt concentrations applied. However, nitrite oxidation was severely affected, especially at 20 g Cl(-) l(-1), in which a complete inhibition was observed. Consequently, high nitrite accumulation occurred. Phosphate removal was also found to be inhibited at the highest salt concentration tested. Complementary experiments have shown that a cascade inhibition effect took place: first, the deterioration of nitrite oxidation resulted in high nitrite concentrations and this in turn resulted in a detrimental effect to polyphosphate-accumulating organisms. By preventing the occurrence of the nitrification process and therefore avoiding the nitrite accumulation, the effect of salt concentrations on the bio-P removal process was shown to be negligible up to 13 g Cl(-) l(-1). Salt concentrations equal to 20 g Cl(-) l(-1) or higher in absence of nitrite also significantly reduced phosphate removal efficiency in the system.

Strength characteristics of aerobic granular sludge.

Aerobic granular sludge has a number of advantages over conventional activated sludge flocs, such as cohesive and strong matrix, fast settling characteristic, high biomass retention and ability to withstand high organic loadings, all aspects leading towards a compact reactor system. Still there are very few studies on the strength of aerobic granules. A procedure that has been used previously for anaerobic granular sludge strength analysis was adapted and used in this study. A new coefficient was introduced, called a stability coefficient (S), to quantify the strength of the aerobic granules. Indicators were also developed based on the strength analysis results, in order to categorize aerobic granules into three levels of strength, i.e. very strong (very stable), strong (stable) and not strong (not stable). The results indicated that aerobic granules grown on acetate were stronger (high density: >150 g T SSL(-1) and low S value: 5%) than granules developed on sewage as influent. A lower value of S indicates a higher stability of the granules.

Behavior of polymeric substrates in an aerobic granular sludge system.

Particulate and slowly biodegradable substrates form an important fraction of industrial wastewater and sewage. To study the influence of suspended solids and colloidal substrate on the morphology and performance of aerobic granular sludge, suspended and soluble starch was used as a model substrate. Degradation was studied using microscopy, micro-electrode measurements, batch experiments and long term laboratory scale reactor operation. Starch was removed by adsorption at the granule surface, followed by hydrolysis and consumption of the hydrolyzed products. Aerobic granules could be maintained on starch as sole influent carbon source, but their structure was filamentous and irregular. It is hypothesized that this is related to the low starch hydrolysis rates, leading to available substrate during the aeration period (extended feast period) and resulting in increased substrate gradients over the granules. The latter induces a less uniform granule development. Starch adsorbed and was consumed at the granule surface instead of being accumulated inside the granules as occurs for soluble substrates. Therefore the simultaneous denitrification efficiencies remained low. Moreover, many protozoa and metazoans were observed in laboratory reactors as well as in pilot- and full-scale Nereda(®) reactors, indicating an important role in the removal of suspended solids too.

Settling behaviour of aerobic granular sludge.

Aerobic granular sludge (AGS) technology has been extensively studied recently to improve sludge settling and behaviour in activated sludge systems. The main advantage is that aerobic granular sludge (AGS) can settle very fast in a reactor or clarifier because AGS is compact and has strong structure. It also has good settleability and a high capacity for biomass retention. Several experimental works have been conducted in this study to observe the settling behaviours of AGS. The study thus has two aims: (1) to compare the settling profile of AGS with other sludge flocs and (2) to observe the influence of mechanical mixing and design of the reactor to the settleability of AGS. The first experimental outcome shows that AGS settles after less than 5 min in a depth of 0.4 m compared to other sludge flocs (from sequencing batch reactor, conventional activated sludge and extended aeration) which takes more than 30 min. This study also shows that the turbulence from the mixing mechanism and shear in the reactor provides an insignificant effect on the AGS settling velocity.

Aerobic granular sludge--state of the art.

In September 2006, preliminary to the IWA biofilm conference, a second workshop about aerobic granular sludge was held in Delft, The Netherlands, of which a summary of the discussion outcomes is given in this paper. The definition of aerobic granular sludge was discussed and complemented with a few additional demands. Further topics were formation and morphology of aerobic granular sludge, modelling and use of the aerobic granular sludge in practice.

Kinetic model of a granular sludge SBR: influences on nutrient removal.

A mathematical model was developed that can be used to describe an aerobic granular sludge reactor, fed with a defined influent, capable of simultaneously removing COD, nitrogen and phosphate in one sequencing batch reactor (SBR). The model described the experimental data from this complex system sufficiently. The effect of process parameters on the nutrient removal rates could therefore be reliably evaluated. The influence of oxygen concentration, temperature, granule diameter, sludge loading rate, and cycle configuration were analyzed. Oxygen penetration depth in combination with the position of the autotrophic biomass played a crucial role in the conversion rates of the different components and thus on overall nutrient removal efficiencies. The ratio between aerobic and anoxic volume in the granule strongly determines the N-removal efficiency as it was shown by model simulations with varying oxygen concentration, temperature, and granule size. The optimum granule diameter for maximum N- and P-removal in the standard case operating conditions (DO 2 mg L(-1), 20 degrees C) was found between 1.2 and 1.4 mm and the optimum COD loading rate was 1.9 kg COD m(-3) day(-1). When all ammonia is oxidized, oxygen diffuses to the core of the granule inhibiting the denitrification process. In order to optimize the process, anoxic phases can be implemented in the SBR-cycle configuration, leading to a more efficient overall N-removal. Phosphate removal efficiency mainly depends on the sludge age; if the SRT exceeds 30 days not enough biomass is removed from the system to keep effluent phosphate concentrations low.

Formation of aerobic granules and conversion processes in an aerobic granular sludge reactor at moderate and low temperatures.

Temperature changes can influence biological processes considerably. To investigate the effect of temperature changes on the conversion processes and the stability of aerobic granular sludge, an aerobic granular sludge sequencing batch reactor (GSBR) was exposed to short-term and long-term temperature changes. Start-up at 8 degrees C resulted in irregular granules that aggregated as soon as aeration was stopped, which caused severe biomass washout and instable operation. The presence of COD during the aerobic phase is considered to be the major reason for this granule instability. Start-up at 20 degrees C and lowering the temperature to 15 degrees C and 8 degrees C did not have any effect on granule stability and biomass could be easily retained in the system. The temperature dependency of nitrification was lower for aerobic granules than usually found for activated sludge. Due to decreased activity in the outer layers of granules at lower temperatures, the oxygen penetration depth could increase, which resulted in a larger aerobic biomass volume, compensating the decreased activity of individual organisms. Consequently the denitrifying capacity of the granules decreased at reduced temperatures, resulting in an overall poorer nitrogen removal capacity. The overall conclusion that can be drawn from the experiments at low temperatures is that start-up in practice should take place preferentially during warm summer periods, while decreased temperatures during winter periods should not be a problem for granule stability and COD and phosphate removal in a granular sludge system. Nitrogen removal efficiencies should be optimized by changes in reactor operation or cycle time during this season.

Effects of oxygen concentration on N-removal in an aerobic granular sludge reactor.

In order to optimise nitrogen removal in an aerobic granular sludge system, short- and long-term effects of decreased oxygen concentrations on the reactor performance were studied. Operation at decreased oxygen concentration is required to obtain efficient N-removal and low aeration energy requirement. A short-term oxygen reduction (from 100% to 50%, 40%, 20% or 10% of the saturation concentration) did not influence the acetate uptake rate. A lower aerobic acetate uptake at lower oxygen concentrations was obviously compensated by anoxic acetate uptake. Nitrogen removal was favoured by decreased oxygen concentrations, reaching a value of 34% for the lowest oxygen concentration tested. Long-term effects were evaluated at two oxygen saturation levels (100% and 40%). Nitrogen removal increased from 8% to 45% when the oxygen saturation was reduced to 40%. However, the granules started to disintegrate and biomass washout occurred. It was impossible to obtain stable granular sludge at this decreased oxygen concentration under applied conditions. A solution to obtain stable aerobic granular sludge at low oxygen concentrations is needed in order to make aerobic granular sludge reactors feasible in practice.

Simultaneous COD, nitrogen, and phosphate removal by aerobic granular sludge.

Aerobic granular sludge technology offers a possibility to design compact wastewater treatment plants based on simultaneous chemical oxygen demand (COD), nitrogen and phosphate removal in one sequencing batch reactor. In earlier studies, it was shown that aerobic granules, cultivated with an aerobic pulse-feeding pattern, were not stable at low dissolved oxygen concentrations. Selection for slow-growing organisms such as phosphate-accumulating organisms (PAO) was shown to be a measure for improved granule stability, particularly at low oxygen concentrations. Moreover, this allows long feeding periods needed for economically feasible full-scale applications. Simultaneous nutrient removal was possible, because of heterotrophic growth inside the granules (denitrifying PAO). At low oxygen saturation (20%) high removal efficiencies were obtained; 100% COD removal, 94% phosphate (P-) removal and 94% total nitrogen (N-) removal (with 100% ammonium removal). Experimental results strongly suggest that P-removal occurs partly by (biologically induced) precipitation. Monitoring the laboratory scale reactors for a long period showed that N-removal efficiency highly depends on the diameter of the granules.

Selection of slow growing organisms as a means for improving aerobic granular sludge stability.

Recently, several groups have showed the occurrence of aerobic granular sludge. The excellent settling characteristics of aerobic granular sludge allow the design of very compact wastewater treatment plants. In laboratory experiments, high oxygen concentrations were needed to obtain stable granulation. However, in order to obtain energy efficient aeration and good denitrification low oxygen concentrations would be required. From earlier research on biofilm morphology, it was learned that slow growing organisms influence the density and stability of biofilms positively. To decrease the growth rate of the organisms in the aerobic granules, easily degradable substrate (e.g. acetate) has to be converted to slowly degradable COD like microbial storage polymers (e.g. PHA). Phosphate or glycogen accumulating bacteria perform this conversion step most efficiently. In this paper it is shown that the selection of such bacteria in aerobic granules indeed led to stable granular sludge, even at low oxygen concentrations.

Aerobic granular sludge technology: an alternative to activated sludge?

Laboratory experiments have shown that it is possible to cultivate aerobic granular sludge in sequencing batch reactors. In order to direct future research needs and the critical points for successful implementation at large scale, a full detailed design of a potential application was made. The design was based on the laboratory results, and two variants of a full-scale sewage treatment plant based on Granular sludge Sequencing Batch Reactors (GSBRs) were evaluated. As a reference a conventional treatment plant based on activated sludge technology was designed for the same case. Based on total annual costs both GSBR variants proved to be more attractive than the reference alternative (7-17% lower costs). From a sensitivity analysis it appeared that the GSBR technology was less sensitive to the land price and more sensitive to a rain weather flow (RWF). This means that the GSBR technology becomes more attractive at lower permissible RWF/DWF ratios and higher land prices. The footprint of the GSBR variants was only 25% compared to the reference. However, the GSBR with primary treatment only cannot meet the present effluent standards for municipal wastewater in The Netherlands, mainly because of a too high suspended solids concentration in the effluent. A growing number of sewage treatment plants in the Netherlands are going to be faced with more stringent effluent standards. In general, activated sludge plants will have to be extended with a post treatment step (e.g. sand filtration) or be transformed into Membrane Bioreactors. In this case a GSBR variant with primary treatment as well as post treatment can be an attractive alternative.