M. Pronk

Effect of Elevated Salt Concentrations on the Aerobic Granular Sludge Process: Linking Microbial Activity with Microbial Community Structure

ABSTRACT The long- and short-term effects of salt on biological nitrogen and phosphorus removal processes were studied in an aerobic granular sludge reactor. The microbial community structure was investigated by PCR-denaturing gradient gel electrophoresis (DGGE) on 16S rRNA and amoA genes. PCR products obtained from genomic DNA and from rRNA after reverse transcription were compared to determine the presence of bacteria as well as the metabolically active fraction of bacteria. Fluorescence in situ hybridization (FISH) was used to validate the PCR-based results and to quantify the dominant bacterial populations. The results demonstrated that ammonium removal efficiency was not affected by salt concentrations up to 33 g/liter NaCl. Conversely, a high accumulation of nitrite was observed above 22 g/liter NaCl, which coincided with the disappearance of Nitrospira sp. Phosphorus removal was severely affected by gradual salt increase. No P release or uptake was observed at steady-state operation at 33 g/liter NaCl, exactly when the polyphosphate-accumulating organisms (PAOs), “Candidatus Accumulibacter phosphatis” bacteria, were no longer detected by PCR-DGGE or FISH. Batch experiments confirmed that P removal still could occur at 30 g/liter NaCl, but the long exposure of the biomass to this salinity level was detrimental for PAOs, which were outcompeted by glycogen-accumulating organisms (GAOs) in the bioreactor. GAOs became the dominant microorganisms at increasing salt concentrations, especially at 33 g/liter NaCl. In the comparative analysis of the diversity (DNA-derived pattern) and the activity (cDNA-derived pattern) of the microbial population, the highly metabolically active microorganisms were observed to be those related to ammonia (Nitrosomonas sp.) and phosphate removal (“Candidatus Accumulibacter”).

Ammonium adsorption in aerobic granular sludge, activated sludge and anammox granules.

The ammonium adsorption properties of aerobic granular sludge, activated sludge and anammox granules have been investigated. During operation of a pilot-scale aerobic granular sludge reactor, a positive relation between the influent ammonium concentration and the ammonium adsorbed was observed. Aerobic granular sludge exhibited much higher adsorption capacity compared to activated sludge and anammox granules. At an equilibrium ammonium concentration of 30 mg N/L, adsorption obtained with activated sludge and anammox granules was around 0.2 mg NH4-N/g VSS, while aerobic granular sludge from lab- and pilot-scale exhibited an adsorption of 1.7 and 0.9 mg NH4-N/g VSS, respectively. No difference in the ammonium adsorption was observed in lab-scale reactors operated at different temperatures (20 and 30 °C). In a lab-scale reactor fed with saline wastewater, we observed that the amount of ammonium adsorbed considerably decreased when the salt concentration increased. The results indicate that adsorption or better ion exchange of ammonium should be incorporated into models for nitrification/denitrification, certainly when aerobic granular sludge is used.

Effect of sludge age on methanogenic and glycogen accumulating organisms in an aerobic granular sludge process fed with methanol and acetate.

The influence of sludge age on granular sludge formation and microbial population dynamics in a methanol‐ and acetate‐fed aerobic granular sludge system operated at 35°C was investigated. During anaerobic feeding of the reactor, methanol was initially converted to methane by methylotrophic methanogens. These methanogens were able to withstand the relatively long aeration periods. Lowering the anaerobic solid retention time (SRT) from 17 to 8 days enabled selective removal of the methanogens and prevented unwanted methane formation. In absence of methanogens, methanol was converted aerobically, while granule formation remained stable. At high SRT values (51 days), γ‐Proteobacteria were responsible for acetate removal through anaerobic uptake and subsequent aerobic growth on storage polymers formed [so called metabolism of glycogen‐accumulating organisms (GAO)]. When lowering the SRT (24 days), Defluviicoccus‐related organisms (cluster II) belonging to the α‐Proteobacteria outcompeted acetate consuming γ‐Proteobacteria at 35°C. DNA from the Defluviicoccus‐related organisms in cluster II was not extracted by the standard DNA extraction method but with liquid nitrogen, which showed to be more effective. Remarkably, the two GAO types of organisms grew separately in two clearly different types of granules. This work further highlights the potential of aerobic granular sludge systems to effectively influence the microbial communities through sludge age control in order to optimize the wastewater treatment processes.

Importance of sediment deposition and denitrification for plant productivity and nutrient retention in various floodplain wetlands

ABSTRACT Questions: Various floodplain communities may differ in their relative abilities to influence water quality through nutrient retention and denitrification. Our main questions were: (1) what is the importance of sediment deposition and denitrification for plant productivity and nutrient retention in floodplains; (2) will rehabilitation of natural floodplain communities (semi-natural grassland, reedbed, woodland, pond) from agricultural grassland affect nutrient retention? Location: Floodplains of two Rhine distributaries (rivers IJssel and Waal), The Netherlands. Methods: Net sedimentation was measured using mats, denitrification in soil cores by acetylene inhibition and bio-mass production by clipping above-ground vegetation in winter and summer. Results: Sediment deposition was a major source of N and P in all floodplain communities. Highest deposition rates were found where water velocity was reduced by vegetation structure (reedbeds) or by a drop in surface elevation (pond). Sediment deposition was not higher in woodlands than in grassland types. Denitrification rates were low in winter but significantly higher in summer. Highest denitrification rates were found in an agricultural grassland (winter and summer) and in the ponds (summer). Plant productivity and nutrient uptake were high in reedbeds, intermediate in agricultural grasslands, ponds and semi-natural grasslands and very low in woodlands (only understorey). All wetlands were N-limited, which could be explained by low N:P ratios in sediment. Conclusions: Considering Rhine water quality: only substantial P-retention is expected because, relative to the annual nutrient loads in the river, the floodplains are important sinks for P, but much less for N. Rehabilitation of agricultural grasslands into ponds or reedbeds will probably be more beneficial for downstream water quality (lower P-concentrations) than into woodlands or semi-natural grasslands.

Controlling effluent suspended solids in the aerobic granular sludge process

The main processes contributing to elevated effluent suspended solids in the full-scale aerobic granular sludge process were studied. The two processes found to be most important were (1) rising of sludge due to degasification of nitrogen gas (produced by denitrification) and (2) wash-out of particles that intrinsically do not settle such as certain fats and foams. A mathematical model was made to describe the process of degasification of nitrogen gas during the feeding phase in an AGS reactor. The process of rising sludge due to degasification could be limited by stripping out the nitrogen gas before starting the settling phase in the process cycle. The wash-out of scum particles could be reduced by introducing a vertical scum baffle in front of the effluent weir, similar to weirs in traditional clarifiers. A full-scale Nereda® reactor in the municipality of Utrecht, The Netherlands, was operated with a nitrogen stripping phase and scum baffles for 9 months at an average biomass concentration of 10 g L-1 and an average granulation grade of 84%. In this period the influent suspended solids concentration was 230±118mgL-1 and the concentration of effluent suspended solids was 7.8±3.8mgL-1.