Investigation of the Fate and Dynamics of Extracellular Polymeric Substances (EPS) during Sludge-Based Photogranulation under Hydrostatic Conditions.

Oxygenic photogranules have received increasing interest due to their ability to treat wastewater without aeration and recover wastewater's chemical energy and solar energy. It has been reported that these photogranules can be produced under both hydrostatic and hydrodynamic conditions, and enrichment of filamentous cyanobacteria is required for this photogranulation to occur. Despite the critical role extracellular polymeric substances (EPS) play in granulation, EPS in photogranulation is yet virtually unknown. Here, we present the fate and dynamics of different fractions of EPS in sludge-based photogranulation under hydrostatic conditions. The study shows that during the transformation of activated sludge into a photogranular biomass, sludge's base-extractable proteins selectively degrade. Strong correlations between base-extracted proteins and the growth of chlorophyll a and chlorophyll a/ b ratio suggest that the bioavailability of this organic nitrogen is linked with selection and enrichment of filamentous cyanobacteria under hydrostatic conditions. The results of soluble and sonication-extractable EPS and microscopy also show that the growth of filamentous cyanobacteria required large amounts of polysaccharide-based EPS for their motility and maintenance. With findings on the progression of photogranulation, the fate and dynamics of EPS, and microscopy on microstructures associated with EPS, we discuss potential mechanisms of photogranulation occurring under hydrostatic conditions.

The importance of filamentous cyanobacteria in the development of oxygenic photogranules.

Microorganisms often respond to their environment by growing as densely packed communities in biofilms, flocs or granules. One major advantage of life in these aggregates is the retention of its community in an ecosystem despite flowing water. We describe here a novel type of granule dominated by filamentous and motile cyanobacteria of the order Oscillatoriales. These bacteria form a mat-like photoactive outer layer around an otherwise unconsolidated core. The spatial organization of the phototrophic layer resembles microbial mats growing on sediments but is spherical. We describe the production of these oxygenic photogranules under static batch conditions, as well as in turbulently mixed bioreactors. Photogranulation defies typically postulated requirements for granulation in biotechnology, i.e., the need for hydrodynamic shear and selective washout. Photogranulation as described here is a robust phenomenon with respect to inoculum characteristics and environmental parameters like carbon sources. A bioprocess using oxygenic photogranules is an attractive candidate for energy-positive wastewater treatment as it biologically couples CO2 and O2 fluxes. As a result, the external supply of oxygen may become obsolete and otherwise released CO2 is fixed by photosynthesis for the production of an organic-rich biofeedstock as a renewable energy source.

Kinetics of nutrient removal and expression of extracellular polymeric substances of the microalgae, Chlorella sp. and Micractinium sp., in wastewater treatment.

Two species of green algae, Chlorella sp. and Micractinium sp., were cultivated in primary effluent wastewater and high-strength wastewater (a mixture of anaerobic digestion centrate and primary effluent) to study nutrient removal and EPS (extracellular polymeric substances) expression during their growth. The high N concentration and P-limited condition in the mixed wastewater (total N=197 mg/L; N/P mass ratio=56) led to about 3 times greater specific N removal rate than the primary effluent set, indicating that algal cells growing in N-rich wastewater had N over-uptake. Both Chlorella and Micractinium grown in the high-strength wastewater also produced larger amounts of protein EPS, possibly accounting for higher N uptake in those cultivation sets. These results suggest that different types of wastewater could cause different nutrient removal kinetics and EPS expression by algae, which may subsequently influence harvesting and anaerobic digestion of their biomass.