The effect of chitosan on the development of granular sludge in upflow anaerobic sludge blanket reactors (UASB) when treating wastewater polluted with the organic solvents ethanol, ethyl acetate, and 1-ethoxy-2-propanol was evaluated. Three UASB reactors were operated for 219 days at ambient temperature with an organic loading rate (OLR) of between 0.3 kg COD m d and 20 kg COD m d. One reactor was operated without the addition of chitosan, while the other two were operated with the addition of chitosan doses of 2.4 mg gVSS two times. The three reactors were all able to treat the OLR tested with COD removal efficiencies greater than 90%. However, the time required to reach stable operation was considerably reduced in the chitosan-assisted reactors. The development of granules in the reactors with chitosan was accelerated and granules larger than 2000 μm were only observed in these reactors. In addition, these granules exhibited better physicochemical characteristics: the mean particle diameter (540 and 613 μm) was approximately two times greater than in the control reactor (300 μm), and the settling velocities exceeded 35 m h. The extracellular polymeric substances (EPS) in the reactors with the chitosan was found to be higher than in the control reactor. The protein-EPS content has been correlated with the granule size. The analyses of the microbial communities, performed through denaturing gradient gel electrophoresis and high-throughput sequencing, revealed that the syntrophic microorganisms belonging to genus Geobacter and the hydrogenotrophic methanogen Methanocorpusculum labreanum were predominant in the granules. Other methanogens like Methanosaeta species were found earlier in the chitosan-assisted reactors than in the control reactor.

UASB showed robustness to treat oxygenated organic solvents under an intermittent feeding pattern with dominance of hydrogenotrophic methanogens.

Quality of the granular sludge developed during the start-up of anaerobic up-flow sludge bed reactors is of crucial importance to ensure the process feasibility of treating industrial wastewater such as those containing solvents. In this study, the microbial granule formation from suspended-growth biomass was investigated in two chitosan-assisted reactors. These reactors operated mimicking industrial sites working with night closures treating a mixture of ethanol, ethyl acetate, and 1-ethoxy-2-propanol. Each reactor operated under different hydrodynamic regimes typical from UASB (R1: < 0.15 m h-1) and EGSB (R2: 3 m h-1). High soluble COD removal efficiencies (> 90%) accompanied by rapid formation of robust anaerobic granules were achieved at both up-flow velocity levels. After three weeks from the start-up, mean size diameters of 475 µm and 354 µm were achieved for R1 and R2, respectively. The performance of the process was found to be stable for the whole operational period of 106 days treating intermittent OLR up to 13 kg COD m-3 d-1. A memory dose of chitosan at day 42 was beneficial to guarantee good quality of the granules by offsetting the negative impact of intermittent water supply on the granular size. Methanocorpusculum was identified as the dominant archaea at both up-flow velocities. Acetobacterium, Geobacter and Desulfovibrio bacteria were also abundant, demonstrating its role on the degradation of light-oxygenated solvents.

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