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The effect of reduced form of sulfur compounds on granular sludge was investigated. Significant release of extracellular polymeric substance (EPS) from the granular sludge occurred in the presence of sulfide and methanethiol according to various concentrations. Granular sludge also showed a rapid increase in turbidity and decrease in diameter in accordance with sulfide concentration during the long-term shaking, suggesting that the strength of the granules was reduced with high-concentration sulfide. A continuous experiment of up-flow anaerobic sludge blanket reactors with different concentrations of sulfide (10, 200, 500 mg-S/L) influence demonstrated that the reactor fed with higher concentration of sulfide allowed more washout of small particle-suspended solid (SS) content and soluble carbohydrate and protein, which were considered as EPS released from biofilm. Finally, the presence of sulfide negatively affected methane production, chemical oxygen demand removal and sludge retention in operational performance.

In this study, eight different submerged aquatic species were screened by batch biochemical methane potential and anaerobic degradability tests to identify a promising/suitable feedstock. Kinetics of the best-screened substrate were studied in a mesophilic semi-continuous experiment. The aquatic species Myriophyllum aquaticum , Egeria densa and Potamogeton perfoliatus showed relatively higher methane yields of over 400 NmL/g-VS (volatile solids). Semi-continuous operation was carried out by feeding E. densa for over 400 days. The achieved results were 33%–53% chemical oxygen demand (COD) reduction and methane yield of 126–231 NmL/g-VS with a short hydraulic retention time (HRT). Additionally, the NH 4 + and PO 4 3− releases from the biomass to water were found to be low (18%–27% and 2.5%–3.9%) throughout the experiment. Hydrolysis was the limiting step in the digestion of E. densa , regardless of changes in HRT (15–45 days). The acid-phase model indicated that the hydrolysis rate constant (k h ) of E. densa was 0.058 one/day, which was one third lower the k h value of food waste, but quite similar to cow manure.

The effect of mixing by siphon flow on anaerobic digestion, sludge distribution and microbial community were examined in parallel experiments using a siphon-mixed reactor (SMR), an unmixed reactor (UMR) and a continuously mixed reactor (CMR). The SMR performed well without the accumulation of fatty acids under COD loading rates varying from 3 to 18 kg/m 3 /day, while the UMR was totally acidified when the loading rate increased to 10 kg/m 3 /day. The methane yield of the SMR was at least 10% higher than that of the UMR, and comparable to that of the CMR. Furthermore, the SMR was found to markedly improve the dispersion of solids and reduce deposit formation compared to the UMR. Besides, during stable operation, the fatty acids level in the effluent of the SMR and UMR was lower than that in the CMR, and the archaeal community structure of the SMR was similar to that of the UMR.