To study the kinetics and physiology of autotrophic denitrifying sulfur bacteria, a steady-state anaerobic master culture reactor (MCR) was operated for over six months under a semi-continuous mode and nitrate limiting conditions using nutrient/mineral/buffer (NMB) medium containing thiosulfate and nitrate. Characteristics of the autotropic denitrifier were investigated through the cumulative gas production volume and rate, measured using an anaerobic respirometer, and through the nitrate, nitrite, and sulfate concentrations within the media. The bio-kinetic parameters were obtained based upon the Monod equation using mixed cultures in the MCR. Nonlinear regression analysis was employed using nitrate depletion and biomass production curves. Although this analysis did not yield exact biokinetic parameter estimates, the following ranges for the parameter values were obtained: μmax =0.12-0.2 hr-1; k=0.3-0.4 hr-1; Ks=3-10mg/L; YNO3=0.4-0.5mg Biomass/mg NO3--N. Inhibition of denitrification occurred when the concentrations of NO3--N, and SO42- reached about 660mg/L and 2,000mg/L, respectively. The autotrophic denitrifying sulfur bacteria were observed to be very sensitive to nitrite but relatively tolerant of nitrate, sulfate, and thiosulfate. Under mixotrophic conditions, denitrification by these bacteria occurred autotrophically; even with as high as 2 g COD, autotrophic denitrification was not significantly affected. The optimal pH and temperature for autotrophic denitrification was about 6.5–7.5 and 33–35 °C, respectively.

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