Interactions of functional bacteria and their contributions to the performance in integrated autotrophic and heterotrophic denitrification

Compared to autotrophic and heterotrophic denitrification process, the Integrated autotrophic and heterotrophic denitrification (IAHD) has wider foreground of applications in the condition where the organic carbon, nitrate and inorganic sulfur compounds usually co-exist in the actual wastewaters. As the most well-known IAHD process, the denitrifying sulfide removal (DSR) could simultaneously convert sulfide, nitrate and organic carbon into sulfur, dinitrogen gas and carbon dioxide, respectively. Thus, systematical metabolic functions and contributions of autotrophic and heterotrophic denitrifiers to the IAHD-DSR performance became an problem demanding to be promptly studied. In this work, three upflow anaerobic sludge bioreactors (UASBs) were individually started up in autotrophic (a-DSR), heterotrophic (h-DSR) and mixotrophic conditions (m-DSR). Then, the operating conditions of each bioreactor were switched to different trophic conditions with low and high sulfide concentrations in the influent (200 and 400 mg/L). The removal efficiencies of sulfide, nitrate and acetate all reached 100% in all three bioreactors throughout the operational stages. However, the sulfur transformation ratio ranged from 34.5% to 39.9% at the low sulfide concentration and from 76.8% to 86.7% at the high sulfide concentration in the mixotrophic conditions. Microbial community structure analyzed by the Illumina sequencing indicated that Thiobacillus, which are autotrophic sulfide-oxidizing, nitrate-reducing bacteria (a-soNRB), was the dominant genus (81.3%) in the a-DSR bioreactor. With respect to the mixotrophic conditions, at low sulfide concentration in the m-DSR bioreactor, Thiobacillus (a-soNRB) and Thauera, which are heterotrophic nitrate-reducing bacteria (hNRB), were the dominant genera, with percentages of 48.8% and 14.9%, respectively. When the sulfide concentration in the influent was doubled, the percentage of Thiobacillus decreased by approximately 9-fold (from 48.8% to 5.4%), and the total percentage of Azoarcus and Pseudomonas, which are heterotrophic sulfide-oxidizing, nitrate-reducing bacteria (h-soNRB), increased by approximately 6-fold (from 10.1% to 59.4%). Therefore, the following interactions between functional groups and their functional mechanisms in the DSR process were proposed: (1) a-soNRB (Thiobacillus) and hNRB (Thauera) worked together to maintain the performance under the low sulfide concentration; (2) h-soNRB (Azoarcus and Pseudomonas) took the place of a-soNRB and worked together with hNRB (Thauera and Allidiomarina) under the high sulfide concentration; and (3) a-soNRB (such as Thiobacillus) were possibly the key bacteria and may have contributed to the low sulfur transformation, and h-soNRB may be responsible for the high sulfur transformation in the DSR process.