Critical Factors Facilitating Candidatus Nitrotoga To Be Prevalent Nitrite-Oxidizing Bacteria in Activated Sludge

Nitrite oxidation is the primary pathway that generates nitrate in engineered systems. However, little is known about the role of a novel nitrite-oxidizing bacteria (NOB) genus Candidatus Nitrotoga in activated sludge systems. To elucidate key factors that impact NOB community composition, laboratory-scale sequencing batch reactors (SBRs) were designed and operated under the same conditions as real wastewater treatment plants to achieve considerable nitrogen removal and similar community; then, different conditions including temperature (T), dissolved oxygen (DO), free nitrous acid (FNA), and free ammonia (FA) were applied. The 16S rRNA gene-based PCR and sequence analysis illustrated that Ca. Nitrotoga were abundant even at ambient temperature, thus further challenging the previous conception of them being solely cold-adapted. Ca. Nitrotoga are less competitive than Nitrospira during oxygen deficiency, indicating its lower affinity to dissolved oxygen. Ca. Nitrotoga are the dominant nitrite oxidizers under regular exposure to FNA and FA due to their relatively higher resistance than other NOB toward these two effective biocides. Therefore, this study demonstrates that Ca. Nitrotoga can play an important role in biological nitrogen removal and also highlights the need for multiple strategies for NOB suppression for the next-generation, shortcut nitrogen removal.