Abstract
Anammox has emerged as a cost-effective and eco-friendly biological treatment technology for high-strength wastewater, and hydrazine (N 2 H 4 ) is a unique intermediate in the anammox metabolism. This study presents the first investigation into the genetic responses of anammox bacteria to elevated N 2 H 4 concentrations, offering critical insights into their potential for sustainable environmental applications. In this scope, anammox cultures were exposed to high levels of N 2 H 4 (up to 3 g/L) over a short-term period to evaluate their nitrogen treatment capacity and transcriptional responses. The results indicated that anammox activity continued at N 2 H 4 concentrations of 1.88 g/L or less. However, acute N 2 H 4 exposure significantly downregulated key genes, such as acetyl-CoA synthase beta and delta subunits, hydrazine synthase, hydrazine dehydrogenase, and hydroxylamine oxidoreductase, except for AAA family ATPase. Overall, high exogenous N 2 H 4 concentrations severely constrained the metabolism and survival of anammox bacteria at a molecular level. Understanding the genetic responses of anammox bacteria to elevated N 2 H 4 concentrations is crucial for optimizing their application in further anammox-based technologies. Future studies should focus on improving the resilience of anammox bacteria to high N 2 H 4 concentrations, thereby broadening their applicability in engineered wastewater treatment and biotechnological processes while maintaining system stability and efficiency.