Understanding the Effect of Free Nitrous Acid on Biofilms

Free nitrous acid (FNA, i.e., HNO2) has been recently applied to biofilm control in wastewater management. The mechanism triggering biofilm detachment upon exposure to FNA still remains largely unknown. In this work, we aim to prove that FNA induces biofilm dispersal via extracellular polymeric matrix breakdown and cell lysis. Biofilms formed by a model organism, Pseudomonas aeruginosa PAO1, were treated with FNA at concentrations ranging from 0.2 to 15 mg N/L for 24 h (conditions typically used in applications). The biofilms and suspended biomass were monitored both before and after FNA treatment using a range of methods including optical density measurements, viability assays, confocal laser scanning microscopy, and atomic force microscopy. It was revealed that FNA treatment caused substantial and concentration-dependent biofilm detachment. The addition of a reactive nitrogen species (RNS) scavenger, that is, 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazolinel-oxy-1-3-oxide, substantially reduced biofilm dispersal, suggesting that the nitrosative decomposition species of HNO2 (i.e., RNS, e.g., center dot NO + center dot NO2) were mainly responsible for the effects. The study provides insight into and support for the use of FNA for biofilm control in wastewater treatment.

Automated summary

This study aims to prove that FNA induces biofilm dispersal by breaking down the extracellular polymeric matrix and causing cell lysis. The results show that FNA treatment leads to significant biofilm detachment, and the nitrosative decomposition species of HNO2 are mainly responsible for this effect.