Nitrogen Source Stabilization of Quorum Sensing in the Pseudomonas aeruginosa Bioaugmentation Strain SD-1

Abstract

Pseudomonas aeruginosa SD-1 is efficient at degrading aromatic compounds and can therefore contribute to the bioremediation of wastewater. P. aeruginosa uses quorum sensing (QS) to regulate the production of numerous secreted "public goods." In wastewater bioaugmentation applications, there are myriad nitrogen sources, and we queried whether various nitrogen sources impact the stabilities of both QS and the bacterial populations. In a laboratory strain of P. aeruginosa, PAO1, the absence of a nitrogen source has been shown to destabilize these populations through the emergence of QS mutant "cheaters." We tested the ability of SD-1 to grow in casein broth, a condition that requires QS for growth, when the nitrogen source with either NH 4 Cl, NaNO 3 , or NaNO 2 or with no added nitrogen source. There was great variability in susceptibility to invasion by QS mutant cheaters and, by extension, the stability of the SD-1 population. When grown with NH 4 Cl as an extra nitrogen source, no population collapse was observed; by contrast, twothirds of cultures grown in the presence of NaNO 2 collapsed. In the populations that collapsed, the frequency of social cheaters exceeded 40%. NaNO 3 and NaNO 2 directly favor QS mutants of P. aeruginosa SD-1. Although the mechanism by which these nitrogen sources act is not clear, these data indicate that the metabolism of nitrogen can affect the stability of bacterial populations, an important observation for continuing industrial applications with this species.IMPORTANCE Bioaugmentation as a method to help remediate wastewater pollutant streams holds significant potential to enhance traditional methods of treatment. Addition of microbes that can catabolize organic pollutants can be an effective method to remove several toxic compounds. Such bioaugmented strains of bacteria have been shown to be susceptible to competition from the microbiota that are present in wastewater streams, limiting their potential effectiveness. Here, we show that bioaugmentation strains of bacteria might also be susceptible to invasion by social cheaters and that the nitrogen sources available in the wastewater might influence the ability of cheaters to overtake the bioaugmentation strains. Our results imply that control over the nitrogen sources in a wastewater stream or selective addition of certain nitrogen sources could help stabilize bioaugmentation strains of bacteria.KEYWORDS quorum sensing, social cheating, bioaugmentation, long-term stabilization, inorganic nitrogen B ioaugmentation, the use of selected microbial strains in activated sludge to strengthen their abilities for pollutant removal (1), has significant advantages in the degradation of toxic and hazardous compounds. These advantages include cost and