In the bacterium Pseudomonas aeruginosa, the synthesis and secretion of extracellular protease is a typical cooperative behavior regulated by quorum sensing. However, this type of cooperative behavior is easily exploited by other individuals who do not synthesize public goods, which is known as the "tragedy of the commons". Here P. aeruginosa was inoculated into casein media with different nitrogen salts added. In casein broth, protease (a type of public good) is necessary for bacterial growth. After 30 days of sequential transfer, some groups propagated stably and avoided "tragedy of the commons". The evolved cooperators who continued to synthesize protease were isolated from these stable groups. By comparing the characteristics of quorum sensing in these cooperators, an identical evolutionary pattern was found. A variety of cooperative behaviors regulated by quorum sensing, such as the synthesis and secretion of protease and signals, were significantly reduced during the process of evolution. Such reductions improved the efficiency of cooperation, helping to prevent cheating. In addition, the production of pyocyanin, which is regulated by the RhlIR system, increased during the process of evolution, possibly due to its role in stabilizing the cooperation. This study contributes towards our understanding of the evolution of quorum sensing of P. aeruginosa.

To investigate whether pollution from antibiotic resistance genes (ARGs) could be affected by bacterial quorum sensing, the oxytetracycline (OTC)-containing manure was fertilized to establish the ARG-polluted soil environment. Under long-term OTC stress, substantial ARGs in the range from 10 to 10 RG/16S rRNA (resistance genes/16S rRNA) were detected in the antibiotics control (AC) group, in which OTC-containing manure was fertilized. Meanwhile, 10 RG/16S rRNA was detected in biological control (BC) group, in which non-OTC-containing manure was fertilized. Subsequently, two typical quorum sensing inhibitors, 4-nitropyridine N-oxide (4-NPO) and 3,4-dibromo-2H-furan-5-one (DBF), were used to treat the ARG-polluted soils. These two groups called 4-NPO treatments (NT) and DBF treatments (FT), respectively. There were no significant differences in bacterial growth and OTC degradation in NT and FT groups, compared to AC group. However, acyl-homoserine lactones such as C-HSL, C-HSL, and C-HSL decreased significantly in both NT and FT groups, compared to AC group. Meanwhile, the abundance of most ARGs decreased dramatically. In FT group, the concentrations of tet(L) and tet(Q) were below the detection limits. It was demonstrated that quorum sensing inhibition could be an effective way to prevent and control the pollution of ARGs in soil.

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