Transboundary intercellular communications between Penicillium and bacterial communities during sludge bulking: Inspirations on quenching fungal dominance

Fungal bulking is caused by the evolution toward a fungi-dominant unbalanced sludge system, which is indeed the phenomenon of fungi competing against bacterial cells. We hypothesized that the cross-kingdom intercellular communication between fungi and bacteria was internal driving force that stimulated fungal bulking. In this study, we identified three signal molecules related to Penicillium fungi bulking under low-pH stress in an activated sludge reactor, which inspired us to propose a sludge bulking prevention strategy using the quorum quenching theory. When pH dropped from 7.0 to 4.5, the abundance of Penicillium increased from 12.5% to 44.8%. However, some functional bacterial genera, such as Nitrosomonas and Sphingopyxis, were washed out from the sludge. The production of quorum-sensing (QS) molecules N-Heptanoyl-L-homoserine lactone (C7-HSL), N-Dodecanoyl-L-homoserine lactone (C12-HSL), and N-Tetradecanoyl-L-homoserine lactone (C14-HSL) was regulated with sludge bulking; especially the response of the latter two was significantly negative to Penicillium blooming (P < 0.05). To test their roles, trace commercial C12-HSL and C14-HSL were added to Penicillium culture, successfully causing 8.3% and 30.2% inhibition of mycelial formation, respectively. They also contributed to the improvement of activated sludge settleability by 6.1% and 39.7%, respectively (represented by sludge volume index). The transcriptome technique further revealed the regulation of the expression of genes in |logFC| >1, involving signal transduction, mycelium synthesis, and metabolic pathways. Our study provided an innovative strategy for controlling fungal bulking from the perspective of microbial transboundary informatics.