In Situ Enrichment of Anammox Bacteria from Pig Farm Anoxic Sludge Through Co-Cultivation with a Quorum-Sensing Functional Strain Pseudomonas aeruginosa

Abstract

Anaerobic ammonium oxidation (anammox), as an efficient and low-carbon method for nitrogen removal from wastewater, faces the challenge of slow enrichment of functional bacteria. In this study, the enrichment of anammox bacteria Candidatus Brocadia was successfully accelerated by co-culturing with the quorum-sensing strain Pseudomonas aeruginosa and anoxic sludge from a pig farm. Experimental results showed that the R2, which had Pseudomonas aeruginosa added, exhibited chemical reaction ratios RS (NO2−-N consumption/NH4+-N consumption) and RP (NO3−-N production/NH4+-N consumption) closer to the theoretical values of the anammox reaction since Phase Ⅱ. Bacterial community analysis indicated that the abundance of Candidatus Brocadia in R2 reached 1.63% in cycle 20, significantly higher than the 0.45% in R1. More quorum-sensing signaling molecules, primarily C6-HSL, were detected in R2. C6-HSL was positively correlated with processes such as the secretion of anammox extracellular polymers (EPS) and the regulation of nitric oxide reductase (Nir), which may explain the reason behind the accelerated increase in the abundance of Candidatus Brocadia through co-culturing. Moreover, the metabolism of the dominant genus Paracoccus within the two groups of reactors also showed positive regulation by C6-HSL, with its abundance trend similar to that of Candidatus Brocadia, jointly completing the nitrogen removal process in the reactors. However, it is still unknown which genera secrete large amounts of C6-HSL after inoculation with Pseudomonas aeruginosa. This research provides a novel and low-cost method for the enrichment of anammox bacteria.