Pseudomonas aeruginosa is a widespread source of hospital-acquired infections and a top priority antibiotic-resistant pathogen as it has developed robust immunity to most traditional antibiotics. Quorum sensing (QS) enables P. aeruginosa to modulate virulence functions and is important for pathogenesis. QS relies on the production and perception of autoinducing chemical signal molecules. Acyl-homoserine lactones are the key autoinducer molecules that mediate P. aeruginosa-associated QS, and N-(3-oxododecanoyl)-L-homoserine lactone (3-O-C12-HSL) and N-butyryl-l-homoserine lactone (C4-HSL) are the two types. This study aimed to identify potential quenching targets of QS pathways that may reduce the chances of resistance developing in P. aeruginosa using co-culture approaches. In co-cultures, Bacillus reduced the production of 3-O-C12-HSL/C4-HSL signal molecules by inactivating acyl- homoserine lactone-based QS to inhibit important virulence factor expression. Moreover, Bacillus is subject to complex crosstalk with other regulatory systems, such as the integrated QS system and the Iqs system. The results showed that blocking one or more QS pathways was insufficient to reduce infection with multidrug resistant P. aeruginosa.