A novel Pbunavirus phage from the Ganges River in biocontrol of multidrug-resistant Pseudomonas aeruginosa in wastewater by downregulation of las/rhl systems and reduction of KPC carbapenemases

Wastewater systems are reservoirs of multidrug-resistant (MDR) pathogens, including Pseudomonas aeruginosa , mirroring those found in the hospital effluents. Effective treatment of raw sewage for MDR P. aeruginosa abatement is critical for safe effluent disposal. This study reports the isolation of a novel phage (ɸPRS-1) within the Pbunavirus genus from a pristine stretch of the Ganges River, targeting the MDR strain of P. aeruginosa . Antibacterial and antibiofilm assays of ɸPRS-1 against P. aeruginosa PAO1 demonstrated robust efficacy, achieving log ₁₀ reductions of 8.14 in planktonic cells, 9.83 in biofilm inhibition, and 7.01 in biofilm disruption after 24 hours. Additionally, ɸPRS-1 effectively disrupted the preformed biofilm of various other MDR P. aeruginosa strains with >5 log ₁₀ reductions. Mechanistically, ɸPRS-1 downregulated quorum-sensing genes and reduced the carbapenemase resistance KPC cluster, validated through qRT-PCR analysis. Evaluation of ɸPRS-1 in raw sewage revealed a 4.2 log ₁₀ CFU/mL reduction in the P. aeruginosa counts, highlighting its efficacy in complex environmental matrices. In vitro, cytotoxicity assays confirmed the phage’s safety. The use of ɸPRS-1 for tackling MDR P. aeruginosa in the wastewater offers a sustainable alternative, potentially reducing reliance on other treatment methods. The findings have attempted to deliver SDGs 3, 6, 11, 14, and 15.

Highlights

• A novel phage (ɸPRS-1) from an untapped location of the Ganges River

• ɸPRS-1 reduces planktonic cells of MDR P. aeruginosa by 8.14 log ₁₀ in 24 hours

• Antibiofilm potentials: 9.83 log ₁₀ reduction (inhibition) >5 log ₁₀ reduction (disruption)

• Downregulates quorum-sensing genes and reduces KPC resistance in MDR P. aeruginosa

• ɸPRS-1 shows 4.2 log ₁₀ CFU/mL reduction of MDR P. aeruginosa in raw sewage

Graphical Abstract