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 ({phi}PRS-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 {phi}PRS-1 against P. aeruginosa PAO1 demonstrated robust efficacy, achieving log10 reductions of 8.14 in planktonic cells, 9.83 in biofilm inhibition, and 7.01 in biofilm disruption after 24 hours. Additionally, {phi}PRS-1 effectively disrupted the preformed biofilm of various other MDR P. aeruginosa strains with >5 log10 reductions. Mechanistically, {phi}PRS-1 downregulated quorum-sensing genes and reduced the carbapenemase resistance KPC cluster, validated through qRT-PCR analysis. Evaluation of {phi}PRS-1 in raw sewage revealed a 4.2 log10 CFU/mL reduction in the P. aeruginosa counts, highlighting its efficacy in complex environmental matrices. In vitro, cytotoxicity assays confirmed the phages safety. The use of {phi}PRS-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. HighlightsO_LIA novel phage ({phi}PRS-1) from an untapped location of the Ganges River C_LIO_LI{phi}PRS-1 reduces planktonic cells of MDR P. aeruginosa by 8.14 log10 in 24 hours C_LIO_LIAntibiofilm potentials: 9.83 log10 reduction (inhibition) >5 log10 reduction (disruption) C_LIO_LIDownregulates quorum-sensing genes and reduces KPC resistance in MDR P. aeruginosa C_LIO_LI{phi}PRS-1 shows 4.2 log10 CFU/mL reduction of MDR P. aeruginosa in raw sewage C_LI Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=111 SRC="FIGDIR/small/621487v1_ufig1.gif" ALT="Figure 1"> View larger version (49K): org.highwire.dtl.DTLVardef@14f4ac0org.highwire.dtl.DTLVardef@b4a8d9org.highwire.dtl.DTLVardef@b97a6borg.highwire.dtl.DTLVardef@114072c_HPS_FORMAT_FIGEXP M_FIG C_FIG