Standardization of in vitro method of Biofilm formation by Clinical Isolates of Pseudomonas aeruginosa and elucidating its Biofilm-specific Proteins through LC-Orbitrap HRMS-based Peptidomics

ABSTRACT: Pseudomonas aeruginosa has a notorious ability to form biofilms, which often facilitate chronic infections. Despite we lack a consensus methodology for its biofilm quantifica-tion/classification for clinical use. Furthermore, the problem gets exacerbated due to het-erogeneity in its matrix composition due to nutritional and oxygen cues, resulting in phe-notypic variations. Here, we formulated the supplement mix for isolate classification based on biofilm forming capacity and tried to profile multiple clinical isolates of Pseu-domonas aeruginosa that revealed many common and some distinct metabolic and pro-teomic signatures among high, medium, low and non-biofilm producing isolates. We further noted the biofilm degrading and inhibiting effects of some in vogue antibiotics in sub-inhibitory doses. We validated our findings employing growth kinetics, confocal mi-croscopy, scanning electron microscopy, Liquid chromatography-Orbitrap-based high resolution mass spectrometry (LC-HRMS). We noted high level resistance against ceftazidime (43.88%), meropenem (43.17%), imipenem (33.09%), and piperacil-lin-tazobactam (30.94%). Also, we noted, that upon our supplementation, 38.84% isolates were found to be high slime producing, while only 1.14% isolates were non-slime pro-ducing. We noted active anoxic metabolism although P. aeruginosa is a strict aerobe. We noted abundant proteins for cellular adaptation to chemical gradients, including glucose catabolism, arginine and polyamine metabolism, and potential virulence and stress me-diation in high slime producer matrix. This work provides insight toward the potential for metabolic speciation in biofilm and supports the development of biofilm-specific diagnos-tics, and development of futuristic therapeutic modalities.

INSTRUMENT(S):

ORGANISM(S): Pseudomonas Aeruginosa Pao1h2o Bacteria

DISEASE(S): Cystic Fibrosis

SUBMITTER: Ashish Singh  

LAB HEAD: Pradyot Prakash

PROVIDER: PXD057726 | Pride | 2026-05-18

REPOSITORIES: Pride