Silver Lactoferrin as Antimicrobials: Mechanisms of Action and Resistance Assessed by Bacterial Molecular Profiles

A diverse silver-lactoferrin (AgLTF) complex, comprising silver ions (Ag⁺) and silver nanoparticles, displayed a synergistic antibacterial effect while being almost five times more lethal than LTF alone. Gas chromatography-mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry-in linear (LP) and reflectron (RP) positive modes-were used to comprehensively analyze metabolites and proteins profiles of bacteria (Staphylococcus aureus (SA), Pseudomonas aeruginosa (PA) and Enterococcus faecalis (EF)) treated using AgLTF complex versus exclusively Ag⁺. Although both agents resulted in similar metabolic shifts in bacteria, AgLTF significantly triggered the production of sulfides (related to bacterial stress resistance), ethanol, 2-butanol (indicating exhaustion of cell respiration), decanoic acid, and nonane (suggesting ongoing oxidative stress). Keto acids formation and fermentation pathways were enhanced by AgLTF and suppressed by Ag⁺. Furthermore, AgLTF appears to interact with proteins fraction of bacteria in a concentration-dependent manner. EF molecular profiles showed less changes between treated and untreated bacteria. On the other hand, SA and PA proteins and metabolic patterns were the most differentiated from untreated bacteria. In conclusion, our study may provide valuable insights regarding the molecular mechanisms involved in AgLTF antimicrobial action.