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<p>The emergence of multidrug-resistant bacteria poses a significant global health challenge, particularly in intensive care settings. This study aimed to isolate, characterize, and evaluate the antibacterial and antibiofilm potential of lytic bacteriophages targeting clinical multidrug-resistant strains of <i>Klebsiella pneumoniae</i>, <i>Escherichia coli</i> and <i>Pseudomonas aeruginosa</i>. Phages were isolated from municipal wastewater samples collected in Iğdır, Turkey, using the double-layer agar method. The isolated phages MTBB P130581-D, MTBB E830-F, and MTBB K1467-F demonstrated specific lytic activity against their respective bacterial hosts. Transmission electron microscopy revealed morphological characteristics consistent with members of the <i>Caudovirales</i> order, including the <i>Myoviridae</i>, <i>Siphoviridae</i> and <i>Podoviridae</i> families. Phage stability was assessed under varying pH and temperature conditions. MTBB E830-F exhibited the highest pH tolerance, while MTBB K1467-F showed considerable thermal resistance. In vitro assays demonstrated significant reductions in planktonic bacterial populations (0.4–6 log CFU/mL) and biofilm mass, with the most substantial antibiofilm effects observed at lower bacterial dilutions and higher multiplicity of infection. Phage treatment reduced preformed biofilms by up to 90.4% in <i>P. aeruginosa</i>, 78.3% in <i>K. pneumoniae</i>, and 62.5% in <i>E. coli</i>. These findings underscore the potential of phage therapy as an effective strategy for combating multidrug-resistant infections and biofilms in clinical settings. The phages isolated in this study expand the existing phage repertoire and offer promising alternatives to traditional antibiotics, especially against hard-to-treat nosocomial pathogens.</p>

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Bacteriophage Isolation, Characterization and Antibiofilm Effect Against Multidrug Resistant Gram Negative Bacteria Isolated from Intensive Care Units; Therapeutic Approach

  • 期刊论文

  • 发表日期：

    2025年7月29日

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  • Ayşe Karacalı Tunç,
  • Büşra Merve Sarıtaş,
  • Medine Çotak

摘要

The emergence of multidrug-resistant bacteria poses a significant global health challenge, particularly in intensive care settings. This study aimed to isolate, characterize, and evaluate the antibacterial and antibiofilm potential of lytic bacteriophages targeting clinical multidrug-resistant strains of Klebsiella pneumoniae, Escherichia coli and Pseudomonas aeruginosa. Phages were isolated from municipal wastewater samples collected in Iğdır, Turkey, using the double-layer agar method. The isolated phages MTBB P130581-D, MTBB E830-F, and MTBB K1467-F demonstrated specific lytic activity against their respective bacterial hosts. Transmission electron microscopy revealed morphological characteristics consistent with members of the Caudovirales order, including the Myoviridae, Siphoviridae and Podoviridae families. Phage stability was assessed under varying pH and temperature conditions. MTBB E830-F exhibited the highest pH tolerance, while MTBB K1467-F showed considerable thermal resistance. In vitro assays demonstrated significant reductions in planktonic bacterial populations (0.4–6 log CFU/mL) and biofilm mass, with the most substantial antibiofilm effects observed at lower bacterial dilutions and higher multiplicity of infection. Phage treatment reduced preformed biofilms by up to 90.4% in P. aeruginosa, 78.3% in K. pneumoniae, and 62.5% in E. coli. These findings underscore the potential of phage therapy as an effective strategy for combating multidrug-resistant infections and biofilms in clinical settings. The phages isolated in this study expand the existing phage repertoire and offer promising alternatives to traditional antibiotics, especially against hard-to-treat nosocomial pathogens.