Microbiological rationale for alternative strategies to combat infections causedby antibiotic-resistant Pseudomonas aeruginosa

This study aimed to investigate the activity of the drug Pyofag against clinical isolates ofPseudomonas aeruginosa and to evaluate the effectiveness of the combined action of surface-activeantiseptics and bacteriophages. To achieve this aim, classical methods for the isolation andidentification of bacteria were employed. Antibiotic susceptibility of Pseudomonas isolates was determined using the disc diffusion method, while susceptibility to surface-active antiseptics(decamethoxine, benzalkonium chloride, chlorhexidine bigluconate, octenidine dihydrochloride, andpolyhexanide) was assessed using the broth dilution method. The susceptibility of clinical isolates toPyofag was evaluated based on the optical density of bacterial suspensions after 18 hours ofincubation with the preparation. The nature of the combined effect of bacteriophages and antisepticson P. aeruginosa was assessed by calculating the lytic index of the phage on planktonic bacterialforms cultured in media containing sub-bacteriostatic concentrations of antiseptics. The resultsshowed that all 54 isolated clinical strains of P. aeruginosa retained high susceptibility only to reserveantibiotics – colistin (94.4%) and cefiderocol (75.9%). Resistance to other antipseudomonalantibiotics (cefepime, ceftazidime, piperacillin-tazobactam, imipenem, and ciprofloxacin) wasobserved in 96.3%-100% of isolates. However, aminoglycosides (gentamicin, tobramycin, amikacin)and meropenem remained effective against 29.6%-44.4% of strains. Antiseptic agents containingsurface-active compounds demonstrate strong antipseudomonal properties and are capable ofinhibiting bacterial proliferation at concentrations ranging from 16.4-22.5 μg/mL (octenidinedihydrochloride, decamethoxine, chlorhexidine bigluconate) to 65-145.7 μg/mL (polyhexanide,benzalkonium chloride). It was confirmed that decamethoxine, octenidine, and chlorhexidine exhibitsignificantly greater antibacterial activity than polyhexanide and benzalkonium chloride (p < 0.01).The isolated Pseudomonas strains showed high susceptibility to the pharmaceutical preparationPyofag: the lytic activity index (Is) of Pyofag exceeded 0.5 in 70.4% of strains, indicating that 50%of the bacterial population was destroyed during the dynamic interaction between bacterial growthand phage replication. In media containing sub-bacteriostatic concentrations of decamethoxine,chlorhexidine, or octenidine, both susceptible (n = 7, Is = 0.69) and resistant (n = 8, Is = 0.15) strainswere lysed more intensively by the bacteriophage. This was evidenced by an increase in thesusceptibility index to 0.80-0.87 in susceptible strains and to 0.54-0.70 in phage-resistant strains,respectively

 

surface-active antiseptics; bacteriophages; antibiotics; Pyofag; opportunisticmicroorganisms

  1. Green SI, Clark JR, Santos HH, Weesner KE, Salazar KC, Aslam S, et al. A retrospective, observational study of 12 cases of expanded-access customized phage therapy: Production, characteristics, and clinical outcomes. Clin Infect Dis. 2023;77(8):1079–91. DOI: 10.1093/cid/ciad335
  2. Pirnay JP, Djebara S, Steurs G, Griselain J, Cochez C, De Soir S, et al. Personalized bacteriophage therapy outcomes for 100 consecutive cases: A multicentre, multinational, retrospective observational study. Nat Microbiol. 2024;9:1434–53. DOI: 10.1038/s41564-024-01705-x
  3. Cesta N, Pini M, Mulas T, Materazzi A, Ippolito E, Wagemans J, et al. Application of phage therapy in a case of a chronic hip-prosthetic joint infection due to Pseudomonas aeruginosa: An Italian real-life experience and in vitro analysis. Open Forum Infect Dis. 2023;10(2):ofad51. DOI: 10.1093/ofid/ofad051
  4. Young MJ, Hall LML, Merabishvilli M, Pirnay JP, Clark JR, Jones JD. Phage therapy for diabetic foot infection: A case series. Clin Ther. 2023;45(8):797–801. DOI: 10.1016/j.clinthera.2023.06.009
  5. Rahimzadeh Torabi L, Doudi M, Naghavi NS, Monajemi R. Isolation, characterization, and effectiveness of bacteriophage Pɸ-Bw-Ab against XDR Acinetobacter baumannii isolated from nosocomial burn wound infection. Iran J Basic Med Sci. 2021;24(9):1254–63. DOI: 10.22038/ijbms.2021.57772.12850
  6. Racenis K, Lacis J, Rezevska D, Mukane L, Vilde A, Putnins I, et al. Successful bacteriophage-antibiotic combination therapy against multidrug-resistant Pseudomonas aeruginosa left ventricular assist device driveline infection. Viruses. 2023;15(5):1210. DOI: 10.3390/v15051210
  7. Rubezhniak I. Antibacterial activities of cultural filtrates of some strains of micromycete. Biol Syst Theory Innov. 2020;11(2):42–9. DOI: 10.31548/biologiya2020.01.042
  8. Order of the Cabinet of Ministers of Ukraine No. 116-p. National Action Plan to Combat Antimicrobial Resistance [Internet]. 2019 March 6 [cited 2024 December 20]. Available from: https://www.fao.org/faolex/results/details/en/c/LEX-FAOC187629/
  9. EUCAST recommendations version 13 [Internet]. 2023 January 2 [cited 2024 December 20]. Available from: https://www.eucast.org/eucast_news/news_singleview?tx_ttnews%5Btt_news%5D=518&cHash=2509b0db92646dffba041406dcc9f20c
  10.  Band M. An introduction to medical statistics. 4th ed. Oxford: University Press; 2015. 447 P.
  11. Kovalchuk V, Kondratiuk V, McGann P, Jones BT, Fomina N, et al. Temporal evolution of bacterial species and their antimicrobial resistance characteristics in wound infections of war-related injuries in Ukraine from 2014 to 2023. J Hosp Infect. 2024;152:99–104. DOI: 10.1016/j.jhin.2024.06.011
  12. Khan ID, Malik M, Rajmohan KS, Banerjee P, Khan S, Panda PS, et al. Hemophagocytosis secondary to pharyngeal abscess in an immunocompetent patient (case report). Int J Med Med Res. 2018;4(1):41–4. DOI: 10.11603/ijmmr.2413-6077.2018.1.8514
  13. Bahniuk N, Faustova M, Riesbeck K, Prokopchuk Z, Paliy V, Nazarchuk O, et al. The correspondence of the carbapenemase genotype and phenotypic antimicrobial profiles of Pseudomonas aeruginosa. Med Ecol Probl. 2023;27(5–6):45–50. DOI: 10.31718/mep.2023.27.5-6.06
  14. Mudenda S, Daka V, Matafwali SK. World Health Organization AWaRe framework for antibiotic stewardship: Where are we now and where do we need to go? An expert viewpoint. Antimicrob Steward Healthc Epidemiol. 2023;3(1):e84. DOI: 10.1017/ash.2023.164
  15. Nazarchuk O, Nagaichuk V, Bahniuk N, Nazarchuk H, Rymsha O, Dobrovanov O, et al. Susceptibility to antimicrobials of Acinetobacter baumannii and Pseudomonas aeruginosa clinical strains and their blaVIM variants in ICU of regional burn centre. Lek Obz. 2023;72(1):18–23.
  16.  World Health Organization. Global antimicrobial resistance and use surveillance system (GLASS) report [Internet]. 2021 [cited 2024 December 20]. Available from: https://iris.who.int/bitstream/handle/10665/341666/9789240027336-eng.pdf?sequence=1
  17. Denysko Т. Comparative study of antimicrobial properties of biomaterials and dressings based on antiseptics against gram-negative bacteria as pathogens of wound infections. Bull Probl Biol Med. 2024;1(172):357–63. DOI: 10.29254/2077-4214-2024-1-172-357-363
  18. Murugaiyan J, Kumar PA, Rao GS, Iskandar K, Hawser S, Hays JP, et al. Progress in alternative strategies to combat antimicrobial resistance: Focus on antibiotics. Antibiotics. 2022;11(2):200. DOI: 10.3390/antibiotics11020200
  19. Jault P, Leclerc T, Jennes S, Pirnay JP, Que YA, Resch G, et al. Efficacy and tolerability of a cocktail of bacteriophages to treat burn wounds infected by Pseudomonas aeruginosa (PhagoBurn): A randomised, controlled, double-blind phase 1/2 trial. Lancet Infect Dis. 2018;19(1):35–45. DOI: 10.1016/S1473-3099(18)30482-1
  20. Nawaz A, Khalid NA, Zafar S, Majid A, Shahzadi M, Saleem S, et al. Phage therapy as a revolutionary treatment for multidrug-resistant Pseudomonas aeruginosa infections: A narrative review. Microbe. 2024;2:100030. DOI: 10.1016/j.microb.2023.100030
  21. Torres-Barceló C, Hochberg ME. Evolutionary rationale for phages as complements of antibiotics. Trends Microbiol. 2016;24(4):249–56. DOI: 10.1016/j.tim.2015.12.011
  22. Derkach S. Bacteriophages: Current issues of phase preparation and evaluation of their activity. Infect Dis. 2022;1:5–10. DOI: 10.11603/1681-2727.2022.1.13014
  23. Oechslin F, Piccardi P, Mancini S, Gabard J, Moreillon P, Entenza JM, et al. Synergistic interaction between phage therapy and antibiotics clears Pseudomonas Aeruginosa infection in endocarditis and reduces virulence. J Infect Dis. 2017;215(5):703–12. DOI: 10.1093/infdis/jiw632
  24. Cui L, Watanabe S, Miyanaga K, Kiga K, Sasahara T, Aiba Y, et al. A comprehensive review on phage therapy and phage-based drug development. Antibiotics. 2024;13(9):870. DOI: 10.3390/antibiotics13090870