Comparison of the in vitro efficacy of commercial bacteriophage cocktails and isolated bacteriophage vB_Pa01 against carbapenem resistant nosocomial Pseudomonas aeruginosa

Abstract

The result of a crucial rise in the prevalence of antibiotic resistance of bacteria, and the development of an inadequate number of new antibiotics, over the last decade a marked increase in interest in the studies of phages has been observed. It was aimed to determine the commercial preparation including Pyo-bacteriophage and Intesti- bacteriophage and compare their efficiency with isolated bacteriophage vB_Pa01 against carbapenem resistant Pseudomonas aeruginosa. Susceptibility to the bacteriophages present in each cocktail was tested using the spot test. A total of 126 carbapenem-resistant P. aeruginosa isolates were included in this test. Bacteriophage susceptibilities of each isolate were tested by spot test method. Confluent, semi confluent, opaque lysis was considered a sensitivity. The absence of any lysis is reported as resistance. The lytic activity of the bacteriophage was found 46% (58/126) Pyo- bacteriophage and 36.5% (46/126) Intesti-bacteriophage and 63.5% (80/126) vB_Pa01 bacteriophage. The most effective bacteriophage preparation was the vB_Pa01 bacteriophage (63.5%) and the least effective one was Intesti-bacteriophage (36.5%). 95 (75.4%) of 126 carbapenem resistant. This is the first study that investigated two commercial bacteriophage cocktails against carbapenem resistant P. aeruginosa. The higher efficacy of vB_Pa01 bacteriophage is promising for creating an alternative preparation especially in the treatment of infections caused by MDR pathogens that are difficult to treat.

Keywords

• Bacteriophage
• carbapenem resistant Pseudomonas aeruginosa
• commercial bacteriophage cocktail
• lytic activity.

References

1. [1] Azimi L, Lari AR. Colistin-resistant Pseudomonas aeruginosa clinical strains with defective biofilm formation. GMS Hyg Infect Control. 2019; 14(12). [CrossRef]
2. [2] Doi Y. Treatment Options for Carbapenem-resistant Gram-negative Bacterial Infections. Clin Infect Dis. 2019; 69(l7): 565-575. [CrossRef]
3. [3] Petrosillo N, Giannella M, Antonelli M, Antonini M, Barsic B, Belancic L, Inkaya AC, Pascaleb GD, Grillia E, Tumbarellof M, Akova M. Clinical experience of colistin-glycopeptide combination in critically ill patients infected with Gram-negative bacteria. Antimicrob Agents Chemother. 2014; 58(2): 851-858. [CrossRef]
4. [4] Lyon J. Phage Therapy's Role in Combating Antibiotic-Resistant Pathogens. Jama. 2017; 318(18): 1746-1748.[CrossRef]
5. [5] Wong D, Nielsen TB, Bonomo RA, Pantapalangkoor P, Luna B, Spellberg B. Clinical and Pathophysiological Overview of Acinetobacter Infections: a Century of Challenges. Clin Microbiol Rev. 2017; 30(1): 409-447. [CrossRef]
6. [6] Sulakvelidze A, Alavidze Z, Morris JG. Bacteriophage therapy. Antimicrob Agents Chemother. 2001; 45(3): 649-659. [CrossRef]
7. [7] Golkar Z, Bagasra O, Pace DG. Bacteriophage therapy: a potential solution for the antibiotic resistance crisis. J Infect Dev Ctries. 2014; 8(2): 129-136. [CrossRef]
8. [8] Wittebole X, De Roock S, Opal SM. A historical overview of bacteriophage therapy as an alternative to antibiotics for the treatment of bacterial pathogens. Virulence. 2014; 5(1): 226-235. [CrossRef]

9. [9] Clokie MRJ, Millard AD, Letarov AV, Heaphy S. Phages in nature. Bacteriophage. 2011; 1:31–45. [CrossRef]
10. [10] Viertel TM, Ritter K, Horz HP. Viruses versus bacteria—novel approaches to phage therapy as a tool against multidrug-resistant pathogens. J Antimicrob Chemother. 2014; 69:2326–2336. [CrossRef]
11. [11] WHO publishes list of bacteria for which new antibiotics are urgently needed 2017. http://www.who.int/mediacentre/news/releases/2017/bacteria-antibiotics-needed/en/, (accessed February 15).
12. [12] Luong T, Salabarria AC, Roach DR. Phage Therapy in the Resistance Era: Where Do We Stand and Where Are We Going? Clin Ther. 2020; 42(9): 1659-1680. [CrossRef]
13. [13] Gundogdu A, Bolkvadze D, Kilic H. In vitro Effectiveness of Commercial Bacteriophage Cocktails on Diverse Extended-Spectrum Beta-Lactamase Producing Escherichia coli Strains. Front Microbiol. 2016; 7(1761). [CrossRef]
14. [14] Sybesma W, Zbinden R, Chanishvili N, Kutateladze M, Chkhotua A, Ujmajuridze A, Mehnert U, Kessler TM.Bacteriophages as Potential Treatment for Urinary Tract Infections. Front Microbiol. 2016; 7(465). [CrossRef]
15. [15] Fitzgerald-Hughes D, Bolkvadze D, Balarjishvili N, Leshkasheli L, Ryan M, Burke L, Stevens N, Humphreys H, Kutateladze M. Susceptibility of extended-spectrum- β-lactamase-producing Escherichia coli to commercially available and laboratory-isolated bacteriophages. J Antimicrob Chemother. 2013; 69(4): 1148-1150. [CrossRef]
16. [16] Bernasconi OJ, Donà V, Tinguely R, Endimiani A. In vitro activity of three commercial bacteriophage cocktails against multidrug-resistant Escherichia coli and Proteus spp. strains of human and non-human origin. J Glob Antimicrob Resist. 2017; 8: 179-185. [CrossRef]
17. [17] Kropinski AM, Mazzocco A, Waddell TE, Lingohr E, Johnson RP. Enumeration of bacteriophages by double agar overlay plaque assay. Methods Mol Biol (Clifton, NJ). 2009; 501:69-76.
18. [18] Oliveira H, Pinto G, Oliveira A, Oliveira C, Faustino MA, Briers Y, Domingues L, Azeredo J. Characterization and genome sequencing of a Citrobacter freundii phage CfP1 harboring a lysin active against multidrug-resistant isolates. Appl Microbiol Biotechnol. 2016; 100(24): 10543-10553. [CrossRef]
19. [19] Adams MH. Bacteriophages: Interscience publishers; 1959.
20. [20] Merabishvili M, Pirnay J-P, Verbeken G, Chanishvili N, Tediashvili M, Lashkhi N, Glonti T, Krylov V, Mast J, Parys LV, Lavigne R, Volckaert G, Mattheus W, Verween G, De Corte P, Rose T, Jennes S, Zizi M, De Vos D,Vaneechoutte M. Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PloS one. 2009; 4(3): 4944-4955. [CrossRef]
21. [21] Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual: Cold spring harbor laboratory press; 1989.
22. [22] Sasikala D, Srinivasan P. Characterization of potential lytic bacteriophage against Vibrio alginolyticus and its therapeutic implications on biofilm dispersal. Microb Pathog. 2016; 101: 24-35. [CrossRef]
23. [23] Garbe J, Wesche A, Bunk B, Kazmierczak M, Selezska K, Rohde C, Sikorski J, Rohde M, Jahn D, Schobert M. Characterization of JG024, a Pseudomonas aeruginosa PB1-like broad host range phage under simulated infection conditions. BMC Microbiol. 2010; 10(1): 301-311. [CrossRef]