YENİ İZOLE EDILEN VB_K1 BAKTERİYOFAJININ İZOLASYONU VE ESBL POZİTİF KLEBSİELLA TÜRLERİ ÜZERİNE DUYARLILIĞININ ARAŞTIRILMASI

Yıl 2021, , 515 - 523, 27.09.2021

Hilal Basak Erol , Banu Kaşkatepe

https://doi.org/10.33483/jfpau.963979

Öz

Amaç: Klebsiella ile ilişkili enfeksiyonların tedavisinde kullanılan antimikrobiyallerin etkinliğindeki azalma, alternatif tedavi strateji arayışlarını gerektirmiştir. Bu çalışma Klebsiella türlerine özgü litik karakterde bakteriyofaj izolasyonunun sağlanmasını ve alternatif antimikrobiyal ajan olarak kullanım potansiyelinin araştırılmasını amaçlamaktadır.

Gereç ve Yöntem: Ankara deresinden alınan su örneklerinden faj izolasyonu için, Genişlemiş spektrumlu beta laktamaz (GSBL) üreticisi bir adet Klebsiella spp. suşu konak bakteri olarak kullanıldı. Faj zenginleştirme sonrası, olası bakteriyofaj varlığını belirlemek için spot test yöntemi uygulandı. Litik bakteriyofaj varlığı, spot testi pozitif numunelere çift tabaka agar yöntemi uygulanarak doğrulandı. Bakteriyofajın duyarlılığı, 38 klinik GSBL pozitif Klebsiella spp. suşu kullanılarak in vitro spot test ile belirlendi.

Sonuç ve Tartışma: İlk taramada petride 1.00 mm çapında görünür plaklar üreten vB_K1 bakteriyofajı izole edildi. GSBL pozitif Klebsiella spp. suşlarının bakteriyofaj duyarlılığı %73.7 olarak belirlendi. vB_K1 bakteriyofajının Klebsiella suşları üzerinde çok etkili olduğu kanıtlanmıştır. Bununla birlikte, karakterize edilen bakteriyofajın in vitro bakteriyofaj duyarlılığı cesaret verici bir gelişmedir.

Anahtar Kelimeler

Bakteriyofaj, direnç, ESBL, Klebsiella spp.

ISOLATION OF NEWLY ISOLATED VB_K1 BACTERIOPHAGE AND INVESTIGATION OF SUSCEPTIBILITY ON ESBL POSITIVE KLEBSIELLA SPP. STRAINS

Öz

Objective: The decrease in the efficacy of antimicrobials in the treatment of Klebsiella-related infections necessitated the search for alternative treatment strategies. This study aims to provide isolation of lytic bacteriophage specific to Klebsiella species and to investigate its potential for use as alternative antimicrobial agent.

Material and Method: One Extended spectrum beta lactamase (ESBL) producer Klebsiella strain was used as host bacteria and water samples were collected from river in Ankara for bacteriophage isolation. Spot test method was applied to determine the possible presence of bacteriophage after phage enrichment. To confirm the presence of the lytic bacteriophage, double layer agar method was applied to spot test positive samples. The susceptibility of the bacteriophage was determined using in vitro spot test. 38 clinical ESBL positive Klebsiella spp. strains were used for this analysis.

Result and Discussion: In the initial screening, the vB_K1 bacteriophage producing visible plaques with a diameter of 1.00 mm was isolated in the petri dish. The susceptibility of ESBL positive Klebsiella spp. strains to this bacteriophage was determined as 73.7%. It was proved that vB_K1 bacteriophage is very effective to Klebsiella spp. strains. However, in vitro bacteriophage susceptibility of characterized bacteriophage is encouraging development.

Anahtar Kelimeler

Bacteriophage, ESBL, Klebsiella spp., resistance

Kaynakça

• 1. Podschun, R., & Ullmann, U. (1998). Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clinical microbiology reviews, 11(4), 589-603. doi:10.1128/cmr.11.4.589
• 2. Moxon, C. A., & Paulus, S. (2016). Beta-lactamases in Enterobacteriaceae infections in children. J Infect, 72 Suppl, S41-49. doi:10.1016/j.jinf.2016.04.021
• 3. Bitsori, M., & Galanakis, E. (2019). Treatment of Urinary Tract Infections Caused by ESBL-producing Escherichia coli or Klebsiella pneumoniae. The Pediatric Infectious Disease Journal, 38(12), e332-e335. doi:10.1097/inf.0000000000002487
• 4. Peleg, A. Y., & Hooper, D. C. (2010). Hospital-acquired infections due to gram-negative bacteria. New England Journal of Medicine, 362(19), 1804-1813.
• 5. Golkar, Z., Bagasra, O., & Pace, D. G. (2014). Bacteriophage therapy: a potential solution for the antibiotic resistance crisis. J Infect Dev Ctries, 8(2), 129-136. doi:10.3855/jidc.3573
• 6. Chanishvili, N. (2012). A literature review of the practical application of bacteriophage research. New York: Nova Biomedical Books.
• 7. Bruttin, A., & Brüssow, H. (2005). Human volunteers receiving Escherichia coli phage T4 orally: a safety test of phage therapy. Antimicrobial agents and chemotherapy, 49(7), 2874-2878. doi:10.1128/AAC.49.7.2874-2878.2005
• 8. Domingo-Calap, P., & Delgado-Martinez, J. (2018). Bacteriophages: Protagonists of a Post-Antibiotic Era. Antibiotics (Basel), 7(3), 1-16. doi:10.3390/antibiotics7030066
• 9. Kazi, M., & Annapure, U. S. (2016). Bacteriophage biocontrol of foodborne pathogens. Journal of food science and technology, 53(3), 1355-1362. doi:10.1007/s13197-015-1996-8
• 10. Guttman, B., Raya, R., & Kutter, E. (2005). Basic phage biology. Bacteriophages: Biology and applications, 4.
• 11. Harper, D. R., Parracho, H. M. R. T., Walker, J., Sharp, R., Hughes, G., Werthén, M., . . . Morales, S. (2014). Bacteriophages and Biofilms. Antibiotics, 3(3), 270-284. doi:10.3390/antibiotics3030270
• 12. Kochetkova, V. A., Mamontov, A. S., Moskovtseva, R. L., Erastova, E. I., Trofimov, E. I., Popov, M. I., & Dzhubalieva, S. K. (1989). Phagotherapy of postoperative suppurative-inflammatory complications in patients with neoplasms. Sov Med(6), 23-26.
• 13. Domingo-Calap, P., Beamud, B., Mora-Quilis, L., González-Candelas, F., & Sanjuán, R. (2020). Isolation and Characterization of Two Klebsiella pneumoniae Phages Encoding Divergent Depolymerases. International journal of molecular sciences, 21(9), 3160. doi:10.3390/ijms21093160
• 14. Karamoddini, M. K., Fazli-Bazzaz, B. S., Emamipour, F., Ghannad, M. S., Jahanshahi, A. R., Saed, N., & Sahebkar, A. (2011a). Antibacterial Efficacy of Lytic Bacteriophages against Antibiotic-Resistant <i>Klebsiella</i> Species. TheScientificWorldJOURNAL, 11, 370161. doi:10.1100/tsw.2011.114
• 15. Kumari, S., Harjai, K., & Chhibber, S. (2011). Bacteriophage versus antimicrobial agents for the treatment of murine burn wound infection caused by Klebsiella pneumoniae B5055. J Med Microbiol, 60(Pt 2), 205-210. doi:10.1099/jmm.0.018580-0
• 16. Wintachai, P., Naknaen, A., Thammaphet, J., Pomwised, R., Phaonakrop, N., Roytrakul, S., & Smith, D. R. (2020). Characterization of extended-spectrum-β-lactamase producing Klebsiella pneumoniae phage KP1801 and evaluation of therapeutic efficacy in vitro and in vivo. Scientific Reports, 10(1), 11803. doi:10.1038/s41598-020-68702-y
• 17. Adams, M. H. (1959). Bacteriophages: Interscience publishers.
• 18. Merabishvili, M., Pirnay, J.-P., Verbeken, G., Chanishvili, N., Tediashvili, M., Lashkhi, N., . . . Vaneechoutte, M. (2009). Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PLoS One, 4(3), e4944-e4944. doi:10.1371/journal.pone.0004944
• 19. Oliveira, H., Pinto, G., Oliveira, A., Oliveira, C., Faustino, M. A., Briers, Y., . . . Azeredo, J. (2016). Characterization and genome sequencing of a Citrobacter freundii phage CfP1 harboring a lysin active against multidrug-resistant isolates. Applied Microbiology and Biotechnology, 100(24), 10543-10553. doi:10.1007/s00253-016-7858-0
• 20. Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: a laboratory manual: Cold spring harbor laboratory press.
• 21. Sasikala, D., & Srinivasan, P. (2016). Characterization of potential lytic bacteriophage against Vibrio alginolyticus and its therapeutic implications on biofilm dispersal. Microb Pathog, 101, 24-35. doi:10.1016/j.micpath.2016.10.017
• 22. Knezevic, P., Kostanjsek, R., Obreht, D., & Petrovic, O. (2009). Isolation of Pseudomonas aeruginosa Specific Phages with Broad Activity Spectra. Current Microbiology, 59(2), 173. doi:10.1007/s00284-009-9417-8
• 23. Herridge, W. P., Shibu, P., O'Shea, J., Brook, T. C., & Hoyles, L. (2020). Bacteriophages of Klebsiella spp., their diversity and potential therapeutic uses. J Med Microbiol, 69(2), 176-194. doi:10.1099/jmm.0.001141
• 24. Harper, D. R. (2018). Criteria for Selecting Suitable Infectious Diseases for Phage Therapy. Viruses, 10(4), 177. doi:10.3390/v10040177
• 25. Townsend, E. M., Kelly, L., Gannon, L., Muscatt, G., Dunstan, R., Michniewski, S., . . . Jameson, E. (2021). Isolation and Characterization of Klebsiella Phages for Phage Therapy. Phage (New Rochelle), 2(1), 26-42. doi:10.1089/phage.2020.0046
• 26. Karumidze, N., Kusradze, I., Rigvava, S., Goderdzishvili, M., Rajakumar, K., & Alavidze, Z. (2013). Isolation and Characterisation of Lytic Bacteriophages of Klebsiella pneumoniae and Klebsiella oxytoca. Current Microbiology, 66(3), 251-258. doi:10.1007/s00284-012-0264-7
• 27. Karamoddini, M. K., Fazli-Bazzaz, B. S., Emamipour, F., Ghannad, M. S., Jahanshahi, A. R., Saed, N., & Sahebkar, A. (2011b). Antibacterial efficacy of lytic bacteriophages against antibiotic-resistant Klebsiella species. TheScientificWorldJOURNAL, 11, 1332-1340. doi:10.1100/tsw.2011.114
• 28. Drulis-Kawa, Z., Mackiewicz, P., Kęsik-Szeloch, A., Maciaszczyk-Dziubinska, E., Weber-Dąbrowska, B., Dorotkiewicz-Jach, A., . . . Kropinski, A. M. (2011). Isolation and characterisation of KP34—a novel φKMV-like bacteriophage for Klebsiella pneumoniae. Applied Microbiology and Biotechnology, 90(4), 1333-1345. doi:10.1007/s00253-011-3149-y