vB-Ea-5: a lytic bacteriophage against multi-drug-resistant Enterobacter aerogenes
-
Fatemeh Habibinava
,
Mohammad Reza Zolfaghari
,
Mohsen Zargar
,
Salehe Sabouri Shahrbabak
,
Mohammad Soleimani
Abstract
Background and Objectives: Multi-drug-resistant Enterobacter aerogenes is associated with various infectious diseases that cannot be easily treated by antibiotics. However, bacteriophages have potential therapeutic applications in the control of multi-drug-resistant bacteria. In this study, we aimed to isolate and characterize of a lytic bacteriophage that can lyse specifically the multi-drug-resistant (MDR) E. aerogenes.
Materials and Methods: Lytic bacteriophage was isolated from Qaem hospital wastewater and characterized morphologically and genetically. Next-generation sequencing was used to complete genome analysis of the isolated bacteriophage.
Results: Based on the transmission electron microscopy feature, the isolated bacteriophage (vB-Ea-5) belongs to the family Myoviridae. vB-Ea-5 had a latent period of 25 minutes, a burst size of 13 PFU/ml, and a burst time of 40 min. Genome sequencing revealed that vB-Ea-5 has a 135324 bp genome with 41.41% GC content. The vB-Ea-5 genome codes 212 ORFs 90 of which were categorized into several functional classes such as DNA replication and modification, transcriptional regulation, packaging, structural proteins, and a host lysis protein (Holin). No antibiotic resistance and toxin genes were detected in the genome. SDS-PAGE of vB-Ea-5 proteins exhibited three major and four minor bands with a molecular weight ranging from 18 to 50 kD.
Conclusion: Our study suggests vB-Ea-5 as a potential candidate for phage therapy against MDR E. aerogenes infections.
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28. Manohar P, Tamhankar AJ, Lundborg CS, Nachimuthu R. Therapeutic characterization and efficacy of bacteriophage cocktails infecting Escherichia coli, Klebsiella pneumoniae, and Enterobacter Species. Front Microbiol 2019;10:574.
29. Xie XT, Kropinski AM, Tapscott B, Weese JS, Turner PV. Prevalence of fecal viruses and bacteriophage in Canadian farmed mink (Neovison vison). Microbiologyopen 2019;8(1):e00622.
30. Zhao J, Zhang Z, Tian C, Chen X, Hu L, Wei X, et al. Characterizing the biology of lytic bacteriophage vB_EaeM_φEap-3 infecting multidrug-resistant Enterobacter aerogenes. Front Microbiol 2019;10:420.
31. Dalmasso M, Strain R, Neve H, Franz CM, Cousin FJ, Ross RP, et al. Three new Escherichia coli phages from the human gut show promising potential for phage therapy. PLoS One 2016;11(6): e0156773.
32. Royer S, Morais AP, da Fonseca Batistão DW. Phage therapy as strategy to face post-antibiotic era: a guide to beginners and experts. Arch Microbiol 2021: 10.1007/s00203-020-02167-5.
33. Cahill J, Young R (2019). Phage lysis: multiple genes for multiple barriers. ADV VIRUS RES. 103: Elsevier; pp. 33-70.
34. Kreuzer KN, Brister JR. Initiation of bacteriophage T4 DNA replication and replication fork dynamics: a review in the Virology Journal series on bacteriophage T4 and its relatives. Virol J 2010; 7:358.
35. Rao VB, Black LW. Structure and assembly of bacteriophage T4 head. Virol J 2010; 7:356.
2. Davin-Regli A. Enterobacter aerogenes and Enterobacter cloacae; versatile bacterial pathogens confronting antibiotic treatment. Front Microbiol 2015;6:392.
3. Pirisi A. Phage therapy—advantages over antibiotics? Lancet 2000;356:1418.
4. Tacconelli E, Carrara E, Savoldi A, Harbarth S, Mendelson M, Monnet DL, et al. Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect Dis 2018;18:318-327.
5. Carlton RM. Phage therapy: past history and future prospects. Arch Immunol Ther Exp (Warsz) 1999; 47:267-274.
6. Kostyrka G, Sankaran N. From obstacle to lynchpin: the evolution of the role of bacteriophage lysogeny in defining and understanding viruses. Notes Rec 2020;74: 10.1098/rsnr.2019.0033.
7. Moghadam MT, Amirmozafari N, Shariati A, Hallajzadeh M, Mirkalantari S, Khoshbayan A, et al. How phages overcome the challenges of drug resistant bacteria in clinical infections. Infect Drug Resist 2020;13:45-61.
8. Morrisette T, Kebriaei R, Lev KL, Morales S, Rybak MJ. Bacteriophage therapeutics: a primer for clinicians on phage-antibiotic combinations. Pharmacotherapy 2020;40:153-168.
9. Parmar KM, Hathi ZJ, Dafale NA. Control of multidrug-resistant gene flow in the environment through bacteriophage intervention. Appl Biochem Biotechnol 2017;181:1007-1029.
10. Verthé K, Possemiers S, Boon N, Vaneechoutte M, Verstraete W. Stability and activity of an Enterobacter aerogenes-specific bacteriophage under simulated gastro-intestinal conditions. Appl Microbiol Biotechnol 2004;65:465-472.
11. Mishra CK, Choi TJ, Kang SC. Isolation and characterization of a bacteriophage F20 virulent to Enterobacter aerogenes. J Gen Virol 2012;93:2310-2314.
12. Li E, Wei X, Ma Y, Yin Z, Li H, Lin W, et al. Isolation and characterization of a bacteriophage phiEap-2 infecting multidrug resistant Enterobacter aerogenes. Sci Rep 2016;6:28338.
13. Zhao J, Zhang Z, Tian C, Chen X, Jiang A, Feng R, et al. Characterizing the biology of lytic bacteriophage vB_EaeM_φEap-3 infecting multidrug-resistant Enterobacter aerogenes. Front Microbiol 2019;10:420.
14. Lin DM, Koskella B, Lin HC. Phage therapy: An alternative to antibiotics in the age of multi-drug resistance. World J Gastrointest Pharmacol Ther 2017;8:162-173.
15. Biochemical Tests for the Identification of Aerobic Bacteria (2010). Clinical Microbiology Procedures Handbook, 3rd Edition: American Society of Microbiology.
16. Mohammadi Bandari N, Zargar M, Keyvani H, Talebi M, Zolfaghari MR. Antibiotic resistance among Klebsiella pneumoniae, molecular detection and expression level of blaKPC and blaGES genes by real-time PCR. Jundishapur J Microbiol 2019; 12 (10); e93070.
17. Shahrbabak SS, Khodabandehlou Z, Shahverdi AR, Skurnik M, Ackermann H-W, Varjosalo M, et al. Isolation, characterization and complete genome sequence of PhaxI: a phage of Escherichia coli O157: H7. Microbiology (Reading) 2013;159:1629-1638.
18. Birge EA (2000). T4 bacteriophage as a model genetic system. Bacterial and Bacteriophage Genetics: Springer; pp. 171-214.
19. Brenner S, Horne RW. A negative staining method for high resolution electron microscopy of viruses. Biochim Biophys Acta 1959;34:103-110.
20. Arndt D, Grant JR, Marcu A, Sajed T, Pon A, Liang Y, et al. PHASTER: a better, faster version of the PHAST phage search tool.Nucleic Acids Res 2016;44:W16-21.
21. Besemer J, Lomsadze A, Borodovsky M. GeneMarkS: a self-training method for prediction of gene starts in microbial genomes. Implications for finding sequence motifs in regulatory regions. Nucleic Acids Res 2001;29:2607-2618.
22. Carrias A, Welch TJ, Waldbieser GC, Mead DA, Terhune JS, Liles MR. Comparative genomic analysis of bacteriophages specific to the channel catfish pathogen Edwardsiella ictaluri. Virol J 2011;8:6.
23. Mercer R, Nguyen O, Ou Q, McMullen L, Gänzle MG.Functional analysis of genes comprising the locus of heat resistance in Escherichia coli. Appl Environ Microbiol 2017;83(20):e01400-17.
24. Xin B, Zheng J, Liu H, Li J, Ruan L, Peng D, et al. Thusin, a novel two-component lantibiotic with potent antimicrobial activity against several Gram-positive pathogens. Front Microbiol 2016;7:1115.
25. McCusker MP, Ferreira DA, Cooney D, Alves BM, Fanning S, Pagès J-M, et al. Modulation of antimicrobial resistance in clinical isolates of Enterobacter aerogenes: A strategy combining antibiotics and chemosensitisers. J Glob Antimicrob Resist 2018;16:187-198.
26. Ackermann H-W (2009). Phage classification and characterization. Bacteriophages: Springer; pp. 127-140.
27. Lopes A, Tavares P, Petit M-A, Guérois R, Zinn-Justin S. Automated classification of tailed bacteriophages according to their neck organization. BMC Genomics 2014;15:1027.
28. Manohar P, Tamhankar AJ, Lundborg CS, Nachimuthu R. Therapeutic characterization and efficacy of bacteriophage cocktails infecting Escherichia coli, Klebsiella pneumoniae, and Enterobacter Species. Front Microbiol 2019;10:574.
29. Xie XT, Kropinski AM, Tapscott B, Weese JS, Turner PV. Prevalence of fecal viruses and bacteriophage in Canadian farmed mink (Neovison vison). Microbiologyopen 2019;8(1):e00622.
30. Zhao J, Zhang Z, Tian C, Chen X, Hu L, Wei X, et al. Characterizing the biology of lytic bacteriophage vB_EaeM_φEap-3 infecting multidrug-resistant Enterobacter aerogenes. Front Microbiol 2019;10:420.
31. Dalmasso M, Strain R, Neve H, Franz CM, Cousin FJ, Ross RP, et al. Three new Escherichia coli phages from the human gut show promising potential for phage therapy. PLoS One 2016;11(6): e0156773.
32. Royer S, Morais AP, da Fonseca Batistão DW. Phage therapy as strategy to face post-antibiotic era: a guide to beginners and experts. Arch Microbiol 2021: 10.1007/s00203-020-02167-5.
33. Cahill J, Young R (2019). Phage lysis: multiple genes for multiple barriers. ADV VIRUS RES. 103: Elsevier; pp. 33-70.
34. Kreuzer KN, Brister JR. Initiation of bacteriophage T4 DNA replication and replication fork dynamics: a review in the Virology Journal series on bacteriophage T4 and its relatives. Virol J 2010; 7:358.
35. Rao VB, Black LW. Structure and assembly of bacteriophage T4 head. Virol J 2010; 7:356.
| Files | ||
| Issue | Vol 13 No 2 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v13i2.5984 | |
| Keywords | ||
| Drug resistance; Bacteriophages; Waste water; Enterobacter aerogenes; Myoviridae | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Habibinava F, Zolfaghari MR, Zargar M, Sabouri Shahrbabak S, Soleimani M. vB-Ea-5: a lytic bacteriophage against multi-drug-resistant Enterobacter aerogenes. Iran J Microbiol. 2021;13(2):225-234.
