Biological characteristics and anti-biofilm activity of a lytic phage against vancomycin-resistant Enterococcus faecium
-
Forough Goodarzi
,
Masoumeh Hallajzadeh
,
Mohammad Sholeh
,
Malihe Talebi
,
Vahid Pirhajati Mahabadi
,
Nour Amirmozafari
Abstract
Background and Objectives: An important leading cause of the emergence of vancomycin-resistant enterococci, especially Enterococcus faecium, is the inefficiency of antibiotics in the elimination of drug-resistant pathogens. Consequently, the need for alternative treatments is more necessary than ever.
Materials and Methods: A highly effective bacteriophage against vancomycin-resistant E. faecium called vB-EfmS-S2 was isolated from hospital sewage. The biological properties of phage S2 and its effect on biofilm structures were determined.
Results: Phage S2 was specifically capable of lysing a wide range of clinical E. faecium isolates. According to Electron microscopy observations, the phage S2 belonged to the Siphoviridea family. Suitable pH spectra for phage survival was 5-11, at which the phage showed 100% activity. The optimal temperature for phage growth was 30-45°C, with the highest growth at 37°C. Based on one-step growth curve results, the latent period of phage S2 was 14 min with a burst size of 200 PFU/ml. The phage S2 was also able to tolerate bile at concentrations of 1 and 2% and required Mg2+ for an effective infection cycle. Biofilms were significantly inhibited and disrupted in the presence of the phage.
Conclusion: According to the results, phage S2 could potentially be an alternative for the elimination and control of vancomycin-resistant E. faecium biofilm.
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6. Khalifa L, Gelman D, Shlezinger M, Dessal AL, Coppenhagen-Glazer S, Beyth N, et al. Defeating antibiotic-and phage-resistant Enterococcus faecalis using a phage cocktail in vitro and in a clot model. Front Microbiol 2018;9:326.
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8. Paganelli FL, Huebner J, Singh KV, Zhang X, van Schaik W, Wobser D, et al. Genome-wide screening identifies phosphotransferase system permease BepA to be involved in Enterococcus faecium endocarditis and biofilm formation. J Infect Dis 2016;214:189-195.
9. Donlan RM. Biofilm formation: a clinically relevant microbiological process. Clin Infect Dis 2001;33:1387-1392.
10. Akpaka PE, Kissoon S, Wilson C, Jayaratne P, Smith A, Golding GR. Molecular characterization of vancomycin-resistant Enterococcus faecium isolates from Bermuda. PLoS One 2017;12(3):e0171317.
11. Ch’ng JH, Chong KKL, Lam LN, Wong JJ, Kline KA. Biofilm-associated infection by enterococci. Nat Rev Microbiol 2019;17:82-94.
12. Lewis K. Riddle of biofilm resistance. Antimicrob Agents Chemother 2001;45:999-1007.
13. Harada LK, Silva EC, Campos WF, Del Fiol FS, Vila M, Dąbrowska K, et al. Biotechnological applications of bacteriophages: state of the art. Microbiol Res 2018;212-213:38-58.
14. Shlezinger M, Coppenhagen-Glazer S, Gelman D, Beyth N, Hazan R. Eradication of vancomycin-resistant enterococci by combining phage and vancomycin. Viruses 2019;11:954.
15. Principi N, Silvestri E, Esposito S. Advantages and limitations of bacteriophages for the treatment of bacterial infections. Front Pharmacol 2019;10:513.
16. Sattari-Maraji A, Jabalameli F, Farahani NN, Beigverdi R, Emaneini M. Antimicrobial resistance pattern, virulence determinants and molecular analysis of Enterococcus faecium isolated from children infections in Iran. BMC Microbiol 2019;19:156.
17. Clinical and Laboratory Standards Institute [CLSI] (2017). Performance Standards for Antimicrobial Susceptibility Testing M02-A12 M-A, and M11-A8, 27th Ed, Philadelphia, PA: Clinical and Laboratory Standards Institute.
18. Talebi M, Pourshafiei MR, Oskouei M, Eshraghi SS. Molecular analysis of vanHAX element in vancomycin resistant enterococci isolated from hospitalized patients in Tehran. Iran Biomed J 2008;12:223-228.
19. Gong P, Cheng M, Li X, Jiang H, Yu C, Kahaer N, et al. Characterization of Enterococcus faecium bacteriophage IME-EFm5 and its endolysin LysEFm5. Virology 2016;492:11-20.
20. Wang Y, Wang W, Lv Y, Zheng W, Mi Z, Pei G, et al. Characterization and complete genome sequence analysis of novel bacteriophage IME-EFm1 infecting Enterococcus faecium. J Gen Virol 2014;95:2565-2575.
21. Hallajzadeh M, Mojtahedi A, Amirmozafari N, Pirhajati Mahabadi V. Characterizing a lytic bacteriophage infecting methicillin-resistant Staphylococcus aureus (MRSA) isolated from burn patients. Arch Clin Infect Dis 2020;15(1):e91634.
22. Kutter E (2009). Phage host range and efficiency of plating. In: Bacteriophages, methods and protocols. Eds, MR Clokie, AM Kropinski. Springer, 1st ed. Switzerland AG, pp. 141-149.
23. Chen X, Guo J, Liu Y, Chai S, Ma R, Munguntsetseg B. Characterization and adsorption of a Lactobacillus plantarum virulent phage. J Dairy Sci 2019;102:3879-3886.
24. Raza T, Andleeb S, Ullah SR, Jamal M, Mehmood K, Ali M. Isolation and characterization of a phage to control vancomycin resistant Enterococcus faecium. Open Life Sci 2018;13:553-560.
25. Kumar L, Cox CR, Sarkar SK. Matrix metalloprotease-1 inhibits and disrupts Enterococcus faecalis biofilms. PLoS One 2019;14(1):e0210218.
26. Adriaenssens EM, Wittmann J, Kuhn JH, Turner D, Sullivan MB, Dutilh BE, et al. Taxonomy of prokaryotic viruses: 2017 update from the ICTV bacterial and archaeal viruses Subcommittee. Arch Virol 2018;163:1125-1129.
27. Golkar Z, Bagasra O, Pace DG. Bacteriophage therapy: a potential solution for the antibiotic resistance crisis. J Infect Dev Ctries 2014;8:129-136.
28. Li B, Webster TJ. Bacteria antibiotic resistance: new challenges and opportunities for implant‐associated orthopedic infections. J Orthop Res 2018;36:22-32.
29. Higuita NIA, Huycke MM (2014). Enterococcal disease, epidemiology, and implications for treatment. In: Enterococci: from commensals to leading causes of drug resistant infection. Massachusetts eye and ear, Last updated, Boston, pp. 65-99.
30. Li J, Shi H, Zhao C, Hao Y, He Y, Sun Y. Complete genome sequence of the siphoviral bacteriophage Ec-ZZ2, which is capable of lysing Enterococcus faecium. Genome Announc 2016;4(6):e01167-16.
31. Bibi Z, Abbas Z, Rehman SU. The phage P. E1 isolated from hospital sewage reduces the growth of Escherichia coli. Biocontrol Sci Technol 2016;26:181-188.
32. Jończyk E, Kłak M, Międzybrodzki R, Górski A. The influence of external factors on bacteriophages-review. Folia Microbiol (Praha) 2011;56:191-200.
33. Nobrega FL, Costa AR, Santos JF, Siliakus MF, Van Lent JW, Kengen SW, et al. Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine. Sci Rep 2016;6:39235.
34. Rahmat Ullah S, Andleeb S, Raza T, Jamal M, Mehmood K. Effectiveness of a lytic phage SRG1 against vancomycin-resistant Enterococcus faecalis in compost and soil. Biomed Res Int 2017;2017:9351017.
35. Marcó MB, Reinheimer JA, Quiberoni A. Phage adsorption to Lactobacillus plantarum: influence of physiological and environmental factors. Int J Food Microbiol 2010;138:270-275.
36. Khanorkar SV (2011). Insights in Physiology. 1st ed. Jaypee Brothers Medical Pub. New Delhi.
37. Ma Y, Pacan JC, Wang Q, Xu Y, Huang X, Korenevsky A, et al. Microencapsulation of bacteriophage felix O1 into chitosan-alginate microspheres for oral delivery. Appl Environ Microbiol 2008;74:4799-4805.
38. Ramirez K, Cazarez-Montoya C, Lopez-Moreno HS, Castro-del Campo N. Bacteriophage cocktail for biocontrol of Escherichia coli O157: H7: stability and potential allergenicity study. PLoS One 2018;13(5):e0195023.
39. Zhang W, Mi Z, Yin X, Fan H, An X, Zhang Z, et al. Characterization of Enterococcus faecalis phage IME-EF1 and its endolysin. PLoS One 2013;8(11):e80435.
40. Szafrański SP, Winkel A, Stiesch M. The use of bacteriophages to biocontrol oral biofilms. J Biotechnol 2017;250:29-44.
41. Limoli DH, Jones CJ, Wozniak DJ. Bacterial extracellular polysaccharides in biofilm formation and function. Microbiol Spectr 2015;3:10.1128/microbiolspec.MB-0011-2014.
42. Azeredo J, Sutherland IW. The use of phages for the removal of infectious biofilms. Curr Pharm Biotechnol 2008;9:261-266.
43. Melo LDR, Ferreira R, Costa AR, Oliveira H, Azeredo J. Efficacy and safety assessment of two enterococci phages in an in vitro biofilm wound model. Sci Rep 2019;9:6643.
44. Abedon ST. Ecology of anti-biofilm agents II: bacteriophage exploitation and biocontrol of biofilm bacteria. Pharmaceuticals (Basel) 2015;8:559-589.
45. Chan BK, Abedon ST. Bacteriophages and their enzymes in biofilm control. Curr Pharm Des 2015;21:85-99.
2. Bin-Asif H, Ali SA (2019). The genus Enterococcus and its associated virulent factors. In: Microorganism. Eds, M Blumenberg, M Shaaban, A Elgaml. IntechOpen, 1st ed, London, pp. 109-130.
3. Uchiyama J, Takemura I, Satoh M, Kato S, Ujihara T, Akechi K, et al. Improved adsorption of an Enterococcus faecalis bacteriophage ΦEF24C with a spontaneous point mutation. PLoS One 2011;6(10):e26648.
4. Cui P, Feng L, Zhang L, He J, An T, Fu X, et al. Antimicrobial resistance, virulence genes, and biofilm formation capacity among Enterococcus species from Yaks in Aba Tibetan autonomous prefecture, China. Front Microbiol 2020;11:1250.
5. Khalifa L, Shlezinger M, Beyth S, Houri-Haddad Y, Coppenhagen-Glazer S, Beyth N, et al. Phage therapy against Enterococcus faecalis in dental root canals. J Oral Microbiol 2016;8:32157.
6. Khalifa L, Gelman D, Shlezinger M, Dessal AL, Coppenhagen-Glazer S, Beyth N, et al. Defeating antibiotic-and phage-resistant Enterococcus faecalis using a phage cocktail in vitro and in a clot model. Front Microbiol 2018;9:326.
7. Castillo-Rojas G, Mazari-Hiríart M, de León SP, Amieva-Fernández RI, Agis-Juárez RA, Huebner J, et al. Comparison of Enterococcus faecium and Enterococcus faecalis strains isolated from water and clinical samples: antimicrobial susceptibility and genetic relationships. PLoS One 2013;8(4):e59491.
8. Paganelli FL, Huebner J, Singh KV, Zhang X, van Schaik W, Wobser D, et al. Genome-wide screening identifies phosphotransferase system permease BepA to be involved in Enterococcus faecium endocarditis and biofilm formation. J Infect Dis 2016;214:189-195.
9. Donlan RM. Biofilm formation: a clinically relevant microbiological process. Clin Infect Dis 2001;33:1387-1392.
10. Akpaka PE, Kissoon S, Wilson C, Jayaratne P, Smith A, Golding GR. Molecular characterization of vancomycin-resistant Enterococcus faecium isolates from Bermuda. PLoS One 2017;12(3):e0171317.
11. Ch’ng JH, Chong KKL, Lam LN, Wong JJ, Kline KA. Biofilm-associated infection by enterococci. Nat Rev Microbiol 2019;17:82-94.
12. Lewis K. Riddle of biofilm resistance. Antimicrob Agents Chemother 2001;45:999-1007.
13. Harada LK, Silva EC, Campos WF, Del Fiol FS, Vila M, Dąbrowska K, et al. Biotechnological applications of bacteriophages: state of the art. Microbiol Res 2018;212-213:38-58.
14. Shlezinger M, Coppenhagen-Glazer S, Gelman D, Beyth N, Hazan R. Eradication of vancomycin-resistant enterococci by combining phage and vancomycin. Viruses 2019;11:954.
15. Principi N, Silvestri E, Esposito S. Advantages and limitations of bacteriophages for the treatment of bacterial infections. Front Pharmacol 2019;10:513.
16. Sattari-Maraji A, Jabalameli F, Farahani NN, Beigverdi R, Emaneini M. Antimicrobial resistance pattern, virulence determinants and molecular analysis of Enterococcus faecium isolated from children infections in Iran. BMC Microbiol 2019;19:156.
17. Clinical and Laboratory Standards Institute [CLSI] (2017). Performance Standards for Antimicrobial Susceptibility Testing M02-A12 M-A, and M11-A8, 27th Ed, Philadelphia, PA: Clinical and Laboratory Standards Institute.
18. Talebi M, Pourshafiei MR, Oskouei M, Eshraghi SS. Molecular analysis of vanHAX element in vancomycin resistant enterococci isolated from hospitalized patients in Tehran. Iran Biomed J 2008;12:223-228.
19. Gong P, Cheng M, Li X, Jiang H, Yu C, Kahaer N, et al. Characterization of Enterococcus faecium bacteriophage IME-EFm5 and its endolysin LysEFm5. Virology 2016;492:11-20.
20. Wang Y, Wang W, Lv Y, Zheng W, Mi Z, Pei G, et al. Characterization and complete genome sequence analysis of novel bacteriophage IME-EFm1 infecting Enterococcus faecium. J Gen Virol 2014;95:2565-2575.
21. Hallajzadeh M, Mojtahedi A, Amirmozafari N, Pirhajati Mahabadi V. Characterizing a lytic bacteriophage infecting methicillin-resistant Staphylococcus aureus (MRSA) isolated from burn patients. Arch Clin Infect Dis 2020;15(1):e91634.
22. Kutter E (2009). Phage host range and efficiency of plating. In: Bacteriophages, methods and protocols. Eds, MR Clokie, AM Kropinski. Springer, 1st ed. Switzerland AG, pp. 141-149.
23. Chen X, Guo J, Liu Y, Chai S, Ma R, Munguntsetseg B. Characterization and adsorption of a Lactobacillus plantarum virulent phage. J Dairy Sci 2019;102:3879-3886.
24. Raza T, Andleeb S, Ullah SR, Jamal M, Mehmood K, Ali M. Isolation and characterization of a phage to control vancomycin resistant Enterococcus faecium. Open Life Sci 2018;13:553-560.
25. Kumar L, Cox CR, Sarkar SK. Matrix metalloprotease-1 inhibits and disrupts Enterococcus faecalis biofilms. PLoS One 2019;14(1):e0210218.
26. Adriaenssens EM, Wittmann J, Kuhn JH, Turner D, Sullivan MB, Dutilh BE, et al. Taxonomy of prokaryotic viruses: 2017 update from the ICTV bacterial and archaeal viruses Subcommittee. Arch Virol 2018;163:1125-1129.
27. Golkar Z, Bagasra O, Pace DG. Bacteriophage therapy: a potential solution for the antibiotic resistance crisis. J Infect Dev Ctries 2014;8:129-136.
28. Li B, Webster TJ. Bacteria antibiotic resistance: new challenges and opportunities for implant‐associated orthopedic infections. J Orthop Res 2018;36:22-32.
29. Higuita NIA, Huycke MM (2014). Enterococcal disease, epidemiology, and implications for treatment. In: Enterococci: from commensals to leading causes of drug resistant infection. Massachusetts eye and ear, Last updated, Boston, pp. 65-99.
30. Li J, Shi H, Zhao C, Hao Y, He Y, Sun Y. Complete genome sequence of the siphoviral bacteriophage Ec-ZZ2, which is capable of lysing Enterococcus faecium. Genome Announc 2016;4(6):e01167-16.
31. Bibi Z, Abbas Z, Rehman SU. The phage P. E1 isolated from hospital sewage reduces the growth of Escherichia coli. Biocontrol Sci Technol 2016;26:181-188.
32. Jończyk E, Kłak M, Międzybrodzki R, Górski A. The influence of external factors on bacteriophages-review. Folia Microbiol (Praha) 2011;56:191-200.
33. Nobrega FL, Costa AR, Santos JF, Siliakus MF, Van Lent JW, Kengen SW, et al. Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine. Sci Rep 2016;6:39235.
34. Rahmat Ullah S, Andleeb S, Raza T, Jamal M, Mehmood K. Effectiveness of a lytic phage SRG1 against vancomycin-resistant Enterococcus faecalis in compost and soil. Biomed Res Int 2017;2017:9351017.
35. Marcó MB, Reinheimer JA, Quiberoni A. Phage adsorption to Lactobacillus plantarum: influence of physiological and environmental factors. Int J Food Microbiol 2010;138:270-275.
36. Khanorkar SV (2011). Insights in Physiology. 1st ed. Jaypee Brothers Medical Pub. New Delhi.
37. Ma Y, Pacan JC, Wang Q, Xu Y, Huang X, Korenevsky A, et al. Microencapsulation of bacteriophage felix O1 into chitosan-alginate microspheres for oral delivery. Appl Environ Microbiol 2008;74:4799-4805.
38. Ramirez K, Cazarez-Montoya C, Lopez-Moreno HS, Castro-del Campo N. Bacteriophage cocktail for biocontrol of Escherichia coli O157: H7: stability and potential allergenicity study. PLoS One 2018;13(5):e0195023.
39. Zhang W, Mi Z, Yin X, Fan H, An X, Zhang Z, et al. Characterization of Enterococcus faecalis phage IME-EF1 and its endolysin. PLoS One 2013;8(11):e80435.
40. Szafrański SP, Winkel A, Stiesch M. The use of bacteriophages to biocontrol oral biofilms. J Biotechnol 2017;250:29-44.
41. Limoli DH, Jones CJ, Wozniak DJ. Bacterial extracellular polysaccharides in biofilm formation and function. Microbiol Spectr 2015;3:10.1128/microbiolspec.MB-0011-2014.
42. Azeredo J, Sutherland IW. The use of phages for the removal of infectious biofilms. Curr Pharm Biotechnol 2008;9:261-266.
43. Melo LDR, Ferreira R, Costa AR, Oliveira H, Azeredo J. Efficacy and safety assessment of two enterococci phages in an in vitro biofilm wound model. Sci Rep 2019;9:6643.
44. Abedon ST. Ecology of anti-biofilm agents II: bacteriophage exploitation and biocontrol of biofilm bacteria. Pharmaceuticals (Basel) 2015;8:559-589.
45. Chan BK, Abedon ST. Bacteriophages and their enzymes in biofilm control. Curr Pharm Des 2015;21:85-99.
| Files | ||
| Issue | Vol 13 No 5 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v13i5.7436 | |
| Keywords | ||
| Enterococcus faecium; Vancomycin-resistant Enterococcus faecium; Phage therapy; Antibiotic-resistance; Biofilm | ||
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How to Cite
1.
Goodarzi F, Hallajzadeh M, Sholeh M, Talebi M, Pirhajati Mahabadi V, Amirmozafari N. Biological characteristics and anti-biofilm activity of a lytic phage against vancomycin-resistant Enterococcus faecium. Iran J Microbiol. 2021;13(5):691-702.
