Isolation and characterization of bacteriophages from wastewater sources on Enterococcus spp. isolated from clinical samples
Abstract
Background and Objectives: In recent decades, enterococcal resistance to antimicrobials has greatly increased. Furthermore, these chemicals include several side effects on the patients. Since no reports are available of the bacteriophages' effects on eukaryotic cells, they can be good solutions for multidrug-resistant bacterial problems. Therefore, the major aim of this study was to isolate bacteriophages from wastewaters on clinical antibiotic-resistant enterococci.
Materials and Methods: Clinical bacteria were isolated, then enterococcal isolates were identified using different methods. The antibiotic resistance scheme of the enterococcal isolates was assessed. The bacterial isolates were exposed to wastewater samples containing potential bacteriophages. Technically, isolated bacteriophages were studied by electron microscopy.
Results: Isolated bacteria were verified as Enterococcus faecium. Results showed that bacteriophages could easily be isolated from wastewater sources. The isolated bacteriophages were effective on E. faecium as well as Streptococcus dysgalactiae. Furthermore, these bacteriophages were challenged with five other bacteria (ATCC) with no visible effects. In general, the isolated bacteriophages belonged to the Myoviridae, Siphoviridae, and Inoviridae families.
Conclusion: Further studies on bacteriophages and their efficacy on enterococcal strains could increase the treatment possibility of enterococcal infections. Due to these bacteriophages' effects on Streptococcus strains, bacteriophages may be used to treat streptococcal infections as well.
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12. 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.
13. Rose T, Verbeken G, Vos DD, Merabishvili M, Vaneechoutte M, Lavigne R, et al. Experimental phage therapy of burn wound infection: difficult first steps. Int J Burns Trauma 2014;4:66-73.
14. Loc-Carrillo C, Abedon ST. Pros and cons of phage therapy. Bacteriophage 2011;1:111-114.
15. Mazaheri Nezhad Fard R, Barton MD, Heuzenroeder MW. Novel bacteriophages in Enterococcus spp. Curr Microbiol 2010;60:400-406.
16. Larsson MC, Karlsson E, Woksepp H, Frölander K, Mårtensson A, Rashed F, et al. Rapid identification of pneumococci, enterococci, beta-haemolytic streptococci and S. aureus from positive blood cultures enabling early reports. BMC Infect Dis 2014;14:146.
17. Adeniji OO, Sibanda T, Okoh AI. Recreational water quality status of the Kidd's beach as determined by its physicochemical and bacteriological quality parameters. Heliyon 2019;5(6):e01893.
18. Mohanty S, Jose S, Singhal R, Sood S, Dhawan B, Das BK, et al. Species prevalence and antimicrobial susceptibility of enterococci isolated in a tertiary care hospital of north India. Southeast Asian J Trop Med Public Health 2005;36:962-965.
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21. Chatterjee A, Willett JLE, Nguyen UT, Monogue B, Palmer KL, Dunny GM, et al. Parallel genomics uncover novel enterococcal-bacteriophage interactions. mBio 2020;11(2):e03120-19.
22. Asadollahi P, Razavi S, Asadollahi K, Pourshafie MR, Talebi M. Rise of antibiotic resistance in clinical enterococcal isolates during 2001–2016 in Iran: a review. New Microbes New Infect 2018;26:92-99.
23. Aghaee BL, Mirzaei MK, Alikhani MY, Mojtahedi A. Sewage and sewage-contaminated environments are the most prominent sources to isolate phages against Pseudomonas aeruginosa. BMC Microbiol 2021;21:132.
24. Maal KB, Delfan AS, Salmanizadeh S. Isolation and identification of two novel Escherichia coli bacteriophages and their application in wastewater treatment and coliform's phage therapy. Jundishapur J Microbiol 2015;8(3):e14945.
25. Menon ND, Kumar MS, Satheesh Babu TG, Bose S, Vijayakumar G, Baswe M, et al. A novel N4-Like bacteriophage isolated from a wastewater source in South India with activity against several multidrug-resistant clinical Pseudomonas aeruginosaisolates. mSphere 2021;6(1):e01215-20.
26. Arredondo-Alonso S, Top J, McNally A, Puranen S, Pesonen M, Pensar J, et al. Plasmids shaped the recent emergence of the major nosocomial pathogen Enterococcus faecium. mBio 2020;11(1):e03284-19.
27. Bhardwaj SB, Mehta M, Sood S, Sharma J. Isolation of a novel phage and targeting biofilms of drug-resistant oral enterococci. J Glob Infect Dis 2020;12:11-15.
28. Horiuchi T, Sakka M, Hayashi A, Shimada T, Kimura T, Sakka K. Complete genome sequence of bacteriophage BC-611 specifically infecting Enterococcus faecalis strain NP-10011. J Virol 2012;86:9538-9539.
29. Wandro S, Oliver A, Gallagher T, Weihe C, England W, Martiny JBH, et al. Predictable molecular adaptation of coevolving Enterococcus faecium and lytic phage EfV12-phi1. Front Microbiol 2019;9:3192.
30. Kowalska JD, Kazimierczak J, Sowinska PM, Wojcik EA, Siwicki AK, Dastych J. Growing trend of fighting infections in aquaculture environment—opportunities and challenges of phage therapy. Antibiotics (Basel) 2020;9:301.
31. Romero-Calle D, GuimarãesBenevides R, Góes-Neto A, Billington C. Bacteriophages as alternatives to antibiotics in clinical care. Antibiotics (Basel) 2019;8:138.
2. Arias CA, Murray BE. The rise of the Enterococcus: beyond vancomycin resistance. Nat Rev Microbiol 2012;10:266-278.
3. Sievert DM, Ricks P, Edwards JR, Schneider A, Patel J, Srinivasan A, et al. Antimicrobial-resistant pathogens associated with healthcare-associated infections: summary of data reported to the National Healthcare Safety Network at the Centers for Disease Control and Prevention, 2009–2010. Infect Control Hosp Epidemiol 2013;34:1-4.
4. Harwood VJ, Delahoya NC, Ulrich RM, Kramer MF, Whitlock JE, Garey JR, et al. Molecular confirmation of Enterococcus faecalis and E. faecium from clinical, faecal and environmental sources. Lett Appl Microbiol 2004;38:476-482.
5. Domig KJ, Mayer HK, Kneifel W. Methods used for the isolation, enumeration, characterization and identification of Enterococcus spp.: 2. pheno and genotypic criteria. Int J Food Microbiol 2003;88:165-188.
6. Bortolaia V, Guardabassi L (2015). Zoonotic transmission of antimicrobial resistant enterococci: a threat to public health or an overemphasisedrisk? In: Zoonoses-infections affecting humans and animals. Ed, A Sing. Springer, 1st ed. Dordrecht, Netherlands, pp. 407-431.
7. Kataoka Y, Umino Y, Ochi H, Harada K, Sawada T. Antimicrobial susceptibility of enterococcal species isolated from antibiotic-treated dogs and cats. J Vet Med Sci 2014;76:1399-1402.
8. Ghosh A, Dowd SE, Zurek L. Dogs leaving the ICU carry a very large multi-drug resistant enterococcal population with capacity for biofilm formation and horizontal gene transfer. PLoS One 2011;6(7):e22451.
9. Gilmore MS, Lebreton F, van Schaik W. Genomic transition of enterococci from gut commensals to leading causes of multidrug-resistant hospital infection in the antibiotic era. Curr Opin Microbiol 2013;16:10-16.
10. Chang S, Sievert DM, Hageman JC, Boulton ML, Tenover FC, Downes FP, et al. Infection with vancomycin-resistant Staphylococcus aureus containing the vanA resistance gene. N Engl J Med 2003;348:1342-1347.
11. Clokie MR, Millard AD, Letarov AV, Heaphy S. Phages in nature. Bacteriophage 2011;1:31-45.
12. 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.
13. Rose T, Verbeken G, Vos DD, Merabishvili M, Vaneechoutte M, Lavigne R, et al. Experimental phage therapy of burn wound infection: difficult first steps. Int J Burns Trauma 2014;4:66-73.
14. Loc-Carrillo C, Abedon ST. Pros and cons of phage therapy. Bacteriophage 2011;1:111-114.
15. Mazaheri Nezhad Fard R, Barton MD, Heuzenroeder MW. Novel bacteriophages in Enterococcus spp. Curr Microbiol 2010;60:400-406.
16. Larsson MC, Karlsson E, Woksepp H, Frölander K, Mårtensson A, Rashed F, et al. Rapid identification of pneumococci, enterococci, beta-haemolytic streptococci and S. aureus from positive blood cultures enabling early reports. BMC Infect Dis 2014;14:146.
17. Adeniji OO, Sibanda T, Okoh AI. Recreational water quality status of the Kidd's beach as determined by its physicochemical and bacteriological quality parameters. Heliyon 2019;5(6):e01893.
18. Mohanty S, Jose S, Singhal R, Sood S, Dhawan B, Das BK, et al. Species prevalence and antimicrobial susceptibility of enterococci isolated in a tertiary care hospital of north India. Southeast Asian J Trop Med Public Health 2005;36:962-965.
19. Karna A, Baral R, Khanal B. Characterization of clinical isolates of enterococci with special reference to glycopeptide susceptibility at a tertiary care center of eastern Nepal. Int J Microbiol 2019;2019:7936156.
20. Li X, Xing J, Li B, Wang P, Liu J. Use of tuf as a target for sequence-based identification of gram-positive cocci of the genus enterococcus, streptococcus, coagulase-negative staphylococcus, and lactococcus. Ann Clin Microbiol Antimicrob 2012;11:31.
21. Chatterjee A, Willett JLE, Nguyen UT, Monogue B, Palmer KL, Dunny GM, et al. Parallel genomics uncover novel enterococcal-bacteriophage interactions. mBio 2020;11(2):e03120-19.
22. Asadollahi P, Razavi S, Asadollahi K, Pourshafie MR, Talebi M. Rise of antibiotic resistance in clinical enterococcal isolates during 2001–2016 in Iran: a review. New Microbes New Infect 2018;26:92-99.
23. Aghaee BL, Mirzaei MK, Alikhani MY, Mojtahedi A. Sewage and sewage-contaminated environments are the most prominent sources to isolate phages against Pseudomonas aeruginosa. BMC Microbiol 2021;21:132.
24. Maal KB, Delfan AS, Salmanizadeh S. Isolation and identification of two novel Escherichia coli bacteriophages and their application in wastewater treatment and coliform's phage therapy. Jundishapur J Microbiol 2015;8(3):e14945.
25. Menon ND, Kumar MS, Satheesh Babu TG, Bose S, Vijayakumar G, Baswe M, et al. A novel N4-Like bacteriophage isolated from a wastewater source in South India with activity against several multidrug-resistant clinical Pseudomonas aeruginosaisolates. mSphere 2021;6(1):e01215-20.
26. Arredondo-Alonso S, Top J, McNally A, Puranen S, Pesonen M, Pensar J, et al. Plasmids shaped the recent emergence of the major nosocomial pathogen Enterococcus faecium. mBio 2020;11(1):e03284-19.
27. Bhardwaj SB, Mehta M, Sood S, Sharma J. Isolation of a novel phage and targeting biofilms of drug-resistant oral enterococci. J Glob Infect Dis 2020;12:11-15.
28. Horiuchi T, Sakka M, Hayashi A, Shimada T, Kimura T, Sakka K. Complete genome sequence of bacteriophage BC-611 specifically infecting Enterococcus faecalis strain NP-10011. J Virol 2012;86:9538-9539.
29. Wandro S, Oliver A, Gallagher T, Weihe C, England W, Martiny JBH, et al. Predictable molecular adaptation of coevolving Enterococcus faecium and lytic phage EfV12-phi1. Front Microbiol 2019;9:3192.
30. Kowalska JD, Kazimierczak J, Sowinska PM, Wojcik EA, Siwicki AK, Dastych J. Growing trend of fighting infections in aquaculture environment—opportunities and challenges of phage therapy. Antibiotics (Basel) 2020;9:301.
31. Romero-Calle D, GuimarãesBenevides R, Góes-Neto A, Billington C. Bacteriophages as alternatives to antibiotics in clinical care. Antibiotics (Basel) 2019;8:138.
| Files | ||
| Issue | Vol 13 No 5 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v13i5.7434 | |
| Keywords | ||
| Enterococcus; Bacteriophages; Antibiotics; Wastewaters | ||
| Rights and permissions | |
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Elahi Y, Nowroozi J, Mazaheri Nezhad Fard R. Isolation and characterization of bacteriophages from wastewater sources on Enterococcus spp. isolated from clinical samples. Iran J Microbiol. 2021;13(5):671-677.
