The morphological and biological characteristics of a virulent PI phage isolated from slaughterhouse sewage in Shiraz, Iran

  • Sepideh Khalatbari-Limaki Department of Food Hygiene and Public Health, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
  • Saeid Hosseinzadeh Mail Department of Food Hygiene and Public Health, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
  • Seyed Shahram Shekarforoush Department of Food Hygiene and Public Health, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
  • Enayat Berizi ORCID Department of Food Hygiene and Quality Control, Nutrition Research Center, School of Nutrition and Food Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
Keywords:
Escherichia coli O157:H7;, Bacteriophage;, Siphoviridae

Abstract

Background and Objectives: Foodborne pathogens are among the serious problems all around the world and thus a novel and natural strategy to control and to inhibit such pathogens is highly demanded nowadays. The aim of this study was to isolate a specific bacteriophage of Escherichia coli O157:H7 from sewage in Fars province, Iran to determine its morphological and antimicrobial activities.
Materials and Methods: In order to isolate the bacteriophage of E. coli O157:H7, 10 samples of slaughterhouse wastewaters were used. Double-Layer Agar method was employed to isolate the bacteriophage. To identify the fine structure of the bacteriophage, electron microscope was employed. Host range and antibacterial activity of the phage was also investigated, in vitro.
Results: The morphological and biological characteristics of a virulent Siphoviridae phage, PI, are reported. It was found that infection of E. coli O157:H7 strains with this specific bacteriophage produce clear plaques. In the one-step growth analysis, it was confirmed that the phage has been characterized with a very short rise period (around 15 min), an average burst size of 193 PFU/cell, high infectivity and potent lytic action. The bacteriolytic activity of PI was also investigated, in vitro. It was also clarified that at the MOI of 100, 10 and 1, the phage rapidly lysed the bacterial cells within 0.5 or 2 h.
Conclusion: These results indicate that the phage PI is a newly discovered phage against E. coli O157:H7 in Iran which may be recommended to use as bio-control purposes.

References

1. Riley LW, Remis RS, Helgerson SD, McGee HB, Wells JG, Davis BR, et al. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med 1983; 308:681-685.
2. Todd W, Dundas S. The management of VTEC O157 infection. Int J Food Microbiol 2001; 66:103-110.
3. Sperandio V, Nguyen Y. Enterohemorrhagic E. coli (EHEC) pathogenesis. Front Cell Infect Microbiol 2012; 2:90.
4. Lee H, Ku HJ, Lee DH, Kim YT, Shin H, Ryu S, et al. Characterization and genomic study of the novel bacteriophage HY01 infecting both Escherichia coli O157: H7 and Shigella flexneri: potential as a biocontrol agent in food. PLoS One 2016; 11(12):e0168985.
5. Sillankorva SM, Oliveira H, Azeredo J. Bacteriophages and their role in food safety. Int J Microbiol 2012; 2012:863945.
6. Carlton RM, Noordman WH, Biswas B, de Meester ED, Loessner MJ. Bacteriophage P100 for control of Listeria monocytogenes in foods: genome sequence, bioinformatic analyses, oral toxicity study, and application. Regul Toxicol Pharmacol 2005; 43:301-312.
7. Amarillas L, Rubí-Rangel L, Chaidez C, González-Robles A, Lightbourn-Rojas L, León-Félix J. Isolation and characterization of phiLLS, a novel phage with potential biocontrol agent against multidrug-resistant Escherichia coli. Front Microbiol 2017; 8:1355.
8. Lu Z, Breidt F. Escherichia coli O157: H7 bacteriophage Φ241 isolated from an industrial cucumber fermentation at high acidity and salinity. Front Microbiol 2015; 6:67.
9. Twort FW. An investigation on the nature of ultra-microscopic viruses. Acta Kravsi 1961;2:37-40.
10. Haq IU, Chaudhry WN, Akhtar MN, Andleeb S, Qadri I. Bacteriophages and their implications on future biotechnology: a review. Virol J 2012; 9:9.
11. Safwat Mohamed D, Farouk Ahmed E, Mohamed Mahmoud A, Abd El-Baky RM, John J. Isolation and evaluation of cocktail phages for the control of multidrug-resistant Escherichia coli serotype O104: H4 and E. coli O157: H7 isolates causing diarrhea. FEMS Microbiol Lett 2018; 365(2). doi: 10.1093/femsle/fnx275.
12. Premarathne J, Thung T, New C, Huat J, Basri DF, Rukayadi Y, et al. Distribution of bacteriophages in food and environment samples. Int Food Res J 2017; 24:888-896.
13. Callaway TR, Edrington TS, Brabban AD, Anderson RC, Rossman ML, Engler MJ, et al. Bacteriophage isolated from feedlot cattle can reduce Escherichia coli O157: H7 populations in ruminant gastrointestinal tracts. Foodborne Pathog Dis 2008; 5:183-191.
14. Sulakvelidze A. Using lytic bacteriophages to eliminate or significantly reduce contamination of food by foodborne bacterial pathogens. J Sci Food Agric 2013; 93:3137-3146.
15. Hagens S, Loessner MJ. Application of bacteriophages for detection and control of foodborne pathogens. Appl Microbiol Biotechnol 2007; 76:513-519.
16. Poullain V, Gandon S, Brockhurst MA, Buckling A, Hochberg ME. The evolution of specificity in evolving and coevolving antagonistic interactions between a bacteria and its phage. Evolution 2008; 62:1-11.
17. Adibi M, Mobasher N, Ghasemi Y, Mohkam M, Mobasher MA. Isolation, purification and identification of E. coli O157 phage for medical purposes. Trends Pharmacol Sci 2017; 3:43-48.
18. Oliveira A, Sillankorva S, Quinta R, Henriques A, Sereno R, Azeredo J. Isolation and characterization of bacteriophages for avian pathogenic E. coli strains. J Appl Microbiol 2009; 106: 1919-1927.
19. Yıldırım Z, Sakin T, Çoban F. Isolation of anti-Escherichia coli O157: H7 bacteriophages and determination of their host ranges. TURJAF 2018; 6:1200-1208.
20. Manohar P, Tamhankar AJ, Lundborg CS, Ramesh N. Isolation, characterization and in vivo efficacy of Escherichia phage myPSH1131. PLoS One 2018; 13(10):e0206278.
21. Mirzaei MK, Nilsson AS. Isolation of phages for phage therapy: a comparison of spot tests and efficiency of plating analyses for determination of host range and efficacy. PLoS One 2015; 10(3):e0118557.
22. Park M, Lee J-H, Shin H, Kim M, Choi J, Kang D-H, et al. Characterization and comparative genomic analysis of a novel bacteriophage, SFP10, simultaneously inhibiting both Salmonella enterica and Escherichia coli O157: H7. Appl Environ Microbiol 2012; 78:58-69.
23. Lee HS, Choi S, Shin H, Lee J-H, Choi SH. Vibrio vulnificus bacteriophage SSP002 as a possible biocontrol agent. Appl Environ Microbiol 2014; 80:515-524.
24. Casey E, Mahony J, Neve H, Noben J-P, Dal Bello F, van Sinderen D. Novel phage group infecting Lactobacillus delbrueckii subsp. lactis, as revealed by genomic and proteomic analysis of bacteriophage Ldl1. Appl Environ Microbiol 2015; 81:1319-1326.
25. Lee J-H, Bai J, Shin H, Kim Y, Park B, Heu S, et al. A novel bacteriophage targeting Cronobacter sakazakii is a potential biocontrol agent in foods. Appl Environ Microbiol 2015; 82:192-201.
26. Parisien A, Allain B, Zhang J, Mandeville R, Lan C. Novel alternatives to antibiotics: bacteriophages, bacterial cell wall hydrolases, and antimicrobial peptides. J Appl Microbiol 2008; 104:1-13.
27. Peng Q, Yuan Y. Characterization of a newly isolated phage infecting pathogenic Escherichia coli and analysis of its mosaic structural genes. Sci Rep 2018; 8:8086.
28. Hyman P, Abedon ST (2010). Bacteriophage host range and bacterial resistance, in Advances in Applied Microbiology. Elsevier; pp: 217-248.
29. Lee Y-D, J Park -H. Characterization and application of phages isolated from sewage for reduction of Escherichia coli O157: H7 IN biofilm. LWT-Food Sci Technol 2015; 60:571-577.
30. Topka G, Bloch S, Nejman-Faleńczyk B, Gąsior T, Jurczak-Kurek A, Necel A, et al. Characterization of bacteriophage vB-EcoS-95, isolated from urban sewage and revealing extremely rapid lytic development. Front Microbiol 2019; 9:3326.
31. Abedon ST, Yin J (2009). Bacteriophage plaques: theory and analysis, in Bacteriophages. Springer. pp: 161-174.
32. Fauquet CM, Mayo MA, Maniloff J, Desselberger U, Ball LA (2005). Virus taxonomy: VIIIth report of the International Committee on Taxonomy of Viruses. Academic Press.
33. Hong Y, Pan Y, Ebner P. Development of bacteriophage treatments to reduce E. coli O157: H7 contamination of beef products and produce. J Anim Sci 2014; 92:1366-1377.
34. Wilson SGS, Parker M, Collier LH (1990). Topley and Wilson's Principles of Bacteriology, Virology and Immunology. Edward Arnold.
35. Drulis-Kawa Z, Majkowska-Skrobek1G, Maciejewska B. Bacteriophages and phage-derived proteins – application approaches. Curr Med Chem 2015; 22: 1757-1773.
36. Raya RR, Varey P, Oot RA, Dyen MR, Callaway TR, Edrington TS, et al. Isolation and characterization of a new T-even bacteriophage, CEV1, and determination of its potential to reduce Escherichia coli O157: H7 levels in sheep. Appl Environ Microbiol 2006; 72:6405-6410.
37. Viscardi M, Perugini AG, Auriemma C, Capuano F, Morabito S, Kim K-P, et al. Isolation and characterisation of two novel coliphages with high potential to control antibiotic-resistant pathogenic Escherichia coli (EHEC and EPEC). Int J Antimicrob Agents 2008; 31:152-157.
38. Viazis S, Akhtar M, Feirtag J, Brabban A, Diez-Gonzalez F. Isolation and characterization of lytic bacteriophages against enterohaemorrhagic Escherichia coli. J Appl Microbiol 2011; 110:1323-1331.
39. Kutter E (2009). Phage host range and efficiency of plating, in Bacteriophages. Springer. pp: 141-149.
40. Litt PK, Jaroni D. Isolation and physiomorphological characterization of Escherichia coli O157: H7-infecting bacteriophages recovered from beef cattle operations. Int J Microbiol 2017; 2017:7013236.
41. Gallet R, Kannoly S, Wang N. Effects of bacteriophage traits on plaque formation. BMC Microbiol 2011; 11:181.
42. Khakhum N, Yordpratum U, Wongratanacheewin R. Bacteriophages and their medical applications. Srinagarind Med J 2010; 25:47-53.
Published
2020-12-16
How to Cite
1.
Khalatbari-Limaki S, Hosseinzadeh S, Shekarforoush SS, Berizi E. The morphological and biological characteristics of a virulent PI phage isolated from slaughterhouse sewage in Shiraz, Iran. Iran J Microbiol. 12(6):616-624.
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