Skip to main content

Isolation, characterization, and genomic analysis of the novel T4-like bacteriophage ΦCJ20

Abstract

Pathogenic Escherichia coli infections have been consistently reported annually. The basic characteristics and genome of the newly isolated ΦCJ20 from swine feces was analyzed. To determine basic characteristics, dotting assays and double-layer agar assays were conducted. Bacteriophage particles were analyzed via transmission electron microscopy. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis was performed to determine the sizes of major structural proteins. The complete genome of the phage was analyzed. Bacteriophage particles were identified as Myoviridae, with a head measuring 110.57 ± 1.89 nm and a contractile tail measuring 107.97 ± 3.20 nm and were found to infect E. coli. Major structural proteins of ΦCJ20 showed two well-pronounced bands of approximately 53.6 and 70.9 kDa. The genome size of ΦCJ20 was 169,884 bp, and 118 of 307 open reading frames were annotated. This study provides a baseline for the development of E. coli infection treatment strategies.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Abbasifar R. Biocontrol of Cronobacter spp. using Bacteriophage in Infant Formula. PhD thesis, The University of Guelph, Ontario, Canada (2013)

  2. Ackermann HW. Frequency of morphological phage descriptions in the year 2000. Archives of Virology. 146: 843-857 (2001)

    CAS  Article  Google Scholar 

  3. Ackermann HW. 5500 Phages examined in the electron microscope. Archives of Virology. 152: 227-243 (2007)

    CAS  Article  Google Scholar 

  4. Al-Gallas N, Bahri O, Bouratbeen A, Ben Haasen A, Ben Aissa R. Prevalence, phenotyping, and molecular epidemiology. American Journal of Tropical Medicine and Hygiene. 77: 571-582 (2007)

    Article  Google Scholar 

  5. Bandara N, Jo J, Ryu S, Kim KP. Bacteriophages BCP1-1 and BCP8-2 require divalent cations for efficient control of Bacillus cereus in fermented foods. Food Microbiology. 31: 9-16 (2012)

    CAS  Article  Google Scholar 

  6. Centers for Disease Control and Prevention (CDC). Outbreak of E. coli Infections Linked to Romaine Lettuce. Available from: https://www.cdc.gov/ecoli/2019/o157h7-11-19/. Accessed Jan. 15, 2020.

  7. Chang Y. Bacteriophage-Derived Endolysins Applied as Potent Biocontrol Agents to Enhance Food Safety. Microorganisms. 8: 724 (2020)

    CAS  Article  Google Scholar 

  8. Chang HW, Kim KH. Comparative genomic analysis of bacteriophage EP23 infecting Shigella sonnei and Escherichia coli. The Journal of Microbiology. 49: 927-34 (2011)

    CAS  Article  Google Scholar 

  9. Chen M, Zhang L, Abdelgader SA, Yu L, Xu J, Yao H, Lu C, Zhang W. Alterations in gp37 Expand the Host Range of a T4-Like Phage. Applied and Environmental Microbiology. 83:e01576-17 (2017)

    PubMed  PubMed Central  Google Scholar 

  10. De Souza GM, Neto E, da Silva AM, Iacia M, Rodrigues MVP, Cataneli Pereira V, Winkelstroter LK. Comparative Study Of Genetic Diversity, Virulence Genotype, Biofilm Formation And Antimicrobial Resistance Of Uropathogenic Escherichia coli (UPEC) Isolated From Nosocomial And Community Acquired Urinary Tract Infections. Infection and Drug Resistance. 12: 3595-3606 (2019)

    Article  Google Scholar 

  11. Duclohier H. Antimicrobial Peptides and Peptaibols, Substitutes for Conventional Antibiotics. Current Pharmaceutical Design. 16: 3212-3223 (2010)

    CAS  Article  Google Scholar 

  12. Fischer S, Kittler S, Klein G, Glunder G. Impact of a Single Phage and a Phage Cocktail Application in Broilers on Reduction of Campylobacter jejuni and Development of Resistance. PLOS ONE. 8: e78543 (2013)

    CAS  Article  Google Scholar 

  13. Goodridge LD. Bacteriophages for managing Shigella in various clinical and non-clinical settings. Bacteriophage 3: e25098 (2013)

    Article  Google Scholar 

  14. Goodridge L, Gallaccio A, Griffiths MW. Morphological, host range, and genetic characterization of two coliphages. Applied and Environmental Microbiology. 69: 5364-71 (2003)

    CAS  Article  Google Scholar 

  15. Jang J, Hur HG, Sadowsky MJ, Byappanahalli MN, Yan T, Ishii S. Environmental Escherichia coli: ecology and public health implications-a review. Journal of Applied Microbiology. 123: 570-581 (2017)

    CAS  Article  Google Scholar 

  16. Kakasis A, Panitsa G. Bacteriophage therapy as an alternative treatment for human infections. A comprehensive review. International Journal of Antimicrobial Agents. 53: 16–21 (2019)

  17. Kaliniene L, Klausa V, Truncaite L. Low-temperature T4-like coliphages vB_EcoM-VR5, vB_EcoM-VR7 and vB_EcoM-VR20. Archives of Virology. 155: 871-880 (2010)

    CAS  Article  Google Scholar 

  18. Kim KH, Chang HW, Nam YD, Roh SW, Bae JW. Phenotypic characterization and genomic analysis of the Shigella sonnei bacteriophage SP18. Journal of Microbiology. 48: 213-222 (2010)

    CAS  Article  Google Scholar 

  19. Kim J, Kim GH, Lee NG, Lee JS, Yoon SS. Whole-Genome Sequencing and Genomic Analysis of a Virulent Bacteriophage Infecting Bacillus cereus. Intervirology. 61: 272-280 (2019)

    Article  Google Scholar 

  20. Kutter E, Sulakvelidze A. 1st ed. Bacteriophages: biology and applications. CRC Press, Boca Raton, Florida, USA. pp. 29–66 (2004)

  21. Lee H, Ku HJ, Lee DH, Kim YT, Shin H, Ryu S, Lee JH. 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. 11: e0168985 (2016)

    Article  Google Scholar 

  22. Lowe TM, Chan PP. tRNAscan-SE On-line: integrating search and context for analysis of transfer RNA genes. Nucleic Acids Research. 44: W54–7 (2016)

  23. Miroshnikov KA, Faizullina NM, Sykilinda NN, Mesyanzhinov VV. Properties of the endolytic transglycosylase encoded by gene 144 of Pseudomonas aeruginosa bacteriophage phiKZ. Biochemistry (Moscow). 71: 300-305 (2006)

    CAS  Article  Google Scholar 

  24. 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. 10: e0118557 (2015)

    Article  Google Scholar 

  25. Muhlen S, Ramming I, Pils MC, Koeppel M, Glaser J, Leong J, Flieger A, Stecher B, Dersch P. Identification of Antibiotics That Diminish Disease in a Murine Model of Enterohemorrhagic Escherichia coli Infection. Antimicrobial Agents and Chemotherapy. 64: e02159-19 (2020)

    Article  Google Scholar 

  26. Nataro JP, Kaper JB. Diarrheagenic Escherichia coli. Clinical Microbiology Reviews. 11: 142-201 (1998)

    CAS  Article  Google Scholar 

  27. Necel A, Bloch S, Nejman-Falenczyk B, Grabski M, Topka G, Dydecka A, Kosznik-Kwasnicka K, Grabowski L, Jurczak-Kurek A, Wolkowicz T, Wegrzyn G, Wegrzyn A. Characterization of a bacteriophage, vB_Eco4M-7, that effectively infects many Escherichia coli O157 strains. Scientific Reports. 10: 3743 (2020)

    CAS  Article  Google Scholar 

  28. Oliveira H, Melo LD, Santos SB, Nobrega FL, Ferreira EC, Cerca N, Azeredo J, Kluskens LD. Molecular aspects and comparative genomics of bacteriophage endolysins. Journal of Virology. 87: 4558-4570 (2013)

    CAS  Article  Google Scholar 

  29. Perera MN, Abuladze T, Li MR, Woolston J, Sulakvelidze A. Bacteriophage cocktail significantly reduces or eliminates Listeria monocytogenes contamination on lettuce, apples, cheese, smoked salmon and frozen foods. Food Microbiology. 52: 42-48 (2015)

    CAS  Article  Google Scholar 

  30. Romero-Calle D, Benevides RG, Goes-Neto A, Billington C. Bacteriophages as Alternatives to Antibiotics in Clinical Care. Antibiotics (Basel). 8: 138 (2019)

  31. Sidjabat HE, Paterson DL. Multidrug-resistant Escherichia coli in Asia: epidemiology and management. Expert Review of Anti-Infective Therapy 13: 575-591 (2015)

    CAS  Article  Google Scholar 

  32. Yang S, Sadekuzzaman M, Ha SD. Treatment with lauric arginate ethyl ester and commercial bacteriophage, alone or in combination, inhibits Listeria monocytogenes in chicken breast tissue. Food Control. 78: 57-63 (2017)

    CAS  Article  Google Scholar 

  33. Zhang C, Li WL, Liu WH, Zou L, Yan C, Lu K, Ren HY. T4-Like Phage Bp7, a Potential Antimicrobial Agent for Controlling Drug-Resistant Escherichia coli in Chickens. Applied and Environmental Microbiology. 79: 5559-5565 (2013)

    CAS  Article  Google Scholar 

  34. Zhang C, Wang Y, Sun H, Ren H. Multiple-site mutations of phage Bp7 endolysin improves its activities against target bacteria. Virologica Sinica. 30: 386-395 (2015)

    CAS  Article  Google Scholar 

Download references

Acknowledgments

This work was supported by research grants from CJ CheilJedang Institute of Biotechnology (Grant No. CD-30-19-01-0002).

Author information

Affiliations

Authors

Corresponding author

Correspondence to Sung-Sik Yoon.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 56 KB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Kim, J., Chae, J.P., Kim, GH. et al. Isolation, characterization, and genomic analysis of the novel T4-like bacteriophage ΦCJ20. Food Sci Biotechnol 30, 735–744 (2021). https://doi.org/10.1007/s10068-021-00906-y

Download citation

Keywords

  • Escherichia coli
  • Bacteriophage
  • Genome annotation
  • Antibiotics substitution
  • Food safety