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Characterization of a bacteriophage, isolated from a cow with mastitis, that is lytic against Staphylococcus aureus strains

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Abstract

Methicillin-resistant strains of Staphylococcus aureus (MRSA) are now the most commonly reported antibiotic-resistant bacterium in clinical settings. Therefore, there is an urgent need to develop novel antibacterial agents to control this pathogen. Bacteriophage therapy is a potential alternative treatment for MRSA infections. The objective of this study was characterization of a novel virulent bacteriophage (MSA6) isolated from a cow with mastitis. Electron microscopy showed its resemblance to members of the family Myoviridae, with an isometric head (66 nm) and a long contractile tail (173 nm). The genome of phage MSA6 was tested by pulsed-field gel electrophoresis and estimated to be about 143 kb. It exhibited rapid adsorption (>82% in 5 min), a short latent period (15 min) and a relatively small burst size (23 PFU/cell). Isolated phage was capable of infecting a wide spectrum of staphylococcal strains of both human and bovine origin. The results of this investigation indicate that MSA6 is similar to other bacteriophages belonging to the family Myoviridae (Twort, K, G1, 812) that have been successfully used in bacteriophage therapy.

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References

  1. Ackermann HW (1998) Tailed bacteriophages: the order caudovirales. Adv Virus Res 51:135–201

    Article  PubMed  CAS  Google Scholar 

  2. Becker SC, Dong S, Foster-Frey J, Pritchard DG, Donovan DM (2009) LysK CHAP endopeptidase domain is required for lysis of live staphylococcal cells. FEMS Microbiol Lett 294:52–60

    Article  PubMed  CAS  Google Scholar 

  3. Chang HC, Chen CR, Lin JW, Shen GH, Chang KM, Tseng YH, Weng SF (2005) Isolation and characterization of novel giant Stenotrophomonas maltophilia phage φSMA5. Appl Environ Microbiol 71:1387–1393

    Article  PubMed  CAS  Google Scholar 

  4. Chibani-Chennoufi S, Dillmann ML, Marvin-Guy L, Rami-Shojaei S, Brüssow H (2004) Lactobacillus plantarum bacteriophage LP65: a new member of the SPO1-like genus of the family myoviridae. J Bacteriol 186:7069–7083

    Article  PubMed  CAS  Google Scholar 

  5. Clokie MRJ, Kropinski AM (2009) Bacteriophages: methods and protocols: molecular and applied aspects, vol 2. Humana Press, Totowa, NJ

    Google Scholar 

  6. Eyer L, Pantůček R, Zdráhal Z, Konečná H, Kašpárek P, Růžičková V, Hernychová L, Preisler J, Doškař J (2007) Structural protein analysis of the polyvalent staphylococcal bacteriophage 812. Proteomics 7:64–72

    Article  PubMed  CAS  Google Scholar 

  7. García P, Martínez B, Obeso JM, Lavigne R, Lurz R, Rodríguez A (2009) Functional genomic analysis of two Staphylococcus aureus phages isolated from the dairy environment. Appl Environ Microbiol 75:7663–7673

    Article  PubMed  Google Scholar 

  8. Gill JJ, Sabour PM, Leslie KE, Griffiths MW (2006) Bovine whey proteins inhibit the interaction of Staphylococcus aureus and bacteriophage K. J Appl Microbiol 101:377–386

    Article  PubMed  CAS  Google Scholar 

  9. Gupta R, Prasad Y (2011) Efficacy of polyvalent bacteriophage P-27/HP to control multidrug resistant Staphylococcus aureus associated with human infections. Curr Microbiol 62:255–260

    Article  PubMed  CAS  Google Scholar 

  10. Hoshiba H, Uchiyama J, Kato S, Ujihara T, Muraoka A, Daibata M, Wakiguchi H, Matsuzaki S (2010) Isolation and characterization of a novel Staphylococcus aureus bacteriophage, φMR25, and its therapeutic potential. Arch Virol 155:545–552

    Article  PubMed  CAS  Google Scholar 

  11. Huang LH, Farnet CM, Ehrlich KC, Ehrlich M (1982) Digestion of highly modified bacteriophage DNA by restriction endonucleases. Nucleic Acids Res 10:1579–1591

    Article  PubMed  CAS  Google Scholar 

  12. Klumpp J, Dorscht J, Lurz R, Bielmann R, Wieland M, Zimmer M, Calendar R, Loessner MJ (2008) The terminally redundant, nonpermuted genome of Listeria bacteriophage A511: a model for the SPO1-like Myoviruses of Gram-Positive bacteria. J Bacteriol 190:5753–5765

    Article  PubMed  CAS  Google Scholar 

  13. Lavigne R, Darius P, Summer EJ, Seto D, Mahadevan P, Nilsson AS, Ackermann HW, Kropinski AM (2009) Classification of Myoviridae bacteriophages using protein sequence similarity. BMC Microbiol 9:224. doi:10.1186/1471-2180-9-224

    Article  PubMed  Google Scholar 

  14. Lu Z, Breidt F Jr, Fleming HP, Altermann E, Klaenhammer TR (2003) Isolation and characterization of Lactobacillus plantarum bacteriophage, φJL-1, from a cucumber fermentation. Int J Food Microbiol 84:225–235

    Article  PubMed  CAS  Google Scholar 

  15. Ma YL, Lu CP (2008) Isolation and identification of a bacteriophage capable of infecting Streptococcus suis type 2 strains. Vet Microbiol 132:340–347

    Article  PubMed  CAS  Google Scholar 

  16. Małek W, Wdowiak-Wróbel S, Bartosik M, Konopa G, Narajczyk M (2009) Characterization of phages virulent for Robinia pseudoacacia Rhizobia. Curr Microbiol 59:187–192

    Article  PubMed  Google Scholar 

  17. Merabishvili M, Pirnay JP, Verbeken G, Chanishvili N, Tediashvili M, Lashkhi N, Glonti T, Krylov V, Mast J, Van Parys L, Lavigne R, Volckaert G, Mattheus W, Verween G, De Corte P, Rose T, Jennes S, Zizi M, De Vos D, Vaneechoutte M (2009) Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PLoS ONE 4:e4944. doi:10.1371/journal.pone.0004944

    Article  PubMed  Google Scholar 

  18. Międzyborski R, Fortuna W, Weber-Dąbrowska B, Górski A (2007) Phage therapy of staphylococcal infections (including MRSA) may be less expensive than antibiotic treatment. Postepy Hig Med Dosw 61:461–465

    Google Scholar 

  19. O’Flaherty S, Coffey A, Edwards R, Meaney W, Fitzgerald GF, Ross RP (2004) Genome of staphylococcal phage K: a new lineage of Myoviridae infecting Gram-positive bacteria with a low G+C content. J Bacteriol 186:2862–2871

    Article  PubMed  Google Scholar 

  20. O’Flaherty S, Ross RP, Flynn J, Meaney WJ, Fitzgerald GF, Coffey A (2005) Isolation and characterization of two anti-staphylococcal bacteriophages specific for pathogenic Staphylococcus aureus associated with bovine infections. Lett Appl Microbiol 41:482–486

    Article  PubMed  Google Scholar 

  21. O’Flaherty S, Ross RP, Meaney W, Fitzgerald GF, Elbreki MF, Coffey A (2005) Potential of the polyvalent anti-Staphylococcus bacteriophage K for control of antibiotic-resistant staphylococci from hospitals. Appl Environ Microbiol 71:1836–1842

    Article  PubMed  Google Scholar 

  22. Pantůček R, Rosypalová A, Doškař J, Kailerová J, Růžičková V, Borecká P, Snopková S, Horváth R, Götz F, Rosypal S (1998) The polyvalent staphylococcal phage φ812: its host-range mutants and related phages. Virology 246:241–252

    Article  PubMed  Google Scholar 

  23. Pantůček R, Doškař J, Růžičková V, Kašpárek P, Oráčová E, Kvardová V, Rosypal S (2004) Identification of bacteriophage types and their carriage in Staphylococcus aureus. Arch Virol 149:1689–1703

    Article  PubMed  Google Scholar 

  24. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y

    Google Scholar 

  25. Shao Y, Wang IN (2008) Bacteriophage adsorption rate and optimal lysis time. Genetics 180:471–482

    Article  PubMed  Google Scholar 

  26. Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Anal Chem 68:850–858

    Article  PubMed  CAS  Google Scholar 

  27. Slanetz LW, Jawetz E (1941) Isolation and Characteristics of bacteriophages for staphylococci of bovine mastitis. J Bacteriol 41:447–455

    PubMed  CAS  Google Scholar 

  28. Son JS, Kim EB, Lee SJ, Jun SY, Yoon SJ, Kang SH, Choi YJ (2010) Characterization of Staphylococcus aureus derived from bovine mastitis and isolation of two lytic bacteriophages. J Gen Appl Microbiol 56:347–353

    Article  PubMed  CAS  Google Scholar 

  29. Štyriak I, Pristaš P, Javorský P (2000) Lack of GATC sites in the genome of Streptococcus bovis bacteriophage F4. Res Microbiol 151:285–289

    Article  PubMed  Google Scholar 

  30. Uchiyama J, Rashe M, Maeda Y, Takemura I, Sugihara S, Akechi K, Muraoka A, Wakiguchi H, Matsuzaki S (2007) Isolation and characterization of a novel Enterococcus faecalis bacteriophage φEF24C as a therapeutic candidate. FEMS Microbiol Lett 278:200–206

    Article  Google Scholar 

  31. Yang H, Liang L, Lin S, Jia S (2010) Isolation and characterization of a virulent bacteriophage AB1 of Acinetobacter baumannii. BMC Microbiol 10:131. http://www.biomedcentral.com/1471-2180/10/131

  32. Yoon SS, Barrangou-Poueys R, Breidt F Jr, Fleming HP (2007) Detection and characterization of a lytic Pediococcus bacteriophage from the fermenting cucumber brine. J Microbiol Biotechnol 17:262–270

    PubMed  CAS  Google Scholar 

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Acknowledgments

The electron microscopy studies were performed in the Laboratory of Electron Microscopy, Nencki Institute of Experimental Biology, Warsaw, Poland (Electron microscope JEM 1400, installed within the project sponsored by EU Structural Funds). We wish to thank Prof. E. Trafny from the Military Institute of Hygiene and Epidemiology in Warsaw, Dr. E. Puacz from the Medical University of Lublin, and Dr. J. Rola from the National Veterinary Institute in Pulawy for providing the bacterial strains. This work was supported by grant OR00004208 from the Ministry of Science and Higher Education, Republic of Poland.

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Correspondence to Magdalena Kwiatek.

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Kwiatek, M., Parasion, S., Mizak, L. et al. Characterization of a bacteriophage, isolated from a cow with mastitis, that is lytic against Staphylococcus aureus strains. Arch Virol 157, 225–234 (2012). https://doi.org/10.1007/s00705-011-1160-3

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