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Isolation and characterization of two lytic bacteriophages against Staphylococcus aureus from India: newer therapeutic agents against Bovine mastitis

  • M. Y. Ganaie
  • S. Qureshi
  • Z. Kashoo
  • S. A. Wani
  • M. I. Hussain
  • R. Kumar
  • R. Maqbool
  • P. Sikander
  • M. S. Banday
  • W. A. Malla
  • P. Mondal
  • R. I. N. Khan
Original Article

Abstract

Bovine mastitis causes severe economic losses to dairy farmers. Staphylococcus aureus, is one of the most important pathogen implicated in etiology of clinical and subclinical mastitis in bovines. In view of increasing antimicrobial resistance alternatives to antibiotic therapy are much needed. The present decade has witnessed a renewed interest in phage based therapeutics and diagnostics. The present study, describes isolation and characterization of two lytic phages SAJK-IND and MSP against Staphylococcus aureus having a potential to be used in therapy against mastitis. SAJK-IND and MSP phages belonged to Myoviridae and Podoviridae families, respectively. TEM imaging of the two phages revealed an iscosahedral head. MSP phage has a short non contractile tail. SAJK-IND and MSP have a burst size of 44 ± 3 and 25 ± 5 PFU/ infected cell, respectively. SAJK-IND and MSP phages revealed ̴ 12 and ̴16 proteins, respectively on SDS-PAGE analysis. The lytic activity of the phages was specific for Staphylococcus aureus. SAJK-IND revealed 100% lytic activity against several strains of Staphylococcus aureus isolated from mastitis milk samples whereas, MSP had only 40% lytic activity. SAJK-IND phage genome was sequenced, assembled and deposited in Genbank under accession no MG010123.

Keywords

Staphylococcus aureus Bacteriophages Mastitis Multiplicity of infection 

Notes

Acknowledgements

The authors are thankful to the Centre for Cellular and Molecular biology Platforms, Bengaluru for electron microscopy. Authors would like to acknowledge the facilities utilized under ICAR (NAE), project niche area of excellence in Anaerobic Bacteriology in Division of Veterinary Microbiology & Immunology.

Compliance with ethical standards

Conflict of interests

Authors declare that there are no conflicts of interest.

References

  1. Abishek MR, Mishra A (2010) Purification and activity of phage induced lysin against mastitogenic strains of Staphylococcus aureus. Biotechnol Int 3:18–25Google Scholar
  2. Arndt D, Grant J, Marcu A, Sajed T, Pon A, Liang Y, Wishart DS (2016) PHASTER: a better, faster version of the PHAST phage search tool. Nucleic Acids Res 44(W1):W16–W21.  https://doi.org/10.1093/nar/gkw387 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Boulanger P (2009) Purification of bacteriophages and SDS-PAGE analysis of phage structural proteins from ghost particles. Methods Mol Biol 502:227–238CrossRefGoogle Scholar
  4. Brakstad OG, Aasbakk K, Maeland JA (1992) Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. J Clin Microbiol 30:1654–166.0PubMedPubMedCentralGoogle Scholar
  5. Capparelli R, Nocerino N, Lanzetta R et al (2010) Bacteriophage-resistant Staphylococcus aureus mutant confers broad immunity against staphylococcal infection in mice. PLoS One 5:117–120CrossRefGoogle Scholar
  6. Chandra M, Thakur S, Narang D, Saxena HM (2011) Isolation of a bacteriophage against Salmonella Dublin and determination of its physical resistance under varied in vitro conditions. Afr J Microbiol Res 5:2044–2047Google Scholar
  7. Chang HC, Chen CR, Lin JW (2005) Isolation and characterization of novel giant Stenotrophomonas maltophilia phage phiSMA5. Appl Environ Microbiol 71:1387–1393CrossRefGoogle Scholar
  8. Eyer L, Pantucek R, Zdrahal Z (2007) Structural protein analysis of the polyvalent staphylococcal bacteriophage 812. Proteomics 7:64–72CrossRefGoogle 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 62:255–260CrossRefGoogle Scholar
  10. Hermoso JA, Garcia JL, Garcia P (2007) Taking aim on bacterial pathogens: from phage therapy to enzybiotics. Curr Opin Microbiol 10:461–472CrossRefGoogle Scholar
  11. Kaur S, Harjai K, Chhibber S (2012) Methicillin-resistant Staphylococcus aureus phage plaque size enhancement using sublethal concentrations of antibiotics. Appl Environ Microbiol 78:8227–8233CrossRefGoogle Scholar
  12. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  13. Li L, Zhang Z (2014) Isolation and characterization of a virulent bacteriophage SPW specific for Staphylococcus aureus isolated from bovine mastitis of lactating dairy cattle. Mol Bio Rep.  https://doi.org/10.1007/s11033-014-3457-2 CrossRefGoogle Scholar
  14. Lin SH, Liao YC (2013) CISA: Contig integrator for sequence assembly of bacterial genomes. PLoS One 8:e60843.  https://doi.org/10.1371/journal.pone.0060843 CrossRefPubMedPubMedCentralGoogle Scholar
  15. Lu Z, Breidt JR F, Fleming HP, Altermann E, Klaenhammer TR (2003). Isolation and characterization of a Lactobacillus plantarum bacteriophage, phiJL-1, from a cucumber fermentation. Int J Food Microbiol 84: 225–235.  https://doi.org/10.1016/S0168-1605(03)00111-9 CrossRefGoogle Scholar
  16. McAdam PR, Holmes A, Templeton KE (2011) Adaptive evolution of Staphylococcus aureus during chronic endobronchial infection of a cystic fibrosis patient. PLoS One 6:1–5CrossRefGoogle Scholar
  17. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis A, Martelli GP, Mayo MA, Summers MD (1995) Virus Taxonomy. Sixth Report of The international committee on taxonomy of viruses. Springer-Verlag, Vienna & New YorkGoogle Scholar
  18. O’Flaherty S, Ross RP, Meaney W (2005) Potential of the polyvalent anti-Staphylococcus bacteriophage K for control of antibiotic-resistant staphylococci from hospitals. Appl Environ Microbiol 71:1836–18420CrossRefGoogle Scholar
  19. Owens J, Barton MD (2013) The isolation and characterization of Campylobacter jejuni bacteriophages from free range and indoor poultry. Vet Microbiol 162:144–150CrossRefGoogle Scholar
  20. Petty NK, Evans TJ, Fineran PC (2007) Biotechnological exploitation of bacteriophage research. Trends Biotechnol 25:7–15CrossRefGoogle Scholar
  21. Quinn PJ, Carter ME, Markey B et al (1994) Clinical veterinary microbiology. Wolfe Publishing, LondonGoogle Scholar
  22. Sangha KK, Kumar BVS, Agrawal RKD, Verma R (2014) Proteomic characterization of lytic bacteriophages of Staphylococcus aureus isolated from sewage affluent of India. International Scholarly Research Notices 2014:1–6.  https://doi.org/10.1155/2014/265298 CrossRefGoogle Scholar
  23. Saxena, HM, Raj S (2018). A novel immunotherapy of Brucellosis in cows monitored non invasively through a specific biomarker.  https://doi.org/10.1371/journal.pntd.0006393 CrossRefGoogle Scholar
  24. Synnott AJ, Kuang Y, Kurimoto M (2009) Isolation from sewage influent and characterization of novel Staphylococcus aureus bacteriophages with wide host ranges and potent lytic capabilities. Appl Environ Microbiol 75:4483–4490CrossRefGoogle Scholar
  25. Wang G, Hindler JF, Ward KW et al (2006) Increased vancomycin MICs for Staphylococcus aureus clinical isolates from a university hospital during a 5-year period. J Clin Microbiol 44:3883–3886.  https://doi.org/10.1128/JCM.01388-06 CrossRefPubMedPubMedCentralGoogle Scholar
  26. Zhou Y, Liang Y, Lynch KH, Dennis JJ, Wishart DS (2011) PHAST: a fast phage search tool. Nucleic Acids Res 39:W347–W352CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • M. Y. Ganaie
    • 1
  • S. Qureshi
    • 1
  • Z. Kashoo
    • 1
  • S. A. Wani
    • 1
  • M. I. Hussain
    • 1
  • R. Kumar
    • 2
  • R. Maqbool
    • 1
  • P. Sikander
    • 1
  • M. S. Banday
    • 3
  • W. A. Malla
    • 2
  • P. Mondal
    • 2
  • R. I. N. Khan
    • 2
  1. 1.Divison of Veterinary Microbiology & Immunology, FVSC & A.H, Shuhama (Aulesteng)SKUAST-KashmirSrinagarIndia
  2. 2.Computational Biology & Genomics Facility, Department of Veterinary BiotechnologyICAR-IVRIBareillyIndia
  3. 3.Department of PharmacologyGovernment Medical CollegeSrinagarIndia

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