Isolation of Phages and Study of their In vitro Efficacy on Staphylococcus aureus Isolates Originating from Bovine Subclinical Mastitis

DOI: 10.18805/IJAR.B-4331    | Article Id: B-4331 | Page : 754-758
Citation :- Isolation of Phages and Study of their In vitro Efficacy on Staphylococcus aureus Isolates Originating from Bovine Subclinical Mastitis.Indian Journal of Animal Research.2022.(56):754-758
A.S. Srujana, J. Sonalika, D.S. Akhila, M.R. Juliet, P. Sheela akhila@nitte.edu.in
Address : Nitte University Centre for Science Education and Research, (Deemed to be University), Deralakatte, Mangaluru-575 018, Karnataka, India. 
Submitted Date : 9-10-2020
Accepted Date : 11-01-2021


Background: India leads the global market in milk production. However, bovine mastitis, which is the mammary gland inflammation in dairy cattle characterized by physical, chemical, bacteriological changes in milk results in commercial losses. Staphylococcus aureus, is the major causative agent. The treatment of mastitis caused by this pathogen is mainly by antibiotics. Emphasizing on the one health concept, phage therapy is an appropriate alternative to antibiotic. The present study was aimed to isolate and characterize bacteriophages against Staphylococcus aureus associated with bovine mastitis.
Methods: Thirty two isolates of S. aureus obtained from the milk of mastitis cows were characterized by phenotypic and genotypic methods. Antibiotic susceptibility of the isolates was carried out by disk diffusion assay. Milk and cow shed wastewater were used for phage isolation. Phages were characterized by host susceptibility and RAPD assay.
Result: Nineteen phages were isolated from the cowshed waste water. All the milk samples showed negative for the presence of phage. The phage 24 (A2) which had the broadest host range, was selected for the CFU drop assay. The phage was able to clear the lawn of S. aureus culture when grown on agar at different time points thus indicating that topical application of this phage would be a potential strategy to control S. aureus infection leading to mastitis. This study provides a basis to continue the exploration of the potential of PSA2 phage as a candidate for the treatment of Staphylococcal mastitis.


Bovine mastitis Phages Staphylococcus aureus


  1. Aguayo-Reyes, A., Quezada-Aguiluz., M., Mella, S., Riedel, G., Opazo-Capurro, A., Bello-Toledo, H., Domínguez, M. and González-Rocha, G. (2018). Molecular basis of methicillin-resistance in Staphylococcus aureus. Revista Chilena de Infectologica. 35(1): 7-14.
  2. Basdew, I.H. and Laing, M.D. (2011). Mini-Review: Biological control of bovine mastitis using bacteriophage therapy. Science against microbial pathogens: communicating current research and technological advances. World Scientific Singapore. 386-93.
  3. Bauer, A.W., Kirby, W.M.M., Sherris, J.C. and Turck, M. (1966). Antibiotic susceptibility testing by a standardized single disk method. Microbiology: A Centenary Perspective. ASM Press Wash. DC USA 40-45.
  4. Ben Said, M., Abbassi, M.S., Bianchini, V., Sghaier, S., Cremonesi, P., Romanò, A., Gualdi, V., Hassen, A. and Luini, M.V. (2016). Genetic characterization and antimicrobial resistance of Staphylococcus aureus isolated from bovine milk in Tunisia. Letters in Applied Microbiology. 63: 473-481. doi: 10.1111/lam.12672.
  5. Breyne, K., Honaker, R.W., Hobbs, Z., Richter, M., Zaczek, M., Spangler, T., Steenbrugge, J., Lu, R., Kinkhabwala, A., Marchon, B., Meyer, E. and Mokres, L. (2017). Efficacy and safety of a bovine-associated Staphylococcus aureus phage cocktail in a murine model of mastitis, Frontiers in Microbiology. 28: | https://doi.org/10.3389/fmicb.2017.02348.
  6. Ganaie, M.Y. Qureschi, S., Kashoo, Z., Wani, S.A., Hussain, M.I., Kumar, R. Maqbool, R., Sikander, P., Banday, M.S., Malla, W.A., Mondal, P. and Khan, R.I.N. (2018). Isolation and characterization of two lytic bacteriophages against Staphylococcus aureus from India: newer therapeutic agents against bovine mastitis. Veterinary Research Communications. 42: 289-295.
  7. Gomes, F. and Henriques, M. (2016). Control of bovine mastitis: Old and recent therapeutic approaches. Current Microbiology. 72: 377-382. doi: 10.1007/s00284-015-0958-8
  8. Gutiérrez, D., Martín Platero, A.M., Rodríguez, A., Martínez Bueno, M., García, P. and Martínez. (2011). Typing of bacteriophages by randomly amplified polymorphic DNA (RAPD) PCR to assess genetic diversity. FEMS Microbiology Letters. https://doi.org/10.1111/j.1574-6968.2011.02342.
  9. Korpysa-Dzirba, W.E. and Osek, J. (2011). Identification of genes encoding classical staphylococcal enterotoxins in Staphylococcus aureus isolated from raw milk. Bulletin of the Veterinary Institute in PULAWY. 55(1): 55-8.
  10. Kwiatek, M., Parasion, S., Mizak, L., Gryko, R., Bartoszcze, M. and Kocik, J. (2012). Characterization of a bacteriophage isolated from a cow with mastitis, that is lytic against Staphylococcus aureus strains. Archives of Virology. 157(2): 225-34.
  11. Labrie, S.J., Samson, J.E. and Moineau, S. (2010). Bacteriophage resistance mechanisms. Nature Reviews Microbiology. 8(5): 317.
  12. Madougou, A.M., Douny, C., Moula, N., Scippo, M.L., Delcenserie, V., Daube, G., Hamani, M. and Korsak, N. (2019). Survey on the presence of antibiotic residues in raw milk samples from six sites of the dairy pool of Niamey, Niger. Veterinary World. 12(12): 1970-1974.
  13. Magro, G., Biffani, S., Minozzi, G., Ehricht, R., Monecke, S., Luini, V. and Piccinini, R. (2017). Virulence genes of S. aureus from dairy cow mastitis and contagiousness risk. Toxins. 9: 195. doi: 10.3390/toxins9060195.
  14. Matsuzaki, S., Rashel, M., Uchiyama, J., Sakurai, S., Ujihara, T., Kuroda, M., Ikeuchi, M., Tani, T., Fujieda, M., Wakiguchi, H. and Imai, S. (2005). Bacteriophage therapy: a revitalized therapy against bacterial infectious diseases. Journal of Infection and Chemotherapy. 1: 11(5): 211-9.
  15. McDougal, L.K., Fosheim, G.E., Nicholson, A., Bulens, S.N., Limbago, B.M., Shearer, J.E., Summers, A.O. and Patel, J.B. (2010). Emergence of resistance among USA300 methicillin resistant Staphylococcus aureus isolates causing invasive disease in the United States. Antimicrobial Agents and Chemotherapy. 54(9): 3804-11.
  16. Sahebekhtiari, N., Nochi, Z., Eslampour, M., Dabiri, H., Bolfion, M., Taherikalani, M., Khoramian, B., Zali, M. and Emaneini, M. (2011). Characterization of Staphylococcus aureus strains isolated from raw milk of bovine subclinical mastitis in Tehran and Mashhad. Acta Microbiologica et Immunologica Hungarica. 58(2): 113- 21.
  17. Shukla, S. and Hirpurkar, S.D. (2011). Recovery status of bacteriophages of different livestock farms of Veterinary College, Adhartal, Jabalpur, India. Veterinary World. 4(3): 117-9.
  18. Su, M.T., Venkatesh, T.V. and Bodmer, R. (1998). Large and small scale preparation of bacteriophage l lysate and DNA. Biotechniques. 25: 44-45.
  19. Wang, J., Zhao, F., Sun, H., Wang, Q., Zhang, C., Liu, W., Zou, L., Pan, Q. and Ren, H. (2019). Isolation and characterization of the Staphylococcus aureus bacteriophage vB_SauS_ SA2. AIMS Microbiology. 5(3): 285-307.

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