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February, 2018 >>

Use of lytic bacteriophages in controlling multi drug resistant staphylococcus aureus

Madiha Gohar, Hafiz Arslan Ali, Muhammad Qamar Saeed  ( Institute of Pure and Applied Biology, Microbiology Division, Bahauddin Zakariya University, Multan, Pakistan. )

Abstract

Madam, due to increasing multidrug resistant (MDR) bacterial pathogens, clinicians are facing limitations in choice of antibiotics to treat infections. Global reports project mortality rates due to MDR infections to exceed those from cancer until 2050.1,2 Use of lytic bacteriophages offers a promising alternative. Unlike antibiotics, they are very specific to their host which means their systemic use does not harm normal flora. Staphylococcus aureus (S. aureus) is a deadly pathogen causing several different pathologies ranging from topical infections to life-threatening systemic ones such as meningitis.3 There are worldwide efforts to develop credible phage therapies to control S. aureus infections.
4 We are isolating and characterizing bacteriophages specific to S. aureus. Sewage samples were collected from different hospitals and enriched in S. aureus as follows: Samples were centrifuged at 4500 RPM for 10 minutes and filtered in 0.2m filters.  Then, 10 ml filtrate was added in 2x Lauria-Bertani (LB) broth and 0.5 ml log-phase S. aureus and cultured at 37°C for 10 hours. They were centrifuged at 180 RPM and filtered as mentioned above. Procedure was repeated thrice. After final enrichment, filtrate was serially diluted until 10-20. Afterwards, 100ml of each dilution was mixed into 300ml of log-phase S. aureus and incubated at 37°C for 20 minutes. LB medium containing 0.8% agar was mixed in this culture and this mix was spread on LB agar plates for 24 hours. Appearance of visible plaques indicated presence of specific phage. Each plaque represented one type of phage. These plaques were collected using sterile microtips and inserted in 1.5ml tubes containing LB broth with 1% chloroform. Tubes were vortexed to dissolve phages in the medium. They were filtered and stored at -20°C until further use. Each isolated plaque was again enriched as mentioned above and tested in plaque forming assay against MDR S. aureus.
Two of our phage isolates, JPh-11 and MG-23, showed antibacterial activity against broad range of MDR S. aureus isolates. These bacterial isolates were isolated from burn, accidental and surgical wounds. Details of these isolates and phage activity of on them is given in the Table.



Our results indicate the potential of phage in treating skin infections caused by S. aureus. These phages should be tested in animal models of skin infections to document their therapeutic potential.

Disclaimer: None.
Conflict of Interest: None.
Funding Sources: Our study is funded as a Research Grant from Bahauddin Zakaraya University, Multan.

References

1. WHO. Tackling antibiotic resistance from a food safety perspective in Europe. Copenhagen: World Health Organization; 2011. [Online] [Cited 2017 Jul 05]. Available from URL: http://www.euro.who.int/en/publications/abstracts/tackling-antibiotic-resistance-from-a-food-safety-perspective-in-europe.
2. Tong SY, Davis JS, Eichenberger E, Holland TL, Fowler VG. Staphylococcus aureus Infections: Epidemiology, Pathophysiology, Clinical Manifestations, and Management. Clin Microbiol Rev 2015; 28: 603-61
3. Liu GY. Molecular Pathogenesis of Staphylococcus aureus Infection. Pediatr Res 2009; 65: 71-7.
4.  Zhang Q, Xing S, Sun Q, Pei G, Cheng S, Liu Y, et al. Characterization and complete genome sequence analysis of a novel virulent Siphoviridae phage against Staphylococcus aureus isolated from bovine mastitis in Xinjiang, China. Virus Genes 2017; 53: 464-76.


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