All efforts were made to minimize the suffering of animals. Bacterial strains and phage used S. aureus ATCC 43300(MRSA) and S. aureus ATCC 29213(MSSA) from ATCC, Mannasse, USA were used in this study. These two strains were used to study the bacterial adherence, invasion and cytotoxicity Repotrectinib on cultured murine epithelial cells. However, S. aureus 43300 was used to establish
the nasal colonisation in BALB/c mice. Clinical isolates of S. aureus were procured from Post-graduate Institute of Medical Education and Research (PGIMER), Chandigarh, India. The strains were isolated from clinical specimens (nasal screening swabs, blood, pus, soft tissue, wound swabs, respiratory samples and body fluids) collected from both in-patient and as out-patient subjects. The strains were identified on the basis of Gram reaction, growth on mannitol salt agar (MSA), catalase activity, SB525334 supplier and coagulase test. Methicillin resistance was determined using cefoxitin disk on Mueller-Hinton agar (Oxoid) followed by determination of MICs of oxacillin for these strains as recommended by Clinical and Laboratory Standards Institute (CLSI) [15]. A total of thirty four MRSA isolates were selected, numbered sequentially as MRSA 01 to MRSA
34 (clearly depicting their source) and stored in glycerol at −80°C. These strains were used for determining the lytic spectrum/host range of the isolated phage. S. aureus specific
selleck kinase inhibitor bacteriophage, MR-10, which had been isolated and characterized in our laboratory was used in the present study [13]. This phage was selected as it showed a broad host range against four standard strains of S. aureus [S. aureus ATCC 43300(MRSA), S. aureus ATCC 29213(MSSA), S. aureus ATCC 25923(MSSA) and S. aureus ATCC 33591(MRSA)] as well as was effective against 32/34 clinical MRSA isolates (data depicting the host range of MR-10 is included in Additional file 1: Table S1). Animals used BALB/c female mice, 4–6 weeks old weighing 20–25 g were used in this study. The animals were obtained from Central Animal House, Panjab University, Chandigarh. The animals were kept in well aerated rooms and given antibiotic free diet (Hindustan Lever, Rolziracetam Mumbai) and water ad libitum. Isolation and culturing of murine nasal epithelial cells (NEC) This was performed according to the method of Grubb et al. [16]. Nasal septum was dissected from five mice and washed with Dulbecco modified Eagles Medium (DMEM) with 100 μg/ml streptomycin. The septum was homogenized and centrifuged at 2000 rpm for 10 min. The nasal tissue was re-suspended in dissociation medium (10 mM HEPES- streptomycin-DMEM) overnight at 4°C. Next day, the tissue suspension was again centrifuged and suspended in isolation media (145 mM NaCl, 4.