METHODOLOGY
Materials and methods Actinomycetes used in this study were isolated from Dr. Ram Manohar Lohia Hospital (RMLH) and RML Park in Lucknow, U.P India. Actinomycetes were isolated by spread plate technique on actinomycetes isolation agar. After serial dilution method, colonies of Actinomycetes were selected for screening. Isolated colonies were preserved on actinomycetes isolation agar (Glycerol based media) and stored at -20°C. The sub culturing of Actinomycetes was done by streaking method on actinomycetes isolation agar and incubated at 28°C for 4-5 days. Screening of Actinomycetes for Antimicrobial activity: The screening process were completed by two methodsPrimary and Secondary Screening:
In primary screening the antimicrobial activity of pure isolates were determined by perpendicular streak method on actinomycetes isolation agar. The test organisms used were; Staphylococcus aureus, Escherichia coli and Pseudomonas aeuginosa. Secondary screening was performed by agar well diffusion method against the standard test organisms. Characterization of Actinomycetes: The Actinomycetes selected after secondary screening, were characterized by morphological and biochemical methods. Morphological characterizations were done by microscopic method. The microscopic characterization was done by cover slip culture method (Kawato and Sinobu, 1979).