(McNeil and Brown, 1994). The need for new, safe and effective antimicrobial agent is the major challenge to the pharmaceutical industry now a days, especially with the obvious increase in opportunistic infections in the immune compromised host via and multiple drug resistant strains (Bhatnagar and Kim, 2010). As there is geographic variation in Indian soil type and their contents, hence it is quite likely that the distribution of antibiotic producing actinomycetes is also variable. Therefore, exploration of unexplored ecosystems for actinomycetes is highly necessary for the discovery of novel bioactive metabolites. The objectives were to isolate and screen actinomycetes from different soil samples (Gwalior) for bioactive compound production and in vitro test for inhibition activity of these bioactive compounds against pathogenic bacterial strains.