Two hundred and twenty seven Actinomycete isolates were screened from soil/compost/manure samples collected from six different places of Nepal (Itahari, Jhapa, Ilam, Kirtipur and Damauli). From these isolates, thirty-five potential Actinomycetes were recovered as antibacterial isolates on the basis of their inhibitory effect against the test organisms in agar medium. The antimicrobial activities were performed by cross streak plate, agar-cup assay, disc diffusion and gradient plate techniques. The test organisms were: E. coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25932), Vibrio cholerae, Bacillus cereus, Bacillus subtilis, Klebsiella sp., Salmonella typhimurium, Salmonella enteritidis, Shigella dysentery, Proteus sp., Micrococcus sp., and Saccharomuces sp.. Most of the active isolates were active against gram-positive bacteria particularly S. aureus . Five out of eleven active isolates belonged to the genera Streptomyces ( S. albovinaceus, S. fulvoviolaceus, S. cylindrosporus, S. libani and S. melanogenes ). three belonged to Micromonospora ( M melanosporea and two of them M. chalcea ), and two belonged to Thermoactinomycetes and one to Nocardia rugosa . The antibiotics recovered from Streptomyces fulvoviolaceus and S. melanogenes were 5.80 mg/100 ml of broth for DHI2 and 4.90 mg/100 ml of broth for CKI2 respectively. Maximum zone of inhibition was 24 mm for S. aureus and 14 mm for E. coli , given by these purified compounds.