Objective: To isolate and make a comparative study of marine sediments actinomycetes from Pulicat estuary, Muttukadu estuary and Ennore estuary, TamilNadu, India. Methods: A unique selective enrichment procedure has resulted in the isolation and identification a total of 304 actinomycetes colonies which were isolated from different stations of marine soil sediments in Pulicat estuary, Muttukadu estuary and Ennore estuary, TamilNadu, India. Results: Among them, 277 isolates were morphologically distinct on the basis of spore mass colour, aerial and substrate mycelium formation and production of diffusible pigment. The majority (60%; 162 isolates) were assigned to the genus Streptomyces. (35%; 104 isolates) were assigned to the genus Actinopolyspora, (5%; 11 isolates) were assigned to the genus Nocardiodes. Conclusions: The present study concluded that the physiological characteristics of actinomycetes Streptomyces, Actinopolyspora and Nocardiodes varied by available nutrients in the medium and the physical conditions.