The aim of the present study was to assess the antimicrobial and larvicidal activity of Streptomyces sp.VITPK9, isolated from brine spring habitat of Manipur, India. The isolate was identified by molecular taxonomy and phylogeny of the isolate revealed that it belonged to the genus Streptomyces and designated as Streptomyces sp.VITPK9 (JN689333). Blast search of 16S rDNA sequence (1300m bases) of the isolate with sequences available in the GenBank (NCBI) database showed 97% similarity with Streptomyces pseudogriseolus (X80827). The ethyl acetate (EA) extract of Streptomyces sp.VITPK9 showed highest zone of inhibition (24mm) against Candida albicans and (17 mm) against P. aeruginosa. Larvicidal efficacy was studied and found to be high after 24h exposure to EA extract (1000ppm), the mortality rate was observed as A. subpictus (LC50=831.78, r2=0.930), C. tritaeniorhynchus (LC50=489.21, r2=0.923), C. gelidus (LC50=151.29, r2=0.831), R. microplus (LC50=369.21, r2=0.917) and H. bispinosa (LC50=424.55, r2=0.943). The purification of the active principles responsible for the biological activity is under process.