The aim of the present study was to assess the acaricidal and larvicidal property of marine actinobacterial compound (2S,5R,6R)-2-hydroxy-3,5,6- trimethyloctan-4- one extracted and isolated from Streptomyces sp. VITDDK3 tested against the larvae of Rhipicephalus (Boophilus) microplus Canestrini (Acari: Ixodidae), Anopheles subpictus Grassi and Culex quinquefasciatus Say (Diptera: Culicidae). The isolate was taxonomically characterised, identified and designated as Streptomyces sp. VITDDK3. The crude compound was loaded on silica gel column and eluted with chloroform-methanol-water. The purity of the compound isolated was analysed by thin layer chromatography using chloroform and methanol as the solvent system and confirmed by high-performance liquid chromatography. The structure of the purified compound was established from infrared, ultraviolet, 1H-nuclear magnetic resonance (NMR), 13C-NMR and mass spectral data. The chemical shift assignments obtained for the aliphatic compound from 1H-NMR corresponding to the molecular formula C11H22O2. Bioassay-guided fractionation led to the isolation of compound, which was identified as (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one. In the present study, Streptomyces sp. VITDDK3 crude extract and different fractions were tested against the larvae of parasites at a concentration of 1,000 ppm. Those fractions showing 100% mortality in 24 h alone was selected for further column chromatographic separation. The purified compound (2S,5R,6R)-2-hydroxy-3,5,6- trimethyloctan-4-one was tested in the concentrations of 250, 200, 150, 100 and 50 ppm and observed the per cent mortality of 100, 88, 62, 50 and 36 against R. microplus; 100, 100, 87, 62 and 39 against A. subpitcus; and 100, 94, 79, 51 and 33 against C. quinquefasciatus, respectively. The crude extract showed parasitic effects after 24 h of exposure at 1,000 ppm, and parasite mortality was observed against the larvae of R. microplus (LC50=88.74 ppm; r2=0.865) against the larvae of A. subpictus (LC50=162.59 ppm; r2=0.817) and against C. quinquefasciatus (LC 50=120.15 ppm; r2=0.782), respectively. The maximum efficacy was observed in purified marine actinobacterial compound (2S,5R,6R)-2-hydroxy- 3,5,6-trimethyloctan-4-one with LC50 and r2 values against the larvae of R. microplus (94.49 ppm; 0.982) and A. subpictus (69.65 ppm; 0.906) and against C. quinquefasciatus (82.82 ppm; 0.957), respectively. The control (distilled water) showed nil mortality in the concurrent assay. ©Springer-Verlag 2012.