Mosquitoes prefer diverse water bodies for egg laying and larval survival. Present study was performed with an objective to characterize physico-chemical properties and microbial profiling of breeding habitat water bodies of Anopheles subpictus mosquitoes. A field survey was accomplished to check the presence of An. subpictus larvae to record per dip larval density in various breeding habitats throughout the year. Physico-chemical and bacteriological properties in relation to mosquito oviposition were assessed. Dissolved oxygen content, pH and alkalinity were found to have major impacts and ponderosity on the prevalence of An. subpictus larvae. Larval density showed significant positive correlation with dissolved oxygen content of water and significant negative correlation with pH and alkalinity of habitat water. Comparatively higher population (cfu/mL) of Bacillus spp. competent with starch hydrolyzing and nitrate reducing properties were recorded all the breeding habitat water bodies of An. subpictus. Higher amplitude of anopheline larvae was portrayed during monsoon and post-monsoon season in clear water with an inclining trend to high dissolved oxygen content and neutral pH. B. cereus, B. megaterium, B. subtilis and B. tequilensis prevalent in all habitat water bodies were marked as oviposition attractants of gravid An. subpictus mosquitoes. Microbial population played key roles in the modulation of physico-chemical parameters of habitat water with a view to enhance its acceptability by gravid mosquitoes in relation to their oviposition. Better understanding of the interactions along with the control of oviposition attractant bacterial strains from mosquito breeding habitats might contribute to the vector management programme.