The present study reports the physiochemical and biological parameters of water and identification of organisms present in Kolavai Lake, Tamil Nadu, India. The lake is heavily polluted due to the dumping of municipal and agriculture wastes from surrounding localities.  Water samples were collected from different places across the lake and examined. Eleven microorganisms were identified by primary and secondary biochemical assays and two bacterial samples were sequenced with 16S rRNA regimen to identify the species. Obtained results were compared with standard values. The new bacterial species identified belonged to taxa Brevundimonas nasdae and Microbacterium trichothecenolyticum which are uncommon in this area.