In the present study, we analysed variations in bacterial community structure along a salinity gradient in a tropical monsoonal estuary (Cochin estuary, CE), on the southwest coast of India, using Illumina nextgeneration sequencing (NGS). Water samples were collected from eight different locations thrice a year, to assess the variability in the bacterial community structure and to determine the physico-chemical factors in uencing the bacterial diversity. Proteobacteria was the most dominant phyla in the estuary followed by Bacteroidetes, Cyanobacteria, Actinobacteria, and Firmicutes. Statistical analysis indicated signi cant variations in bacterial communities between freshwater, mesohaline and euryhaline regions, as well as between the monsoon (wet) and non-monsoon (dry) periods. Non-metric multidimensional scaling (NMDS) analysis demonstrated that the bacterial communities cluster according to different salinity regimes of the estuary. Canonical Correspondence analysis (CCA) showed a clear spatial and temporal variation in the distribution of bacterial communities in the CE. Abundance of Betaproteobacteria was high in the freshwater regions, while Gammaproteobacteria, Alphaproteobacteria and Epsilonproteobactera were more abundant in mesohaline and euryhaline regions of the estuary.Correlagram based on Pearson correlation analysis demonstrated the impact of different physicochemical variables on the distribution of dominant phyla, class and genera. Spatial and temporal variations in bacterial community structure could be due to regional variations in environmental conditions imparted by allochthonous inputs, monsoonal rainfall, and tidal in uence.