This chapter aims to assess the trend and status as well as seasonal and spatial distribution of water quality in the Tonle Sap Lake (TSL) basin based on the observation from 1995 to 2010. Cluster analysis showed that the water quality in TSL and tributaries in both upstream and downstream rivers varied seasonally. Nutrients, suspended solids, chloride, and sodium in TSL show higher concentration in the late dry and early wet seasons than during the late wet and early dry season. A significant increase was detected in several water quality parameters, including major ions, electrical conductivity, and nutrients in TSL and the tributaries, indicating the water quality degradation caused by increasing anthropogenic stress over the period. Consequently, water quality in TSL at least until 2010 was not compliant with the recommended guidelines regarding human health, aquatic life, and eutrophication. This was the case especially during the late dry season (i.e., March to April) and the early wet season (i.e., May to June) possibly due to the arrival of early monsoon rain (e.g., runoff), low water depth, stagnant water, high temperature, and increasing human activities (e.g., fisheries).