Ami River is a tributary of Rapti River, and this study was carried out along serpentine length of Ami River, which travels 126 Kmlength at Siddharth Nagar, Sant Kabir Nagar, and Gorakhpur District. To assess the water quality status of Ami River water and ground water which is adjoining to river basin and selected five stations along length of Ami River and to observed the monthly variation of surface and ground water. The surface water samples collected from Mid-stream of Ami River and Ground water samples collected from both side of mid-stream to 1 Km catchment area of Ami River at five stations such as S1 (Kore Kutwa), S2 (Khalilabad), S3 (Adilapar), S4 (Unwal-Jarlahi), S5 (Sohagaura) see Figure 1. And also another ground water sample (Control Point) collected away from catchment area of Ami River. The control point which is not affected from any water bodies i.e. River, Lakes, Ponds, etc. The observed data of Surface and Ground Water compare with IS 10500-2012 and Control Point to interpreted result of water pollution due industrial and domestic discharge into Ami River. The study was carried out during January 2016 to February 2016 and all collected samples subjected to test for Physico-chemical and biological analysis. The Ami River water quality at KoreKutwa indicates same as ground water quality, but without treatment cannot use for any purposes. Ami River water and ground water lose their natural quality is observed at Khalilabad, where paper mill effluents discharge and Adilapar where GIDA effluents discharge into Ami River. The fury of Industrial pollution is also realized at downstream stations such as Unwal-Jarlahi and Sohagaura. The ground water quality of these locations reached at very worst condition. B.O.D. data indicates that the ground water facing high organic load due to river pollution and pollution is beyond limit that river never adopt self-purification capacity, see Figure 4. The biological analysis of Ami River and ground water with respect to total coliform/100ml at all five stations has reveal that river water is unfit for direct use of any purposes and ground water is unhealthy for drinking and other domestic use, see Figure 4. The other parameters such as pH, Turbidity, Hardness (Figure 2), Chloride, Alkalinity, Acidity (Figure 3), and Total Dissolved Solids (Figure 4) also confirmed that high deterioration in Ami River water and ground water quality. With view of some parameter like Hardness, Chloride, and TDS has confirm that ground water is more polluted than river water, at some ground water locations like Khalilabad, Unwal-Jarlahi, and Sohagaura.