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The River Lambourn rises near the village of Lambourn in the chalk of the Berkshire Downs at an altitude of about 152m. It is 26km long and flows in a south-easterly direction to Newbury where it joins the River Kennet. It is part of the Thames river system. There is one important tributary, the Winterbourne Stream, which flows into the Lambourn just upstream of Newbury. The sampling site is located at Bagnor in the town of Newbury at an altitude of about 80m, about 5km above the confluence with the Kennet. The underlying bedrock is chalk, with overlying river gravels.
The catchment is mostly rural, with mixed farming as the main industry, and there are extensive deciduous woodlands on the catchment boundary. Ten kilometres from the source, the river receives input from East Shefford sewage works. Water quality in the Lambourn is good; the river is classified as General Quality Assessment (GQA) biological class 'b' and chemical class 'A'.