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Contamination of water systems can not only entail high risks to human health but can also result in economic losses due to closure of beaches and shellfish harvesting areas. Understanding the origin of fecal pollution at locations where shellfish are grown is essential in assessing associated health risks—as well as the determining actions necessary to remedy the problem. The aim of this work is to identify the species-specific source(s) of fecal contamination impacting waters overlying the shellfisheries in the Blackwater Estuary, East Anglia, UK. Over a twelve-month period, water samples were taken from above the oysters and from a variety of upstream points considered to be likely sources of fecal microorganism, together with oyster samples, and the number of fecal streptococci and E. coli were determined. Transition from low to high tide significantly decreased the concentration of fecal streptococci in waters overlying the oyster beds, indicative of a freshwater input of fecal pollution in oyster bed waters. In 12 months, the number of E. coli remained constant throughout, while fecal streptococci numbers were generally higher in the winter months. Analyses of upstream samples identified a sewage outfall to be the main source of E. coli to the oyster beds, with additional fecal streptococci from agricultural sources. The findings may assist in developing approaches for assessing the risks to shellfishery industries of various fecal inputs into an estuary, which could then help local governmental authorities address the problem.