This poster shows the results of an analysis of freshwater biodiversity significance in the Lake Ontario and Upper St. Lawrence River watershed. A Biodiversity Significance index was calculated using data on fish and mussels present in watersheds that are tributary to Lake Ontario and Upper St. Lawrence River. Methods were modified from Staton and Mandrak (2006) who conducted a similar analysis for Great Lakes tributary watersheds in Ontario. These methods were modified and applied to watersheds in New York to create a binational assessment of fish and mussel diversity in Lake Ontario and Upper St. Lawrence tributaries. Data were combined from several sources to create a list of native fish and mussel species known to occur in each tertiary watershed (Ontario) and 8-digit HUC watershed (New York).