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This chapter addresses the methodologies for assessing the potential ecological effects of transgenic fish before they actually enter unconfined environments. A strategy for identifying the most important data is outlined, and methods for generating them are presented. The following are also discussed: types of information needed to accurately characterize potential receiving environments for the transgenic fish; phenotypic characteristics of transgenic fish that will influence their interaction with ecosystem processes and components; experimental approaches for assessing transgenic fish phenotypes and their ecological impact; and major sources of scientific uncertainty influencing empirical assessments.