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A compelling reason to conduct learning progression research in ecology is that almost all students will learn about ecology during their formal schooling. Thus, we embarked on a study to investigate how students develop an understanding of the interdependent relationships in ecosystems. We collected data from 807 middle and high school students in a large, diverse, urban school district and analyzed the data using Item Response Theory. The ecosystems assessment items developed through this study are of immediate use in teachers formative assessment practice, and the learning progression can make an impact on how teachers think about how students’ understanding of ecosystems. Thus, we created sound products that can be used immediately in biology classrooms across the country.