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The purpose of this study was to establish the nature and extent of scientific literacy (SL) themes coverage in Zambian national high school biology curriculum. The three data sources are biology textbooks, biology syllabi, and grade twelve national biology examination papers for a five-year period (2000–2004). These data sources were analyzed using the framework and procedure developed by Chiappetta, Fillman, and Sethna. The framework has four themes of SL namely: Science as a body of knowledge; Science as a way of investigating; science as a way of knowing; and Interaction between science, technology and society. The results show that the biology textbooks and syllabi content objectives emphasized basic knowledge of science while the biology examination papers, and the syllabi aims and assessment objectives emphasized science as a way of knowing. The interaction between science, technology and society theme was the least represented in the biology course. The results also suggest lack of curriculum and instructional validity in biology examinations with respect to the four themes of SL. The inadequate coverage of interaction of science, technology and society theme in the curriculum materials may be a barrier to a better preparation of scientifically literate citizens. Some recommendations have been made.