Scientific Literacy is an important goal of biology education. Challenge based learning (CBL) has been defined as a new learning strategy that builds on the problem-based learning practice, in which students work on real- world problems in collaborative teams. CBL can be implemented along with online interaction to help improve students’ communication skills and provide assistance if students are facing difficulties in finding a proper solution to the challenges addressed. This study employed a quasi-experimental pretest-posttest non-equivalent control group design. The research was conducted at Biology Department, Universitas Negeri Malang. Research samples consisted of 31 students learned with challenge based learning (the experimental group), 29 students learned with problem based learning (control group 1), and 31 students learned with the presentation-discussion strategy (control group 2). Research data was collected using a scientific literacy test. Data on the students’ scientific literacy was analyzed using ANCOVA with 5% significance level. The analysis was performed to investigate the difference in scientific literacy of students learned with three learning strategies. Further analysis was conducted using Paired-t test and Cohen-d to examine the significance of the students’ scientific literacy improvement and to determine the size effect of each learning strategy. The results showed that all three teaching methods had a significant effect on the students’ scientific literacy with a large size effect, but the CBL and PBL students’ scores could increase significantly higher than that of the presentation-discussion group of students. The result of the LSD test indicated that CBL and PBL did not differ significantly. Both learning strategies had a similar effect on the students’ scientific literacy.

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