This study investigated students’ performance in two objective test formats in evaluating biology academic achievement in secondary schools in Ondo State, Nigeria. It adopted descriptive research of the survey type. One hundred and fifty male(150) and 150 female senior secondary school class two students (SS2) were selected from 10 secondary schools using stratified sampling. Multiple-choice and two-tier multiple-choice biology achievement test formats were the instruments for data collection. Data collected were subjected to mean, standard deviation and t- test statistics. The results of three hypotheses tested at 0.05 alpha level showed that: significant difference existed in performance of students in multiple-choice and two-tier multiple–choice biology achievement test formats; significant difference existed between performance of male and female students in multiple–choice test format, but no significant difference exists between male and female performance in two–tier multiple – choice test format. Based on findings of this study, it was recommended that teachers and examining bodies should utilize two-tier multiple-choice test items as supplement to  multiple-choice test to prevent students from guess work and inculcating habit of reading to understand, think deeply and apply learnt principles, concepts, and knowledge of topic contents to solve daily human problems.

Keywords: Multiple-choice, two-tier multiple choice, test formats, evaluating biology achievement.

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