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Studies show that retrieval practices such as homework assignments that are completed during the encoding phase of learning benefit knowledge acquisition and retention. In addition, desirable difficulties, which are strategies that intentionally create a greater challenge during initial learning to enhance encoding and retrieval pathways, also benefit learning long term. Our objective was to determine whether weekly homework questions intended to create desirable difficulties by requiring higher-order cognitive skills (HOCS) benefited students’ long-term retention of physiology concepts compared to questions designed to require lower-order cognitive skills (LOCS). Undergraduate students in a junior-level animal physiology course were presented information during weekly laboratory periods, and then required to complete retrieval practices in the form of online homework assignments 5 d after each lab. Homework questions were formatted per Bloom’s Taxonomy to require HOCS (i.e., level 4 or 5) or LOCS (i.e., level 1 or 2). Information retention was assessed the next week via performance on an in-class quiz and again at semesters’ end via performance on a final practical exam. We observed no differences in performance on the in-class quiz or final practical exam between students randomly assigned to complete homework with HOCS questions compared to LOCS questions. However, students that received homework with HOCS questions had decreased (P < 0.05) performance scores on 9 out of the 11 homework assignments compared to those receiving homework with LOCS questions. These findings indicate that desirable difficulties were not created by our HOCS homework questions because students receiving these more difficult retrieval practices did not achieve equal success on them. As a result, this attempt to create variations in cognitive demand did not enhance retention of knowledge in this study [ABSTRACT FROM AUTHOR]