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It has long been recognized that bats and birds contain less DNA in their genomes than their non-flying relatives. It has been suggested that this relates to the high metabolic demands of powered flight, a notion that is supported by the fact that pterosaurs also appear to have exhibited small genomes. Given the long-standing interest in this question, it is surprising that almost no data have been presented regarding genome size diversity among megabats (family Pteropodidae). The present study provides genome size estimates for 43 species of megabats in an effort to fill this gap and to test the hypothesis that all bats, and not just microbats, possess small genomes. Intriguingly, megabats appear to be even more constrained in terms of genome size than the members of other bat families.