Induction of sexual reproduction in Brachionus plicatilis (Monogononta, Rotifera) by a density-dependent chemical cue Abstract-Induction of mixis (sexual reproduction) in rotifers of the genus Brachionus is believed to be triggered by a chemical that is released into the water and accumulates at high population densities. However, direct and conclusive evidence for this hypothesis is thus far lacking. In this study, two mass cultures of the rotifer Brachionus plicatilis were monitored as they grew from low to high population densities. Conditioned water was prepared daily from these cultures, and females were exposed in a bioassay, which consisted of juvenile Brachionus, cultured individually in large volumes that would normally suppress mixis. Conditioned water induced mixis in the bioassay at rates comparable to those found in the mass cultures. Both in bioassay and mass cultures, mixis was essentially absent in the beginning of the experiment, when population densities were very low. The first mictic females appeared at densities of 0.1 females ml Ϫ1 , and their proportion increased rapidly as the populations grew to Ͼ1 female ml Ϫ1 . The maximum rates of mixis in the bioassay were highly significant and reached 51% of those observed in the mass cultures. These results strongly support the hypothesis that mixis in Brachionus plicatilis is induced by a density-dependent chemical cue.