Cholinesterase activity has been investigated in Rotifera Bdelloidea (Philodina roseola, Philodina tubercolata, Rotaria rotatoria and other unidentified species) by histochemical methods and in vivo observations. Parallel histological studies have been carried out. The enzyme specificity was tested by employing different substrates and inhibitors. The effects in vivo of tubocurarin, bungarotoxin and acetylcholine were also observed. Acetylcholinesterase activity is localized in the nervous and muscular tissues, in sensory organs and in all the ciliated cells. Secretory cells (subcerebral, salivary and pedal glands) and gonad cells (nuclei of the syncytial vitellarium and follicular layer, oocytes and eggs) show both acetyl- and butyrylcholinesterase activities. The effects in vivo of cholinesterase inhibitors, as well as those of tubocurarin, bungarotoxin and acetylcholine, are consistent with the histochemical results, indicating a cholinergic system of transmission and acetylcholinesterase, as well as butyrylcholinesterase, activity.