Using a large-scale open-channel flume, the swimming ability and behaviour of individual adult European eel Anguilla anguilla and river lamprey Lampetra fluviatilis, species that exhibit anguilliform locomotion, were quantified under complex hydraulic conditions created by a 0·2–0·3 m high under- or overshot weir during four discharge regimes. Fishes were allowed to approach the weirs from both up- and downstream. All fishes passed the undershot weir, independent of discharge and direction of movement, and under high flow (mean ±s.e. 194·63 ± 6·48 l s−1) moved upstream against velocities that ranged between 1·75 and 2·12 m s−1, suggesting greater maximum swimming capability than previously reported. In comparison, passage efficiency during upstream movement was lower for the overshot weir for both L. fluviatilis and A. anguilla. Downstream moving A. anguilla took longer to pass the over- than undershot weir. This study describes a methodology to attain realistic measures of swimming ability and behavioural performance required to develop multispecies fish passage criteria.