Summary. A regimented electroejaculation protocol (120 electrical stimulations; 10-30 V) was used to collect semen and characterize ejaculate quality from 9 adult, free-ranging African elephants under anaesthesia. Eight of the 9 ejaculates contained high concentrations of progressively motile spermatozoa. The overall mean ejaculate volume, sperm concentration/ml ejaculate, sperm motility, sperm status and ejaculate pH were 93·3 ml, 2408·6 × 106 spermatozoa/ml, 70%, 3·9 and 7·4, respectively. A high percentage (mean 77·5%) of spermatozoa within each ejaculate was morphologically normal. Of the aberrant spermatozoa, 72% had a cytoplasmic droplet defect. When sperm viability was tested in vitro at 37°C, sperm motility rating declined by at least half of the initial assessment within 3·5 h of semen collection. Generally, spermatozoa maintained motility in vitro for < 6 h. Serum testosterone ranged from 1·4 to 8·2 ng/ml in 4 males evaluated in the morning (07:30–08:00 h). In 4 of the 5 bulls assessed in the afternoon (15:00–18:00 h), testosterone levels were <0·9 ng/ml. The remaining bull, evaluated at 16:00 h, had exceptionally high testosterone concentrations (peak 25·6 ng/ml) and a preputial discharge potentially indicative of 'musth'. The present study demonstrates that high quality semen can be collected consistently from the African elephant and that striking differences exist in serum testosterone amongst free-ranging males which may be due, in part, to a diurnal rhythm.