In this study, semen was obtained by rectal probe electrostimulation (RPE) from the Sulawesi crested black macaque (Macaca nigra). Three experimental series were conducted. First, semen was collected from four animals anesthetized with either tiletamine-zolazepam (Telazol®) or ket-amine-HCl (Vetalar®) (five collections from each animal with each drug). Because of greater muscle relaxation and analgesia, we found tiletaminezolazepam to be an attractive alternative to ketamine-HCl as an anesthetic agent for RPE in M. nigra. Second, semen was collected from another four animals at stimulation frequencies of either 30 Hz or 60 Hz (five collections from each animal at each frequency). There were no significant differences in sperm number, in percentage of sperm with progressive motility, in the current required for sample recovery between tiletamine-zolazepam or ketamine-HCl anesthesia, or between a 30 Hz or 60 Hz stimulation frequency. Third, to check for retrograde sperm loss, the bladders of four animals were emptied, flushed with sterile saline, and then infused with TALP-Hepes medium. After RPE, sperm numbers in the bladder were compared with those in the ejaculate. Although sperm were recovered from the bladder [1.6 (± 0.9) × 10] (mean ± SEM), the numbers were significantly less (P < 0.05) than those in the ejaculate [49 (± 18) × 10]. The percentage of sperm with normal morphology in these samples was high (96.8 ± 1.0%). The average sperm number in the 84 samples collected for this study was 33.8 (± 4.1) × 10. In preliminary experiments, we found that M. nigra sperm will fertilize rhesus monkey oocytes (Macaca mulatta) in vitro. © 1992 Wiley-Liss, Inc.