Investigation into antemortem semen collection methods in reptiles is required to counteract anthropogenic induced reptile biodiversity loss and to further develop assisted reproductive technologies in reptiles. The purpose of this study was to evaluate whether repeated electroejaculation under anesthesia could be done safely in the veiled chameleon (Chamaeleo calyptratus). Electroejaculation under anesthesia (alfaxalone 15 mg/kg intravenously) was performed once weekly over a 4-wk period in 12 Chamaeleo calyptratus. External vent appearance and cloacoscopy was performed prior to and after electroejaculation to assess the effect of the electroejaculation on the external and internal anatomy. Hematological and chemistry values were evaluated prior to the study and afterwards. Semen collection was successful in 50% of the electroejaculation events. Cloacoscopy images scores were significantly higher between the pre- and postelectroejaculation events for each week. No significant difference was observed between study week 1 prior to electroejaculation and study week 4 after electroejaculation in vent external morphology (P = 0.157), and internal cloacal scoring (P = 0.058). A significant increase in the white blood cell counts (P = 0.015), absolute heterophil counts (P = 0.02), absolute monocyte counts (P = 0.003), and albumin (P = 0.038) was observed between the pre- and postejaculation sampling periods. Aspartate aminotransferase (P = 0.001) and potassium (P = 0.036) decreased significantly between the pre- and postsampling periods. Vent image scores were significantly higher (P = 0.046) between the pre- and postsampling images collected during week 1; however, there were no significant differences noted between pre- and postsample images for weeks 2 (P= 0.063), 3(P= 0.083), or 4 (P= 0.083). Serial electroejaculation procedures under alfaxalone anesthesia can be performed safely in veiled chameleons. Although changes were observed in the complete blood count and biochemistry results, these changes were determined to not be clinically significant. The results of this study demonstrate that antemortem assisted reproductive techniques such as electroejaculation can be safely used repeatedly.