This study evaluated the effects of habituation and electro-ejaculation techniques on semen quality in merino-type rams in South Africa, aiming to refine the electro-ejaculation method to improve semen quality. Parameters assessed included semen volume, sperm motility, sperm viability, sperm concentration, sperm viscosity, sperm consistency, and sperm abnormalities. Forty mature (six- to eight-tooth) merino-type rams, approximately two and a half to four years old, were randomly assigned to four groups: habituated with a 3-second rest (H3) (n = 10), non-habituated with a 3-second rest (N3) (n = 10), habituated with a 10-second rest (H10) (n = 10), and non-habituated with a 10-second rest (N10) (n = 10). Each group underwent a two-week habituation period to human presence before electro-ejaculation using a commercial device. Analysed semen samples showed no significant differences in semen volume (0.93–1.28 mL), sperm motility (75.2%–83.1%), sperm viability (73.1%–85.5%), sperm concentration score (3.45–3.83), sperm viscosity score (3.5–4.00), or sperm consistency score (3.5–4.0). However, the H3 group had a higher percentage of sperm abnormalities (12.3%) compared to the N3 (6.11%) and H10 (6.7%) groups, indicating that a shorter rest interval may negatively impact semen quality. The study concludes that extending the rest interval to ten seconds can enhance semen parameters.