This article describes the semen characteristics from different collection methods between captive and confiscated Malayan pangolins, Manis javanica. Semen was collected from 15 pangolins; two captive and 13 confiscated individuals at the mean weight of 9.36 ± 1.94 kg. The three semen collection methods employed were electroejaculation, rectal massage and a combination of both techniques. The semen characteristics (mean ± standard deviation) of the Malayan pangolin are volume (73.75 ± 144.57 µL), pH (7.63 ± 0.53), spermatozoa concentration (997.19 ± 728.98 × 106 /mL), total motility (59.60% ± 30.00%), progressive motility (48.95% ± 30.93%), mass motility (3.50 ± 1.50) and live spermatozoa (80.25% ± 13.45%). There was no significant difference in semen characteristics between the three collection methods. The percentages of live spermatozoa were significantly different, suggesting better samples from captive compared to confiscated animals. However, there was no significant difference in spermatozoa kinetics between the captive and confiscated samples, suggesting the potential of utilizing confiscated individuals for gamete recovery to conserve the genetic pool of pangolins. All three methods of semen collection were successfully performed in pangolins and should be considered; however, electroejaculation remains the most consistent method of obtaining semen from the species.