Cryogenic protocols have been successful in storing spermatozoa collected from African elephants (Loxodonta africana). However, these same protocols and modifications of these protocols have failed to preserve spermatozoa collected from Asian elephants (Elephas maximus) [Buice et al., 1995, Proceedings of the Association of Zoo Veterinary Technicians, Baltimore, MD; O’Brien et al., 1997, Second International Elephant Research Symposium, Springfield, MO]. Because the success or failure of cryogenic freezing may rely on differences in membrane composition, a postcryogenic comparison of the membrane fatty acid composition of spermatozoa isolated from African and Asian elephants was studied. The spermatozoa of African elephants possessed significantly higher levels of docosahexaenoic acid (22:6, n-3) and docosapentaenoic acid (22:5, n-6) as compared to the spermatozoa of Asian elephants. Meanwhile, the spermatozoa of Asian elephants had higher levels of myristic acid (14:0), arachidonic acid (20:4, n-6), and docosatetraenoic acid (22:4, n-6) as compared to the spermatozoa of African elephants. The most abundant membrane fatty acid was docosahexaenoic acid (22:6, n-3). The percentage of membrane docosahexaenoic acid (22:6, n-3) in the spermatozoa of African elephants was 68.13 ± 0.52 as compared to 42.88 ± 0.87 in the spermatozoa of Asian elephants (t = 31.48, P ≤ 0.0001). Zoo Biol 19:461–473, 2000. © 2000 Wiley-Liss, Inc.