Technical note Enriching the captive elephant population genetic pool through artificial insemination with frozen-thawed semen collected in the wild T.B. Hildebrandt a, *, R. Hermes a , J. Saragusty a , R. Potier b , H.M. Schwammer c , F. Balfanz d , H. Vielgrader d , B. Baker e , P. Bartels f , F. Göritz a a Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany b Zooparc de Beauval, Cher, France c Tiergarten Schoenbrunn, Vienna, Austria d Tierärztliche Ordination Tiergarten Schoenbrunn, Vienna, Austria e Pittsburgh Zoo and PPG Aquarium, Pittsburgh, Pennsylvania, USA f Wildlife Biological Resource Centre/BioBank SA, National Zoological Gardens of SA, National Research Foundation, Pretoria, Republic of South Africa Received 23 March 2012; received in revised form 6 June 2012; accepted 10 June 2012 Abstract The first successful AI in an elephant was reported in 1998, using fresh semen. Since then almost 40 calves have been produced through AI in both Asian and African elephants worldwide. Following these successes, with the objective of enriching the captive population with genetic material from the wild, we evaluated the possibility of using frozen-thawed semen collected from wild bulls for AI in captivity. Semen, collected from a 36-yr-old wild African savanna elephant (Loxodonta africana) in South Africa was frozen using the directional freezing technique. This frozen-thawed semen was used for four inseminations over two consecutive days, two before and two after ovulation, in a 26-yr-old female African savanna elephant in Austria. Insemination dose of 1200  10 6 cells per AI with 61% motility resulted in pregnancy, which was confirmed through ultrasound examination 75, 110 and 141 days after the AI procedure. This represents the first successful AI using wild bull frozen-thawed semen in elephants. The incorporation of AI with frozen-thawed semen into the assisted reproduction toolbox opens the way to preserve and transport semen between distant individuals in captivity or, as was done in this study, between wild and captive populations, without the need to transport stressed or potentially disease-carrying animals or to remove animals from the wild. In addition, cryopreserved spermatozoa, in combination with AI, are useful methods to extend the reproductive lifespan of individuals beyond their biological lifespan and an important tool for genetic diversity management and phenotype selection in these endangered mammals. © 2012 Elsevier Inc. All rights reserved. Keywords: Genome bank; Reproduction; African elephant; Directional freezing; Cryopreservation; Loxodonta; Elephas 1. Introduction The order Proboscidea contains only one family of living animals—the elephantidae, which includes the genera Loxodonta with two species—African savanna elephant (Loxodonta africana) and African forest ele- phant (Loxodonta cyclotis), and Elephas with a single species—Asian elephant (Elephas maximus). Survival of all three elephant species in their native habitats is threatened, particularly that of the Asian elephant that is classified as an endangered species in the Interna- tional Union for Conservation of Nature and Natural * Corresponding author. Tel.: +49 30 5168 220; fax: +49 30 5126 104. E-mail address: hildebrandt@izw-berlin.de (T.B. Hildebrandt). Available online at www.sciencedirect.com Theriogenology 78 (2012) 1398–1404 www.theriojournal.com 0093-691X/$ – see front matter © 2012 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.theriogenology.2012.06.014