• Chelonoidis;
  • captive and wild-born lineage identification;
  • Galapagos;
  • tortoise;
  • microsatellites

Genetic tools have become a critical complement to traditional approaches for meeting short- and long-term goals of ex situ conservation programs. The San Diego Zoo (SDZ) harbors a collection of wild-born and captive-born Galápagos giant tortoises (n = 22) of uncertain species designation and unknown genealogical relationships. Here, we used mitochondrial DNA haplotypic data and nuclear microsatellite genotypic data to identify the evolutionary lineage of wild-born and captive-born tortoises of unknown ancestry, to infer levels of relatedness among founders and captive-born tortoises, and assess putative pedigree relationships assigned by the SDZ studbook. Assignment tests revealed that 12 wild-born and five captive-born tortoises represent five different species from Isabela Island and one species from Santa Cruz Island, only five of which were consistent with current studbook designations. Three wild-born and one captive-born tortoise were of mixed ancestry. In addition, kinship analyses revealed two significant first-order relationship pairs between wild-born and captive-born tortoises, four second-order relationships (half-sibling) between wild-born and captive tortoises (full-sibs or parent-offspring), and one second-order relationship between two captive-born tortoises. Of particular note, we also reconstructed a first-order relationship between two wild-born individuals, violating the founder assumption. Overall, our results contribute to a worldwide effort in identifying genetically important Galápagos tortoises currently in captivity while revealing closely related founders, reconstructing genealogical relationships, and providing detailed management recommendations for the SDZ tortoises. Zoo Biol 31:107;–120, 2012. © 2011 Wiley Periodicals, Inc.