To determine the impact of infectious agents on Galapagos tortoises, we conducted health assessments of five species from the islands of Santa Cruz, Isabela, and Española. We performed health evaluations of up to 454 animals and PCR testing for pathogens known to be relevant in other tortoise species. Through this screening, we identified two novel sequences of adenoviruses (ADVntST-1 and ADVntST-2) and four of herpesviruses (HVntST-1, HVntST-2, HVntST-3 and HVntST-4).