A herpetological survey was conducted at Pulau Besar, Melaka from 10 to 12 June 2008, resulting in the discovery of five species of amphibians, nine species of lizards (including one new species of the gekkonid Cyrtodactylus) and one snake species. This report constitutes the first checklist of the herpetofauna of Pulau Besar and the Water Islands Archipelago.