Testing for Batrachochytrium dendrobatidis (Bd)

Samples used to test for the presence of Bd were collected by swabbing at least 25 times on the ventral surfaces and rear foot following Brem et al. (2007). The swabs were stored in sterile Eppendorf vials, which were refrigerated at - 15ºC. Bd detection was carried out by using the PCR approach described by Annis et al. (2004). DNA was extracted from each swab using Prepman Ultra (Applied Biosystems). All sampled were handled with sterile latex gloves maintaining biosecurity measures to avoid Bd spread between sites and individuals (Aguirre & Lampo 2006).