The lack of Bd detection in previous field surveys of wild amphibians and these newly reported Bd-positive animals suggest the presence of Bd in Madagascar is a recent phenomenon and not yet widespread. Bd-related die-offs have not been documented and infection prevalence is expected to still be extremely low in wild populations, if currently affected. The conditions necessary to result in Bd establishment in amphibian populations Echinacoside following exposure are poorly understood. Infection with as little as one Bd zoospore can result in chytridiomycosis, and affected amphibians have been observed to release 68 infectious Bd particles per minute when in an aquatic environment. Ganoderic-acid-F Therefore, the detection of Bd in H. betsileo and H. alboguttatus is especially concerning because these species breed in both permanent and temporary water bodies and an outbreak in wild populations may both promote extended environmental persistence and facilitate indirect transmission to nearby aquatic species, increasing the opportunity for pathogen establishment. The spread of Bd can occur rapidly following introduction to a naive region, estimated as much as 25�C282 km/y, and the data presented herein provides impetus to quickly reevaluate the presence of Bd in Madagascar. The confirmation of Bd in amphibians exported from Madagascar presents an opportunity to intervene prior to the first confirmed outbreak in wild populations �C an outbreak with potentially irreparable ecological consequences. It is no longer questionable whether or not Bd will become introduced to Madagascar; it is now a tangible threat. Survival of the country’s amphibians now requires an efficient network of proactive surveillance and rapid response to quickly identify additional introduction events and minimize exposure to wild populations, because pathogen eradication is considered implausible following establishment. The provenance of Bd detected in this investigation remains an enigma, especially considering the absence of commercial amphibian trade into Madagascar, suggesting a more insidious mechanism is responsible for the introduction. Accordingly, Bd may continue to arrive in Madagascar and creep closer towards establishment until the true introduction pathway is identified, targeted and controlled. Early detection now provides the opportunity to interrupt pathogen establishment, but if not acted upon with haste, disease-associated ecological decline in Madagascar may soon become inescapable. Following regulatory clearance for importation into the USA, the shipment was collected from the airport and immediately transported to a small greenhouse specifically constructed to provide a controlled area for receipt and sampling of these animals. This structure had no previous exposure to amphibians and all interior surfaces were first washed with a 10% bleach solution to minimize any potential risk of domestic Bd contamination. The shipment remained sealed for the duration of transport from Madagascar to the USA, and was not opened until first secured inside this sampling location to further prevent opportunities for contamination. Amphibians arrived inside two wooden crates insulated with 1/ 40 Styrofoam.