Cook Kimberly J, Voyles Jamie, Kenny Heather V, Pope Karen L, Piovia-Scott Jonah
School of Biological Sciences, Washington State University, Vancouver, Vancouver, WA 98686, USA.
Dis Aquat Organ. 2018 Jul 4;129(2):159-164. doi: 10.3354/dao03238.
The ability to isolate and purify pathogens is important for the study of infectious disease. A protocol for isolating Batrachochytrium dendrobatidis (Bd), a lethal pathogen of amphibians, has been available for over a decade, but the method relies on sacrificing infected animals. We validated a non-lethal protocol for Bd isolation that uses biopsy punches from toe webbing to collect skin samples from live amphibians in remote field locations. We successfully isolated Bd from the Cascades frog Rana cascadae and found a positive association between Bd infection and probability of Bd growth in culture. Recapture rates of sampled animals suggest that our isolation protocol did not affect frog survival. The ability to collect isolates from live animals will facilitate investigations of the biology of Bd and enhance amphibian conservation efforts.
分离和纯化病原体的能力对于传染病研究至关重要。一种用于分离两栖动物致死病原体——蛙壶菌（Batrachochytrium dendrobatidis，简称Bd）的方案已经存在了十多年，但该方法依赖于处死受感染动物。我们验证了一种用于分离Bd的非致死性方案，该方案使用来自趾蹼的活检打孔器从偏远野外地点的活体两栖动物身上采集皮肤样本。我们成功地从喀斯喀特林蛙（Rana cascadae）中分离出了Bd，并发现Bd感染与培养物中Bd生长的可能性之间存在正相关。采样动物的重新捕获率表明我们的分离方案不会影响青蛙的存活。从活体动物中收集分离株的能力将有助于对Bd生物学的研究，并加强两栖动物的保护工作。
