McMahon Taegan A, Rohr Jason R
University of Tampa, Tampa, FL, USA,
Ecohealth. 2014 Jun;11(2):164-7. doi: 10.1007/s10393-014-0908-0. Epub 2014 Feb 12.
Batrachochytrium dendrobatidis (Bd) has been implicated in hundreds of amphibian declines and is the focus of a vast amount of research. Despite this, there is no reported efficient way to assess Bd viability. Discriminating between live and dead Bd would help determine the dose of live Bd zoospores and whether factors have lethal or sublethal effects on Bd. We tested whether trypan blue, a common stain to discriminate live and dead cells, could be used to assess Bd viability. We show that the proportion of live zoospores (zoospores that excluded the trypan blue dye) matched the proportion of known live zoospores added to cultures. In contrast, all of the zoosporangia stages of Bd stained blue. These results demonstrate that trypan blue can be used to determine the viability of Bd zoospores but not zoosporangia. We recommend using trypan blue to report the number of live zoospores to which hosts are exposed.
蛙壶菌（Batrachochytrium dendrobatidis，简称Bd）已被认为与数百种两栖动物数量减少有关，并且是大量研究的焦点。尽管如此，尚无报道称有有效的方法来评估Bd的活力。区分活的和死的Bd有助于确定活的Bd游动孢子的剂量，以及某些因素对Bd是否具有致死或亚致死效应。我们测试了一种用于区分活细胞和死细胞的常用染料——台盼蓝，是否可用于评估Bd的活力。我们发现，活游动孢子（即排斥台盼蓝染料的游动孢子）的比例与添加到培养物中的已知活游动孢子的比例相符。相比之下，Bd的所有游动孢子囊阶段均被染成蓝色。这些结果表明，台盼蓝可用于确定Bd游动孢子的活力，但不能用于确定游动孢子囊的活力。我们建议使用台盼蓝来报告宿主接触到的活游动孢子数量。
