Environmental DNA samples
We collected 500 ml of water every 40 m around the perimeter of each
site. Due to varying lake sizes, the amount of water collected varied
between sites (Table 1). We split each water sample in half and filtered
duplicate 250 ml samples using separate filter cups for comparison
testing (Fig. 2). The amount of water collected was 5x the amount of
water used for detection of Bd at a site in 2015 (Kamoroff & Goldberg,
2017). At the end of each sample collection, we filtered duplicate
200-250 ml samples of distilled water for a negative control (Fig. 2).
We filtered all water samples using a single-use 47 mm diameter 0.45 µm
cellulous nitrate filter membrane (Sterlitech, location) and a
polypropylene vacuum flask with a rubber stopper fixed to a hand or
motorized pump. To remove the filter membrane, we used single use
forceps or forceps soaked in a 50% commercial bleach solution and then
rinsed in distilled water. All personnel wore single use disposable
latex gloves during sample collection and changed gloves prior to
handling filter membranes.