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.