Noninvasive Method for a Statewide Survey of Eastern Hellbenders Cryptobranchus alleganiensis Using Environmental DNA

Table 2

Detection of Hellbender cyt b gene eDNA in multiple Kentucky watersheds using 2 L samples as well as using ethanol precipitation of DNA resulting from three 1 L water samples denoted as 3E.

Location	Site number	Liters	Hellbender cyt b eDNA +/−
Four rivers	1-1	3E	+
Four rivers	1-2	3E	+
Four rivers	1-3	3E	+
Licking	2-1	3E	+
Licking	2-2	3E	+
Licking	2-3	3E	+
Salt	3-1	2	−
Salt	3-2	2	+
Salt	3-3	2	+
Salt	3-4	2	−
Salt	3-5	2	+
Upper green	4-1	3E	+
Upper green	4-2	3E	+
Upper green	4-3	3E	+
Upper green	4-4	3E	+
Upper green	4-5	3E	+
Upper green	4-6	3E	−
Upper green	4-7	3E	+
Upper green	4-8	3E	+
Upper green	4-9	3E	+
Upper green	4-10	3E	+
Upper green	4-11	3E	−
Upper green/river number 1 system	5-1	2	+
Upper green/river number 1 system	5-2	2	+
Upper green/river number 1 system	5-3	2	+
Upper green/river number 1 system	5-4	2	+
Upper green/river number 1 system	5-5	2	+
Cooler blank		3E	−
Environmental negative control		2	−