Tigers in North Dakota; Tiger Salamander (Ambystoma tigrinum) eDNA Surveying for Presence in Cattle Ponds

Abstract

Environmental DNA (eDNA) allows for efficient and convenient data collection using DNA segments to confirm species presence. There is little literature using eDNA to specifically examine tiger salamander presence in North Dakota. I sampled 8 different cattle ponds in Western North Dakota twice each using a set eDNA collection method from the United States Department of Agriculture. After collecting water samples, the filters were sent to Jonah Ventures for DNA extraction and metabarcoding. Samples were incubated, extracted, ran through a first round of Polymerase Chain Reaction (PCR) inspected on agarose gel. Then the amplicons-product of an amplified or replicated piece of DNA, were cleaned, a second round of PCR followed, then the amplicons were cleaned again. Finally sequencing and bioinformatics took place. Salamanders (genus Ambystoma) were detected at 5 of the 8 sites. Average single season Bayesian occupancy point estimate was 0.655 {+/-} (.165), with a detection probability of 0.735{+/-} (.156) over the two site visits. Of the covariates including elevation, surface area, and air temperature, air temperature on visit 2 had the best model but still was not significant. There was no significant correlation between any of the covariates and the naive detections. This method was effective in detecting the genus Ambystoma, however more work and greater sample site variation could elucidate impacts of species level presence absence data.