One of the deadliest roads in North America for species at risk fragments a marsh-lake ecosystem. To reduce road mortality, stakeholders installed >5 km of exclusion fencing along a southwestern Ontario, Canada, causeway in 2008. Between 2012 culverts were installed to provide safe crossings. We evaluated the success of these mitigation strategies by 1) comparing results of road surveys conducted 5 years before and 5 years after fencing installation; and 2) monitoring use of culverts by turtles using motion-activated cameras at culvert openings and stationary antennas placed to detect movements of passive integrated transponder (PIT)-tagged turtles (68 Blanding's turtles [Emydoidea blandingii] and 30 spotted turtles [Clemmys guttata]). We also radio-tracked 30 Blanding's turtles to measure culvert use in relation to home ranges. Turtle and snake abundance was 89% and 53% lower, respectively, in completely fenced road sections than in unfenced sections; abundance was 6% and 10% higher, respectively, between partially fenced and unfenced sections. After mitigation, locations where we found reptiles on the road were associated with fence ends, underscoring the importance of fence integrity and ineffectiveness of partial fencing as a mitigation strategy. We confirmed use of culverts by Blanding's turtles, northern map turtles (Graptemys geographica), snapping turtles (Chelydra serpentina), and midland painted turtles (Chrysemys picta). Through radio-tracking, we determined that male and female Blanding's turtles home ranges overlapped with different segments of the causeway. We recommend that stakeholders emphasize ensuring fence integrity and continuity, limiting impact of edge effects, and conducting a comprehensive monitoring program. Ó 2017 The Wildlife Society.

There is very little on the affinity of the human immunoreactive ouabainlike substance (OLS) to individual a-isoforms of Na+,K+-ATPase. The present study addresses this issue by comparing ouabain and OLS binding to dog kidney a1, rabbit kidney a, and porcine cerebral cortex a3 Na+,K+-ATPase. OLS was initially isolated by solid phase extraction from human serum using C18 columns. The extract was further purified by reverse phase HPLC in an acetonitrile/water (containing 0.1% TFA) step-up gradient (16-80%). In this system, two distinct ouabain immunoreactive peaks were resolved. Peak I demonstrated a polarity identical with that of authentic ouabain. In contrast, peak II was relatively non-polar and eluted later in the run. The final step in the purification of OLS involved immuno-affinity chromatography of peak I using a specific sepharose immobilized mouse monoclonal anti-ouabain antiserum. Dose response curves (range 0-100 nmol/I) for ouabain with canine a, and porcine a3 Na+,K+-ATPase showed similar inhibitory profiles (IC50=15 nmol/I), whilst rabbit a1 Na+,K+-ATPase was relatively insensitive to ouabain and purified peak I OLS. Two fold serial dilution of Peak I OLS, with subsequent analysis by canine and porcine Na+,K+-ATPase inhibition assays and RIA, demonstrated strong positive correlations between OLS determined by RIA and both canine (y=0.945x-2.532, r2=0.977) and porcine (y=0.428x-1.685; r2=0.993) Na+,K+-ATPase assays. The difference in the respective slopes suggests, however, that peak I OLS has a greater affinity for the canine derived enzyme compared to the porcine. In conclusion, these data suggest that like authentic ouabain, peak I OLS is a-isoform and species selective. (Hypertens Res 2000; 23 Suppl: 545-550)