. The Canadian field-naturalist. Natural history; Sciences naturelles. 440 The Canadian Field-Naturalist Vol. 120. Barbed-wired enclosure Figure 2: Experimental design for Cougar detection, a passive method combining a baited hair-trap (central post and barbed-wired enclosure) protocol and a camera-trap (upper left comer, strapped on a tree). A volume of IS^iL contained 100-500 ng template DNA, Ix reaction buffer, mmol (miUimoles) MgCl2, mmol each primer, 250 mmol dNTPs and 1 U Taq polymerase. After initial incubation at 94°C for 2 min- utes, 40 cycles of PCR were performed at 94°C for
. The Canadian field-naturalist. Natural history; Sciences naturelles. 440 The Canadian Field-Naturalist Vol. 120. Barbed-wired enclosure Figure 2: Experimental design for Cougar detection, a passive method combining a baited hair-trap (central post and barbed-wired enclosure) protocol and a camera-trap (upper left comer, strapped on a tree). A volume of IS^iL contained 100-500 ng template DNA, Ix reaction buffer, mmol (miUimoles) MgCl2, mmol each primer, 250 mmol dNTPs and 1 U Taq polymerase. After initial incubation at 94°C for 2 min- utes, 40 cycles of PCR were performed at 94°C for 1 minute, 53°C for minutes, 72°C for minutes, and 72°C for 10 minutes. Results were then visualized on a agarose gel and compared to 20 species from the six families cited above. A second amplification of 600 bp of a specific region of the 16S mtDNA was then conducted according to the protocol detailed by Mills et al. (2000). Restriction enzymes Haelll, Hpall, and Rsal then digested PCR products at 37°C during 12 hours, and results were visualized on agarose gel. Since restriction enzymes produce species-specific patterns, it was then possible to distinguish Cougar from other felid species. Results Baited hair traps A total of 207 samples was collected from hair traps (Table 1). MtDNA analyses conducted on hair samples collected in 2003 revealed that two samples collected in FNP were Cougar hair. The first hair sam- ple was found near the main road entering the park Table 1. Number of hair samples collected per location and per year for 2003-2006. Location Sampling periods 2003 2004 2005 2006 KNP 0 6 15 7 FNP 29 34 49 23 CBHNP 1 3 17 16 MIRAM _* - 6 1 Total 30 43 87 47 *Baited hair-traps and camera-traps were installed in the Miramichi (MIRAM) area during the fall of 2004. (, Kinnie Brook), in a young Red Spruce {Picea rubens) and Balsam Fir {Abies balsamea) stand (6 to 12 m high, canopy closure of 70%). Ground cover mostly consisted of Sphagnum spp
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