Identifying the best control method for problematic wildlife is an ever present issue in wildlife management. Popular control methods have ranged from lethal techniques, extirpating the animal, to multiple non-lethal methods focused on deterring undesired behavior. In the past, lethal methods were the preferred choice. However, with increased awareness of the need for biodiversity conservation, new management methods focus on non-lethal control, with emphasis on exploiting aspects of naturally occurring organismal behaviors and ecology. Over the past decade, technological advances in extraction method’s and equipment have also developed new techniques providing a broader range of information about species biology for management use. One of the most well documented conflicts between wildlife and humans is that of the wolf. Using advanced technology and new techniques, we investigated the implication of using chemosensory signals in canid urine to modify behavior as a possible non-lethal alternative in large predator management. Here we used the SBSE method coupled with improved GC/MS equipment to analyze the volatile organic compounds in the urine of four canid species, gray wolf (Canis lupus), red wolf (Canis rufus), wolf-dog hybrids (Canis familiaris) and the domestic dog (Canis familiaris) in order to create working urinary profiles. The extraction method identified several compounds also seen in the urinary profiles of other large predators. In addition, similarities and differences were also noted between taxa and the sexes, and these can be further explored in future studies. Two identified urinary compounds, acetophenone and methyl propyl sulfide, were selected for further behavioral evaluation. We focused on these compounds and their influence as chemosensory signals triggering urine marking events in both the gray wolf and red wolf. Behavioral observations of the effects of these two chemicals indicated they elicited responses from captive wolves. At each of the three study sites, the combination of these chemicals produced urine-marking events along the territory boundary by dominant animals. As a result, the investigation focused on what triggered the urine-marking events, the chemicals themselves, their combination, or the breakdown of the chemicals producing other odorants. It was found that there was no significant degradation of the chemicals over time and environmental conditions produced no significant breakdown of the acetophenone prior to the addition of methyl propyl sulfide. This posed a number of new questions and illustrated the need for additional behavioral studies. The results of this study analyzing chemosensory signals in canid urine, provides biologists with new information to aid in the development of new non-lethal management strategies for handling problematic wildlife as well as providing useful information for future research involving reproduction, predator/prey dynamics, territory maintenance, and a host of other studies focusing on animal ecology in association with chemosensory signaling.

While studying packs of the eastern timber wolf in Algonquin Provincial Park in Ontario, DNA profiles at 8 microsatellite loci and the mitochondrial control region were found to be similar to those of the red wolf, C. rufus. Based on this it was suggested that both the red wolf and the eastern timber wolf have a common origin, evolving in North America, with the coyote diverging from them 150,000-300,000 years ago and with neither having any recent connection with the gray wolf that evolved in Eurasia. It was further proposed, that the eastern timber wolf retain its original species designation of C. lycaon instead of the present status of a subspecies of the gray wolf. Four "types" or "races" of wolves have been previously described in Ontario. Using DNA profiles, assignment tests identified four groups, which were typified by animals in Algonquin Provincial Park, Pukaskwa National Park, Frontenac Axis and those north of Lake Superior. The tests indicate that Frontenac animals are hybrids between the western coyote and C. lycaon and represent the eastern coyote. Pukaskwa maintains a small wolf population, which is genetically closer to the gray wolves of the Northwest Territories than the surrounding C. lycaon. These may represent an isolated remnant population of the original "Ontario type" (C. lupus). Animals north of Lake Superior were identified as C. lycaon, but represent products of hybridization between C. lycaon and C. lupus. Currently within Ontario, Algonquin Park contains the largest protected area of the eastern timber wolf. DNA profiles, including Y-linked microsatellite loci were used to establish maternity, paternity and kin relationships for 102 animals from 24 packs over a 12-year period. A complex pack structure was identified. A pack is not composed simply of an unrelated breeding pair and their offspring and subordinates appear to enter pack systems through adoption, pack splitting, dispersal and immigration. Relatively high genetic structuring was found between the Park animals and the "Tweed" wolves to the southeast suggesting introgression of coyote genetic material is not a present concern to the integrity of park animals. Evidence of gene flow with animals to the west, northeast and northwest coupled with the high genetic diversity, suggest that the Park animals are not an island population, but the southern part of a larger metapopulation of C. lycaon. Increased interest in the relationship of the red and eastern wolves led to the investigation of a gene in the major histocompatibility complex. Allelic variation in the exon 2 region of the DLA-DQA1 locus was analysed for gray wolves, red wolves, the eastern timber wolf and the western coyote. Twelve alleles were identified, seven of which were previously characterized in dogs. Non-synonomous nucleotide substitutions was 3.0 times higher than the synonomous changes, indicative of strong positive selection. These data provide baselines for the determination of allele frequencies and their distribution across the geographical range of the four species in North America. The results in this thesis have sparked numerous debates with respect to the protection of the wolves in Algonquin Provincial Park and reintroduction of wolves into Northeastern United States. The data support the idea that the C. lycaon population in Ontario is relatively large, numbering in the thousands rather than the hundreds. Concern for the conservation of wolves in Ontario should be directed at the declining numbers of gray wolves present in Ontario. / Thesis / Master of Science (MSc)

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