David Paetkau

Microsatellite analysis of population structure in Canadian polar bears

Attempts to study the genetic population structure of large mammals are often hampered by the low levels of genetic variation observed in these species. Polar bears have particularly low levels of genetic variation with the result that their genetic population structure has been intractable. We describe the use of eight hypervariable microsatellite loci to study the genetic relationships between four Canadian polar bear populations: the northern Beaufort Sea, southern Beaufort Sea, western Hudson Bay, and Davis Strait ‐ Labrador Sea. These markers detected considerable genetic variation, with average heterozygosity near 60% within each population. Interpopulation differences in allele frequency distribution were significant between all pairs of populations, including two adjacent populations in the Beaufort Sea. Measures of genetic distance reflect the geographic distribution of populations, but also suggest patterns of gene flow which are not obvious from geography and may reflect movement patterns of these animals. Distribution of variation is sufficiently different between the Beaufort Sea populations and the two more eastern ones that the region of origin for a given sample can be predicted based on its expected genotype frequency using an assignment test. These data indicate that gene flow between local populations is restricted despite the long‐distance seasonal movements undertaken by polar bears.

Microsatellite analysis of genetic variation in black bear populations

Measuring levels of genetic variation is an important aspect of conservation genetics The informativeness of such measurements is related to the variability of the genetic markers used; a particular concern in species, such as bears, which are characterized by low levels of genetic variation resulting from low population densities and small effective population sizes We describe the development of microsatellite analysis in bears and its use in assessing interpopulation differences in genetic variation in black bears from three Canadian National Parks These markers are highly variable and allowed identification of dramatic differences in both distribution and amount of variation between populations Low levels of variation were observed in a population from the Island of Newfoundland The significance of interpopulation differences in variability was tested using a likelihood ratio test of estimates of θ= 4Neu.

NONINVASIVE GENETIC SAMPLING TOOLS FOR WILDLIFE BIOLOGISTS: A REVIEW OF APPLICATIONS AND RECOMMENDATIONS FOR ACCURATE DATA COLLECTION

Abstract Noninvasive genetic sampling provides great potential for research and management applications in wildlife biology. Researchers can obtain DNA from a variety of sources including hair, feces, urine, feathers, shed skin, saliva, and egg shells without handling or observing animals. These samples can then be used to identify the presence of rare or elusive species, count and identify individuals, determine gender, and identify diet items, or samples can be used to evaluate genetic diversity, population structure, and mating system. We review the recent advancements and techniques used for identifying species, individuals, and gender. We also address the potential pitfalls of noninvasive genetic sampling and provide recommendations for laboratory- and field-based methods to improve the reliability and accuracy of data collected from noninvasive genetic samples.

Gene flow between insular, coastal and interior populations of brown bears in Alaska

The brown bears of coastal Alaska have been recently regarded as comprising from one to three distinct genetic groups. We sampled brown bears from each of the regions for which hypotheses of genetic uniqueness have been made, including the bears of the Kodiak Archipelago and the bears of Admiralty, Baranof and Chichagof (ABC) Islands in southeast Alaska. These samples were analysed with a suite of nuclear microsatellite markers. The ‘big brown bears’ of coastal Alaska were found to be part of the continuous continental distribution of brown bears, and not genetically isolated from the physically smaller ‘grizzly bears’ of the interior. By contrast, Kodiak brown bears appear to have experienced little or no genetic exchange with continental populations in recent generations. The bears of the ABC Islands, which have previously been shown to undergo little or no female‐mediated gene flow with mainland populations, were found not to be genetically isolated from mainland bears. The data from the four insular populations indicate that female and male dispersal can be reduced or eliminated by water barriers of 2–4 km and 7km in width, respectively.

Genetic structure of the world’s polar bear populations

We studied genetic structure in polar bear (Ursus maritimus) populations by typing a sample of 473 individuals spanning the species distribution at 16 highly variable microsatellite loci. No genetic discontinuities were found that would be consistent with evolutionarily significant periods of isolation between groups. Direct comparison of movement data and genetic data from the Canadian Arctic revealed a highly significant correlation. Genetic data generally supported existing population (management unit) designations, although there were two cases where genetic data failed to differentiate between pairs of populations previously resolved by movement data. A sharp contrast was found between the minimal genetic structure observed among populations surrounding the polar basin and the presence of several marked genetic discontinuities in the Canadian Arctic. The discontinuities in the Canadian Arctic caused the appearance of four genetic clusters of polar bear populations. These clusters vary in total estimated population size from 100 to over 10 000, and the smallest may merit a relatively conservative management strategy in consideration of its apparent isolation. We suggest that the observed pattern of genetic discontinuities has developed in response to differences in the seasonal distribution and pattern of sea ice habitat and the effects of these differences on the distribution and abundance of seals.

Demography and Genetic Structure of a Recovering Grizzly Bear Population

Abstract Grizzly bears (brown bears; Ursus arctos) are imperiled in the southern extent of their range worldwide. The threatened population in northwestern Montana, USA, has been managed for recovery since 1975; yet, no rigorous data were available to monitor program success. We used data from a large noninvasive genetic sampling effort conducted in 2004 and 33 years of physical captures to assess abundance, distribution, and genetic health of this population. We combined data from our 3 sampling methods (hair trap, bear rub, and physical capture) to construct individual bear encounter histories for use in Huggins–Pledger closed mark–recapture models. Our population estimate, N̂ = 765 (95% CI = 715–831) was more than double the existing estimate derived from sightings of females with young. Based on our results, the estimated known, human-caused mortality rate in 2004 was 4.6% (95% CI = 4.2–4.9%), slightly above the 4% considered sustainable; however, the high proportion of female mortalities raises concern. We used location data from telemetry, confirmed sightings, and genetic sampling to estimate occupied habitat. We found that grizzly bears occupied 33,480 km2 in the Northern Continental Divide Ecosystem (NCDE) during 1994–2007, including 10,340 km2 beyond the Recovery Zone. We used factorial correspondence analysis to identify potential barriers to gene flow within this population. Our results suggested that genetic interchange recently increased in areas with low gene flow in the past; however, we also detected evidence of incipient fragmentation across the major transportation corridor in this ecosystem. Our results suggest that the NCDE population is faring better than previously thought, and they highlight the need for a more rigorous monitoring program.

Grizzly Bear Density in Glacier National Park, Montana

Abstract We present the first rigorous estimate of grizzly bear (Ursus arctos) population density and distribution in and around Glacier National Park (GNP), Montana, USA. We used genetic analysis to identify individual bears from hair samples collected via 2 concurrent sampling methods: 1) systematically distributed, baited, barbed-wire hair traps and 2) unbaited bear rub trees found along trails. We used Huggins closed mixture models in Program MARK to estimate total population size and developed a method to account for heterogeneity caused by unequal access to rub trees. We corrected our estimate for lack of geographic closure using a new method that utilizes information from radiocollared bears and the distribution of bears captured with DNA sampling. Adjusted for closure, the average number of grizzly bears in our study area was 240.7 (95% CI = 202–303) in 1998 and 240.6 (95% CI = 205–304) in 2000. Average grizzly bear density was 30 bears/1,000 km2, with 2.4 times more bears detected per hair trap inside than outside GNP. We provide baseline information important for managing one of the few remaining populations of grizzlies in the contiguous United States.

Estimating marten Martes americana population size using hair capture and genetic tagging

We tested non-invasive genetic methods for estimating the abundance of marten Martes americana using baited glue-patch traps to pull hair samples from individual animals. We divided our 800–km2 study area into 3 × 3 km cells and put one hair trap in each cell. We trapped 309 sites for an average of 15 days each between 15 January and 14 March 1997. Based on tracks in snow and hair morphology, we captured hair from marten, red squirrels Tamiasciurus hudsonicus, flying squirrels Glaucomys sabrinus, short or long-tailed weasels Mustela erminea and M.frenata, and several unidentified mouse and vole species. Of 309 sites, 58% collected a marten hair sample while 8% of sites removed weasel hair. When roots were embedded in adhesive, a xylene wash was used to remove them before extracting DNA. All marten samples were genotyped at six microsatellite loci to identify individuals. Xylene-washed samples yielded similar genotyping success to samples that had never been exposed to xylene, and genotyping success increased with the number of hairs in the sample. Genetic data allowed 139 samples to be assigned to 88 individual marten, constituting 124 capture events during the four trapping sessions. The population estimate for our study area was 213 (95% Cl: 148–348) and the average capture probability was 0.15. The density of marten in our study area was 0.33/km2 when inhospitable habitat was removed from the calculation. We believe hair sampling and genetic analysis could be used to measure population distribution, trend and size for marten, and perhaps also for other carnivores.

An empirical test of DNA mark-recapture sampling strategies for grizzly bears

Abstract Despite the widespread use of DNA mark–recapture for estimation of grizzly bear (Ursus arctos) population size, there have been no designed experiments of DNA sampling strategies. We designed a large-scale study (8,820 km2) in the foothills of Alberta, Canada, to test sampling strategies associated with the hair snag DNA method. The main sampling method for this project used a traditional design in which bait sites were moved within 180 7 x 7 km grid cells for 4 2-week sampling sessions in the spring of 2004. However, we also tested other strategies concurrently with the traditional design. We sampled fixed sites within each cell to test the utility of moving sites compared to the less-expensive method of not moving sites. We also placed a second, lower strand of barbed wire on bait sites to see if this could identify cubs, which are not typically sampled by the usual knee-height strand of barbed wire. We compared summary statistics, capture probability variation, population estimates, and the precision of population estimates for each design. The moved-sites designs captured more bears each session, captured more individual bears (especially females), and displayed population estimates that were 15–25% higher for females. Estimates for males were similar between designs. These results suggest that the moved-sites designs were more efficient in sampling the entire population at the 7 x 7 km grid cell size. These results highlight the need for all bears to have adequate trap encounter opportunities to ensure unbiased estimates. It also demonstrates the utility of collecting enhanced data sets to test and optimize DNA sampling strategies.

Limited effect of anthropogenic habitat fragmentation on molecular diversity in a rain forest skink, Gnypetoscincus queenslandiae.

To examine the effects of recent habitat fragmentation, we assayed genetic diversity in a rain forest endemic lizard, the prickly forest skink (Gnypetoscincus queenslandiae), from seven forest fragments and five sites in continuous forest on the Atherton tableland of northeastern Queensland, Australia. The rain forest in this region was fragmented by logging and clearing for dairy farms in the early 1900s and most forest fragments studied have been isolated for 50–80 years or nine to 12 skink generations. We genotyped 411 individuals at nine microsatellite DNA loci and found fewer alleles per locus in prickly forest skinks from small rain forest fragments and a lower ratio of allele number to allele size range in forest fragments than in continuous forest, indicative of a decrease in effective population size. In contrast, and as expected for populations with small neighbourhood sizes, neither heterozygosity nor variance in allele size differed between fragments and sites in continuous forests. Considering measures of among population differentiation, there was no increase in FST among fragments and a significant isolation by distance pattern was identified across all 12 sites. However, the relationship between genetic (FST) and geographical distance was significantly stronger for continuous forest sites than for fragments, consistent with disruption of gene flow among the latter. The observed changes in genetic diversity within and among populations are small, but in the direction predicted by the theory of genetic erosion in recently fragmented populations. The results also illustrate the inherent difficulty in detecting genetic consequences of recent habitat fragmentation, even in genetically variable species, and especially when effective population size and dispersal rates are low.