The conservation of large carnivores is a formidable challenge for biodiversity conservation. Using a data set on the past and current status of brown bears (Ursus arctos), Eurasian lynx (Lynx lynx), gray wolves (Canis lupus), and wolverines (Gulo gulo) in European countries, we show that roughly one-third of mainland Europe hosts at least one large carnivore species, with stable or increasing abundance in most cases in 21st-century records. The reasons for this overall conservation success include protective legislation, supportive public opinion, and a variety of practices making coexistence between large carnivores and people possible. The European situation reveals that large carnivores and people can share the same landscape.

/pdf/recovery-of-large-carnivores-in-europe-s-modern-human-49oq04a76u.pdf

Recovery of large carnivores in Europe’s modern human-dominated landscapes

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.

/pdf/noninvasive-genetic-sampling-tools-for-wildlife-biologists-a-1vuc2fervk.pdf

Noninvasive genetic sampling tools for wildlife biologists: a review of applications and recommendations for accurate data collection

Museums and other natural history collections (NHC) worldwide house millions of specimens. With the advent of molecular genetic approaches these collections have become the source of many fascinating population studies in conservation genetics that contrast historical with present-day genetic diversity. Recent developments in molecular genetics and genomics and the associated statistical tools have opened up the further possibility of studying evolutionary change directly. As we discuss here, we believe that NHC specimens provide a largely underutilized resource for such investigations. However, because DNA extracted from NHC samples is degraded, analyses of such samples are technically demanding and many potential pitfalls exist. Thus, we propose a set of guidelines that outline the steps necessary to begin genetic investigations using specimens from NHC.

/pdf/back-to-the-future-museum-specimens-in-population-genetics-ifzt2u4g9s.pdf

Back to the future: museum specimens in population genetics.

Noninvasive genetic approaches continue to improve studies in molecular ecology, conservation genetics and related disciplines such as forensics and epidemiology. Noninvasive sampling allows genetic studies without disturbing or even seeing the target individuals. Although noninvasive genetic sampling has been used for wildlife studies since the 1990s, technological advances continue to make noninvasive approaches among the most used and rapidly advancing areas in genetics. Here, we review recent advances in noninvasive genetics and how they allow us to address important research and management questions thanks to improved techniques for DNA extraction, preservation, amplification and data analysis. We show that many advances come from the fields of forensics, human health and domestic animal health science, and suggest that molecular ecologists explore literature from these fields. Finally, we discuss how the combination of advances in each step of a noninvasive genetics study, along with fruitful areas for future research, will continually increase the power and role of noninvasive genetics in molecular ecology and conservation genetics.

https://www.fs.fed.us/rm/pubs_other/rmrs_2009_beja_pereira_a001.pdf

Advancing ecological understandings through technological transformations in noninvasive genetics.

Noninvasive genetic methods can be used to estimate animal abundances and offer several ad- vantages over conventional methods. Few attempts have been made, however, to evaluate the accuracy and precision of the estimates. We compared four methods of estimating population size based on fecal sampling. Two methods used rarefaction indices and two were based on capture-mark-recapture (CMR) estimators, one combining genetic and field data. Volunteer hunters and others collected 1904 fecal samples over 2 consecutive years in a large area containing a well-studied population of brown bears (Ursus arctos). On our 49,000-km 2 study area in south-central Sweden, population size estimates ranged from 378 to 572 bears in 2001 and 273 to 433 bears in 2002, depending on the method of estimation used. The estimates from the best model in the program MARK appeared to be the most accurate, based on the minimum population size estimate from radio-marked bears in a subsection of our sampling area. In addition, MARK models included heterogeneity and temporal variation in detection probabilities, which appeared to be present in our samples. All methods, though, incorrectly suggested a biased sex ratio, probably because of sex differences in detection probabilities and low overall detection probabilities. The population size of elusive animals can be estimated reliably over large areas with noninvasive genetic methods, but we stress the importance of an adequate and well-distributed sampling effort. In cases of biased sampling, calibration with independent estimates may be necessary. We rec- ommend that this noninvasive genetic approach, using the MARK models, be used in the future in areas where sufficient numbers of volunteers can be mobilized.

/pdf/estimating-population-size-of-elusive-animals-with-dna-from-3cysp2hx1r.pdf

Estimating Population Size of Elusive Animals with DNA from Hunter-Collected Feces: Four Methods for Brown Bears

The southern Norwegian wolverine (Gulo gulo) population was considered functionally extinct in the 1960s but has partly recovered in recent years. Proper management of this population is highly dependent on reliable estimates of critical population parameters such as population size, sex ratio, immigration rate, and reproductive contribution from immigrants. We report on a large-scale population monitoring project assessing these parameters through genetic tagging of individuals, with feces as the source of DNA. Sixty-eight different individuals were detected among 147 successfully genotyped samples collected in 2000 and 2001. Sixty of these individuals were represented in the 2001 sample, which may be considered a minimum estimate of the population size. Almost 50% of these animals were sampled only once, however, indicating that the true population size may be markedly higher. Accordingly, a capture-recapture estimate based on the observed resampling rates suggested a population size of 89 wolverines (95% confidence interval (CI) = 74-104), which is approximately 35% higher than an estimate of 64 obtained from the number of active natal dens (95% CI = 46-95; p = 0.08). Indirect estimates of dispersal distances inferred from mother-offspring relationships sug- gested that wolverine males have the ability to disperse up to 500 km, a distance exceeding anything previously reported in the literature. Dispersal distances of more than 100 km were detected for females. Bayesian clus- tering analysis and subsequent assessment of individual relationships suggest that immigrants from northern Scandinavia have contributed and still contribute to the southern Norwegian gene pool, counteracting genetic erosion and reducing the risk of inbreeding depression. Additional sampling efforts will be undertaken during the coming years to allow for observations of population trends, immigration rate, and reproductive variance among individuals. Such data will provide an important basis for the design of an appropriate conservation plan for this small and vulnerable population.

/pdf/colonization-history-and-noninvasive-monitoring-of-a-2on6yxt63y.pdf

Colonization History and Noninvasive Monitoring of a Reestablished Wolverine Population

Knowledge of the wolverine Gulo gulo mating system is limited. In this study, we use 20 microsatellite loci for paternity testing in 145 wolverine offspring with known mothers. Samples were collected during > 10 years in two Scandinavian populations, mainly in connection with radio-telemetry studies and as part of long-term population monitoring. In total, 51% of the offspring were assigned a father. Our results demonstrate that the wolverine exhibits a polygamous mating system as some males were shown to produce offspring with more than one female in a single year. Females often reproduced with the same male in subsequent breeding years, but sometimes changed their partner, potentially as a consequence of a change in the territory-holding male in the area. In the majority of litters, siblings were unambiguously assigned the same father, indicating that multiple paternity is rare. Of 23 breeding pairs, for which telemetry data were available, 20 had overlapping home ranges, suggesting that pair formation generally is consistent with the territories held by wolverine males and females.

/pdf/paternity-and-mating-system-in-wolverines-gulo-gulo-uo8by3y4op.pdf

Paternity and mating system in wolverines Gulo gulo

Colonization history and non-invasive monitoring of a re-established wolverine (Gulo gulo) population.

The wolverine is an endangered carnivore that in northwestern Europe is restricted to the mountain range along the border between Sweden and Norway. The Scandinavian wolverine population experienced a severe decline in numbers due to human persecution during the 20th century, although with legislative protection the population has recently implied that the population slowly has started to recover (current population size estimate of 800 individuals). In the mid 1990s, wolverines appeared in two new and isolated areas east of the mountain range, in the forest landscape close to the Gulf of Bothnia. Using non-invasive, DNA-based monitoring, we show here that these new subpopulations were likely founded by as few as 2 and 2–4 individuals, respectively, and that little, if any, genetic contact with the main population since colonisation has been established. A high degree of genetic similarity among individuals in the two areas indicates inbreeding. We estimate the minimum number of wolverines known to be alive in these areas during the period of 2001–2005 to 5 and 17, respectively, with one subpopulation showing decreasing (currently 2) numbers and the other increasing (10). For the somewhat larger population, we infer a tentative pedigree from relatedness values and parentage tests, which indicates the occurrence of brother–sister matings. This study illustrates the usefulness of non-invasive monitoring in the management of endangered carnivore populations.

DNA-based monitoring of two newly founded Scandinavian wolverine populations

Recently, a two-step PCR approach, referred to as multiplex pre-amplification, was proposed to improve microsatellite amplification from non-invasive samples such as faecal DNA. Here, we compare this new approach to standard PCR with respect to amplification success and genotyping error rates in microsatellite analysis (18 markers) of wolverine faecal DNA (48 extracts initially shown to contain amplifiable DNA). The multiplex pre-amplification approach was clearly advantageous both in terms of successful PCR amplifications (91% vs. 80%) and allelic dropout rate (2.4% vs. 12.5%). However, dropouts were to a high extent repeated in all second-step amplifications following multiplex pre-amplification, indicative of being generated during the initial PCR. Analysing more than one PCR from the initial multiplex PCR product may thus be of limited value. We instead suggest to perform two initial multiplex PCRs and to analyse a single second-step PCR from each of them. This was tested for 22 extracts at 18 loci and proved to be an effective way to obtaining a correct genotype.

A test of the multiplex pre-amplification approach in microsatellite genotyping of wolverine faecal DNA

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