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 suggested 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 clustering 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. Historia de Colonización y Monitoreo No Invasivo de una Población Reestablecida de Gulo gulo Resumen: La población sureña de Gulo gulo se consideraba funcionalmente extinta en la década de 1960 pero se ha recuperado parcialmente en años recientes. El manejo adecuado de esta población depende, en gran medida, de estimaciones confiables de parámetros poblacionales críticos como por ejemplo el tamaño poblacional, la proporción de sexos, la tasa de inmigración y la contribución reproductiva de inmigrantes. Reportamos un proyecto de monitoreo poblacional a gran escala que evaluó estos parámetros por medio del marcaje genético de individuos, con heces como la fuente de ADN. Se detectaron 68 individuos diferentes entre 147 muestras de genotipos identificados exitosamente y colectadas en 2000 y 2001. Sesenta de estos individuos estuvieron representados en la muestra de 2001, lo que puede considerarse como una estimación §Current Noninvasive Monitoring of Wolverines 677 ...

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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.

Large carnivore feeding ecology plays a crucial role for management and conservation for predators and their prey. One of the keys to this kind of research is to identify the species composition in the predator diet, for example, prey determination from scat content. DNA‐based methods applied to detect prey in predators’ scats are viable alternatives to traditional macroscopic approaches, showing an increased reliability and higher prey detection rate. Here, we developed a molecular method for prey species identification in wolf (Canis lupus) scats using multiple species‐specific marker loci on the cytochrome b gene for 18 target species. The final panel consisted of 80 assays, with a minimum of four markers per target species, and that amplified specifically when using a high‐throughput Nanofluidic array technology (Fluidigm Inc.). As a practical example, we applied the method to identify target prey species DNA in 80 wolf scats collected in Sweden. Depending on the number of amplifying markers required to obtain a positive species call in a scat, the success in determining at least one prey species from the scats ranged from 44% to 92%. Although we highlight the need to evaluate the optimal number of markers for sensitive target species detection, the developed method is a fast and cost‐efficient tool for prey identification in wolf scats and it also has the potential to be further developed and applied to other areas and large carnivores as well.