EVALUATION OF THE EFFICIENCY OF SAMPLING AND PRESERVING METHODS FOR NON-INVASIVE SAMPLES WHEN BROWN BEAR POPULATION-GENETIC STUDIES ARE CONDUCTING

Abstract

The possibilities of using non-invasively sampled biological material to study the genetic diversity and status of the brown bear population in the European North of Russia are discussed. The effectiveness of sampling and preservation methods is evaluated. Fieldwork including non-invasive collection of biological material was carried out in the study area in spring, summer and autumn 2014–2016. The studied area covers the southeast of the Murmansk Region and the Republic of Karelia. 393 samples of bear feces were collected. We conducted DNA extraction from samples and genotyping with 8 microsatellite markers in the DNA laboratory of the NIBIO Svanhovd Research Station (Svanvik, Norway). 48 brown bear individuals were successfully identified. The influence of several factors (the preservation method, age of the collected sample, diet content and sun exposure) on the amplification success was studied. When we analyzed all factors together, only diet content had a significant effect on the amplification success (p <0.05). The presence of berries in feces was found to have a positive effect on DNA and increased the effectiveness of genotyping. The analysis of each factor independently showed that both the ‘freshness’ of the sample and the method of its preservation significantly influenced the result of genotyping. Fresh feces and samples preserved in STOOL solution were the most suitable for genetic analysis. According to these results, recommendations for using noninvasively sampled biomaterial in population genetic studies are given.

Keywords

• brown bear
• ursus arctos l.
• feces
• non-invasive sampling
• method
• preservation
• diet
• dna extraction
• str analysis.