High efficiency protocol of DNA extraction from Micromys minutus mandibles from owl pellets: a tool for molecular research of cryptic mammal species

Abstract

Owl pellets have high potential as a source of DNA. However, this noninvasive method of collecting DNA is rarely used, and its methodological aspects are poorly understood. We investigated the methodology for DNA extraction and amplification from owl pellets containing the smallest European rodent—the Harvest mouse Micromys minutus—as an example. We used mandibles identified in owl pellets for mitochondrial and nuclear DNA amplification. For DNA extraction, we tested two commercial protocols and utilized a protocol being a combination of two commercial kits which ensured high efficiency of DNA extraction. Additionally, we recorded that the amount of DNA was five times higher in extracts from teeth as compared to DNA extracts from jawbones derived from the same mandible. The quantity of DNA was significantly positively correlated with biological sample weight; however, the age of the pellet remains had an impact on the level of inhibition. We recorded inhibition in 40 % of mtDNA extracts derived from pellets older than 150 months, whereas in DNA extracts from pellets younger than 80 months, we did not observe a negative impact of inhibition on PCR efficiency. The amplification success rate was 89.9 % for the mitochondrial fragment and 39.4 % in the case of the nuclear fragment. We observed partial degradation of DNA evidenced by the fact that the longest fragments that we were able to amplify in the case of mtDNA were 450 and 200 bp for nuDNA. The study shows that pellets can be considered as a source of DNA and have high potential for molecular research in the case of threatened species and species that are difficult to study using standard field techniques.

Sexual dimorphism, asymmetry, and the effect of reproduction on pelvis bone in the bank vole, Myodes glareolus

“…The results of our study provide also an opportunity (however, not tested through the paper) to establish sexual reproductive status of the bank vole, captured by diurnal raptor and owls (Raczyński and Ruprecht 1974;Zalewski 1996). Advanced molecular techniques are currently being used to confirm species diagnostics and even the sex structure of consumed prey by raptors (Buś et al 2013), and added classical osteological methods will enlarge information on the reproductive status of females.…”

Small mammal diversity in Semi-deciduous Seasonal Forest of the southernmost Brazilian Pampa: the importance of owl pellets for rapid inventories in human-changing ecosystems

“…Despite the material being fragmentary, it is possible to achieve great taxonomic resolution through comparison with specimens housed in scientific collections and the literature. Furthermore, molecular analysis could also be used to elucidate the taxonomic identity of several taxa with poorly resolved taxonomies (Buś et al, 2014;Rocha et al, 2015). An increase in morphological studies is desirable to help in the identification of the material retrieved from owl pellets.…”

Long Eared Owls (Asio otus Linnaeus, 1758) as Field-Assistants in an Integrative Taxonomy Survey of a Peculiar Microtus savii (Rodentia, Cricetidae) Population

“…As suggested by some authors [19,[53][54][55], the analysis of small mammal remains recovered from raptor pellets offers many resources for several kind of studies. Our results confirm the presence of M. savii in Umbria, as previously reported by Gaggi and Paci [56], whereas no solid data were found to support the presence of M. brachycercus, which shows the northernmost distribution near the regional border.…”