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. 2024 Jan 5;51(1):76.
doi: 10.1007/s11033-023-09010-2.

Application of real-time PCR for the identification of the endangered species Galemys pyrenaicus through faecal samples

Adriana Ripa  1 , José A Díaz-Caballero  2   3 , María Jesús Palacios-González  2 , Antonio Espinosa  1 , Juan Luis García-Zapata  4 , José Luis Fernández-Garcia  5
Affiliations

Application of real-time PCR for the identification of the endangered species Galemys pyrenaicus through faecal samples

Adriana Ripa et al. Mol Biol Rep. .

Abstract

Background: Currently, many micromammals are important targets for study. The endangered Galemys pyrenaicus is an outstanding example. Globally, their populations have suffered a substantial decline in last 20 years. In the surveyed area, the capture of desman is legally forbidden due to the high conservation concerns. Reason by non-invasive sampling through faeces is proposed for its monitoring. Furthermore, the confusion between faeces from desman and Mediterranean water shrews must be considered. Thus, the aim of this study was focused on developing RT-PCR assays to determine the presence of Galemys pyrenaicus and N. a. anomalus from non-invasive samples.

Methods and results: The study was conducted in the mountains of the System Central of Extremadura (Spain). A total of 186 samples were collected from 2018 to 2021 by experts where historically reported and/or our previous studies confirmed their presence. RT-PCR assays using hydrolysis probes were designed to detect genetic material from both desman and Mediterranean water shrews and its specificity was confirmed. The reliability of the method was further assessed by PCR sequencing of mitochondrial Cyb and d-loop, resulting fully compatible with the RT-PCR approach. Intraspecific phylogenetic relationship was reported to improve knowledge about mtDNA variability in the desman from the Central System.

Conclusions: We demonstrated that RT-PCR gives a gold opportunity to further map the species using faeces which minimizes disturbance and reports both population status and individual presence. Cost-effective RT-PCR combined with field-collected faeces allows us to better investigate the full range of occurrence of the species.

Keywords: Conservation; Galemys pyrenaicus; Identification; Non-Invasive samples; RT-PCR.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1
Fig. 1
Original Network of the Shortest Median-Joining trees. Empty dots indicate sequences from Gene Bank except one (see text). The network draws two opposites branches for Clades A and B according to Igea [8] and Querejeta [2] (A and B). GPA, GPV and GPJ indicate sequences of the populations located in the Ambroz Valley, La Vera and Jerte Valley, respectively (populations from Tajo basin, Extremadura autonomous community, Spain). In this last case node size is proportional to sequence number
See this image and copyright information in PMC

References

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