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Who is who? High Resolution Melting analysis to discern between hare species using non-invasive sampling

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Abstract

Identification of the species is a crucial step in many ecological studies. Sometimes, this could become a challenge, because of animal elusive behavior, low density population or for sympatric species leaving similar signs that are impossible to discriminate based only on their morphology. Here, we set up non-invasive molecular method to discern between the Italian hare (Lepus corsicanus) and the European hare (Lepus europaeus) using High Resolution Melting assay on fecal DNA, for the first time on these species. The Italian hare is endemic of the Central-Southern Italy and Sicily and it is classified vulnerable by the International Union for Conservation of Nature. Our procedure could be a useful tool to help conservation and management strategies, mainly in areas where the Italian hare and the European hare live in sympatry. The 9.5% of the peninsular range distribution of the European hare is took up by the range of the Italian hare. Our workflow allows sure species discrimination, rapidly and inexpensively (in one day at least 36 samples could be processed at costs of about 259 euros, including both DNA extractions and HRM run), also when large numbers of samples have to be processed. Moreover, our method could be widely applicable to other Lepus and/or mammalian species with similar concerns, by small adjustments to the protocol and its further validation, focusing on primes and corresponding HRM annealing temperature.

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Acknowledgements

We thank the Cilento, Vallo di Diano e Alburni National Park (Salerno, Italy) for facilities. We are also grateful to Emeritus Dr. S.P. Goyal and one anonymous review for their helpful suggestions.

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Authors and Affiliations

  1. Department of Biology, University of Naples Federico II, Via Cinthia 26, 80126, Naples, Italy

    Maria Buglione, Simona Petrelli, Tommaso Notomista & Domenico Fulgione

  2. Istituto Di Gestione Della Fauna (IGF), Via M. da Caravaggio 143, 80126, Naples, Italy

    Gabriele de Filippo

  3. Ente Parco Nazionale del Cilento, Vallo di Diano e Alburni, Via F. Palumbo 18, 84078, Vallo della Lucania, Salerno, Italy

    Romano Gregorio

Authors
  1. Maria Buglione
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  2. Simona Petrelli
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  3. Tommaso Notomista
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  4. Gabriele de Filippo
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  5. Romano Gregorio
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  6. Domenico Fulgione
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Contributions

Conceptualization: Maria Buglione and Domenico Fulgione; Methodology: Maria Buglione, Domenico Fulgione; Sample collection: Maria Buglione, Domenico Fulgione, Simona Petrelli, Tommaso Notomista and Gabriele de Filippo; Formal analysis: Maria Buglione and Domenico Fulgione; Investigation: Maria Buglione, Domenico Fulgione and Simona Petrelli; Resources: Domenico Fulgione, Maria Buglione, Simona Petrelli, Tommaso Notomista, Gabriele de Filippo and Romano Gregorio; Supervision: Maria Buglione and Domenico Fulgione; Writing – original draft: Maria Buglione and Domenico Fulgione; Writing – review & editing: Maria Buglione, Domenico Fulgione, Simona Petrelli, Tommaso Notomista and Gabriele de Filippo and Romano Gregorio.

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Correspondence to Domenico Fulgione.

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Buglione, M., Petrelli, S., Notomista, T. et al. Who is who? High Resolution Melting analysis to discern between hare species using non-invasive sampling. Conservation Genet Resour 12, 727–732 (2020). https://doi.org/10.1007/s12686-020-01153-9

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