Latest version published by Biodiversity Data Journal on 13 February 2024 Biodiversity Data Journal

Stoneflies of Medvednica Nature Park: genetic diversity and morphological variability

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Researchers should cite this work as follows:

Kermek D, Pischiutta N, Hlebec D (2024): croplm. v1.2. Biodiversity Data Journal. Dataset/Occurrence.


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The publisher and rights holder of this work is Biodiversity Data Journal. This work is licensed under a Creative Commons Attribution Non Commercial (CC-BY-NC) 4.0 License.

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This resource has been registered with GBIF, and assigned the following GBIF UUID: f5b16e64-610a-4fcf-b46d-3efb068caaa1.  Biodiversity Data Journal publishes this resource, and is itself registered in GBIF as a data publisher endorsed by Participant Node Managers Committee.


Occurrence; Specimen; stoneflies; Medvednica Mt.; Croatia; biodiversity


Who created the resource:

Dora Kermek
Faculty of Science, University of Zagreb
Nikola Pischiutta
Faculty of Science, University of Zagreb
Dora Hlebec
Faculty of Science, University of Zagreb

Who can answer questions about the resource:

Dora Hlebec
Faculty of Science, University of Zagreb
Horvatovac 102a
10000 Zagreb

Who filled in the metadata:

Dora Hlebec
Faculty of Science, University of Zagreb
Horvatovac 102a
10000 Zagreb

Who else was associated with the resource:

Iva Mihoci
Croatian Natural History Museum
Demetrova 1
10000 Zagreb
Geographic Coverage

Croatia (Medvednica Mt.)

Bounding Coordinates South West [-90, -180], North East [90, 180]
Taxonomic Coverage


Order  Plecoptera (Stoneflies)
Temporal Coverage
Start Date / End Date 2021-06-17 / 2022-03-04
Project Data

Medvednica Nature Park, located near Croatia's capital and largest city, Zagreb, experiences significant urbanization, which strongly impacts the fauna within this ecosystem. Surprisingly, despite the area's diverse (micro)habitats, including the discovery of several rare species previously unknown in the Croatian fauna, comprehensive field research on any group of organisms, coupled with an integrative approach, has never been conducted in this region. In this study, we present the first insights into the genetic and morphological diversity of stoneflies, one of the most endangered groups of organisms overall. Phylogenetic reconstructions and species delineation methods revealed significant intraspecific haplotype variation in most examined species (e.g., Brachyptera seticornis, Isoperla grammatica, and Leuctra braueri), except for Leuctra prima. Additionally, our study has identified isolated populations that merit further in-depth investigation concerning morphology, genetics, and ecology. These findings contribute valuable knowledge to the conservation of stoneflies in this area.

Title Utilizing public sequence databases to identify genetic diversity of stoneflies in Medvednica Nature Park
Identifier Dora Kermek, Nikola Pischiutta
Funding Croatian Science Foundation
Study Area Description Medvednica Mt., Croatia
Design Description Project provides the first insight into the genetic and morphological diversity of stoneflies in the Medvednica Nature Park, thereby contributing to the faunal knowledge of the area. Additionally, it establishes phylogenetic and phylogeographical relationships between populations of widely distributed species in Europe: Brachyptera seticornis, Isoperla grammatica, Leuctra braueri, and Leuctra prima. These relationships enhance understanding of diversification patterns and the biogeographic history of these selected stonefly species, emphasizing populations that warrant further detailed morphological, genetic, and ecological investigation.

The personnel involved in the project:

Sampling Methods

Adults were collected using an entomological net while larvae were collected by hand under stones and branches.

Study Extent Specimens were collected at eight sampling sites in Medvednica Nature Park.
Quality Control All collected individuals were preserved in ethanol. Morphological determination of adults was conducted using a ZEISS SteREO Discovery V20 stereo microscope along with various identification keys. Due to the lack of identification keys the larvae were not determined to the species level. Larval identification was carried out by comparing their sequences with sequences available in databases.

Method step description:

  1. Total genomic DNA was isolated from the tissues of one to three legs of adults and larvae using the GenElute Mammalian Genomic DNA Miniprep Kit. Chromatograms were analyzed and edited in Geneious Prime. Misread nucleotides were manually corrected. The presence of stop codons in all sequences was checked using Mesquite. Poor quality sequences and those containing stop codons were excluded from further analyses. For phylogeographic analyses, we selected species with a broader distribution and a larger number of COI sequences available in the BOLD and GenBank databases.
Collection Data
Collection Name Collection Plecoptera Sivec&Hlebec
Specimen preservation methods Alcohol
Curatorial Units Between 488 and 512 vial
Additional Metadata