Endemism and hidden diversity of Habrophlebia, Eaton, 1881 in Algeria's Northeastern Mountains (Ephemeroptera, Leptophlebiidae)

Endemism and hidden diversity of Habrophlebia, Eaton, 1881 in Algeria’s Northeastern Mountains (Ephemeroptera, Leptophlebiidae) Boudjéma Samraoui1,2, Laurent Vuataz3,4, Michel Sartori3,4 1 Biology Department, University Badji Mokhtar, Annaba, Algeria 2 Laboratoire de Conservation des Zones Humides, Université 8 mai 1945, Guelma, Algeria 3 Department of Zoology, State Museum of Natural Sciences (Naturéum), Palais de Rumine,Place Riponne 6, CH-1005 Lausanne, Switzerland 4 Department of Ecology and Evolution, University of Lausanne (UNIL), CH-1015 Lausanne, Switzerland https://zoobank.org/866C37ED-D606-406B-953E-062621C7AED7 Corresponding author: Boudjéma Samraoui ([email protected]) Academic editor: Jean-Luc Gattolliat ♦ Received 8 September 2025 ♦ Accepted 10 November 2025 ♦ Published 27 November 2025 Abstract Northeastern Algeria has recently emerged as a hotspot of freshwater biodiversity and a refugium for both mountainous cold stenothermic and Afrotropical relict species. Intensive surveys of mayflies uncovered striking cryptic diversity within the genus Habrophlebia Eaton, 1881 (Ephemeroptera, Leptophlebiidae). By integrating molecular and morphological evidence, we describe five new species (H. callensis sp. nov., H. ghora sp. nov., H. seybouse sp. nov., H. annaba sp. nov., and H. edough sp. nov.). Our findings increase the number of recognised Habrophlebia species in the Maghreb from nine to fourteen, all of which are endemic to the region. The results further suggest that northeastern Algeria constitutes a major diversification centre for Habrophlebia within the Maghreb and, more broadly, across the Mediterranean basin. Furthermore, distributional patterns support an east–west divide within the Maghreb, most likely shaped by distinct Quaternary glacial refugia. Finally, we provide an updated identification key to the nymphs of Habrophlebia species in the western Palaearctic. Key Words Biogeography, COI, freshwater biodiversity, Maghreb, mayflies, new species, taxonomy Introduction Mayflies, belonging to the order Ephemeroptera, comprise approximately 4,000 species, 500 genera, and 40 families (Sartori and Brittain 2015). Among these families, Leptophlebiidae is one of the most diverse, encompassing eight subfamilies and around 810 species, collectively accounting for about 20% of all known mayfly species (Sartori and Gattolliat 2023). However, within the Palaearctic region, Leptophlebiidae exhibits relatively low diversity, with only 60 species recorded across four subfamilies (Peters and Edmunds 1970; Sartori and Gattolliat 2023). The subfamily Habrophlebiinae includes 25 species in this region, nine of which belong to the genus Habrophlebia Eaton, 1881. In the Maghreb, Habrophlebia is represented by six species: two (H. hassainae and H. djurdjurensis) have been described from Algeria, while the remaining four (H. vaillantorum, H. dakki, H. abietis, and H. linae) are currently known only from Morocco. Earlier records of H. fusca in the region (Thomas 1998) need confirmation, while the presence of H. consiglioi, originally described from Sardinia and later reported in Tunisia (Zrelli et al. 2011a), has proven to be unfounded. Of particular interest is the close genetic relationship between the two Algerian species, with a minimum interspecific COI divergence of 2.1% (El Alami et al. 2023), suggesting recent reproductive isolation (Hebert et al. 2003), as well as the occurrence of numerous hybrid populations in the Rif (El Alami et al. 2025). Alpine Entomology 9 2025, 113–151 | DOI 10.3897/alpento.9.171490 Copyright Boudjéma Samraoui et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 114 The Maghreb, forming part of the Mediterranean Basin, is one of the world’s biodiversity hotspots (Myers et al. 2000). Freshwater biodiversity in the region is unevenly distributed and primarily concentrated in coastal areas. In northeast Algeria and northwest Tunisia, the Numidia region harbours a wide range of wetlands, including mountainous streams and rivers that support a rich diversity of waterbirds, fish, and amphibians, many endemic to the region (Samraoui and Samraoui 2008; Ben Hassine and Escoriza 2017; Dufresnes et al. 2019). These ecosystems also sustain a diverse assemblage of aquatic insects and zooplankton of varied biogeographic origins (Samraoui et al. 1998). Lowland wetlands harbour Afrotropical relict species such as the dragonflies Urothemis edwardsii and Acisoma inflatum (Samraoui et al. 1993); the montane zones act as refugia for cold-stenothermic taxa, including the aeshnid Aeshna cyanea (Samraoui and Alfarhan 2015) and Pyrrhosoma nymphula (Korbaa et al. 2014). Following the pioneering studies of Eaton (1899) and Lestage (1925), research on Algerian mayflies stagnated for much of the twentieth century, before resuming in its final decades and culminating in the synthesis by Thomas (1998). Since the early 21st century, there has been renewed interest in the taxonomy and ecology of Algerian mayflies, resulting in a number of new publications (e.g. Soldán et al. 2005). More recently, several additions to the Algerian mayfly checklist have been made (Benhadji et al. 2018, 2019; Kechemir et al. 2020; Samraoui et al. 2021a, 2021b; Kaltenbach et al. 2022), along with reports of numerous yet undescribed species (Labdaoui et al. 2024; Hezil et al. 2025). The aim of this study is to investigate the diversity of the genus Habrophlebia in northeastern Algeria by integrating molecular (COI) and morphological data, with the objectives of providing preliminary ecological information on species distribution, habitats, and phenology, and offering biogeographical insights into the region’s role in the diversification of the genus. Material and methods Material for this study was collected from rivers and streams across northeastern Algeria between 2018 and 2022 (Fig. 1). The study area, as well as the sampling protocol for nymphs, are described in detail by Samraoui et al. (2021a, 2021b). Adult specimens were captured along stream margins using a hand net. Morphological analyses Specimens were observed under a Leica MZ12 and M205C stereomicroscope. Nymphs were dissected in Cellosolve (2-ethoxyethanol); mouthparts, legs and the abdomen were mounted on microscopic slides with Euparal® as the medium. Microscopic pictures were taken using an Olympus BX51 microscope coupled with an Olympus SC50 camera; pictures were enhanced with the stacking software Olympus Stream Basic ver. 2.3.2. and Adobe Photoshop ver. 21.2.2. Pictures of habitus were made using a Canon EOS 6D camera and the Visionary Digital Passport imaging system (formerly available and distributed by Dun Inc., Virginia), and processed with Adobe Photoshop Lightroom and Helicon Focus ver. 5.3. For scanning electron microscopy (SEM) imaging, eggs were dehydrated in pure ethanol, coated with 12 nm platinum and SEM pictures were performed at Lausanne University with a FEI Quanta FEG 250 at 10 kV with a WD ranging between 10.1 and 12.1 mm. Final figure plates were assembled in Adobe Photoshop Elements 2022 ver. 20.0. Molecular analyses To complement our morphological investigations, we analyzed a 658 bp fragment of the mitochondrial cytochrome oxidase subunit 1 gene (COI). The dataset comprises the 91 West Palearctic Habrophlebia COI sequences from El Alami et al. (2025) and 27 newly generated sequences from Algerian specimens (Table 1), for a total of 118 sequences. DNA from the new specimens was extracted using the non-destructive method outlined in Vuataz et al. (2011). PCR amplification, purification, and sequencing followed the protocol of El Alami et al. (2022a). Forward and reverse reads were assembled and edited in CodonCode Aligner ver. 12.0.1 (CodonCode Corporation, Dedham, MA). Two sequences from Habroleptoides Schönemund, 1929 were retrieved from GenBank and used as outgroups. All sequences were aligned in MAFFT (Katoh et al. 2019) via Jalview ver. 2.11.4.1 (Waterhouse et al. 2009) with default settings. The number of parsimony-informative sites was calculated in MEGA ver. 10.2.4 (Kumar et al. 2018; Stecher et al. 2020). To examine and visualize COI evolutionary divergence, we used pairwise genetic distances and gene tree approaches. Pairwise distances were calculated in R ver. 4.5.1 (R Core Team 2025) using the dist.dna function from the ape ver. 5.8.1 package (Paradis and Schliep 2019), with the raw model and the pairwise.deletion option to obtain uncorrected p-distances (see Srivathsan and Meier 2012), excluding missing data pairwise. Mean, minimum, and maximum distances within and between putative COI species (hereafter Molecular Operational Taxonomic Units, MOTUs), were calculated using the ddply function from the plyr ver. 1.8.9 package (Wickham 2011). The 97 previously published COI sequences were assigned to MOTUs following the 12 clades defined in Fig. 13 of El Alami et al. (2025), while the 27 sequences generated in this study were assigned based on the species delimitation analyses described below. Before reconstructing the COI gene tree, the best evolutionary model (GTR + Γ + I) was selected using the second-order Akaike information criterion (AICc; Hurvich and Tsai 1989) in JmodelTest ver. 2.1.10 (Darriba et al. 2012) with three substitution schemes, six gamma categories, and default settings for other parameters. Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 115 Table 1. Details of newly sequenced specimens used in this study, including species name, specimen catalogue number, developmental stage, collection locality with GPS coordinates and date, and GenBank accession numbers. All specimens are paratypes (GenSeq Nomenclature: genseq-2 COI). Species Specimen catalogue nb Stage Locality GPS coordinates Date GenBank ID Habrophlebia annaba sp. nov. GBIFCH00658258 nymph Edough, O. Agab 36°54'44"N, 7°39'35"E 1.VI.2022 PX283157 Habrophlebia annaba sp. nov. GBIFCH01118439 nymph Edough, O. Zouz 36°55'54"N, 7°40'03"E 20.III.2022 PX283166 Habrophlebia annaba sp. nov. GBIFCH01118440 male imago Edough, O. Agab 36°54'44"N, 7°39'35"E 1.V.2021 PX283167 Habrophlebia annaba sp. nov. GBIFCH01314437 nymph Edough, O. Agab 36°54'44"N, 7°39'35"E 10.II.2022 PX283177 Habrophlebia annaba sp. nov. GBIFCH01314677 nymph Edough, O. Agab 36°54'44"N, 7°39'35"E 1.V.2021 PX283178 Habrophlebia annaba sp. nov. GBIFCH01314689 nymph Edough, O. Agab 36°54'44"N, 7°39'35"E 26.IV.2021 PX283179 Habrophlebia callensis sp. nov. GBIFCH00763554 nymph Hadada, Dar Essalem 36°53'10"N, 8°37'2"E 1.VIII.2019 PX283160 Habrophlebia callensis sp. nov. GBIFCH00763555 nymph Hadada, O. Ftitich 36°54'01"N, 8°37'05"E 11.VII.2019 PX283161 Habrophlebia callensis sp. nov. GBIFCH01118444 nymph Hadada, O. Ftitich 36°54'01"N, 8°37'05"E 12.III.2020 PX283171 Habrophlebia edough sp. nov. GBIFCH00654947 nymph Edough, O. Agab 36°54'44"N, 7°39'35"E 9.IX.2021 PX283153 Habrophlebia edough sp. nov. GBIFCH00654948 nymph Edough, O. Agab 36°54'44"N, 7°39'35"E 18.VIII.2021 PX283154 Habrophlebia edough sp. nov. GBIFCH01118441 nymph Edough, O. Agab 36°54'44"N, 7°39'35"E 18.VIII.2021 PX283168 Habrophlebia ghora sp. nov. GBIFCH00673045 nymph Ghora, O. Guitna inférieur 36°37'5"N, 8°20'46"E 1.V.2019 PX283158 Habrophlebia ghora sp. nov. GBIFCH00673115 nymph Ghora, O. Louar supérieur 36°37'1"N, 8°22'47"E 1.V.2019 PX283159 Habrophlebia ghora sp. nov. GBIFCH00763556 nymph Ghora, O. Louar supérieur 36°37'1"N, 8°22'47"E 6.VI.2019 PX283162 Habrophlebia ghora sp. nov. GBIFCH01118437 nymph Menzel Beldi 36°36'59"N, 8°21'51"E 15.V.2021 PX283164 Habrophlebia ghora sp. nov. GBIFCH01118438 male imago Menzel Beldi 36°36'59"N, 8°21'51"E 15.V.2021 PX283165 Habrophlebia ghora sp. nov. GBIFCH01118442 nymph El Ayoun 36°50'14"N, 8°36'4"E 15.V.2021 PX283169 Habrophlebia ghora sp. nov. GBIFCH01118443 male imago El Ayoun 36°50'14"N, 8°36'4"E 9.V.2021 PX283170 Habrophlebia ghora sp. nov. GBIFCH01118445 male imago Ghora, O. Louar supérieur 36°37'1"N, 8°22'47"E 15.V.2021 PX283172 Habrophlebia ghora sp. nov. GBIFCH01118446 male imago Ghora, O. Guitna inférieur 36°37'5"N, 8°20'46"E 7.VI.2020 PX283173 Habrophlebia ghora sp. nov. GBIFCH01118447 nymph Ghora, O. Louar supérieur 36°37'1"N, 8°22'47"E 10.V.2021 PX283174 Habrophlebia ghora sp. nov. GBIFCH01118448 male imago Ghora, O. Louar inférieur 36°37'2"N, 8°22'49"E 15.V.2021 PX283175 Habrophlebia ghora sp. nov. GBIFCH01118449 nymph Ghora, O. Guitna supérieur 36°37'0"N, 8°21'19"E 26.IV.2022 PX283176 Habrophlebia seybouse sp. nov. GBIFCH00658256 nymph Collo, O. Tizagban 37°00'39"N, 6°22'31"E 21.I.2022 PX283155 Habrophlebia seybouse sp. nov. GBIFCH00658257 nymph Collo, O. Tizagban 37°00'39"N, 6°22'31"E 21.I.2022 PX283156 Habrophlebia seybouse sp. nov. GBIFCH01118436 male imago Selaoua, O. Cherf 36°20'54"N 7°16'2"E 6.VI.2020 PX283163 alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 116 To account for different substitution rates among COI codon positions, the dataset was partitioned into positions 1 + 2 and position 3. Bayesian inference was performed in BEAST ver. 1.10.4 (Suchard et al. 2018) on the CIPRES Science Gateway (Miller et al. 2010). The input file was generated in BEAUTi ver. 1.10.4 (Suchard et al. 2018), incorporating the selected model and partition scheme. A relaxed molecular clock (uncorrelated lognormal) and a UPGMA starting tree were used, with default values for other parameters. Two independent Markov chain Monte Carlo (MCMC) runs of 100 million generations each were performed, sampling every 1000 generations. Convergence was assessed visually in Tracer ver. 1.7.2 (Rambaut et al. 2018). Log and tree files from the two runs were combined in LogCombiner ver. 1.10.4 (Suchard et al. 2018) after discarding the first 10% of trees as burn-in, ensuring all parameters had effective sample sizes (ESS) > 200. The maximum clade credibility tree was produced in TreeAnnotator ver. 1.10.4 (Suchard et al. 2018), with node heights set to common ancestor heights and default settings for other parameters. The tree was visualized and edited in iTOL ver. 7.2.1 (Letunic and Bork 2024). Finally, we applied three contrasting single-locus species delimitation methods to the COI dataset: the distance-based ASAP (Assemble Species by Automatic Partitioning; Puillandre et al. 2020) and the tree-based mPTP (multi-rate Poisson Tree Processes; Kapli et al. 2017) and GMYC (General Mixed Yule-Coalescent; Pons et al. 2006; Fujisawa and Barraclough 2013) approaches. The ASAP analysis was performed in ASAPy ver. 1.0, downloaded from the iTaxoTools platform (Vences et al. 2021; https://itaxotools.org). Genetic distances were calculated from the COI alignment using simple p-distances, with all other settings left at default. The best partition was selected based on the lowest asap-score. Since two partitions had the same lowest scores, we chose the one with a threshold distance closest to 3% divergence, which aligns with previously reported intrato inter-species thresholds in Ephemeroptera (Gattolliat et al. 2015). The mPTP analysis was conducted using the webserver https://mptp.h-its. org/#/tree with the BEAST tree generated earlier as input, and the command line execution “mptp --ml --multi”. For the GMYC approach, the ultrametric tree was generated in BEAST following the same procedure as above, but using a reduced dataset that excluded outgroups and pruned haplotypes (see Talavera et al. 2013) with Collapsetypes ver. 4.6 (Chesters 2013). The best-fit evolutionary model (GTR + Γ + I) was re-estimated for this reduced dataset using the same method as before. GMYC was then implemented in R with the SPLITS package ver. 1.0.20 (Ezard et al. 2009), using the single-threshold model, which has been shown to outperform the multiple-threshold version (Fujisawa and Barraclough 2013). Abbreviations MZL Muséum Cantonal des Sciences Naturelles (Naturéum), Lausanne (Switzerland). Figure 1. Location map of northeastern Algeria showing the distribution of the newly described Habrophlebia species. Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 117 Results Systematics Descriptions Habrophlebia ghora Sartori, Vuataz & Samraoui, sp. nov. https://zoobank.org/213C2984-EAF5-4BF4-B2D1-20D7A809E083 Figs 2–6 Habrophlebia consiglioi sensu Zrelli et al. (2011a), nec Biancheri 1959. Habrophlebia spK, Samraoui et al. 2021b; Habrophlebia sp. 11, Hezil et al. 2025. Material examined. Holotype: one nymph in ethanol (GBIFCH00654989), Algeria, Wilaya of El Tarf, Ghora, Oued Louar supérieur, 36°37'1"N, 8°22'47"E, 594 m, 1.V.2019, B. Samraoui leg., MZL. Paratypes. Algeria, Wilaya of El Tarf, Ghora, Oued Louar supérieur, same data as holotype, 135 nymphs in ethanol (GBIFCH00672312), 1 nymph on slide (GBIFCH00673115); same locality as holotype, 6.VI.2019, 1 nymph on slide (GBIFCH00763556); same locality as holotype, 10.V.2021, 20 nymphs in ethanol (GBIFCH00892922), 1 nymph on slide (GBIFCH01118447); same locality as holotype, 15.V.2021, 9 nymphs in ethanol (GBIFCH00892917), 21 male imagos in ethanol (GBIFCH00892924), 1 male imago in ethanol (GBIFCH01118445) • Wilaya of El Tarf, Ghora, Oued Louar inférieur, 36°37'2"N, 8°22'49"E, 589 m, 15.V.2021, 7 male imagos in ethanol (GBIFCH01118448, GBIFCH01123083) • Wilaya of El Tarf, Ghora, Oued Guitna supérieur, 36°37'0"N, 8°21'19"E, 584 m, 26.IV.2022; 1 nymph on slide (GBIFCH01118449), 10 nymphs in ethanol (GBIFCH01123084) • Wilaya of El Tarf, Ghora, Oued Guitna inférieur, 36°37'5"N, 8°20'46"E, 476 m, 1.V.2019, 1 nymph on slide (GBIFCH00673045); same locality, 1.VI.2020, 12 male imagos in ethanol (GBIFCH00892916); same locality, 7.VI.2020, 28 male, 1 female imagos (GBIFCH00892915), 1 male imago (GBIFCH01118446); same locality, 6.VI.2021, 1 nymph in ethanol (GBIFCH01123082); same locality, 26.IV.2022, 1 nymph in ethanol (GBIFCH00892921); same locality, 5.V.2022, 1 male imago in ethanol (GBIFCH00892919); same locality, 12.V.2022, 1 female imago in ethanol (GBIFCH00892918); same locality, 18.V.2022, 3 male, 4 female imagos (GBIFCH00892914); same locality, 21.V.2022, 2 male imagos in ethanol (GBIFCH00892920); same locality, 30.V.2022, 1 male, 3 female imagos in ethanol (GBIFCH00892913). • Wilaya of El Tarf, El Ayoun; 36°50'14"N, 8°36'4"E, 263 m, 9.V.2021, 1 male imago in ethanol (GBIFCH01118443), 4 nymphs, 22 male imagos in ethanol (GBIFCH00654970); same locality, 15.V.2021, 1 nymph on slide (GBIFCH01118442), 7 nymphs in ethanol (GBIFCH00892923, GBIFCH00654971) • Wilaya of El Tarf, Menzel Beldi; 36°36'59"N, 8°21'51"E, 590 m, 15.V.2021, 5 nymphs, 7 male imagos in ethanol (GBIFCH00654986), 1 male imago in ethanol (GBIFCH01118438), 1 nymph on slide (GBIFCH01118437). All B. Samraoui leg., MZL. Other material. Algeria, Wilaya of El Tarf, Ghora, Oued Louar supérieur, 36°37'1"N, 8°22'47"E, 594 m, 21.I.2019, 3 nymphs (GBIFCH00834912); same locality, 16.IV.2019, 4 nymphs (GBIFCH00834904); same locality, 1.V.2019, 81 nymphs (GBIFCH00672312-GBIFCH00672313); same locality, 6.VI.2019, 50 nymphs (GBIFCH00672308); same locality, 1.VII.2019, 10 nymphs (GBIFCH00835008); same locality, 22.I.2020, 7 nymphs (GBIFCH00835007); same locality, 18.IV.2018, 15 nymphs (GBIFCH00889986). • Wilaya of El Tarf, Ghora, Oued Louar inférieur, 36°37'3"N, 8°22'49"E, 589 m, 18.IV.2018, 1 nymph (GBIFCH00889509); same locality, 6.VI.2019, 45 nymphs (GBIFCH00672306); same locality, 1.VII.2019, 3 imagos (GBIFCH00835071), 10 nymphs (GBIFCH00835082). • Wilaya of El Tarf, Ghora, Oued Guitna supérieur, 36°37'0"N, 8°21'19"E, 584 m, 14.VI.2018, 15 nymphs (GBIFCH00889962); same locality, 6.VI.2019, 15 nymphs (GBIFCH00834830); same locality, 1.VII.2019, 3 imagos (GBIFCH00834537). • Wilaya of El Tarf, Ghora, Oued Guitna inférieur, 36°37'5"N, 8°20'46"E, 476 m, 1.V.2019, 8 nymphs (GBIFCH00672315). • Wilaya of El Tarf, Ghora, Oued Guitoun, 36°37'0"N, 8°19'34"E, 293 m, 14.VI.2018, 2 nymphs (GBIFCH00889505). • Wilaya of El Tarf, Kherrata, Oued Bougous, 36°38'2"N, 8°18'25"E, 262 m, 18.IV.2018, 4 nymphs (GBIFCH00889513). • Wilaya of El Tarf, Zitoun Meftah, Bayadh, 36°38'0"N, 8°21'55"E, 282 m, 28.III.2018, 2 nymphs (GBIFCH00889989); same locality, 1.VII.2019, 3 nymphs (GBIFCH00834516). • Wilaya of El Tarf, Ghora, Oued Houadfia, 36°36'0"N, 8°23'29"E, 455 m, 5.VI.2018, 21 nymphs (GBIFCH00835076-GBIFCH00889532); same locality, 13.III.2019, 1 nymph (GBIFCH00672309); same locality, 19.V.2019, 60 nymphs (GBIFCH00672310), one nymph on slide (GBIFCH01225270); same locality, 6.VI.2019, 25 nymphs (GBIFCH00834910). • Wilaya of El Tarf, Oued Bougous, 36°39'0"N, 8°19'59"E, 610 m, 28.III.2018, 2 nymphs (GBIFCH00889521). • Wilaya of El Tarf, Oued Besbassa, 36°39'0"N, 8°20'1"E, 233 m, 28.III.2018, 14 nymphs (GBIFCH00889982). • Wilaya of El Tarf, Oued Khemissa, 36°39'0"N, 8°21'57"E, 237 m, 28.III.2018, 7 nymphs (GBIFCH00889540). • Wilaya of El Tarf, Oued Oudaï Smid, 36°40'0"N, 8°22'49"E, 202 m, 28.III.2018, 7 nymphs (GBIFCH00889987). All in ethanol, B. Samraoui leg., MZL. Tunisia, Jendouba Governorate, Aïn Draham, Oued Ennour, 36°48'0"N, 8°39'31"E, 418 m, 28.IV.2010, 1 nymph in ethanol (GBIFCH00281533); same locality, 25.IV.2009, 1 nymph on slide (GBIFCH01225269); S. Zrelli leg., MZL. Etymology. The name “Ghora” is derived from the Coot (Fulica atra), known locally as “Ghor”, whose conspicuous white frontal shield evokes the image of a snowcapped peak. Mount Ghora (Djebel Ghora), situated on the Algerian–Tunisian border, takes its name from this association and forms part of the Kroumiria massif. alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 118 Figure 2. H. ghora sp. nov., habitus of female (above) and male (below) nymphs (A); labrum (B); left mandible (C); right mandible (D); maxilla (E); hypopharynx, left half (F); labium and submentum (G). Scale bars: 1 mm (A); 50 µm (B, E, F); 100 µm (C, D, G). Descriptions. Nymph. Body length of final instar, excluding caudal filaments, up to 5 mm for male and up to 6 mm for female. Cerci as long as body length. Coloration. General coloration medium brown; light brown between compound eyes and antennae. Upper portion of male eyes light brown to orange (Fig. 2A). Antenna with scape and pedicel greyish brown, filament yellowish. Proand mesonotum medium brown, with light brown maculae, on medium; lateroparapsidal sutures dark brown. Legs light to medium brown; dorsal surface Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 119 Figure 3. H. ghora sp. nov., thoracic structures, pronotum, right half (A); fore femur (B); mid femur (C); hind femur (D); setae on upper face of hind femur (E); hind tibia (F); claw (G). Scale bars: 100 µm (A–D, F); 20 µm (E); 50 µm (G). of femora darker; tibiae and tarsi light brown to yellowish. Abdominal tergites medium brown with a sagittal line paler; distal half of each tergite darker, sometimes with two elongated light maculae medio-anteriorly, on segments I–VI, especially in younger nymphs. Sternites light to medium brown, nervous ganglia well visible. Cerci and paracercus medium brown at base, lighter distally, rather uniform in color, without apparent banding. Head. Labrum rectangular, ca. 1.6× wider than long (Fig. 2B); dorsal surface covered distally with scattered stout setae; anterior margin with a row of stout, long and spatulate setae medially; emargination smooth, U-shaped, flared, with indistinct flat denticles; ventral surface with two bunches of stout setae medially. Mandibles similar to other Habrophlebia species, incisor with three teeth, the lowest on left mandible characterised by distinct basal indentation (Fig. 2C), kinetodontium with 3 teeth, 11–15 long and thin setae below mola of right mandible (Fig. 2D). Maxilla stocky, subapical row of 6–7 pectinate setae (Fig. 2E); maxillary palp with three segments, segment 1 and 2 subequal in length, and longer than segment 3; segment 3 triangular 1.11–1.15× longer than wide at base; all setae on palp segments stout and entire. Hypopharynx with highly developed superlinguae terminated by a membranous digitation (Fig. 2F). Labium with rhomboid glossae, outer margin and apex covered by short, broad setae; paraglossae enlarged laterally, covered with fine and long setae on the dorsal surface; with long, thick setae alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 120 Figure 4. H. ghora sp. nov., abdominal structures, posterior margin of tergite IX (A); tergite VIII (B); tergite VII (C); gill (D). Scale bar: 20 µm. on outer margin; labial palp with three segments, inner margin of segment 1 highly dilated near the middle, about 1.20× longer than the maximum width, segment 2 as long as segment 3, ca. 0.8× length of segment 1; segment 3 ca. 1.75–1.80× longer than wide at base, conical shape and with 7–8 stout and long, simple setae on dorsal surface. Submentum with numerous long, stout and pointed setae laterally; ventral surface with few long, stout and pointed setae (Fig. 2G). Thorax. Pronotum with anterior corners bearing few long and pointed setae laterally, and 6–8 stout and strong setae dorsally. Anterior margin of pronotum with 3–5 stout setae laterally (Fig. 3A). Fore legs femora elongated, ca. 2.65× longer than wide, outer margin and upper surface covered with long, entire and pointed setae (Fig. 3B); tibiae slightly shorter than femora, outer margin with sparse thin and long setae, inner margin with long stout and feathered or entire setae more numerous near apex; tarsi 0.55× length of tibiae, outer margin with sparse long and thin setae, inner margin with long and pointed entire setae. Middle legs similar to fore legs, femora ca. 2.9× longer than wide, dorsal surface of femora with more numerous and slightly longer stout and pointed setae (Fig. 3C); tibiae slightly shorter than femora; tarsi 0.5× length of tibiae. Hind legs with femora 3.4× longer than wide (Fig. 3D), dorsal surface covered with stout, long and short, pointed or rounded and entire (non-feathered) setae more or less in rows (Fig. 3E); hind tibiae as long as hind femora, outer margin with scattered stout, pointed setae; inner margin with stout, pointed and entire setae (Fig. 3F); tarsi 0.45× length of tibiae, outer margin with few long and thin setae, inner margin with long, stout, pointed and entire setae. Claws of all legs slightly hooked, with 12–14 denticles that decrease in size from the apex to the tarsus (Fig. 3G). Abdomen. Posterolateral expansions only on segments VIII and IX. Posterior margin of tergite IX with triangular, broad and narrow, pointed spines, ca. 1.5–2.5× longer than wide at base (Fig. 4A); tergite VIII with small triangular, narrow spines 1.5–2× longer than wide (Fig. 4B); tergite VII with tiny needle-shaped spines (Fig. 4C); tergites VI and above with barely visible spines. Gills present on segments I–VII; all gills elongated with long filaments; dorsal lamella bearing 4–5 filaments, ventral lamella with 2–3 filaments (Fig. 4D). Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 121 Figure 5. H. ghora sp. nov., male imago in lateral view (A); forewing (B); hindwing (C); genitalia in ventral view (D); penis in lateral view (E). Scale bar: 1 mm. Imago. Male imago. Size: body length: ca. 5.5 mm; forewing length: 5.0–5.5 mm; cerci and terminal filament broken. Head dark brown; basal portion of compound eyes greyish, upper portion light brown, scape dark brown, pedicel medium brown, flagellum light brown (Fig. 5A). Thorax. Pronotum dark brown; mesoand metanotum uniformly dark brown, pleurae, coxae, and trochanters medium brown, washed with dark brown; fore femora dark brown, fore tibiae medium brown, tarsi light brown; midand hind legs with femora medium brown, dark brown distally, tibiae and tarsi medium brown. Femur/tib- alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 128 Habrophlebia callensis Sartori, Vuataz & Samraoui, sp. nov. https://zoobank.org/BB3DBAD0-64A9-4E23-B7DC-D00F91A3D31C Figs 12–16 Habrophlebia spH, Samraoui et al. 2021b; Habrophlebia sp. 7, Hezil et al. 2025. Material examined. Holotype: one nymph in ethanol (GBIFCH00902495), Algeria, Wilaya of El Tarf, Hadada, Dar Essalem; 36°53'10"N, 8°37'2"E, 201 m, 11.VII.2019, B. Samraoui leg., MZL. Paratypes. Algeria, Wilaya of El Tarf, Dar Essalem, same data as holotype, 2 nymphs in ethanol (GBIFCH00829846); same locality as holotype, 1.VIII.2019, 1 nymph on slide (GBIFCH00763554), B. Samraoui leg.; same locality as holotype, 18.IV.2019, 12 nymphs in ethanol (GBIFCH00829847), B. Samraoui leg., same locality as holotype, 2.V.2019, 4 nymphs in ethanol (GBIFCH00829845), B. Samraoui leg., same locality as holotype, 25.VI.2019, 1 nymph in ethanol (GBIFCH00829842), B. Samraoui leg.; same locality as holotype, 23.I.2020, 1 nymph in ethanol (GBIFCH00832063), B. Samraoui leg., MZL • Wilaya of El Tarf, Hadada, Chaaba Waera, 36°53'37"N, 8°36'27"E, 113 m, 2.V.2019, 12 nymphs in ethanol (GBIFCH00829837, GBIFCH00829836), B. Samraoui leg., same locality, 9.III.2019, 13 nymphs in ethanol, (GBIFCH00829838), B. Samraoui leg., MZL. • Wilaya of El Tarf, Hadada, Oued Ftitich; 36°54'01"N, 8°37'05"E, 165 m, 18.IV.2019, 6 nymphs in ethanol (GBIFCH00672314), 1 nymph on slide (GBIFCH01231272), B. Samraoui leg.; same locality, 11.VII.2019, 1 nymph on slide (GBIFCH00763555), B. Samraoui leg.; same locality, 23.I.2020, 4 nymphs in ethanol (GBIFCH00832011), B. Samraoui leg.; same locality, 12.III.2020, 2 nymphs in ethanol (GBIFCH00654972), 1 nymph on slide (GBIFCH01118444), B. Samraoui leg.; same locality, 18.V.2022, 2 females (GBIFCH00654974), B. Samraoui leg.; same locality, 25.V.2022, 1 male (GBIFCH01314443), B. Samraoui leg.; same locality, 1.VI.2022, 1 female (GBIFCH00654975), B. Samraoui leg., all MZL. Etymology. The name of El Kala derives from the Arabic al-Qalʿa (“the fortress”), a reference to its strategic role through history. During the French period it was known as La Calle, while in antiquity it was recorded as Thinisa in Numidia. Descriptions. Nymph. Body length of final instar, excluding caudal filaments, up to 6 mm for male and up to 7 mm for female. Cerci as long as body length. Coloration. General coloration medium brown; dark brown between ocelli, whitish in front of the eyes, light brown in front and on the clypeus. Upper portion of male eyes light brown to orange. Antenna with pedicel greyish brown, scape light brown and filament yellowish. Proand mesonotum dark brown, with light brown maculae, on medium and lateral margins; lateroparapsidal sutures dark brown. Legs light to medium brown; dorsal surface of fore femora dark brown, fore tibiae greyish brown; midand hind legs light brown to yellowish, except at apex of femora grayish brown; tarsi light brown to yellowish. Abdominal tergites medium brown with three pairs of lighter maculae, especially well visible on segments I–VI (Fig. 12A); posterolateral angles and posterior margin of tergites I–VIII dark brown. Sternites medium brown to greyish brown, nervous ganglia well visible (Fig. 12B). Cerci and paracercus medium brown at base, lighter distally, dark banding slightly visible every two segments proximally. Head. Labrum rectangular, ca. 1.85× wider than long (Fig. 12C); dorsal surface covered distally with scattered stout setae; anterior margin with a row of stout, long and spatulate setae medially; emargination narrow, U-shaped with flat denticles; ventral surface with two bunches of stout setae medially. Mandibles similar to other Habrophlebia species, incisor with 3 teeth, the lowest of the left mandible with a clear indentation (Fig. 12D), kinetodontium with 3 teeth, 14–15 long and thin setae below mola of right mandible (Fig. 12E). Maxilla stocky, subapical row of 5 or 6 pectinate setae (Fig. 12F); maxillary palp with three segments, segment 1 and 2 subequal in length, and longer than segment 3; segment 3 triangular 1.51– 1.71× longer than wide at base; all setae on palp segments stout and entire. Hypopharynx with highly developed superlinguae terminated by a membranous digitation (Fig. 12G). Labium with rhomboid glossae, outer margin and apex covered by short, broad setae; paraglossae enlarged laterally, covered with fine and long setae on the dorsal surface; with long, thick setae on the outer margin; labial palp with three segments, inner margin of segment 1 highly dilated near the middle, about 1.15–1.20× longer than the maximum width, segments 2 0.85× length of segment 1, segment 3, ca. 0.75× length of segment 1; segment 3 ca. 1.7–1.8× longer than wide at base, conical shape and with 5–7 stout and long, simple setae on dorsal surface. Submentum with numerous long, stout and pointed setae laterally and on ventral surface (Fig. 12H). Thorax. Pronotum with anterior corners bearing numerous long and pointed setae laterally, and dorsally. Anterior margin of pronotum with a row of stout setae (Fig. 13A). Fore legs femora stocky, ca. 2.1× longer than wide, outer margin and upper surface covered with long, entire and pointed setae (Fig. 13B); fore tibiae slightly shorter than femora, outer margin with thin and long setae, inner margin with long stout and feathered or entire setae especially near apex; tarsi 0.55× length of tibiae, outer margin with long and thin setae, inner margin with long and pointed entire setae. Middle legs similar to fore legs, femora ca. 2.5× longer than wide, dorsal surface of femora with numerous and long, stout and pointed setae; tibiae and femora of subequal length; tarsi 0.5× length of tibiae. Hind legs with femora 2.8–3.0× longer than wide (Fig. 13C), dorsal surface covered with stout, long, pointed and entire setae (Fig. 13D); hind tibiae as long as hind femora, outer margin with a row of stout, pointed setae and few thin and long setae; inner margin with stout, pointed and entire Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 129 Figure 12. H. callensis sp. nov., habitus of female nymph in dorsal (A) and ventral position (B); labrum (C); left mandible (D); right mandible (E); maxilla (F); hypopharynx, left half (G); labium and submentum (H). Scale bars: 1 mm (A); 50 µm (B, E, F); 100 µm (C, D, G). setae (Fig. 13E); tarsi 0.4× length of tibiae, outer margin with few long and thin setae, inner margin with long, stout, pointed and entire setae. Claws of all legs slightly hooked, with 17–21 denticles subequal in size (Fig. 13F). Abdomen. Posterolateral expansions only on segments VIII and IX. Posterior margin of tergite IX with triangular, broad, pointed spines, ca. 1.5–2× longer than wide at base (Fig. 14A); tergite VIII with triangular, narrow alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 130 Figure 13. H. callensis sp. nov., thoracic structures, pronotum, right half (A); fore femur (B); hind femur (C); setae on upper face of hind femur (D); hind tibia (E); claw (F). Scale bars: 100 µm (A–C, E); 20 µm (D); 50 µm (F). spines 2× longer than wide (Fig. 14B); from tergite VII to tergite V, spines becoming smaller, needle-shaped (Fig. 14C–E); tergites I–IV with barely visible spines. Gills present on segments I–VII; all gills elongated with long filaments; dorsal lamella bearing 7–8 filaments, ventral lamella with 4–5 filaments (Fig. 14F). Imagos. Male imago in poor condition, only midand hind legs and abdomen preserved. Mid and hind legs coloration as in nymphs, whitish with apex of femur brownish. Femur/tibia/tarsi ratio in mid leg: 1/1.3/0.06/0.05/0.05/0.1; hind leg: 1/1.3/0.1/0.08/0.06/0.1. Midand hind claws dissimilar, one paddle-like and one hooked. Abdomen. Coloration similar to male nymphs. Styliger plate medium brown, dark brown along the margins, first segment of the gonopods medium brown, segments 2 and 3 yellowish brown. Posterior margin of the styliger plate concave, median incision U-shaped (Fig. 15C); segment 1 slightly longer than segments 2 and 3 combined; inner margin of segment 1 with a broad base, and with bulge on the outer margin. Penis lobes rounded and tight together, ventral spine long, thin and slightly curved outwards, reaching the base of the styliger plate (Fig. 15D). Cerci and terminal filament broken. Female imago: body length: 4.0–4.5 mm; forewing length: 4.5–5.0 mm. Forewing transparent, slightly tinted in brown in subcostal field; longitudinal and transversal veins dark brown; pterostigmatic area with ca. 10 oblique and simple transversal veins. MA and MP forks asymmetrical, cubital field with two long and three short intercalary veins (Fig. 15A). Hindwing with rounded costal process approx- Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 131 Figure 14. H. callensis sp. nov., abdominal structures, posterior margin of tergite IX (A); tergite VIII (B); tergite VII (C); tergite VI (D); tergite V (E); gill (F). Scale bar: 20 µm. imately in the middle of the wing; vein Sc long, reaching the costal margin almost at the tip of the wing (Fig. 15B). Eggs ovoid, ca. 200 µm × 90 µm, with longitudinal ribs non-punctuated, the majority running from one pole to the other (Fig. 16A); chorion covered with thin membranous filaments perpendicular to pole axis, sometimes longitudinal. Micropyle in equatorial area (Fig. 16B). Habrophlebia annaba Sartori, Vuataz & Samraoui, sp. nov. https://zoobank.org/FF8685EE-013E-450A-AA5A-83B2D2C470B1 Figs 17–21 Habrophlebia sp. 9, Hezil et al. 2025. Material examined. Holotype: one nymph in ethanol (GBIFCH00654946), Algeria, Wilaya of Annaba, Edough, Oued Agab, 36°54'44"N, 7°39'35"E, 750 m, 5.VI.2021, B. Samraoui leg., MZL. Paratypes. Algeria, Wilaya of Annaba, Edough, Oued Agab, same data as holotype, 1 nymph in ethanol (GBIFCH00654980); same locality as holotype, 26.IV.2021, 1 nymph on slide (GBIFCH01314689); 4.V.2022, 1 female imago in ethanol (GBIFCH00654978); 16.V.2021, 1 nymph, 1 female imago in ethanol (GBIFCH00654977); 1.V.2021, 1 male imago in ethanol (GBIFCH01118440); 5.VI.2021, 1 nymph on slide (GBIFCH01314677); 10.II.2022, 1 nymph in ethanol (GBIFCH00654976), 1 nymph on slide (GBIFCH01314437); 1.VI.2022, 1 nymph on slide (GBIFCH00658258) • Wilaya of Annaba, Edough, Oued Zouz, 36°55'54"N, 7°40'03"E, 550 m, 20.III.2022, 15 nymphs in ethanol (GBIFCH00654982), 1 nymph on slide (GBIFCH01118439); same locality, 25.IV.2022, 1 female imago in ethanol (GBIFCH00654983); all B. Samraoui leg., MZL. Etymology. The name Annaba derives from the Arabic ʿannāb (jujube), in reference to the jujube tree (Ziziphus jujuba), once abundant in the region. In antiquity, the city was known as Hippo Regius, a major centre of Roman alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 132 Figure 15. H. callensis sp. nov., cubital field of male imago (A); hindwing of male imago (B); genitalia in ventral view (C); penis in lateral view (D). Abbreviations: CuA, cubital anterior vein; CuP, cubital posterior vein. North Africa and the episcopal See of Saint Augustine (354–430 CE). Later, it was renamed Bouna in Arabic and was known as Bône during the French period. Descriptions. Nymph. Body length of final instar, excluding caudal filaments, up to 6 mm for male and up to 7 mm for female. Cerci as long as body length. Coloration. General coloration medium brown; dark brown between ocelli, light brown in front and on the clypeus, especially in young nymphs. Upper portion of male eyes red-brick to reddish brown (Fig. 17B). Antenna with pedicel dark brown, scape greyish brown and filament light brown, more or less entirely whitish in young nymphs. Pronotum medium brown, washed with dark brown in middle and anteriorly, lateral margins whitish; mesonotum medium brown, with dark brown maculae; lateroparapsidal sutures dark brown. Legs light to medium brown; femora dark brown in mature nymphs; tibiae medium brown; tarsi light brown to yellowish. Abdominal tergites dark brown to greyish brown, with two lighter maculae medio-anteriorly, especially well visible on segments I–VII (Fig. 17A). Sternites medium brown in the middle, posterior corner underlined with dark brown laterally, thicker in mature nymphs, nervous ganglia well visible. Cerci and paracercus light brown at base, whitish distally, without dark banding. Head. Labrum rectangular, ca. 1.55× wider than long (Fig. 17C); dorsal surface covered distally with scattered stout setae; anterior margin with a row of stout, long and spatulate setae medially; emargination flared, U-shaped with flat denticles; ventral surface with two bunches of stout setae medially. Mandibles similar to other Habrophlebia species, incisor with 3 teeth, the lowest of left mandible with a clear indentation at base (Fig. 17D), kinetodontium with 3 teeth, 10–11 long and thin setae below mola of right mandible (Fig. 17E). Maxilla stocky, subapical row of 7 or 8 pectinate setae (Fig. 17F); maxillary palp with three segments, segment 1 and 2 subequal in length, and longer than segment 3; segment 3 trian- Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 133 Figure 16. H. callensis sp. nov., egg in toto (A); micropyle and details of the chorion (B). gular 1.42–1.70× longer than wide at base; all setae on palp segments stout and entire. Hypopharynx with highly developed superlinguae terminated by a membranous digitation. Labium with rhomboid glossae, outer margin and apex covered by short, broad setae; paraglossae enlarged laterally, covered with fine and long setae on the dorsal surface; with long, thick setae on the outer margin; labial palp with three segments, inner margin of segment 1 highly dilated near the middle, about 1.35× longer than the maximum width, segments 2 slightly longer than segment 3, both ca. 0.65–0.70× length of segment 1; segment 3 ca. 1.41–1.54× longer than wide at base, conical shape and with 8–10 stout and long, simple setae on dorsal surface. Submentum with numerous long, stout and pointed setae laterally; ventral surface with numerous long, stout and pointed setae (Fig. 17G). Thorax. Pronotum with anterior corners bearing few long and pointed setae laterally, and 5–7 stout and strong setae dorsally. Anterior margin of pronotum with numerous stout setae (Fig. 18A). Fore legs femora elongated, ca. 2.45× longer than wide, outer margin and upper surface covered with long, entire and pointed setae (Fig. 18B); fore tibiae subequal in length to femora, outer margin with few thin and long setae, inner margin with several rows of long stout and entire setae; tarsi 0.5× length of tibiae, outer margin with few long and thin setae, inner margin with long and pointed entire setae. Middle legs similar to fore legs, femora ca. 2.6× longer than wide, dorsal surface of femora with more numerous and slightly longer stout and pointed setae; tibiae and femora of subequal length; tarsi 0.5× length of tibiae. Hind legs with femora 2.8× longer than wide (Fig. 18C), dorsal surface covered with stout, long, pointed or blunt and entire (non-feathered) setae (Fig. 18D); hind tibiae as long as hind femora, outer margin with numerous stout, pointed setae; inner margin with stout, pointed and entire setae; tarsi 0.5× length of tibiae, outer margin with long and thin setae, inner margin with long, stout, pointed and entire setae. Claws of all legs slightly hooked, with 15–17 denticles that decrease in size from the apex to the tarsus (Fig. 18E). Abdomen. Posterolateral expansions only on segments VIII and IX. Posterior margin of tergite IX with triangular, broad, pointed spines, ca. 1.5–2× longer than wide at base (Fig. 19A); tergite VIII with triangular, broad spines 1.5–2× longer than wide (Fig. 19B); tergite VII with slightly shorter spines than tergite VIII (Fig. 19C); tergite VI, with short and broad spines (Fig. 19D), and tergite V with minute but broad spines (Fig. 19E); tergites I–IV with barely visible spines. Gills present on segments I– VII; all gills elongated with long filaments; dorsal lamella bearing 8–10 filaments, ventral lamella with 3–6 filaments (Fig. 19F). Imagos. Male imago. Size: body length: 5.5–6.0 mm; forewings missing; cerci and terminal filament broken. Head dark brown; basal portion of compound eyes blackish, upper portion orange brown, scape and pedicel dark brown, flagellum broken. Thorax. Pronotum dark brown; mesoand metanotum uniformly dark brown, pleurae, coxae, and trochanters greyish brown, washed with dark brown; fore femora and fore tibiae dark brown, tarsi light brown, apex of each segment dark brown; midand hind legs with femora light brown, dark brown distally, tibiae medium brown, dark brown at base, and tarsi medium brown, except the last segment light brown. Femur/tibia/tarsi ratio in fore leg: 1/1.4/0.5/0.5/0.3/0.2; mid leg: 1/1.2/0.1/0.07/0.05/0.1; hind leg: 1/1.2/0.1/0.05/0.05/0.15. Fore claws similar, paddle-shaped, midand hind claws dissimilar, one pad- alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 134 Figure 17. H. annaba sp. nov., habitus of mature female nymph (above) and younger nymph (below) (A); habitus of mature male nymph (above) and younger nymph (below) (B); labrum (C); left mandible (D); right mandible (E); maxilla (F); labium and submentum (G). Scale bars: 1 mm (A, B); 50 µm (C, G); 100 µm (D, E, F). dle-like and one hooked. Hindwing with rounded costal process approximately in the middle of the wing; vein Sc long and well separated from the costal vein, reaching the costal margin almost at the tip of the wing (Fig. 20C). Abdomen. Tergites and sternites medium brown, posterior margin of each segment dark brown. Styliger plate medium brown, dark brown along the margins, gonopods broken. Posterior margin of the styliger plate straight and clearly protruding in the middle, median incision U-shaped (Fig. 20D). Penis lobes rounded and tight together, ventral spine long, thin and straight, almost reaching the base of the styliger plate. Cerci and terminal filament broken. Female imago. Size: body length: 6.5–7.0 mm; forewing length: 7.0–7.5 mm. Forewing transparent, slightly Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 135 Figure 18. H. annaba sp. nov., thoracic structures, pronotum, right half (A); fore femur (B); hind femur (C); setae on upper face of hind femur (D); claw (E). Scale bars: 100 µm (A–C); 20 µm (D); 50 µm (E). tinted in brown in subcostal field; longitudinal and transversal veins dark brown; pterostigmatic area with ca. 10 oblique and simple transversal veins. MA and MP forks asymmetrical, cubital field with two long and two short intercalary veins (Fig. 20A). Hindwing with rounded costal process approximately in the middle of the wing; vein Sc long and close to the costal vein, reaching the costal margin before the tip of the wing (Fig. 20B). Eggs ovoid, ca. 180–195 µm × 80–100 µm, with longitudinal ribs non-punctuated, long and entire, most of them running from one pole to the other (Fig. 21A). Ribs relatively broad, ca. 4.5–5.0 µm in width. In some specimens, chorion folded. Micropyle in equatorial area (Fig. 21B). Habrophlebia edough Sartori, Vuataz & Samraoui, sp. nov. https://zoobank.org/9536F790-E5EF-4844-95A1-D3182BBB7434 Figs 22–25 Habrophlebia sp. 17, Hezil et al. 2025. Material examined. Holotype: one nymph in ethanol (GBIFCH00654973), Algeria, Wilaya of Annaba, Edough, Oued Agab, 36°54'44"N, 7°39'35"E, 750 m, 9.IX.2021, B. Samraoui leg., MZL. Paratypes. Algeria, Wilaya of Annaba, Edough, Oued Agab, same data as holotype, 2 nymphs in etha- alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 136 Figure 19. H. annaba sp. nov., abdominal structures, posterior margin of tergite IX (A); tergite VIII (B); tergite VII (C); tergite VI (D); tergite V (E); gill (F). Scale bar: 20 µm. nol (GBIFCH00654981), 1 nymph on slide (GBIFCH00654947); same locality as holotype, 18.VIII.2021, 2 nymphs in ethanol (GBIFCH00654979), 2 nymphs on slide (GBIFCH00654948, GBIFCH01118441). Etymology. The name Edough (Djebel Edough), designating the prominent mountain massif overlooking the city of Annaba, is thought to derive from undocumented Berber (Amazigh) roots. In antiquity, it was referred to as Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 137 Figure 20. H. annaba sp. nov., cubital field of female imago (A); hindwing of female imago (B); hindwing of male imago (C); styliger plate in ventral view (D). Figure 21. H. annaba sp. nov., egg in toto (A); micropyle and details of the chorion (B). Mons Pappua by Procopius who describes it as a Numidian stronghold, and as Mons Giddaba by Saint Augustine. Descriptions. Nymph. Body length of final instar, excluding caudal filaments, up to 7 mm for female; male nymphs unknown. Cerci as long as body length. Coloration. General coloration medium brown; dark brown between ocelli, light brown in front and on the clypeus (Fig. 22A), especially in young nymphs (Fig. 22B). Antenna with pedicel dark brown, scape greyish brown and filament light brown, more or less entirely alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 144 Figure 26. Habrophlebia spp. H. consiglioi (A, C, F) and H. eldae (B, D, E); labrum (A, B); pronotum, right half (C, D); hypopharynx, left half (E, F). Scale bars: 100 µm (A); 50 µm (B–F). genetic Euro-Mediterranean lineages are reported here (Fig. 27). To date, 14 species are known exclusively from the Maghreb, indicating that this region constitutes a major centre of endemism and a potential focus of diversification for the genus. Each Maghrebian Habrophlebia species appears to be geographically restricted: H. annaba sp. nov. and H. edough sp. nov. occur in the Edough Mountains; H. callensis sp. nov. in the El Kala northern hills; H. ghora sp. nov. in the Kroumiria Mountains spanning Algeria and Tunisia; H. seybouse sp. nov. in the Seybouse Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 145 Figure 27. Bayesian (BEAST) maximum clade credibility COI tree of Habrophlebia in the West Palearctic. Branch tips labelled with GBIF codes indicate newly sequenced specimens; all other codes correspond to sequences from El Alami et al. (2025). Outer color ranges represent CO1 clades, delimited either following El Alami et al. (2025) or by morphological characters for the five newly described species (green: Algeria; red: Morocco; blue: Europe). Species names (where available) are placed adjacent to their respective clades, with the newly described species highlighted in bold. The three inner brown rings indicate MOTUs inferred from the ASAP, mPTP, and GMYC delimitation methods. Circles on branches denote Bayesian posterior probabilities > 0.9. Outgroup branches are shown in grey. alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 146 and Collo regions; H. hassainae in north-western Algeria (Benhadji et al. 2018); H. djurdjurensis in the Djurdjura Mountains (Kechemir et al. 2020); H. vaillantorum in the High Atlas, southern Morocco (Thomas et al. 1999); H. dakkii in northern Morocco (El Alami et al. 2023); and H. abietis and H. linae in the Rif region, northern Morocco (El Alami et al. 2025). In contrast to what was observed in the Rif region of Morocco, the Algerian species do not exhibit marked phenotypic heterogeneity, which in the Rif had suggested the occurrence of hybridisation between species pairs (H. dakkii–H. abietis, H. dakkii–H. linae, H. abietis–H. linae). These observations were further supported by the topology of the COI gene tree, in which all three Habrophlebia species from the Rif region, together with their morphologically intermediate forms, were grouped within a single clade (El Alami et al. 2025). The concentration of endemism in the Maghreb is not confined to mayflies (Samraoui et al. 2021c; Kaltenbach et al. 2022; Dambri et al. 2022, 2023) but extends to diverse freshwater taxa, including Odonata (Ferreira et al. 2014; Bouhala et al. 2019), caddisflies (Bemmoussat-Dekkak et al. 2021; Samraoui et al. 2024), stoneflies (Yasri-Cheboubi et al. 2013, 2016), amphipods (Ayati et al. 2019), and crabs (Rouabhi et al. 2025). It also encompasses vertebrates such as the Maghreb bleak Tropidophoxinellus callensis (Guichenot, 1850), the ribbed newt Pleurodeles nebulosus (Guichenot, 1850), and the tree frog Hyla carthaginiensis Dufresnes et al., 2019. In northeastern Algeria, wetlands function as refuges for Afrotropical relicts (Samraoui et al. 1993) and as key wintering and breeding sites for waterbirds (Samraoui and Samraoui 2008), highlighting the region’s global significance for freshwater biodiversity. The pronounced endemism of Maghrebian biota, shaped by complex topography and long-term climatic fluctuations, has produced a characteristic checkerboard distribution (Husemann et al. 2014; Benslama et al. 2010). Interactions between orography, geodynamics, and climate have further promoted microendemism and local radiations through habitat fragmentation and limited dispersal (Veith et al. 2004; Neubauer et al. 2015). Most Habrophlebia species in the Maghreb exhibit highly restricted ranges and can be regarded as true microendemics. They are typically associated with headwater streams at elevations of 200–900 m, usually characterised by moderate current velocity. The remarkable diversity of the genus in this region is likely linked to its narrow ecological tolerance (stenoecy) and limited dispersal capacity relative to other mayfly lineages (Peredo Arce et al. 2021). Adult mayflies generally live only a few hours to days and possess weak flight capacity, restricting their movements primarily to local or catchment scales, although occasional long-distance dispersal events have been documented (Monaghan et al. 2005; Vuataz et al. 2013; Rutschmann et al. 2014). These dispersal limitations enhance the potential for genetic and phenotypic divergence, particularly in fragmented habitats where geographic isolation fosters allopatric speciation within montane refugia (Hewitt 2004). Consequently, Habrophlebia species may serve as reliable bioindicators of relict mountain streams that have remained largely unaffected by human disturbance (Samraoui et al. 2021b). Their persistence in such isolated habitats points to longterm ecological stability and underscores the importance of montane refugia in sustaining freshwater biodiversity across the Maghreb. During the Quaternary glacial periods, the Maghreb served as an important, though long overlooked, refugium for Western Palaearctic biota, with the narrow straits of Gibraltar and Sicily acting as key biogeographical corridors between North Africa and Europe (Husemann et al. 2014). Evidence from diverse taxa, including colubrid snakes (Carranza et al. 2006), freshwater turtles (Fritz et al. 2006), and nymphalid butterflies (Habel et al. 2011, 2017), indicates the presence of two or more glacial refugia within the region. Quaternary glacial and interglacial oscillations likely drove pronounced east to west divergence and regional disjunctions, notably across Kabylie, the Aures, and the Rif, shaping the phylogeographical structure of Maghrebian biota (Beddek et al. 2018; Dufresnes et al. 2019; Machado et al. 2021). Earlier, during the Miocene, alternating marine transgressions and tectonic reorganisation of the Tell and Rif belts, including the development of Numidian Flysch basins and uplift of massifs such as Edough, fragmented freshwater habitats and isolated upland streams (Caby et al. 2001; Thomas et al. 2010; Ben Khalifa et al. 2019). Superimposed Pleistocene humidity and aridity cycles (Gasse and van Campo 1994; Zielhofer et al. 2004) further modified connectivity, alternately promoting dispersal and secondary contact before reinstating isolation, a process recognised as a major driver of diversification in North African freshwater biotas (Habel et al. 2008). Marine incursions may have been particularly influential in shaping the biota of the Edough Mountains, which rise above the Seybouse and Guerbes Senhadja plains and the Fetzara marsh. The massif is a recognised hotspot of Mediterranean North African plant endemism (Véla and Benhouhou 2007), supporting numerous endemic vascular plants (Hamel et al. 2013), the threatened ribbed newt Pleurodeles poireti (Carranza and Wade 2004; Veith et al. 2004; Samraoui et al. 2012), several endemic caddisflies (Samraoui et al. 2024), the springtail genus Edoughnura (Deharveng et al. 2007), and several locally restricted Collembola species (Brahim-Bounab et al. 2020). The Messinian Salinity Crisis, approximately 5.97 to 5.33 million years ago, was marked by the near-complete desiccation of the Mediterranean followed by rapid refilling (Duggen et al. 2003; Rouchy and Caruso 2006; Roveri et al. 2014). The temporary emergence of terrestrial corridors during this period may have enabled range expansions from North Africa to southern Europe (Carranza et al. 2006; Habel et al. 2010; Gattolliat et al. 2015). However, biogeographical links between the Maghreb and southern Europe remain poorly understood. Alpine Entomology 9 2025, 113–151 alpineentomology.pensoft.net 147 The distribution of Habrophlebia species is consistent with the Maghrebian east–west divide, which separates Algerian and Moroccan lineages across several taxa, including the odonates Gomphus lucasi and G. simillimus, mayflies of the genus Centroptilum (Kaltenbach et al. 2022) and Rhithrogena (Zrelli et al. 2011b; Samraoui et al. 2021c; El Alami et al. 2022b), and ribbed newts of the genus Pleurodeles (Veith et al. 2004). The concentration of Habrophlebia diversity in the Maghreb supports the hypothesis that European taxa may have originated from Maghrebian ancestors dispersing northwards across the Strait of Sicily. Alternatively, a reverse colonisation scenario is plausible, with an ancestral lineage entering the Maghreb from Europe and subsequently diversifying in isolation. Such contrasting biogeographical patterns, involving dispersal and in situ diversification, are well documented in both island and continental systems, exemplified by the adaptive radiation of Darwin’s finches in the Galápagos (Grant and Grant 2008). Comparable processes have been reported in New Guinea, where Miocene and Pliocene orogeny triggered extensive arthropod diversification through habitat fragmentation and ecological opportunity (Toussaint et al. 2014). By analogy, the complex topography and climatic oscillations of the Maghreb, combined with intermittent connectivity across the Mediterranean during low sea-level stands, may have facilitated alternating phases of isolation and gene flow, ultimately promoting the diversification of Habrophlebia across this region. In this scenario, the Strait of Sicily, much like the Strait of Gibraltar, would have acted as a biogeographic corridor between North Africa and Europe, with glacial periods and associated sea-level fluctuations providing repeated opportunities for faunal exchange (Husemann et al. 2014). Within Algeria, an east–west disjunction is suggested by the clade H. djurdjurensis and H. hassainae geographically isolated from their sister clade, comprising H. callensis sp. nov. and H. annaba sp. nov. This pattern may reflect broader biogeographic structuring across the Maghreb, where recurrent Pleistocene climatic oscillations, marine transgressions, and habitat discontinuities promoted population isolation and lineage divergence. In this context, the Edough, Numidian (Mount Ghora and Kroumiria), and Kabylie massifs appear to have acted as important refugial zones in eastern Algeria, while the Rif and Middle Atlas fulfilled a similar role in the west, together generating a recurrent east–west phylogeographic break across multiple taxa. Not all Habrophlebia species exhibit restricted ranges. H. ghora sp. nov. and H. seybouse sp. nov. both display relatively wide distributions, similar to other recently described Maghrebian taxa that also occupy broad areas. For example, H. djurdjurensis in Algeria is widespread in streams of the Djurdjura Mountains (Kechemir et al. 2020; Labdaoui et al. 2024), and H. dakkii in northern Morocco, although endemic to the Rif Mountains, is distributed across much of this range (El Alami et al. 2025). Such cases indicate that dispersal is possible when environmental and geographical conditions are favourable. Factors that may promote wider distributions include the presence of continuous or closely spaced headwater networks, riparian corridors that maintain ecological connectivity, and regional hydrological stability, which reduces the likelihood of local extinctions (Bilton et al. 2001; Finn et al. 2011; Sarremejane et al. 2021). These examples suggest that, despite generally limited dispersal capabilities, certain Habrophlebia species can expand beyond isolated refugia when the landscape offers suitable pathways and habitat continuity. In the Collo–Seybouse Highlands and Mount Ghora (Kroumiria) sector, which is inhabited by two closely related species (H. seybouse sp. nov. and H. ghora sp. nov.), potential dispersal pathways include the Oued Seybouse corridor and the intervening hill ranges, such as the Monts of Nechmaya, which may intermittently connect headwater systems. The continuity of such routes is likely influenced by historical climate oscillations, with wetter periods enhancing riparian connectivity and drier phases promoting isolation. This pathway also appears to have been utilised by another, as yet undescribed, mayfly species, Ecdyonurus sp. C (Hezil et al. 2025), suggesting that it may function as a recurrent conduit for faunal exchange across the region. Similarly, the close relationship between H. annaba sp. nov. and H. callensis sp. nov. may be attributed to the extensive wetland complex located between Mount Edough and northern El Kala, which is likely to have provided both ecological connectivity and dispersal pathways that subsequently facilitated speciation. In addition to their underappreciated biogeographical significance, which provides valuable insights into how past climatic oscillations have shaped regional and extra-regional biota (Carrington et al. 2001), North African wetlands deliver essential ecosystem services (Boulton et al. 2016). Yet, they are increasingly threatened by escalating anthropogenic pressures, placing a substantial proportion of Maghrebian freshwater species at risk of extinction (García et al. 2010; Benslimane et al. 2019). A major obstacle to effective conservation is the limited public and institutional recognition of the fundamental role that biodiversity plays in maintaining ecological integrity and supporting human well-being (Aouadi et al. 2022; Samraoui et al. 2025). Bridging this awareness gap is critical for promoting sustainable management and ensuring the long-term resilience of these ecosystems. Acknowledgements We are grateful to M. El Alami, R. Godunko, and J.-L. Gattolliat for their valuable comments and suggestions. We sincerely thank Farrah Samraoui for her constant support and assistance throughout the project. We also thank Sonia Zrelli (Bizerte, Tunisia) for providing access to material, and Céline Stoffel (MZL) for her invaluable help with molecular laboratory work. Scanning electron microscopy was carried out at the Electron Microscopy Facility of the University of Lausanne, with the kind assistance of Antonio Mucciolo. alpineentomology.pensoft.net Samraoui, B. et al.: Diversity and endemism of Habrophlebia Eaton, 1881 148 References Aouadi A, Samraoui F, Talbi A, Souiki L, Samraoui B (2022) Socio-demographic correlates of biodiversity perception: The need for environmental education. Journal of Contemporary Trends and Issues in Education 1: 19–51. https://doi.org/10.55628/jctie.v2i1.15 Ayati K, Hadjab R, Khammar H, Dhaaouadi S, Piscart C, Mahmoudi E (2019) Origin, diversity, and distribution of freshwater epigean amphipods in Maghreb. 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