Expanding the range: First record and redescription of the male of Limnephilus petri Marinković Gospodnetić, 1966 in R. North Macedonia

© 2025 The Author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Expanding the range: First record and redescription of the male of Limnephilus petri Marinković Gospodnetić, 1966 in R. North Macedonia JELENA HINIĆ-JORDANOVSKA1, VALENTINA SLAVEVSKA-STAMENKOVIĆ1, HALIL IBRAHIMI2, MLADEN KUČINIĆ3, ASTRIT BILALLI4, MAJA JORDANOVA1 & AJTEN RUSHITI-DAUTI5 1 Institute of Biology, Faculty of Natural Sciences and Mathematics,“Ss. Cyril and Methodius University in Skopje” Arhimedova 3, 1000 Skopje, Republic of North Macedonia. E-mails: [email protected]; [email protected]; [email protected] https://orcid.org/0000-0002-6194-614; https://orcid.org/0000-0001-9031-5249; https://orcid.org/0000-0002-6272-0738 2 Department of Biology, Faculty of Mathematical and Natural Sciences, University of Prishtina, Mother Theresa Street p.n.,10000 Prishtina, Republic of Kosovo. E-mail: [email protected]; https://orcid.org/0000-0002-4301-4387 3 Department of Biology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10000 Zagreb, Croatia. E-mail: [email protected]; https://orcid.org/0000-0003-2756-2305 4 Faculty of Agribusiness, Haxhi Zeka University of Peja, UÇK Street, 30000 Pejë, Republic of Kosovo. E-mail: [email protected]; https://orcid.org/0000-0003-2820-8009 5 Department of Biology, Faculty of Mathematics and Natural Sciences, University of Tetovo, R. North Macedonia. E-mail: [email protected]; https://orcid.org/0009-0000-7142-4087 *Corresponding author. E-mail: [email protected] Received 16 March 2025 │ Accepted by V. Pešić: 31 March 2025 │ Published online 10 April 2025. Abstract Limnephilus petri, an endemic caddisfly species, is reported for the first time from the Republic of North Macedonia. This discovery expands the known distribution of the species, which has previously been observed in limited geographic areas. In this study, the redescription of the male of L. petri is provided, including detailed morphological features that aid in the species’ identification. The findings contribute significantly to the understanding of the aquatic biodiversity of the region, emphasizing the ecological importance of the species. This study also underscores the importance of continued monitoring and conservation efforts to protect such rare and specialized species. Key words caddisflies, new records, rare species, distribution, taxonomy, Balkan Peninsula. Introduction The genus Limnephilus Leach, 1815 is the most species-rich group within the family Limnephilidae, comprising 223 living and 15 fossil species (Morse, 2025). Many species of this genus are known to Ecologica Montenegrina 84: 15-24 (2025) This journal is available online at: www.biotaxa.org/em https://dx.doi.org/10.37828/em.2025.84.2 LIMNEPHILUS PETRI IN NORTH MACEDONIA 16 have very restricted distribution, while some of them even represent regional endemics (Balibrea et al. 2017). Due to the wide spawn of different ecological demands, species from genus Limnephilus remain one of the most challenging and interesting groups of trichopterans, consistently drawing the attention of caddisfly researchers (Waringer et al. 2012; Oláh et al. 2019; Zang et al. 2022; Spahler 2024). Species of genus Limnephilus count for approximately 4% of the total caddisfly species in the West Palearctic region, where Republic of North Macedonia belongs (Ibrahimi, 2024). Detailed literature review showed that today, the genus Limnephilus in R. North Macedonia is represented with 11 species (Radovanović 1935; 1942; 1953; Botosaneanui 1960; Arsov 1991; Kovachev et al. 1999; Oláh, 2010; Oláh & Kovács, 2013; 2014; Bilalli et al. 2019; SlavevskaStamenković et al. 2021; Musliu et al. 2024). Due to the unique geomorphological features and climate elements (Milevski 2015), R. North Macedonia has a distinct blend of habitats offering a variety of environmental conditions able to meet the different ecological demands of many limnephilids (Zang et al. 2022). This part of the Balkan Peninsula probably hides even more Limnephilus species, yet to be found. The results presented in this paper aim to contribute to the knowledge of the caddisfly diversity in Republic of North Macedonia by presenting the first record of the rare endemic caddisfly species Limnephilus petri Marinković Gospodnetić, 1966. Considering that the original description of the species by Marinković Gospodnetić (1966) contains only the drawing of the lateral view of the genitalia that could easily lead to misidentifications, we redescribe the species offering a detailed description of the genital segments and providing non-genital characters for its identification. Material and methods Study area The Shar Mountain spans from southern Kosovo to northwestern R. North Macedonia and northeastern Albania and is characterized by rugged terrain and sparse vegetation. This mountain is known to be among the highest in the Balkans, with 30 peaks exceeding 2,500 meters above sea level (Jovanovska et al. 2020). The areas of the mountain within Kosovo and R. North Macedonia are designated national parks (Ibrahimi et al. 2023; Hinić - Jordanovska et al. 2024). The sampling site is located at a small mountain stream above the forest zone adjacent to Karanikolicko Lake (42.074913°N, 20.820664°E, 1798 m above sea level, Figure 1). The locality belongs to the 6th ecoregion (Hellenic Western Balkan) according to Illies (1978). Data sampling and processing Adult caddisfly specimens were collected with entomological nets. A single specimen of Limnephilus petri was collected on 01.08.2016. The collected material was preserved in 96% ethanol. Adult specimens were identified under a stereomicroscope using the identification keys of Kumanski (1988) and Malicky (2004). To provide photographs of the male genitalia, Limnephilus petri specimens were cleared using 10% KOH solution. The material is deposited at the Laboratory of Zoology of the Faculty of Natural and Mathematical Sciences, University of Prishtina, Kosovo. Drawings and microscopic photographs were completed by Prof. Halil Ibrahimi, PhD. Results The findings outlined in this paper provide a summary of the recent discovery of the rare endemic Limnephilus petri, noted in R. North Macedonia for the first time. Detailed redescription of the species is provided. Drawings of lateral, dorsal and caudal view of male genitalia of Limnephilus petri are given in Figure 2. Additionally, photo of male genitalia in lateral view is given in Figure 3. Material examined. Lectotype: R. North Macedonia, Shar Mountains, Karanikolicko Lake, 01. VIII. 2016, 1 male, leg. H. Ibrahimi. HINIĆ-JORDANOVSKA ET AL. Ecologica Montenegrina, 84, 2025, 15-24 17 Figure 1. Map of the sampling locality of Limnephilus petri in R. North Macedonia (red square represents the small mountain stream above the forest zone adjacent to Karanikolicko Lake). Male general description. After preserving the specimen in ethanol, antennae, maxillary palps, and legs are light brown in color, while the wings are whitish grey. Forewings without any spots; spur formula 1.3.4; maxillary palps 3-segmented; labial palps 5-segmented; three ocelli, one located between scapi, two of which located dorsally near each compound eye; head and thorax dorsal warts with dark brown setae; dorsally three dark brown setae near each ocellus; above both side two dark brown setae on wart; near scapi seven setae on each side; posterior warts of head with four setae each; paranotal warts with ten dark brown setae and seven pale brown setae on anterior and posterior edges each; left and right warts of mesonotum with twenty dark brown setae each. Forewing length 11–12.5 mm. Male genitalia. In lateral view, segment IX widest (longest) in the middle with narrow (short) dorsal and ventral portions. Preanal appendages (pa) 4/5 (3/4) as long as intermediate appendages (ia), oval, two times as long as tall (1/2 as tall as long), with apex pointed and directed caudad (Figure 2a). Intermediate appendages apically sclerotized, in lateral view long with distal part and apex curved caudad under obtuse angle (Figure 2a). Inferior appendages (inf) broad in lateral view with apex pointed and directed caudad (Figure 2a). In dorsal view, transverse posteromesal spinulose zone (sz) of tergite VIII 1/3 as long as broad, posterior edge sinuous, dorsomesally convex and laterally well rounded (Figure 2b). Phallic parameres (par) extending as long as phallicata apex, each with subapicodorsal row of spines and apicodorsal and ventral tuft of setae (Figure 2d). LIMNEPHILUS PETRI IN NORTH MACEDONIA 18 Figure 2. Drawings of male genitalia of Limnephilus petri a) lateral view; b) dorsal view; c) caudal view; d) parameres. (Abbreviations: sz - spinulose zone; ia - intermediate appendages; pa - preanal appendages; inf - inferior appendages; par - parameres). Discussion Separating Limnephilus petri from other species of the Limnephilus bipunctatus species group While thoroughly analyzing the Limnephilus bipunctatus species group in Turkey, Sipahiler (2024) provided detailed insights into the distinctive morphological characters between the species included. According to the structure of preanal and inferior appendages, it seems that Limnephilus petri is mostly similar to Limnephilus malickyi Sipahiler 1992. At the same time, according to the form of the spinulose zone on the eight abdominal segment, as well as the intermediate appendages, L. petri resembles Limnephilus bipunctatus Curtis 1834. Misidentification issues with L. malickyi cannot occur as it is a stenoendemic species found only on the highest part of the Taurus Mountains (Sipahiler 1992). Limnephilus petri can easily be distinguished from L. malickyi primarily by the shape of intermediate appendages, being more bulbous and with dorsal hump in lateral view in the later species. In addition to this, intermediate appendages in L. petri in dorsal view are broader on their basal half ending basally acuminate in ventral view, whereas in L. malicky they end broadly rounded basally in ventral view. On the other hand, Limnephilus bipunctatus is a part of the Macedonian caddisfly fauna (Radovanović 1953; HINIĆ-JORDANOVSKA ET AL. Ecologica Montenegrina, 84, 2025, 15-24 19 Oláh & Kovács 2013; Slavevska-Stamenković et al. 2021) and inhabits a wide range of habitats from small springs to large streams and lakes (Sipahiler 2024). Since it has already been reported from Shar Mountain (Radovanović 1953; Oláh & Kovács 2013) it is not excluded that its distribution patterns could overlap with those of L. petri. Limnephilus bipunctatus is also reported frequently from areas close to Republic of North Macedonia (Olah & Kovacs, 2013; Ibrahimi & Sejdiu, 2018; Ibrahimi et al. 2014, 2017, 2019). As there are significant similarities in the morphological structure of their male genitalia, the main diagnosable characters of these two species are the following: (1) In dorsal view, the transverse spinulose zone of the eight abdominal segment in both species is narrow (longitudinally short) and the posterior margin has a noticeable median lobe. However, the lateral lobes along the margin are noticeably rounder and more prominent in L. petri than in L. bipunctatus; (2) In lateral view, the apices of the preanal appendages are pointed caudad in L. petri, while in L. bipunctatus these appendages finish with a dull distal margin; (3) In dorsal view the intermediate appendages of L. bipunctatus contain darkly sclerotized inner edges. In lateral view, the inferior appendages of this species are evenly curved caudodorsad to acute apices. The intermediate appendages of L. petri are apically sclerotized, with distal part and apex curved caudad under obtuse angle. (4) In lateral view, the apices of the inferior appendages of L. petri are pointed caudad, while the inferior appendages of L. bipunctatus end with two rounded lobes posterodorsally. (5) Forewings of L. petri whitish grey, without any spots. Forewings of L. bipunctatus spotted and dark brown. Figure 3. a) Lateral view and b) dorsal view of male genitalia of Limnephilus petri. Ecological and distributional notes Limnephilus petri was originally described by Marinković Gospodnetić (1966; 1975) on specimens collected in 1954 from Brodska Reka River in Dragash Municipality (today in Kosovo). The entomological study conducted by Ibrahimi et al. (2015) resulted in recording L. petri near the type locality again more than a half century later. In the meantime, Kumanski (1988) identified some Limnephilus specimens from Rila Mountains in Bulgaria, Kopaonik Mountain in Serbia and Shar Mountain in R. North Macedonia only as Limnephilus cf. petri. Therefore, the presence of this species in Kopaonik, Rila and Shar mountains (R. North Macedonia) remained debatable. Bearing all this in mind, up until now, the only valid location of L. petri occurrence was its type locality. According to the investigations conducted on Shar Mountain in Kosovo (Ibrahimi et al. 2016a, 2016b; Ibrahimi & Vehapi, 2017; Ibrahimi et al. 2019; 2023; Karaouzas et al. 2018), this area is recognized for hosting a significant diversity of rare caddisfly species. However, these aquatic insects LIMNEPHILUS PETRI IN NORTH MACEDONIA 20 within the Macedonian part of this mountain remain insufficiently explored. Trichopterological research on the Macedonian territory of Shar Mountain has been inconsistent, with significant gap years between studies (Radovanović 1953; Kumanski, 1997; Kovachev et al. 1999; Mementi and Janeva 1999; Oláh and Kovác 2013; 2014; Oláh et al. 2017; 2022). Luckily, the entomological investigations on the caddisfly fauna on Shar Mountain were intensified over the last six years when caddisflies were sampled from mountainous streams, springs, rivers, wetlands and glacial lakes on different altitudes situated within and above the forest zone all over the mountain (Hinić - Jordanovska et al. 2024). Yet, L. petri was not recorded in any of these investigations, meaning that although it is present in Shar Mountain (R. North Macedonia) after all, its distribution area is restricted only to the vicinity of Karanikolicko Lake. The species is characterized by very limited flight period as the adult stage of its life cycle can be encountered only during the summer months July – August (Kumanski 1988). This observation perfectly corresponds to the time of the first finding of the species in the spring region of Brodska Reka river in July 1954 (Marinković Gospodnetić 1966; 1975) as well as to the results presented in Ibrahimi et al. (2015) when the species was recorded again around the type locality in July 2010. The collection of the species in August 2016 presented in this paper confirms the flight period of the species. Moreover, the fact that all three recorded occurrences of L. petri (Marinković-Gospodnetić 1966, 1975; Ibrahimi et al. 2015), including the one documented in this study, are from Shar Mountain, establishes this rare caddisfly as a steno-endemic species with a distribution restricted only to Shar Mountain. Based on the observations in this study, the species favors cold, slow-flowing mountain streams at high altitudes. A noteworthy detail is that since the entomological collection of Marinković-Gospodnetić containing all L. petri specimens including the holotype (Marinković-Gospodnetić 1966, 1975,) is no longer available, the newly collected specimen from Shar Mountain becomes the lectotype. Along with the specimens collected from Shar Mountain in Kosovo (Ibrahimi et al. 2015), this lectotype becomes the only available material of this species deposited in the Laboratory of Zoology of the Faculty of Natural and Mathematical Sciences, University of Prishtina, Kosovo. Bearing in mind that only pencil drawing illustrations of the lateral view of the genitalia were available (Marinković-Gospodnetić 1966), this paper provides the complete description of the species including photographs of lateral and dorsal view of male genitalia (Figure 3). Also, it employes the morphological characters of the forewings for separating L. petri from the Limnpehilus bipunctatus species group, since the use of non-genital morphological characters in evaluation of species delimitation was proven to be inevitable element when it comes to this group of aquatic insects (Sganga et al. 2022). Considering all aspects, uncovering new insights into rare species such as L. petri highlights the importance of aquatic biodiversity and entomological studies, particularly in regions that have not been extensively explored. Such efforts enhance conservation initiatives and management strategies, ensuring the protection of these crucial species. Specifically, in R. North Macedonia, there are ongoing efforts to compile updated National lists of strictly protected and protected wild species. While L. petri was already listed as a protected wild species (Official Gazette of the Republic of Macedonia no. 139/2011) although its presence in the country in the past was debatable, the scientific evidence presented in this paper represents the first reliable data supporting its inclusion on the list. Therefore, the first two authors of this paper incorporated L. petri in the updated version of the National Lists of Protected and Strictly protected wild species in R. North Macedonia. In summary, this record serves as an additional contribution to the catalog of caddisfly species in the Republic of North Macedonia, a region that still remains among the least studied in Europe (Radovanović 1935, 1943; Bilalli et al. 2018; Botosaneanu 1960; Pavlovski 1984, 1991; Rimcheska et al. 2015; Arsov 1991; Kumanski 1997; Kumanski & Malicky 1999; Vitecek et al. 20215; Kučinić et al. 2016; Oláh et al. 2018; Hinić et al. 2020; Slavevska-Stamenković et al. 2020; 2021; Hinić et al. 2024; Valladolid et al. 2021; 2022). The species redescription presented in this paper is a valuable contribution that simplifies the identification of this stenoendemic species while emphasizing the conservation significance of the area where it is found. Acknowledgements The first author would like to express her gratitude to prof. Halil Ibrahimi, PhD for generously and selflessly sharing his valuable knowledge and experience in entomological research. HINIĆ-JORDANOVSKA ET AL. 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