Hemiclepsis Vejdovský, 1884

Solijonov, Khayrulla, Utevsky, Serge, Izzatullaev, Zuvayd, Umarov, Farrukh U., Fazliddinov, Firdavs, Shrestkha, Maria & Utevsky, Andriy, 2024, First record of Hemiclepsis marginata (O. F. Müller, 1773) (Hirudinida: Glossiphoniidae) from the Ferghana Valley, Uzbekistan, Ecologica Montenegrina 75, pp. 74-84 : 78-84

Genus: Hemiclepsis Vejdovský, 1884

Hemiclepsis marginata marginata Moore, 1924

Material examined: FVZ 55, Chartak discrit, Namangan region, Ferghana Valley (41°19'24.8"N, 71°50'16.4"E), 9. VI.2020. Kh. Solijonov leg. FVZ 157, Balikkul spring, Chartak discrit, Namangan region, Ferghana Valley (41°19'25.5"N, 71°50'11.3"E), 877 meters above sea level (m.a.s.l.), 9.VII.2020, Kh. Solijonov leg. FVZ 265, Kaynarbulak spring, Balikchi discrit, Andijan region, Ferghana Valley (40°53'09.5"N, 71°50'08.6"E), 410 (m.a.s.l.), 28. VI.2021, Kh. Solijonov leg. FVZ 391, Uchtepa spring, Uychi discrit, Namangan region, Ferghana Valley (40°55'49.3"N, 71°52'49.2"E), 415 (m.a.s.l.), 28. VI.2021 .

Description: Hemiclepsis marginata marginata is a medium-sized leech. The body length of juvenile leeches is 6–15 mm, width is 1–1.4 mm; in adults, it is on average 16–28 mm, width, 4–7 mm. In a calm state, the body has an elongated oval shape, the middle of the body is wider than the anterior end, and it is slightly narrower towards the posterior end. The first 10 annuli at the anterior end are compressed, forming the shape of a head. The edges of the body are somewhat like saw teeth. The dorsal surface of the body is smooth. The body colour is mainly greenish-brown ( Figure 2A ). Juvenile individuals are transparent, slightly green, and become greenish-brown as they mature ( Figure 2E ). On the dorsal side, there are seven rows of pale-yellow spots. The ventral side is whitish, without spots ( Figure 2B ). The width of the posterior sucker is larger than that of the anterior sucker, and most of it protrudes below the ventral side of the body ( Figure 2D ). There are two pairs of eyes, the eyes of the anterior pair are smaller and closer to each other than the eyes of the posterior pair ( Figure 2C ). Due to the transparency of skin cells, the digestive organs of the leeches can be seen with the simple eye. The oesophagus is with 3 pairs of small diverticula. The crop forms 7 pairs of caeca: the 1 st pair is smaller and "butterfly"-shaped; the 2 nd – 6 th pairs are larger and the last 7 th pair forms 4 branches.

Natural habitats. This species lives at a depth of 30–70 cm, in very clear waters at a temperature of 12–16°C, at an altitude of 410–877 meters above sea level ( Figure 3 A-C). Its habitat is characterized by muskgrass, where various vertebrate and invertebrate animals reside, including fish, amphibians, water molluscs, crustaceans, and insect larvae. Our observations indicate that the studied individuals of H. marginata marginata exhibit oxyphilic habitat preferences, residing in oxygen-rich waters of moderately warm springs. The fact that they exclusively inhabit springs suggests that the leech is crenophilous within this particular portion of the subspecies' range. Obviously, H. marginata marginata is sensitive to sudden changes in other environmental factors. Adults are temporary ectoparasites and feed on fish and amphibians by sucking their blood. When they leave their hosts, they can mostly be found under stones and bricks ( Figure 3D ) ( Pazilov & Umarov 2021; Solijonov & Umarov 2022).

We compared the internal and external morphology, reproductive characteristics, and habitat preferences of H. marginata marginata found in Uzbekistan with those of H. marginata asiatica from India. When analysing the body size, it was found that samples from Uzbekistan are larger (6–28 mm) and samples from India are smaller (5–20 mm). When the coloration of living specimens was studied, our specimens were mainly greenish-brown, while leeches were reddish-brown in Kashmir ( Moore 1924; Harding & Moore 1927; Soos 1967; Chandra 1983; Mandal 2015; Praveenraj 2021), Bihar ( Chandra 1991; Mandal 2013), Hargana, Assam, Rajasthan, Maharashtra, and Karnataka ( Mandal 2015), and pinkish white leeches were found in Kashmir, Haryana, Bihar, Assam, Himachal Pradesh, Rajasthan, and Maharashtra ( Mandal 2010, 2013). Annulation consists of 69–70 rings in our specimens, and up to 70–73 in Indian specimens ( Mandal 2013). Leeches in the study area were found to lay up to 96 eggs. Indian leeches lay a maximum of 101 eggs ( Mandal, 2015). While Hemiclepsis marginata asiatica typically inhabits slow-flowing streams in regions of India, our specimens are found in spring habitats. Hosts of Indian leeches are known from the literature to be the albino red-bellied pacu Piaractus brachypomus (Cuvier, 1818) and some molluscs ( Harding & Moore 1927; Praveenraj 2021). In our research, we found that H. marginata marginata is ectoparasitic on the Aral basin snowtrout Schizothorax eurystomus Kessler, 1872 and the marsh frog Pelophylax ridibundus (Pallas, 1771) . As a temporary parasite, it feeds by sucking the host's blood, predominantly on the ventral side. After feeding, the leeches leave their hosts and hide mainly under rocks.

The morphological characters and biology of the leeches examined largely conform the previous descriptions of H. marginata marginata . The reduction of the anterior pair of eyes appears to be common in the Palearctic leeches and should not be attributed exclusively to H. marginata asiatica . Obviously, the two subspecies need a thorough comparative analysis based on a representative number of samples from both ranges.

Phylogeny and genetic differentiation

The new sample clustered within a well-supported clade in the phylogenetic tree based on COI sequences, alongside other samples of H. marginata marginata from the Palearctic, confirming its classification within that subspecies ( Figure 4 ). The minimum spanning network analysis identified the Uzbekistan haplotype as a singleton with a frequency of 1, connected by two mutation steps to the most widespread haplotype found in Eastern Europe and Kazakhstan ( Figure 5 ). This structure suggests a relatively simple genetic makeup in H. marginata marginata . Comparison of 12S sequences from the Uzbekistan sample with the Indian sample, identified as H. marginata asiatica , revealed a significant genetic divergence (0.0562±0.0113) between the two sequences, indicating species- level differences, while the Uzbekistan 12S sequence is identical to the sequence obtained from the French H. marginata , suggesting that both Uzbekistan and French leeches are the same species/subspecies. This finding reinforces our identification of the leeches from the Ferghana Valley as H. marginata marginata . The average evolutionary divergence of COI sequences within the Hemiclepsis marginata clade is 0.0027±0.0010. The genetic distance between the Uzbekistan sample and other COI sequences of the Hemiclepsis marginata clade is 0.0042±0.0025, falling within the range of variability of this clade. Taken together, these results affirm the assignment of the newly sequenced sample to H. marginata marginata .

This study provides conclusive evidence of the presence of H. marginata marginata in Uzbekistan, a country situated at the boundary of the Palearctic and Oriental regions. The phylogenetic analysis placed the COI sequence of the Uzbekistan leech within a strongly supported clade alongside other Palearctic H. marginata samples, indicating its classification as H. marginata marginata . The genetic distance observed between the 12S sequences of the two subspecies, H. marginata marginata and H. marginata asiatica , suggests species-level distinctions. However, further investigation is required to discern morphological differences between these taxa and identify reliable distinguishing features. Moreover, our observations revealed the Uzbekistan leeches parasitizing the Aral basin snowtrout S. eurystomus and the marsh frog P. ridibundus .

Acknowledgements

This research was completed within the project ALM-202303153-01 of Samarkand State University named after Sharof Rashidov. We are grateful to Odiljon Tobirov, doctoral student of Gulistan State University and Kerim Çiçek, professor of Ege University, for their help in developing maps of geographical distribution. Thanks to Ferghana State University Associate Professor Bakhtiyor Sheraliev for his scientific advice on identifying fish species. This research was supported in part by the Academic Sanctuaries Fund created by XTX Markets and Eurizon Fellowship Programme (Grant Agreement #EU-3040).

References

ArcGIS.com (2019) ArcGIS Geographic Information System. ArcGIS Online. https://www.arcgis.com

Baugh, S.C. (1960). Studies on Indian Rhynchobdellid leeches. Parasitology, 50, 287–301. https://doi.org/10.1017/S0031182000025403

Bolbat, A., Bukin, Y. & Kaygorodova, I. (2022). Genome-Based Taxa Delimitation (GBTD): A New Approach. Diversity, 14 (11), 948. https://doi.org/10.3390/d14110948

Bolotov, I.N., Klass, A.L., Kondakov, A.V., Vikhrev, I.V., Bespalaya, Y.V., Gofarov, M.Y., Filippov, B.Y., Bogan, A.E., Lopes-Lima, M., Lunn, Z., Chan, N., Aksenova, O.V., Dvoryankin, G.A., Chapurina, Y.E., Kim, S.K., Kolosova, Y.S., Konopleva, E.S., Lee, J.H., Makhrov, A.A., Palatov, D.M., Sayenko, E.M., Spitsyn, V.M., Sokolova, S.E., Tomilova, A.A., Win, T., Zubrii, N.A. & Vinarski, M.V. (2019) Freshwater mussels house a diverse mussel-associated leech assemblage. Scientific Reports, 9, 16449. https://doi.org/10.1038/s41598-019-52688-3

Bolotov, I.N., Kondakov, A.V., Eliseeva, T.A. et al. (2022) Cryptic taxonomic diversity and highlatitude melanism in the glossiphoniid leech assemblage from the Eurasian Arctic. Scientific Reports, 12, 20630. https://doi.org/10.1038/s41598-022-24989-7

Bolotov, I.N., Vikhrev, I.V., Kondakov, A.V., Konopleva, E.S., Gofarov, M.Yu., Aksenova, O.V. & Tumpeesuwan, S. (2017) New taxa of freshwater mussels (Unionidae) from a species-rich but overlooked evolutionary hotspot in Southeast Asia. Scientific Reports, 7, 1–18.

https://doi.org/10.1038/s41598-017-11957-9

Borda, E. & Siddall, M.E. (2004) Review of the evolution of life history strategies and phylogeny of the Hirudinida (Annelida: Oligochaeta). Lauterbornia, 52, 5–25.

Chandra, M. (1991) The leeches of India. Zoological Survey of India Press, Calcutta, India, 130 pp.

Chandra, M.A. (1983) A check-list of leeches of India. Records of the Zoological Survey of India, 80, 265–290.

Chelladurai, J.E. (1934) On a new Indian leech Hemiclepsis viridis sp. nov. Records of the Indian Museum, 36, 345–352.

Foote, M., Iwama, R., de Carle, D. & Kvist S. (2022) An integrative taxonomic study of the genus Theromyzon (Hirudinea: Glossiphoniidae), with description of a new North American species. Invertebrate Systematics, 36, 631–646.

Govedich, F.R., Moser, W.E., Nakano, T., Bielecki, A., Bain, B.A. & Utevsky, S. (2019) Subclass Hirudinida. In: Rogers, D.Ch. & Thorp, J.H. (Eds.), Thorp and Covich’s Freshwater Invertebrates Volume IV: Keys to Palaearctic Fauna, Academic Press, London, United Kingdom, 491–507.

Guindon, S., Dufayard, J.F., Lefort, V., Anisimova, M., Hordijk, W. & Gascuel, O. (2010) New Algorithms and Methods to Estimate Maximum-Likelihood Phylogenies: Assessing the Performance of PhyML 3.0. Systematic Biology, 59, 307–321.

https://doi.org/10.1093/sysbio/syq010

Harding, W.A. & Moore, J.P. (1927) Hirudinea. The Fauna of British India, including Ceylon and Burma. Taylor and Francis, London, 302 pp.

Jovanović, M., Haring, E., Sattmann, H., Grosser, C. & Pešić, V. (2021). DNA barcoding for species delimitation of the freshwater leech genus Glossiphonia from the Western Balkan (Hirudinea, Glossiphoniidae). Biodiversity Data Journal, 9, e66347.

https://doi.org/10.3897/BDJ.9.e66347

Kalyaanamoorthy, S., Minh, B., Wong, T. et al. (2017) ModelFinder: fast model selection for accurate phylogenetic estimates. Nat Methods, 14, 587–589. https://doi.org/10.1038/nmeth.4285

Katoh, K., Rozewicki, J. & Yamada, K.D. (2019) MAFFT online service: multiple sequence alignment, interactive sequence choice and visualization. Brief Bioinform., 20(4), 1160–1166.

https://doi.org/10.1093/bib/bbx108

Kaygorodova, I.A., Mandzyak, N., Petryaeva, E. & Pronin, N.M. (2014) Genetic diversity of freshwater leeches in Lake Gusinoe (Eastern Siberia, Russia) Scientific World Journal, 619127.

https://doi.org/10.1155/2014/619127

Kearn, G.C. (2005) Leeches, lice and lampreys: a natural history of skin and gill parasites of fishes. Springer, The Netherlands, 446 pp.

Kumar, S., Stecher, G., Li. M., Knyaz, C. & Tamura, K. (2018) MEGA X: molecular Evolutionary Genetics Analysis across computing platforms. Mol Biol Evol., 35(6), 1547–1549.

https://doi.org/10.1093/molbev/msy096

Leerhoei, F. (2020) DNAmark Project (unpublished)

Leigh, J.W. & Bryant, D. (2015) PopART: Full-feature software for haplotype network construction. Methods in Ecologe and Evolution, 1110–1116. https://doi.org/10.1111/2041-210X.12410

Lukin, E.I. (1976) Leeches of fresh and brackish water bodies. Fauna of the USSR. Publishing house Nauka, Leningrad, 484 pp. (in Russian)

Mandal, C.K. (2010) Fauna of Uttarakhand. In: (Ed) Halder K.R. Zoological Survey of India, Kolkata, India, 193–197.

Mandal, C.K. (2013) Hemiclepsis chharwardamensis sp. nov. (Hirudinea: Glossiphonidae) a new species of leech from Jharkhand. Records of the Zoological Survey of India, 113, 153–155.

Mandal, C.K. (2015) Hemiclepsis marginata asiatica the highest egg-laying aquatic leech in the world. Records of the Zoological Survey of India, 115, 317–319.

Minh, B.Q., Nguyen, M. & von Haeseler, A. (2013) Ultrafast approximation for phylogenetic bootstrap. Molecular Biology and Evolution, 30 (5), 1188–1195. https://doi.org/10.1093/molbev/mst024

Moore, J.P. (1924) Notes on some Asiatic leeches (Hirudinea) principally from China, Kashmir, and British India. Proceedings of the Academy of Natural Sciences of Philadelphia, 76, 343–388.

Nesemann, H. & Neubert, E. (1999) Annelida, Clitellata: Branchiobdellida, Acanthobdellea, Hirudinea. In: Schwoerbel, J. & Zwick, P. (Eds.), Süsswasserfauna von Mitteleuropa. Vol. 6/2. Spektrum Akademischer Verlag, Heidelberg, 1–178

Nguyen, L.-T., Schmidt, H.A., von Haeseler, A. & Minh, B.Q. (2015) IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution, 32, 268–274. https://doi.org/10.1093/molbev/msu300

Oka, A. (1910) der Japanischen Hirudineen, mit Diagnosen der Neuen Species. Annotationes Zoologicae Japonenses, 7, 165–183

Oka, A. (1932) Sur une nouvelle espèce de Placobdella, Pl. japonica n. sp. Proceedings of the Imperial Academy, 8, 51–53.

Osmanov, S.O. (1971) Parasites of fishes of Uzbekistan. Fan, Tashkent, 532 pp. (in Russian)

Pazilov, A.P. & Umarov, F.U. (2021) On the ecology and species diversity of the freshwater gastropods of springs in Andijan region, Uzbekistan. Bulletin of the Iraq Natural History Museum, 16 (3), 325–340. https://doi.org/10.26842/binhm.7.2021.16.3.0325

Plotnikov, V. (1907) Glossosiphonidae, Hirudinidae i Herpobdellidae Zoologicheskogo muzeya Akademii Nauk. Ezhegodnik Zoologicheskago muzeia Akademii Nauk, 10, 133–158. (in Russian)

Praveenraj, A., Uma, A., Saravanan, K., Rebecca, G. & Mandal, C.K. (2021) Outbreak of hirudiniasis in aquarium-reared albino red-bellied pacu Piaractus brachypomus. Diseases of Aquatic Organisms, 144, 55–59. https://doi.org/10.3354/dao03569

Rozas, J., Ferrer-Mata, A., Sánchez- DelBarrio, J.C., Guirao-Rico, S., Librado, P., Ramos-Onsins, S. & Sánchez- Gracia, A. (2017) DnaSP 6: DNA sequence polymorphism analysis of large data sets. Molecular biology and evolution, 34, 3299–3302.

https://doi.org/10.1093/molbev/msx248

Sawyer, R.T. (1986) Leech Biology and Behaviour. Oxford University Press, New York, USA, 1046 pp.

Shchegolev, G.G. (1912) Fauna of leeches of Turkestan. Proceedings of the Hydrobiological Station on Glubokoe Lake, 4, 163–192. (in Russian)

Siddall, M.E. & Burreson, E.M. (1998) Phylogeny of leeches (Hirudinea) based on mitochondrial cytochrome c oxidase subunit I. Molecular Phylogenetics and Evolution, 9, 156–62.

https://doi.org/10.1006/mpev.1997.0455

Sket, B. & Trontelj, P. (2008) Global diversity of leeches (Hirudinea) in freshwater. Hydrobiologia, 595, 129–137. https://doi.org/10.1007/s10750-007-9010-8

Solijonov, Kh., Izzatullaev, Z. & Umarova, D. (2023) New record of malacophagous leech of the genus Alboglossiphonia Lukin, 1976 from Fergana Valley, Uzbekistan. Bulletin of the Iraq Natural History Museum, 17 (3), 459-468. https://doi.org/10.26842/binhm.7.2022.17.2.0229

Solijonov, Kh.Kh. & Umarov, F.U. (2022) Ecology of leeches and gastropods of the lower Ak-Buura River, Fergana Valley, Uzbekistan. Bulletin of the Iraq Natural History Museum, 17 (2), 229- 250. https://doi.org/10.26842/binhm.7.2023.17.3.0459

Soós, Á. (1967) On the genus Hemiclepsis Vejdovsky, (1884), with a key and catalogue of the species (Hirudinoidea: Glossiphoniidae). Opuscula Zoologica (Budapest), 7, 233–240.

Tan, E.G. & Liu, X.Q. (2001) One new species of the genus Hemiclepsis (Rhynchobdellida: Glossiphoniidae). Acta Zootaxonomica Sinica, 3, 289–291.

Tessler, M., de Carle, D., Voiklis, M.L., Gresham, O.A., Neumann, J., Cios, S. & Siddall, M.E. (2018) Worms that suck: phylogenetic analysis of Hirudinea solidifies the position of Acanthobdellida and necessitates the dissolution of Rhynchobdellida. Molecular Phylogenetics and Evolution, 127, 129-134. https://doi.org/10.1016/j.ympev.2018.05.001

Utevsky, A. & Utevsky, S. (2018) New Antarctic deep-sea weird leech (Hirudinida: Piscicolidae): morphological features and phylogenetic relationships. Systematic Parasitology, 95 (8-9), 849- 861. https://doi.org/10.1007/s11230-018-9816-y

Xu, Z., Yang, C., Gofarov, M.Y., Eliseeva, T.A., Kondakov, A.V., Yuan, H., Bolotov, I.N. & Yang, D. (2021) A new freshwater leech species from Asian Swamp Eel stocks in China. Parasitology Research 120: 2769–2778. https://doi.org/10.1007/s00436-021-07228-2

Yang, T. (1981) Two new species of parasitic leeches from freshwater fishes in China. Acta Zootaxonomica Sinica, 6, 27–30.

Yang, T. (1996) Annelida Hirudinea. Science Press, Beijing, 259 pp.

Supplementary material

The list of the GenBank sequences, their provenance and authors.

Authors: Khayrulla Solijanov, Serge Utevsky, Zuvayd Izzatullaev, Farrukh U. Umarov, Firdavs Fazliddinov, Maria Shrestkha, Andriy Utevsky

Data type: Excel document

Link: https://www.biotaxa.org/em/article/view/85684/version/81958/80636

Figures

Figure 2. Hemiclepsis marginata marginata: A, dorsal view; B, ventral view; C, eyes; D, posterior sucker; E, juvenile leech (photo by Kh. Solijonov), F, fixed sample (photo by A. Utevsky).

Figure 3. Natural habitats of Hemiclepsis marginata marginata: A, Balikkul spring; B, Kaynarbulak spring; D, Uchtepa spring; D, leeches on a stone (photo by Kh. Solijonov).

Figure 4. Phylogenetic relationships of Hemiclepsis marginata from the Ferghana Valley (Uzbekistan) inferred using the maximum likelihood optimality criterion. The analysis is based on a dataset consisting of the newly generated COI sequence of the Uzbekistan leech and previous GenBank sequences. Both ultrafast bootstrapping and SH-like approximate likelihood-ratio test (SHaLRT) values are provided for branches. The newly generated sequence is highlighted in bold, while the entire clade of H. marginata (H. marginata marginata) is highlighted in blue.

Figure 5. Minimum spanning network of COI haplotypes of Hemiclepsis marginata, including the newly generated unique sequence from Uzbekistan (5). Bars on branches represent mutation steps. Haplotype numbers correspond to those in Table 1.