First record of Gnaphosa jodhpurensis Tikader & Gajbe, 1977 (Araneae, Gnaphosidae) from Iraq and the Middle East

the journal of biodiversity data NOTES ON GEOGRAPHIC DISTRIBUTION 902 Academic editor: Alireza Zamani Received: 14 July 2025 Accepted: 11 September 2025 Published: 3 October 2025 Copyright © The authors. This is an open‑access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International – CC BY 4.0) Abstract. We report Gnaphosa jodhpurensis Tikader & Gajbe, 1977 (Araneae, Gnaphosidae) from Iraq and the Middle East for the first time, based on material collected from Karbala, Babylon, and Dhi Qar provinces. Identification was based on morphological comparison with the original description and further supported by DNA barcoding of the mitochondrial cytochrome c oxidase subunit I gene. Known distribution records of the species across its range are mapped. Key words. Biodiversity, distribution, DNA barcoding, faunistics, ground spiders, taxonomy Al-Mshrfawy TAA, Al-Khazali AM (2025) First record of Gnaphosa jodhpurensis Tikader & Gajbe, 1977 (Ara‑ neae, Gnaphosidae) from Iraq and the Middle East. Check List 21 (5): 902–908. https://doi.org/10.15560/21.5. 902 INTRODUCTION The family Gnaphosidae Banks, 1892, commonly known as ground spiders, is among the most species‑rich and widely distributed within araneomorph spiders, globally comprising 2,500 described species in 153 genera (WSC 2025). These spiders are generally free‑living, with most species being ground‑dwellers, and they inhabit a variety of microhabitats, including leaf litter, soil surfaces, and rock crevices (Jocqué and Dippenaar‑Schoeman 2006). Despite this ecological breadth, the gnaphosid fauna of Iraq remains far less documented than that of neighboring countries, such as Turkey (163 species in 34 genera; Danışman et al. 2025) and Iran (143 species in 40 genera; Zamani 2025). According to the most recent national checklist, 16 species in 11 genera of gnaphosids are known from Iraq (Al‑Khazali et al. 2023). Early records included Nomisia conigera (Spassky, 1941) and Pterotricha arzhantsevi Fomichev, Marusik & Koponen, 2018 (Fomichev et al. 2018), followed by Berlandina mesopotamica Al‑Khazali, 2020 from Dhi Qar (Al‑Khazali 2020), with the female described later (Al‑Khazali and Fomichev 2021). During the two years following the checklist, seven additional species were documented from Iraq, raising the total to 23 species in 15 genera by 2025. These include Cryptodrassus helvolus (O. Pickard‑Cambridge, 1872), Zelotes laetus (O. Pickard‑Cambridge, 1872) (Abdalrazzaq and Najim 2024), Drassodes lapidosus (Walckenaer, 1802) (Zamani et al. 2024), Minosia simeonica Levy, 1995, Synaphosus shirin Ovtsharenko, Levy & Platnick, 1994 (Al‑Khazali 2024) newly recorded from Iraq, and Talanites gilgamesh Al‑Khazali, 2024 (Al‑Khazali 2024) and Zelotes hazarmerdensis Zamani & Marusik, 2024 (Zamani and Marusik 2024) described as new to science. Within Gnaphosidae, the genus Gnaphosa Latreille, 1804 is both species‑rich and taxonomically com‑ plex, with 151 valid species currently recognized worldwide (WSC 2025). In Iraq, only one species has so far been recorded: G. dolosa Herman, 1879 (Al‑Khazali et al. 2023). The present study documents the occurrence of G. jodhpurensis Tikader & Gajbe, 1977 as the second representative of the genus in Iraq, extending its known distribution into the Middle East. Originally described from India and later reported from China (WSC 2025) and Pakistan (Tahir et al. 2019), G. jodhpurensis has not been the subject of an integrative taxonomic study. We document here the first occurrence of G. jodhpurensis in Iraq, based on a combined approach using morphological and molecular evidence. STUDY AREA Specimens were collected from three Iraqi provinces: Karbala (Alwand) and Babylon (Al‑Musayyib) in cen‑ tral Iraq, and Dhi Qar (Al‑Fuhud) in southern Iraq (Figure 1). These areas lie within the Mesopotamian Plain 21 (5) · https://doi.org/10.15560/21.5.902 21 (5): 902–908 First record of Gnaphosa jodhpurensis Tikader & Gajbe, 1977 (Araneae, Gnaphosidae) from Iraq and the Middle East Tuqa A.A. Al-Mshrfawy1, Azhar M. Al-Khazali1 1 Department of Science, College of Basic Education, University of Sumer, Dhi Qar, Iraq Corresponding author: Azhar M. Al-Khazali (azhar[email protected]) Check List 21 (5) · https://doi.org/10.15560/21.5.902 Al-Mshrfawy and Al-Khazali · First record of Gnaphosa jodhpurensis from the Middle East 903 and are characterized by arid to semi‑arid climate conditions. The terrain is flat to gently undulating, with predominantly alluvial soils. Average annual rainfall is less than 200 mm, with peak temperatures exceeding 45°C during the summer months. Vegetation is sparse and dominated by xerophytic and halophytic plants. None of the surveyed locations is designated as a protected area. METHODS Specimen collection and morphological examination. Adult specimens were manually collected from soil surfaces and preserved in 75% ethanol. Morphological examination was conducted using a ste‑ reomicroscope (ED4) equipped with a Nikon digital camera. Epigynes were dissected and cleared in 10% potassium hydroxide (KOH) for examination, and placed in shallow ethanol‑filled dishes lined with white cotton to enhance imaging contrast. Measurements (in mm) were taken dorsally, and leg segment lengths are presented as total length (femur, patella, tibia, metatarsus, tarsus). Spination is based on the right‑side legs. A distribution map was created using SimpleMappr (Shorthouse 2010). Voucher specimens are depos‑ ited at the Arachnid Collection, Department of Science, College of Basic Education, University of Sumer, Iraq. Molecular procedures. DNA was extracted from leg tissue using the Wizard Genomic DNA Purification Kit (Promega, USA), with modifications. Tissue was ground under liquid nitrogen and lysed in Nuclei Lysis Solution at 65°C for 30 min. Proteinase K (17.5 μl, 20 mg/ml) was then added and incubated at 55 °C for 3–12 h. DNA was precipitated in isopropanol, washed with 70% ethanol, and rehydrated in DNA Rehydration Solution. The COI gene was amplified using LCO1490 and HCO2198 primers (Folmer et al. 1994) in 25 μl reactions containing 12.5 μl Master Mix, 1 μl of each primer, 5 μl template DNA, and 5.5 μl nuclease‑free water. PCR cy‑ cling conditions were as follows: initial denaturation at 94 °C for 15 min; 35 cycles of 94 °C for 60 s, 50 °C for 60 s, 72 °C for 90 s; final extension at 72°C for 7 min. PCR products were visualized on 1.5% agarose gels stained with RedSafe and documented under UV. A 100‑bp kb DNA ladder (SMOBIO) was used as a size standard. Phylogenetic analysis. A dataset of 13 published Gnaphosa COI sequences, along with the Iraqi speci‑ men (GenBank accession PV926656.1), was aligned using MUSCLE implemented in MEGA X. Runcinia acuminata (Thorell, 1881) (EU168166), Family Thomasidae, was included as the outgroup. Phylogenetic reconstruc‑ tion was performed using the maximum‑likelihood method under the Tamura–Nei model, selected as the best‑fit substitution model test in MEGA X. Node support was evaluated with 1,000 bootstrap replicates. RESULTS Family Gnaphosidae Banks, 1892 Genus Gnaphosa Latreille, 1804 Gnaphosa jodhpurensis Tikader & Gajbe, 1977 Figures 2–6 For a list of taxonomic references, see the WSC (2025). Figure 1. Known distribution records of Gnaphosa jodhpurensis Tikader & Gajbe, 1977. Squares = previous records; circles = new records from Iraq. Check List 21 (5) · https://doi.org/10.15560/21.5.902 Al-Mshrfawy and Al-Khazali · First record of Gnaphosa jodhpurensis from the Middle East 904 Material examined. IRAQ — Karbala • Alwand; 32.65°N, 044.00°E; elev. 30 m; 26.XII.2024; under building rubble near agricultural land; 1 ♀, UOS‑GJ‑2024‑01, GenBank MK154517.1 — babylon • Al‑Musayyib; 32.78°N, 044.30°E; elev. 28 m; 26.XII.2024; disturbed soil near farmland; 2 ♀, UOS‑GJ‑2024‑02 — Dhi Qar • Al‑Fuhud; 30.95°N, 046.50°E; elev. 17 m; 04.XII.2024; field margins and debris piles; 1 ♀, 1 juvenile ♀, UOS‑GJ‑2024‑03. The Iraqi female specimen was identified as G. jodhpurensis based on morphological similarities in the epigyne with the original type description from India (Figure 2A–C; cf. Tikader and Gajbe 1977). Molecular analysis of the COI gene revealed 99.7% similarity with a reference sequence from Pakistan (MK154238.1), confirming the morphological identification. Redescription. Female. Body length 8.40 mm. Carapace 3.60 mm long, 2.95 mm wide. Abdomen 4.80 mm long, 3.15 mm wide. Carapace narrowing anteriorly; dorsally slightly convex; anteriorly bearing a V‑shaped brown mark (Figure 3A). Sternum and coxae uniformly yellowish brown, finely set with short, brown hairs (Figure 3B). Endites and maxillae dark brown (Figure 3C). Posterior median eyes large and rounded; anterior median eyes smaller and closely set (Figure 3D). Chelicerae dark brown, with serrated keels; margins of keel strongly elevated (Figure 4A, B). Palps and legs reddish brown, without distinct mark‑ ings. Leg measurements (in mm): I 13.80 (3.85, 1.35, 3.00, 3.60, 2.00), II 12.40 (3.90, 1.20, 2.80, 3.10, 1.40), III 12.50 (3.10, 1.00, 3.30, 3.20, 1.90), IV 15.90 (4.40, 1.30, 3.40, 4.50, 2.30). Abdomen elongate‑oval, moderately convex, clothed with fine, greenish setae giving velvety appear‑ ance; dorsum with three longitudinal pairs of chalky‑white sigilla, posterior pair largest, extending toward spinnerets (Figure 4C). Venter pale, sparsely setose, without distinct patterning (Figure 2B). Spinnerets as in Figure 4D. Figure 2. Female Gnaphosa jodhpurensis, habitus. A. Dorsal view. B. Ventral view. Check List 21 (5) · https://doi.org/10.15560/21.5.902 Al-Mshrfawy and Al-Khazali · First record of Gnaphosa jodhpurensis from the Middle East 905 Figure 3. Gnaphosa jodhpurensis, female. A. Prosoma, dorsal view. B. Same, ventral view. C. Chelicerae and mouth parts, ventral view. D. Ocular region, anterodorsal view. Figure 4. Chelicerae and somatic structures of Gnaphosa jodhpurensis female. A. Left chelicera, prolateral view. B. Right chelicera, retrolateral view. C. Abdominal sigilla. D. Spinnerets. Check List 21 (5) · https://doi.org/10.15560/21.5.902 Al-Mshrfawy and Al-Khazali · First record of Gnaphosa jodhpurensis from the Middle East 906 Epigyne as in Figure 5; scape large, about twice as wide as long, with concave posterior margin; anterior hood (Ah) distinctly narrowed behind; receptacles (Re) oval, obliquely elongated at about 40–50° to body axis, directed latero‑posteriorly; glands (Rg) relatively long and slender, running parallel to the receptacles and laterally outward. Male. See Gajbe (2005, 2007). Distribution. The species was originally described from India (Tikader and Gajbe 1977) and subsequently reported from China (Song et al. 1999). Later, Ashfaq et al. (2019) provided molecular evidence supporting its occurrence in Pakistan. The present record from Iraq represents the first confirmed occurrence of G. jodhpurensis in the Middle East, significantly extending the species’ known range westward into the Western Palearctic. Behavioral observation. An unusual behavioral pattern was observed in a female individual of G. jodhpurensis reared in captivity. The spider gathered fine wood shavings and small twigs from the substrate of the rearing container, and subsequently wrapped them in silk. Following the completion of this construc‑ tion, the female positioned herself inside the resulting cocoon‑like tubular structure and remained hidden within it throughout the day. This behavior is interpreted as an adaptive retreat strategy, likely serving func‑ tions of camouflage or predator avoidance (Figure 6B). Barcoding results. The mitochondrial cytochrome c oxidase subunit I (COI) gene was successfully am‑ plified and sequenced from the examined specimen. The resulting 658 bp sequence was analyzed using the BLAST algorithm against the GenBank database. The top match was with G. jodhpurensis (voucher UOS‑ MTSPD‑0118), with a similarity of 99.69% and 100% coverage, corresponding to GenBank accession number MK154517.1. This sequence originates from a published DNA barcode reference library for spiders from Paki‑ stan (Tahir et al. 2019). The high sequence similarity confirms the molecular identity of the Iraqi specimen as G. jodhpurensis and supports the morphological identification. To determine the genetic affiliation of the Iraqi specimen, a phylogenetic tree was constructed using the maximum‑likelihood method based on COI sequences from 13 Gnaphosa species and the outgroup (Runcinia acuminata) (EU168166)). The phylogenetic tree (Figure 7) clearly shows that the Iraqi specimens (in blue) cluster within a strongly supported clade (bootstrap = 99%) alongside two reference sequences of G. jodhpurensis from Pakistan. Figure 5. Epigyne and vulva of Gnaphosa jodhpurensis female. A. Epigyne, ventral view (Ah = anterior hood). B. Vulva, dorsal view (Re = receptacle; Rg = receptacle gland). C. Schematic illustration of epigyne, ventral view. D. Schematic illustration of vulva, dorsal view. Check List 21 (5) · https://doi.org/10.15560/21.5.902 Al-Mshrfawy and Al-Khazali · First record of Gnaphosa jodhpurensis from the Middle East 907 ACKNOWLEDGEMENTS We thank the anonymous reviewers for their insightful comments and suggestions that greatly improved the quality of this manuscript. We sincerely thank the academic editor, Dr. Alireza Zamani, for his valuable guidance and professional support during the editorial process. We are also grateful to the family members of the first author for their kind assistance during the field collection of specimens. ADDITIONAL INFORMATION Conflict of interest The authors declare that no competing interests exist. Ethical statement All applicable ethical standards for the field collection of non‑endangered invertebrate species were fol‑ lowed. No protected or regulated taxa were involved in this study. Funding This study is part of the M.Sc. thesis of T.A. Al‑Mshrfawy, conducted under the supervision of Prof. Azhar M. Al‑Khazali at the University of Sumer, Iraq. The authors declare that no financial support or external funding was received for the completion of this study. Author contributions Conceptualization: AMA. Fieldwork and specimen collection: TAAM. Morphological analysis: AMA. Molecu‑ lar laboratory work: TAAM. Data interpretation: AMA, TAAM. Visualization and mapping: TAAM. Supervision: AMA. Writing – original draft: AMA, TAAM. Writing – review and editing: AMA. Figure 7. Maximum‑likelihood phylo‑ genetic tree based on COI sequences showing placement of the Iraqi specimen (dot blue) among other Gnaphosa species. Bootstrap values >30% shown at nodes. GenBank accession numbers are provided. Figure 6. Retreat behavior of Gnaphosa jodhpurensis in the laboratory. A. Female constructing a tubular retreat using silk and wood shavings. B. Completed retreat with plant debris bound by silk threads. Check List 21 (5) · https://doi.org/10.15560/21.5.902 Al-Mshrfawy and Al-Khazali · First record of Gnaphosa jodhpurensis from the Middle East 908 Author ORCID iDs Azhar M. Al‑Khazali https://orcid.org/0000‑0001‑7268‑696X Tuqa A. Al‑Mshrfawy https://orcid.org/0009‑0004‑0087‑4004 Data availability All data supporting the findings of this study are provided within the main text and the Figure. The COI sequence has been submitted to GenBank under accession number [PV926656.1]. REFERENCES Abdalrazzaq RK, Najim SA (2024) Two new records of gnaphosid spiders (Araneae: Gnaphosidae) from eastern Al‑Hammar marsh south of Iraq. Serket 20 (2): 141–146. Al-Khazali AM (2020) Berlandina mesopotamica sp. nov. (Araneae: Gnaphosidae): new record of the genus in Iraq. Arachnology 18 (5): 444–446. https://doi.org/10.13156/arac.2020.18.5.444 Al-Khazali AM (2024) New species and records of ground spiders (Araneae: Gnaphosidae) from Iraq. Ecologica Montenegrina 77: 12–23. Al-Khazali AM and Fomichev AA (2021) Description of the female of Berlandina mesopotamica (Araneae: Gnaphosidae). Arachno‑ lo gische Mitteilungen 61: 70–72. https://doi.org/10.30963/aramit6111 Al-Khazali AM, Kachel HS, Zamani A, Hussen FS (2023) An updated and annotated checklist of the spiders (Arachnida: Araneae) of Iraq. Zootaxa 5339 (2): 132–158. https://doi.org/10.11646/zootaxa.5339.2.2 Ashfaq M, Blagoev G, Tahir HM, Khan AM, Mukhtar MK, Akhtar S, Butt A, Mansoor S, Hebert PDN (2019) Assembling a DNA barcode reference library for the spiders (Arachnida: Araneae) of Pakistan. PLoS ONE 14 (5): e0217086. https://doi.org/10.1371/ journal.pone.0217086 Danışman T, Kunt KB, Özkütük RS, Coşar İ (2024) The checklist of the spiders of Turkey. Version 2023. http://www.spidersofturkey. info. Accessed on: 2024‑1‑2. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R )1994( DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology Biotechnology 3 5): 294–299. Fomichev AA, Marusik YM, Koponen S (2018) New data on spiders (Arachnida: Araneae) of Iraq. Zoology in the Middle East 64 (4): 329–339. https://doi.org/10.1080/09397140.2018.1484018 Gajbe UA (2005) Studies on some spiders of the family Gnaphosidae (Araneae: Arachnida) from Madhya Pradesh, India. Records of the Zoological Survey of India 105 (3–4): 111–140. Gajbe UA (2007) Araneae: Arachnida. In: Fauna of Madhya Pradesh (including Chhattisgarh). State Fauna Series. Zoological Survey of India, Kolkata 15 1): 419–540. Jocqué R, Dippenaar-Schoeman AS (2006) Spider families of the world. Musée Royal de l’Afrique Central, Tervuren, Belgium, 336 pp. Murphy J (2007) Gnaphosid genera of the world. Volumes 1 and 2. British Arachnological Society, St Neots, Cambridgeshire, United Kingdom, 605 pp. Nieuwenhuys E (2000) Spiders of North‑West Europe. https://ednieuw.home.xs4all.nl/Spiders/spidhome.htm. Accessed on: 2025‑ 09‑04. Shorthouse DP (2010) SimpleMappr, an online tool to produce publication‑quality point maps. http://simplemappr.net. Accessed on: 2025‑07‑23. Song DX, Zhu MS, Chen J (1999) The spiders of China. Hebei Science and Technology Publishing House, Shijiazhuang, China, 640 pp. Tahir HM, Khan SY, Shakeel M, Mukhtar MK, Khan MA, Younas M (2019) Identification of ground spiders through integrative approach from cotton fields of Layyah, Punjab, Pakistan. Pakistan Journal of Zoology 51 (3): 1089–1095. Tikader BK, Gajbe UA (1977) Studies on some spiders of the genera Gnaphosa Latreille and Callilepis Westring (family: Gnaphosi‑ dae) from India. Records of the Zoological Survey of India 73: 43–52. WSC (2025) World spider catalog. Version 23.0. Natural History Museum, Bern. https://wsc.nmbe.ch. Accessed on: 2025‑06‑02. Zamani A (2025) Arthropod diversity in Iran: Class Arachnida Lamarck, 1801 (excl. Acariformes Zakhvatkin, 1952 and Parasitiformes Reuter, 1909). Journal of Insect Biodiversity and Systematics 11 (4): 981–1002. Zamani A (2023) Spiders of Iran – systematics, diversity and distribution. Doctoral thesis, University of Turku, Turku, Finland, 385 pp. Zamani A, Khudhur FA, Marusik YM (2024) New data on spiders (Arachnida: Araneae) of Iraqi Kurdistan, with new species and records. Zootaxa 5492 (2): 260–278. https://doi.org/10.11646/zootaxa.5492.2.6 Zamani A, Marusik YM (2022) On a small collection of spiders (Arachnida, Araneae) from Iraq, with new species and records. Zoo‑ diversity 56 (4): 291–306. https://doi.org/10.15407/zoo2022.04.291