#ZooKeys ZooKeys 1228: 115-126 (2025) DOI: 10.3897/zookeys.1228.137496 Short Communication Barbastella caspica (Chiroptera, Vespertilionidae) in China: first record and complete mitochondrial genome Zhong-Yu Wang'™®, Shamshidin Abduriyim'® 1 College of Life Science, Shihezi University, Shihezi 832003, Xinjiang, China Corresponding author: Shamshidin Abduriyim (shamshidin@shzu.edu.cn) OPEN Qaccess This article is part of: Biology of Pangolins and Bats Academic editor: Wieslaw Bogdanowicz Received: 25 October 2024 Accepted: 11 January 2025 Published: 18 February 2025 ZooBank: https://zoobank.org/ E3C990CC-7911-43EA-9116- 44CB/54C04227 Citation: Wang Z-Y, Abduriyim S (2025) Barbastella caspica (Chiroptera, Vespertilionidae) in China: first record and complete mitochondrial genome. ZooKeys 1228: 115-126. https://doi. org/10.3897/zookeys.1228.137496 Copyright: © Zhong-Yu Wang & Shamshidin Abduriyim. This is an open access article distributed under terms of the Creative Commons Attribution License (Attribution 4.0 International - CC BY 4.0). Abstract The Caspian barbastelle, Barbastella caspica, has spread widely in the Caspian region, Iran, and Central Asia; however, there is no evidence of its occurrence in China so far. During a field investigation, we collected a single specimen of B. caspica in China's Xinjiang Uy- ghur Autonomous Region. At the same time, we obtained the free-flight echolocation calls of the bat. It omitted signals with start frequency of 33.15 + 1.43 kHz, end frequency of 29.82 + 0.40 kHz, frequency of most energy 31.48 + 0.40 kHz, duration of 2.43 + 0.24 ms, and a pulse interval of 246.57 + 9.48 ms, which are probably type-I sounds emitted through the mouth. We also sequenced its entire mitochondrial genome to elucidate the genomic structure and its evolutionary relationships with closely related Barbastella. The mitochon- drial genome of B. caspica spans 16,933 bp, comprising 13 protein-encoding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a displacement loop/control region. Consistent with previous bat mitogenome reports, the majority of mitochondrial genes are encoded on the heavy chain. A phylogenetic analysis based on 13 protein-coding genes revealed that Rhogeessa, Plecotus, and B. caspica formed a clade within Vespertilionidae. Barbastella caspica was found to be a sister species to B. beijingensis and B. leucomelas in phylogenetic trees using the cytochrome b and ND7 gene sequences. This is the first re- port of the mitogenome of a member of the genus Barbastella, as well as the first record of the distribution of B. caspica in China and first documentation of its echolocation calls. Key words: Echolocation calls, phylogenetic analysis, Xinjiang Introduction The Barbastella genus is widely distributed from Northeast Africa to across Eurasia to Taiwan and Japan. Currently, only six species are recognized: B. barbastellus Schreber, 1774, B. beijingensis Zhang et al., 2007, B. caspica Satunin, 1908, B. dar- jelingensis Hodogson, 1855, B. leucomelas Cretzschmar, 1826, and B. pacifica Kru- skop et al., 2019 (https://www.checklistbank.org/). In China, distributional records exist only for B. beijingensis and B. darjelingensis (http://www.sp2000.org.cn/). The Caspian barbastelle, B. caspica, primarily inhabits drier habitats and is occasionally found in caves, crevices, and mines. Its main distribution encom- passes northern Iran, the Caucasus region (Armenia, Azerbaijan, and Dagestan in Russia), Uzbekistan, and Tajikistan (Kruskop 2015). Research on this spe- cies is relatively limited, with a few studies focusing on taxonomic status and 115 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica distribution (Kruskop et al. 2019). Furthermore, genomic studies on species of Barbastella have been lacking, and the phylogenetic position of this genus within the family Vespertilionidae has not been explored. In this study, we used mist nets to capture and ultrasound recording equip- ment to record B. caspica echolocation calls. Furthermore, we conducted a comprehensive assembly and analysis of the complete mitochondrial genome of B. caspica, thus establishing the first genomic resource of Barbastella. Spe- cifically, we analyzed the nucleotide composition of the entire mitochondrial DNA molecule, investigated the codon usage patterns and selective constraints of protein-coding genes (PCGs), and described the secondary structure of each identified tRNA gene. Finally, based on mitochondrial PCGs, cytochrome b (Cytb), and ND7 sequences, we examined the phylogenetic position of Bar- bastella among other representative species of Vespertilionidae and of B. caspi- ca within its genus. On the one hand, the complete assembly of mitochondrial genome markers was a Significant step toward advancing our understanding of the genomic evolutionary biology and systematics of Barbastella species. On the other hand, this study also reported the first documentation of this species in China and the features of echolocation calls during flight. Materials and methods A bat individual was captured using mist nets during a survey of chiropteran re- sources in Yarkand County (37°54'24.75'N, 76°47'2.86'E), Xinjiang Uygur Auton- omous Region of China, in July 2023 (Fig. 1). The specimen (SC230705005) is currently stored at the College of Life Sciences, Shihezi University. Morphological identification revealed that the bat had short, wide ears with the front ends of both ears connected, indicating that it belongs to a species of barbastelles bat, Bar- bastella genus (https://www.checklistbank.org/). A Song Meter SM4BAT FS ultra- sonic recording device (Wildlife Acoustics, USA) was placed next to the mist net to record bat echolocation calls. Subsequently, the recorded echolocation sound waves were analyzed using sound analysis software (Kaleidoscope v. 5.4.8). In the laboratory, total genomic DNAs were extracted from muscle tissues using the Tiancheng Genomic DNA Extraction Kit (Tiangen Biotech, Beijing, China). The mitochondrial genome of B. caspica was amplified using PCR with 11 pairs of custom-designed primers (Suppl. material 1). Products that met quality-control criteria were purified and commercially sequenced. Sequencing data were processed and assembled using SeqMan software (Tamura et al. 2013). The annotation of the mitochondrial genome was performed using the GeSeq organelle genome annotation server (Tillich et al. 2017) (https://chloro- box.mpimp-golm.mpg.de/geseg.html). Annotation refinement and adjustment of start/stop codons were performed using MEGA X (Kumar et al. 2018). The finalized mitochondrial sequence has been deposited in NCBI GenBank under accession number PP963575. Results and discussion The echolocation call of Barbastella caspica is characterized by frequency mod- ulation (FM) (Fig. 2a, b). In free-flight outdoor conditions, the pulses are com- posed of a single harmonic. The peak frequency is notably low, with the highest ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 116 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica 50° N 40° N 30° N 40° E ~ Turanskaya Nizm~- Tarim Basin pa Sample collection site z aes Distribution of Barbastella caspica Yep) TORS A @ 60° E 70° E 80° E Figure 1. Map of Central Asia showing the geographic range of Barbastella caspica (green) and sampling site (red trian- gle) in southern Xinjiang, China. energy peak occurring at 31.48 + 0.40 kHz (Fig. 2c). The frequency bandwidth is narrow, measuring only 5.79 + 1.04 kHz. The initial frequency is at 33.15 + 1.43 kHz and the final frequency is at 29.82 + 0.40 kHz. The pulse duration is relatively short, approximately 2.43 + 0.24 ms, with an interpulse interval of 246.57 + 9.48 ms (Table 1). These characteristics closely resemble the sound waves emitted by other species of Barbastella while foraging (Zhang et al. 2007) and were similar to the type-I sounds of Barbastella species (Denzinger et al. 2001). However, considering that certain species of horseshoe bat consistently emit two different types of sound waves during foraging (Seibert et al. 2015), it is possible that our sound-wave detector failed to capture type-II sounds. Alter- natively, it is likely that B. caspica does not produce this particular sound during foraging or that the frequency of the emitted sound waves is lower than in other Barbastella species. These possibilities should be confirmed in future studies. The mitochondrial genome of B. caspica is a circular DNA molecule with a length of 16,933 base pairs (Fig. 3). The genome encompasses a total of 37 genes, consisting of 13 PCGs, 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and one D-loop region. The size and organization of these mitochondrial genes (Table 2) are consistent with previous reports of other vespertilionid species (Guo et al. 2021; Martinez-Cardenas et al. 2024; Valencia M. et al. 2024). Among the 13 PCGs (11,408 bp), they exhibit similarities with other species of Vespertilionidae, such as being located on the heavy strand except for ND6 (Martinez-Cardenas et al. 2024; Valencia M. et al. 2024). The ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 117 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica Frequency (kHz) Frequency (kHz) ee Time (s) Amplitude (%) Frequency (kHz) Time (ms) Energy (dB) Figure 2. Echolocation calls features of B. caspica in free flight conditions: the spectrogram and waveform with time unit in milliseconds (a), the spectrogram with a time unit of seconds (b) and the energy spectrum (c). Table 1. Echolocation calls features of Barbastella caspica in free-flight conditions. Items Range Mean + SD Initial frequency (kHz) 29.97-34.63 3315-45143 Terminate frequency (kHz) 28.99-30.19 29.82 + 0.40 Frequency bandwidth (kHz) 4.02-7.27 9.79 + 1.04 Main frequency (kHz) 31.07-31.96 31.48 + 0.40 Duration time (ms) 2,.0552.74 2.43 + 0.24 Interval time (ms) 232.29-266.43 246.57 + 9.48 average A+T content of PCGs in mitochondria is 59.92%, ranging from 56.31% (COX1) to 64.73% (ATP8), which is higher than the G+C content (40.08%) of the 13 PCGs. Furthermore, they show similar negative AT skew and CG skew, as well as a high At+G content (60.03%) (Suppl. material 2) (Guo et al. 2021; Martinez-Cardenas et al. 2024; Valencia M. et al. 2024). All PCGs start with ATG or ATA codons and terminate with TAA or truncated T residues, except for the Cytb gene, which terminates with AGA (Table 2). Suppl. material 3 shows the codon counts and RSCU values of B. caspica. The 33 codons are used more frequently (RSCU > 1, Suppl. material 4). The codons AAU-Asn (158), ACA-Thr (133), CCA-Pro (130), ACU-Thr (121), and CUA-Leu (118) are the most frequently used. There are 22 typical tRNA genes, ranging in length from 59 bp (tRNA-Ser1) to 75 bp (tRNA-Leu2). Eight of these genes are located on the L strand, while 14 are on the H strand. In total, they span 1520 bp. Except for tRNA-Ser (Table 2, Suppl. material 5), all these tRNA ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 118 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica Table 2. Composition and organization of the mitochondrial genome of Barbastella caspica. Gene tRNA-Phe 12S Rrna tRNA-Val 16S rRNA tRNA-Leu2 ND1 tRNA-lle tRNA-GIn tRNA-Met ND2 tRNA-Trp tRNA-Ala tRNA-Asn tRNA-Cys tRNA-Tyr COX1 tRNA-Ser2 tRNA-Asp COX2 tRNA-Lys ATP8 ATP6 COX3 tRNA-Gly ND3 tRNA-Arg ND4L ND4 tRNA-His tRNA-Ser1 tRNA-Leu1 ND5 ND6 tRNA-Glu Cytb tRNA-Thr tRNA-Pro D-loop Strand H Zprmy Dy oye yey] ey oy] oy yy yy yy] oY Sy cy yey ee yy yey yy cy] cy] ty Location Size(bp) | StartCodon StopCodon | Anticodon Continuity l=Z2 Le = = GAA 0 72-1031 960 = = = =i 1032-1100 69 7 = TAC 1101-2668 1569 = = = 2669-2743 75 = = TAA -1 2749-3705 957 ATG TAA a 5 AUS = 7 ie 68 . . GAT =i 3770-3843 74 = = TTG a3 3844-3911 68 - a CAT 0 3912-4953 1042 ATA ie = 0 4954-5020 67 = = TCA 0 5028-5095 68 = = TGC vi 5096-5168 73 = = GTT 0 5200-5266 67 = = GCA 31 5267-5332 66 = = GTA 0 5334-6878 1545 ATG TAA = 1 6882-6950 69 = = TGA 3 6958-7024 67 = = GTC 7, 7025-7706 684 ATG TAA = 0 7711-7779 69 = = THT 2 7780-7983 204 ATG TAA = 0 7941-8621 681 ATG TAA - -43 8621-9404 784 ATG > = =i 9404-9472 69 ry x LEG ail 9472-9818 347 ATA TA- = vl 9819-9889 Fl = = TCG 0 9891-10187 297 ATG TAA = 1 10181-11558 1378 ATG Ses = a7 11559-11627 69 = = GTG 11628-11686 59 = = GCT 11688-11758 71 7 = TAG 1 11759-13579 1821 ATA TAA oS 0 13563-14090 528 ATG TAA 7 =|7 14091-14158 68 = ni Ee 14164-15303 1140 ATG AGA = 15304-15375 72 ie = TGT 15373-15441 69 oS - TGG =3 15442-16933 1492 = = = 0 molecules have the classical cloverleaf structure. This phenomenon has been mentioned in previous studies and is common among metazoans (Vivas-Toro et al. 2021; Basaldua et al. 2023). However, more research is needed to deter- mine the functionality of these features in B. caspica. Based on 13 PCG sequences, we successfully constructed the phylogenetic topology of 31 species from the vespertilionid subfamilies Myotinae and Ves- pertilioninae. Consequently, Rhogeessa, Plecotus, Pipistrellus, Glischropus, Hyp- sugo, and Barbastella formed the subfamily Vespertilioninae, with Barbastella being a sister genus to Plecotus (Fig. 4). Consistent with previous results based ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 119 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica z 9 = = z y E Barbastella caspica mitochondrial genome 16,933 bp [J complex | (NADH dehydrogenase) [J complex IV (cytochrome c oxidase) fi ATP synthase i other genes i transfer RNAs i ribosomal RNAs “loop i origin of replication £§ 8 Figure 3. Mitochondrial genome map of B. caspica. The mitochondrial DNA of B. caspica is 16,933 base pairs long, consisting of different segments: 22 blue segments representing tRNA coding regions, 2 red segments corresponding to 12SrRNA and 16SrRNA, 7 yellow segments for ND7, ND2, ND3, ND4L, ND4, ND5, and ND6, 3 pink segments for COX7, COX2, and COX3, 1 purple segment for the Cytb gene, and 1 light red segment for the D-loop region. on the CO/ gene (Chakravarty et al. 2020), we found that Plecotus and Bar- bastella belonged to the same tribe, Plecotini, which also includes four other genera (Wilson and Mittermeier 2019; https://www.checklistbank.org/), imply- ing that more genome-based phylogeny is required to understand the interge- neric evolutionary relationships within the Plecotini. Phylogenetic trees were constructed to elucidate the evolutionary relation- ship of B. caspica with other species of vespertilionids, based on the Cytb and ND1 genes along with all PCGs. Within Barbastella, B. caspica is identified as a distinct species. However, differential topological structures were observed in the phylogenetic trees constructed based on the ND7 and Cytb genes (Fig. 5). ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 120 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica 98 100 100 0.07 KT199099 Myotis petax 100 85 KT199101 Myotis petax 100 NC 022694 = Myotis macrodactylus NC 056111 Myotis ricketti NC 025568 = Myotis davidii MT588108 Myotis blythii 100 100 NC 029346 Myotis myotis MT985382 Myotis bombinus MN122860 Myotis daubentonii 2 100 Myotis NC 034227 = Myotis bechsteinii NC 029422 = =Myotis muricola 100 100 NC 036324 Myotis atacamensis NC 022698 Myotis ikonnikovi 100 NC 060697 Myotis aurascens NC 015828 Myotis formosus NC 036321 Myotis leibii 100 100 NC 036323 Myotis thysanodes 100 NC 036313 Myotis evotis 100 OP345292 Myotis lucifugus MF143484 = =Myotis nigricans 68 NC 036317 Myotis riparius NC 065469 = Submyotodon moupinensis I S u bmyotodon NC 082319 Rhogeessa parvula 100 NC 083922 Rhogeessa mira 100 NC 082320 Rhogeessa Rhogeessa aeneus 100 NC 083923 Rhogeessa genowaysi KR134369 Plecotus macrobullaris 100 Plecotus NC 015484 = Plecotus auritus 83 99 PP963575 Barbastella caspica I B arbast ell, a MN122927 _ Pipistrellus pygmaeus | P. ipist. r ellus 100 zai NC 029191 — Glischropus tylopus | Glischropus MF459671 = Hypsugo alaschanicus I Aypsu ZO NC 044489 = Miniopterus fuliginosus MK177282 = Tadarida latouchei Figure 4. The phylogenetic relationships of the Vespertilionidae based on 13 protein-coding genes using the maxi- mum-likelihood method with 1,000 bootstrap replicates. Tadarida latouchei and Miniopterus fuliginosus were designated as outgroups. Nodes with support values = 80 are indicated. Namely, B. caspica was a sister species to B. leucomelas in the ND7 phyloge- netic tree, but sister to B. beijingensis in the Cytb phylogenetic tree. The pairwise distances (Table 3) shows that the smallest genetic distances (3.7% based on the ND7 gene) are between B. caspica and B. leucomelas. How- ever, the direct pairwise distance between the two species based on the Cytb gene is 13.4%, which is consistent with the results of the phylogenetic tree. The systematic construction of the ND7 phylogenetic tree, as well as the ND1 genetic distances within Barbastella, consistently indicate a close genetic rela- tionship between B. caspica and B. leucomelas, which agrees with the results of Smirnov et al. (2020). In contrast, the findings of the Cytb analyses are conflicting (Fig. 5, Table 3). Phylogenetic tree inconsistencies are common among mammals, especially due to important evolutionary events (Rokas and Chatzimanolis 2008). ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 121 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica PP963575 Barbastella caspica Barbastella caspica PP963575 LC456173 Barbastella caspica Barbastella caspica —_LC456159 LC456172 Barbastella caspica Barbastella caspica LC456158 DQ915030 Barbastella leucomelas Barbastella beijingensis EF534759 Barbastella beijingensis OP393470 OP393469 = Barbastella beijingensis Barbastella leucomelas KU922958 EF534770 —_ Barbastella beijingensis Barbastella leucomelas EF534765 DQ915031 Barbastella barbastellus Barbastella pacifica LC456155 KJ948255 —_ Barbastella barbastellus Barbastella pacifica LC456145 KF218431 Barbastella barbastellus Barbastella barbastellus JQ683212 LC456170 Barbastella pacifica Barbastella barbastellus JQ683211 LC456160 Barbastella pacifica Barbastella barbastellus JQ683180 beseol i Rarhastella sp Barbastella barbastellus JQ683186 NC083922 Rhogeessa mira Rhogeessa mira NC083922 0.06 Figure 5. The Bayesian analyses of phylogenetic relationships of members of the Barbastella genus based on 806 bp ND1 (left) and 1140 bp Cytb (right) sequences using Bayesian-inference (Bl) and maximum-likelihood (ML) methods. Rhogeessa mira is used as the root, and nodes with support values of = 0.7 (BI) and 80 (ML) are labeled. Table 3. ML distances (above the diagonal) and p-distances (below the diagonal) (in %) for ND1 and Cytb sequences of Barbastella caspica. Species B. caspica_ _ B.leucomelas | B. beijingensis _ B. barbastellus |B. pacifica B. caspica r 5.0 / 15.4 12.8,713:6 14.1 / 16.1 16.9 / 16.0 B. leucomelas 3.7/13.4 7 13:27-15.3 14.7 / 16.6 18.2/ 14.9 B. beijingensis | 9.4/12.0 9.6 / 18.0 = 15.4/18.4 17.1/17.6 B. barbastellus | 10.6 / 13.9 10.8 / 14.2 1 ial Salle aS as = 1797152 B. pacifica 12.6 / 13.8 135/13. 12.6/ 14.9 I.3F 17-9 4 These discrepancies can be attributed to factors such as inadequate gene sam- pling, hybridization events, gene introgression, or horizontal transfer. Although these findings provide enough evidence to consider B. caspica as an independent species (Fig. 5, Table 3), our understanding of its evolutionary relationship with other Barbastella species remains limited. Furthermore, the Central Asian species is named B. walteri (Smirnov et al., 2021). Hence, to obtain more comprehensive information, it is necessary to explore the genomic aspects of all species rather than confining our study solely to partial genes. Previous reports have indicated that B. caspica is distributed from the Caucasus region through Iran to Tajikistan (Kruskop 2015), excluding China. This report has expanded our understanding of the geographic distribution of B. caspica. Com- bining these findings with previous research, we infer that the eastern edge of the B. caspica distribution extends to Xinjiang, China. Before this discovery, only two species of barbastelles (B. darjelingensis and B. beijingensis) had been document- ed in China, and B. darjelingensis was found exclusively in Xinjiang. Therefore, our report adds an additional species of barbastelle bats to the Chinese biodiversity. ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 122 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica Conclusions This study highlights the presence of B. caspica in Xinjiang, China, for the first time and presents the first complete assembly of the mitochondrial genome, providing valuable genetic resources for investigating inter- and intraspecific evolutionary relationships. In addition, we describe for the first time free-flight echolocation calls, possibly of type-I] sounds omitted through the mouth. Tak- ing the collection site of our specimen of B. caspica into account, it is neces- sary to conduct further ecological and genetic studies at the population level on a whole distributional scale. Acknowledgements We express our sincere appreciation to subject editor and reviewer for their constructive comments and suggestions on our manuscript. We thank Chai Guanghou and Zhang Yan for their indispensable support during the two-month field sampling period. We acknowledge financial support from the National Natural Science Foundation of China (grant no. 32260328). We thank the edi- tor, Wieslaw Bogdanowicz, and the reviewers for their constructive comments on the manuscript. Additional information Conflict of interest The authors report that they have no conflicts of interest. Ethical statement The Biology Ethics Committee of Shihezi University approved all sample handling and experimental procedures (Approval: 2023-221). All bat treatment procedures were in accordance with the Bat Workers’ Manual (Mitchell-Jones and McLeish 2004). Funding This study received financial support from the National Natural Science Foundation of China (no. 32260328). Author contributions Conceptualization: SA. Data curation: SA. Formal analysis: ZYW. Investigation: ZYW. Methodology: SA. Project administration: SA. Software: ZYW. Funding Acquisition: SA. Writing — original draft: ZYW. Writing — review and editing: SA, ZYW. Author ORCIDs Zhong-Yu Wang ® https://orcid.org/0009-0002-6277-942X Shamshidin Abduriyim © https://orcid.org/0000-0002-7038-077X Data availability The mtDNA sequences we obtained have been deposited in the NCBI GenBank databas- es under accession numbers PP963575. 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Zhang J-S, Han N-J, Jones G, Lin L-K, Zhang J-P, Zhu G-J, Huang D-W, Zhang S-Y (2007) Anew species of Barbastella (Chiroptera: Vespertilionidae) from north China. Journal of Mammalogy 88: 1393-1403. https://doi.org/10.1644/07-MAMM-A-114R2.1 Supplementary material 1 PCR primers designed for mitochondrial genome analysis of B. caspica Authors: Zhong-Yu Wang , Shamshidin Abduriyim Data type: docx Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/zookeys.1228.137496.suppl1 Supplementary material 2 Base composition of the mitogenomes of B. caspica Authors: Zhong-Yu Wang , Shamshidin Abduriyim Data type: docx Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/zookeys.1228.137496.suppl2 ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 125 Zhong-Yu Wang & Shamshidin Abduriyim: Complete mitochondrial genome of Barbastella caspica Supplementary material 3 RSCU values of protein-coding genes in mitochondrial genome of B.caspica Authors: Zhong-Yu Wang , Shamshidin Abduriyim Data type: pdf Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/zookeys.1228.137496.suppl3 Supplementary material 4 Frequency and RSCU values of codon in protein coding genes in the mitogenome of B. caspica Authors: Zhong-Yu Wang , Shamshidin Abduriyim Data type: docx Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/zookeys.1228.137496.suppl4 Supplementary material 5 The secondary structure of tRNA gene Authors: Zhong-Yu Wang , Shamshidin Abduriyim Data type: pdf Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/zookeys.1228.137496.suppl5 ZooKeys 1228: 115-126 (2025), DOI: 10.3897/zookeys.1228.137496 126